TY - JOUR
AU - Marchetti,, Daniela
AB - Abstract Since the introduction in 1990, the solid-phase microextraction (SPME) technology has brought significant progress in many fields of forensic sciences due to the versatility of this fast and solventless alternative to conventional extraction techniques. A systematic review about SPME applications in forensic context from January 1995 to June 2018 was carried out according to systematic review guidelines. The majority of the reviewed articles (40/133) aimed to identify drugs (cannabinoids, cocaine, opiates, amphetamines, simultaneous detection of different drugs of abuse, prescribed drugs); 29 of the 133 articles focused on the investigation of fatalities; 28 of the 133 papers used headspace SPME technique for the identification of markers of chronic alcohol abuse. Sixteen papers involved this technique for the isolation of volatile organic compounds for the human odor profile and 20 concerned forensic applications regarding living people. Solid-phase microextraction was preferably employed in the headspace mode and many kinds of fibers were employed, although polydimethylsiloxane was the most adaptable to many forensic realities. Gas chromatography/mass spectrometry was more frequently used, probably for the well-established coupling with SPME. Most of the papers validated their method to harmonize the scientific approaches of procedures development. Good outcomes are reported on biological material collected from living people as well as on cadaveric samples. The results obtained by most of the studies about alcohol biomarkers on scalp hair have been adopted by the “Society of Hair Testing” to demonstrate abstinence over a pre-defined time period and to assess chronic excessive alcohol consumption. SPME, forensic, systematic review Introduction Forensic toxicology covers important areas of forensic investigations concerning death and poisoning related to use/abuse of drugs and/or crimes performed by the means of drugs and toxic compounds. Laboratories performing toxicology tests aim to detect drugs of abuse and other psychoactive substances in biological matrices to identify possible impairment due to drug use or to detect or exclude drug addiction. Drug testing is then used in many contexts, including crime investigations (crime under the influence of alcohol/drugs), safety and performance monitoring, date rape, driving under the influence, determination of cause of death, workplace surveillance, psychiatric disorders and treatment assessment; overall, it covers important issues in medical legal diagnosis. The legal consequences of the results of a toxicological analysis compel the management of significant and reliable data; hence, the analytical toxicological laboratory should respect some important requirements suggested by guidelines (1, 2). Toxicologists thus are often involved in the investigation of analytical processes that ensure easy, fast, reliable extraction and detection of small amounts of substances in complex matrices, such as biological cadaveric samples. Sample preparation is an essential step in analytical fields for isolating desired components from complex matrices. The first treatment of specimens greatly influences the reliability and accuracy of the analysis of compounds of interest. In the forensic context, this fact is valuable for the large heterogeneity of biological samples and their matrix complexity (such as for post-mortem samples), thus requiring major analytical treatments. Low-cost and risk-free procedures through low solvent consumption, miniaturization, automation, high-throughput performance and online coupling with analytical instruments are the daily goals of analysts. The well-known, widely used, and commonly accepted sample preparation techniques include liquid–liquid extraction (LLE) and solid-phase extraction. Because these procedures are well documented and well defined, they are considered methods of choice by many laboratories. However, many challenging tasks, such as complex matrix applications, on-site and in vivo applications, and determination of matrix-bound and free concentrations of analytes, are not always easily achievable with these classical approaches. Solid-phase microextraction (SPME) has proven to be an important technique for the treatment of biological specimens, and it has been expanding its applications in routine and research forensic toxicological analysis because of the many advantages respect to traditional strategies. Solid-phase microextraction The introduction of SPME by Dr. J. Pawliszyn and colleagues (3) has brought significant progress in the sample preparation area by facilitating time and cost savings in applications, obtaining high performances in terms of quali-quantitative detection of compounds. It was designed with the aim to “address the need for a fast, solvent-free, and field compatible sample preparation method” (4). The inventor of SPME received an “R&D 100” Award in 1994 recognizing its invention as a major advancement in the analytical sciences. Solid-phase microextraction is a solvent-free technique consisting of a certain length of fused silica fiber coated on the outside with an appropriate stationary phase housed in a syringe-like handling device that protects phases and samples. Fibers are able to isolate and concentrate analytes with an adsorption/absorption mechanism; organic substances can then be directly introduced into the injector of the analytical instrument, where the analytes are desorbed and transferred into the column for separation and quantification. All steps of the conventional LLE, which consist of extraction, concentration, sometimes derivatization and transfer to the instrument, are combined into one step and one device, considerably simplifying sample pre-treatment, reducing the time necessary for sample preparation, decreasing the purchase and disposal costs of solvents and improving detection limits. The fundamental concepts of SPME were presented by Pawliszyn (5) in a article describing the mathematical model developed to understand the principal processes of SPME. Briefly, the inventor of this extraction approach explained that SPME can use a small volume of sorbent typically dispersed on the surface of small fibers to isolate and concentrate analytes from the sample matrix, absorbing or adsorbing analytes after contact with the sample until equilibrium is reached in the system. The amount of an analyte extracted by the coating at equilibrium is determined by the magnitude of the analyte partition coefficient between the sample matrix and the coating material. After the extraction step, a syringe-like handling device allows the fibers to insert into an analytical instrument for separation and quantitation of target analytes. SPME is compatible with separation techniques, mainly with gas chromatography (GC), concentrating analytes on the fiber with a rapid delivery to the column and providing linear results for a great number of compounds, with low detection limits. The requirement of solvent desorption for non-volatile or thermally labile compounds has proven challenging for interfacing SPME with liquid-phase separations. Numerous options have been described in the literature, showing that SPME coupled with liquid chromatography (LC) has great potential in the analysis of a wide range of compounds in different matrices, with applications in several different areas of analysis (6). The direct coupling of SPME and mass spectrometry (MS) demonstrated the possibility to improve limits of quantitation, accelerate analysis throughput, and diminish potential matrix effects when compared to direct injection into the mass spectrometer (7). The presented design consisted of minimal alterations to the front end of the instrument and provided better sensitivity, simplicity, speed, wider compound coverage, and higher throughput in comparison to the LC–MS-based approach (6). The only manufacturer of commercially available SPME fibers is Supelco (Bellefonte, PA, USA), which supplies fibers consisting of fused silica (length 1 cm, diameter 110 μm) or StableFlex (a flexible fused-silica core, less breakable, lengths 1 or 2 cm) coated with different single-polymer or mixed-polymer materials. The coatings have different film thicknesses between 7 and 100 μm and can be non-bonded or bonded to the core. They are either in a quasi-liquid state (polydimethylsiloxane [PDMS], polyacrylate [PA], carbowax [CW]) or porous solid particles embedded into a PDMS or a CW layer (polydivinylbenzene, carboxen [CAR], templated resin [TPR]). Proper selection of the fiber depends on the polarity, molecular size and volatility of the analytes. The commercial coatings for SPME fibers have not essentially changed for many years, but some developments have been made for the modality of employment. For analytes with no or very low volatility, the fiber can be directly immersed in a sample solution (direct immersion SPME [DI-SPME]). Since the sample is in direct contact with the fiber, strong acidic or alkaline conditions must be avoided. Generally, the fiber is exposed to the liquid sample at constant temperature between 30 and 100°C and with the stirring of the sample for a certain time, after which it can be directly introduced into the GC or LC injection system. Alternatively, the fiber can be exposed to the vapor phase above the liquid or solid sample (headspace SPME [HS-SPME]) with undeniable advantages of protecting the fiber from damaging by aggressive matrix components and minimizing the adsorption of matrix substances. The extraction kinetics are governed by Henry’s law. If the Henry’s constant of a given substance is high, then the concentration of the compounds in the headspace is also high. Under these conditions, a rapid extraction from the headspace occurs. The headspace version of SPME has the advantage of cleaner extracts and is superior to headspace gas chromatography (HS-GC) because of the accumulation of analytes on the fiber, which can be injected into the GC without an excess of air. On the contrary, the loss of analyte from the fiber for volatile liquids is its main disadvantage against HS-GC. The nature of target analytes (e.g. low volatility) and the complexity of sample matrix with polymer components (e.g. proteins in plasma) determine the level of difficulties in accomplishing a successful extraction, so limiting the application of SPME. Matrix composition and the physicochemical properties of the analytes strongly affect HS-SPME and volatile compounds are extracted faster than semivolatiles since they are at a higher concentration in the headspace, which contributes to faster mass transport rates through the headspace. Different from of the parent drugs, some metabolites are frequently not detected by HS-SPME and DI-SPME has to be preferred. Finally, quantitative analyses can be limited by saturation- and matrix-dependent suppression due to the limited sorbent capacity of the fiber; hence, the employ of deuterated internal standards (ISs) is necessary to achieve reliable quantification. The first applications of this technique aimed to verify the acceptability of the environment, to monitor processes for clinical, forensic, food, flavor, fragrance and drug analyses. Several scientific publications showed the results obtained in inter-laboratory validation studies in which SPME was applied to determine the presence of different organic compounds at parts per trillion concentrations, thus demonstrating its reliability for quantitative analysis (8). Since the first developments, many new different tools have been studied and commercialized, and a review on the possible applications in the forensic context of various kinds of SPME was published in 2012 (9). The principle of SPME was extended to extraction in coated capillaries (in-tube SPME), to solid-phase dynamic extraction by internally coated syringes (SPDE), and to stir-bar sorptive extraction methods. In-tube SPME, using segments of commercial GC capillaries for highly efficient extraction by repeated aspiration-ejection cycles of the liquid sample was developed for application with LC–MS (10). Headspace extraction on fibers or needles (SPDE) represents the first commercially available inside-needle device for headspace analysis using GC–MS. This is the most advantageous version of SPME because of very pure extracts and the availability of automatic samplers. An alternative approach is the total vaporization of the sample (TV-SPME) so that analytes partition directly between the vapor phase and the SPME fiber (11). The sensitivity of TV-SPME is higher than that of traditional techniques, and the sampling selectivity is increased with respect to conventional devices as well. Aim of the systematic review SPME is actually under investigation worldwide for its usefulness in the analysis of a wide variety of compounds. As new classes of compounds are investigated and new challenges arise, the methods are adapted to accommodate them. For these peculiarities, in the present systematic review, we aim to evaluate the use of SPME as an analytical technique in the forensic context. Methods A systematic review using a computerized search of English and non-English articles was performed using the National Library of Medicine’s Medline and TOXNET databases from 1 January 1995, to 30 June 2018. The searches were refined using the following MeSH terms and text words: SPME OR solid phase micro extraction: AND forensic OR legal medicine; AND forensic toxicology. AND post mortem OR postmortem OR post-mortem. AND drivers OR driver. AND analytical toxicology. AND drugs of abuse OR recreational drugs. No filters for special study design were used. The methods of the systematic review and exclusion criteria were specified in advance according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement (12). Titles and abstracts were independently screened by the authors based on the following exclusion criteria: duplicate papers reviews articles not written in English studies on animals studies on seizure materials studies using different micro-techniques (microextraction on packed sorbent, dispersive liquid–liquid extraction, liquid-phase microextraction, etc.) articles not about forensic issues environmental studies When a reference was considered potentially relevant, the full text was independently read by the two authors. Papers about studies with spiked specimens were then excluded. Additional data were identified through reference lists. Disagreements between reviewers were resolved by discussion. Results Selection of the studies Figure 1 represents the flow chart developed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement (12). Figure 1 Open in new tabDownload slide Flow chart of the computerized systematic review according to the PRISMA statement. Figure 1 Open in new tabDownload slide Flow chart of the computerized systematic review according to the PRISMA statement. A total of 1031 peer-reviewed articles were recognized as potentially suitable for inclusion. Article titles and abstracts were scanned for relevance, and 855 articles were then discarded. Most were duplicates. The full text of the remaining 176 works was examined in detail, and 50 of them were excluded. A total of 126 articles were included in the systematic review. Seven additional papers were identified through reference lists. Five tables report in chronological sequences the 133 studies included in the systematic review categorized by the application of SPME in forensic context. Literature reviewed The 133 articles reviewed (13–145) were divided on the basis of the application of SPME in the forensic context as follows: identification of markers of alcohol abuse and volatile organic compounds (VOCs) for the human odor profile; applications in the forensic toxicology investigation concerning death and living people; analytical studies for the detection of xenobiotics in biological samples. The papers were summarized in five main tables divided by author/year, population, samples investigated (matrix), analytical procedures, validation parameters, and main results. Two of the 133 papers have been considered twice in the tables due to SPME application either with living or dead people (67, 71). Two additional articles reported results about living people that were not distinguishable from those about dead people (104, 105). In the supplementary online material, tables are completed by analytical parameters. Overall, 75 of the 133 studies reviewed were published in important analytical journals, while the remaining were issued in forensic journals. The majority of the reviewed articles involved application of SPME as an analytical tool for the identification of drugs (cannabinoids, COC, opiates, amphetamines [APs], simultaneous detection of different drugs of abuse, prescribed drugs) (Table 5, 40 articles), followed by SPME in the investigation of fatalities (Table 3, 29 articles) and the HS-SPME GC–MS technique for the identification of markers of chronic alcohol abuse (Table 1, 28 papers). Sixteen papers involved the SPME technique for the identification of VOCs for the human odor profile (Table 2), and 20 concerned SPME in forensic applications regarding living people (Table 4). Table 1 Headspace SPME GC–MS technique for the identification of markers of alcohol abuse Ref. Author/year Population Matrix Substances identified Results 13 Pragst et al./2000 3 C and 3 T (children) Scalp hair (segmentation n.r.) Ethyl palmitate, ethyl oleate, ethyl stearate Ethyl palmitate, ethyl oleate and ethyl stearate were qualitatively identified in hair of alcoholics as useful markers by the method proposed. 5 B, 5 SD, 5 T Volatile compounds (such as alcohols, ketones, ...) Ethanol itself was found in almost every hair samples, mostly due to absorption from surrounding air, hence it cannot be used as a marker of alcohol consumption. 21 C FAEE were detected in all alcoholics but concentrations differ strongly from case to case. 14 Pragst et al./2001 10 SD Scalp and body hair (segmentation n.r.) ∑FAEE Ethyl palmitate was always detected at concentration clearly below that of alcoholics. 10 T Only traces of ethyl palmitate were sometimes detected. 15 Auwarter et al./2001 19 A, 10 C, 13 SD, 5 T Scalp hair (1–12 segments: 1–2.5 cm in the proximal region) ∑FAEE Mean of ∑FAEE concentration performed on 0–6 cm. No correlation between the reported daily alcohol consumption and hair concentration. Routes of incorporation widely discussed: in most cases, segmental concentration increases from proximal to distal demonstrating incorporation mainly from sebum. Large individual differences were described, but FAEE concentrations are considered useful for the diagnosis of excessive alcohol use also in postmortem. 16 Hartwig et al./2003 n.r. C, 75 (B + SD + T) Scalp hair in full length (segmentation in some case) ∑FAEE Traces of esters found in 49 hair care products, but no influence on the concentration of FAEE; false-positive results can be obtained with daily treatment with hair lotion containing ethanol. 17 Hartwig et al./2003 18 C, 5 SD, 1 T Scalp hair in full length and body hair ∑FAEE FAEE are deposited in all kinds of human hair after alcohol consumption; their concentration in body hair varies between different sites in the same subject. Cutoff n.r. ∑FAEE always lower than 0.8 ng/mg for SD and T. 18 Yegles et al./2004 10 A, 4 SD, 3 T, 11 C Scalp hair (proximal segment 0–6 cm) ∑FAEE A cutoff value of ∑FAEE ≥1 ng/mg and/or a positive EtG result in hair may evidence excessive alcohol consumption. No significant correlation between EtG and FAEE was shown. Segmental analysis of some of the specimens did not reveal the same distribution for EtG compared to FAEE in hair. No chronological accordance compared to the self-reported alcohol consumption was observed for both parameters. 19 Pragst et al./2004 13 T, 16 SD, 10 C Skin and sebum ∑ FAEE SQ was used as a natural reference substance to which FAEEs were related for the variable amounts of lipids. The ratio FAEE/SQ was calculated. The evaluation of abstinence from this ratio is not possible. The comparison with the self-reported ethanol amounts consumed the week before and during the test gave no good correlation (R2 = 0.42). FAEE concentration in skin surface lipids can be used for medium-term retrospective detection of heavy drinking. 20 Wurst et al./2004 18 A (14 M, 4 F, age 24–55) Scalp hair (segmentation n.r.) ∑FAEE ROC curve for ∑FAEE indicated sensitivity =100% and specificity = 90% for a cutoff of 0.29 ng/mg. By using a cutoff of 0.4 ng/mg, ∑FAEE identified 94.4% correctly. ∑FAEE and ethyl palmitate were significantly associated as were ∑FAEE and PEth. A cutoff value of 0.4 ng/mg for ∑FAEE allows to distinguish teetotallers/moderate/social drinkers from alcoholics. Comparison with other markers disclosed a good qualitative agreement between ∑FAEE, CDT and Peth. 10 SD, 10 T 21 Auwarter et al./2004 13 T – 16 SD–12 C–9 n.r. (unclear alcohol anamnesis) Scalp hair (0–6 cm segmentation) ∑FAEE SQ enables a control of the lipid content of hair and a correction of ∑FAEE in cases with deviations from the usual lipid content in a similar way as creatinine in urine. However, the relative concentration ∑FAEE/SQ cannot completely replace the absolute one. 22 De Giovanni et al./2007 12 B, 10 SD, 10 T Scalp hair (segmentation when hair length higher than 5 cm) ∑FAEE A partial overlapping between the 3 groups; 83% of alcoholics showed FAEE higher than 0.5 ng/mg; variation between segments in the same sample was observed. Special attention to the interpretation of results is suggested for the possible external contamination. 23 Schulz et al./2008 20 drivers and a volunteer Serum Eugenol Eugenol detected in 5/20 samples with positive BAC, ranging from 12.1 to 172.3 ng/mL related to amount and type of beverages and time of collection. 24 Schulz et al./2008 50 drivers and a volunteer Serum Anethole Anethol detected in 9/50 samples with concentrations ranging from 5.4 to 17.6 ng/mL related to amount and type of beverages and time of collection. 25 Schulz et al./2009 100 drivers and a volunteer Serum Menthone, isomenthone, neomenthol, menthol Menthone and isomenthone detected in 8/100 serum samples with positive BAC; neomenthol detected in 35 cases; menthol detected in 59 cases. Concentrations were related to amount and type of beverages and time of collection. 26 Kulaga et al./2009 324 (225 F, 96 M, 3 n.r.) Age 18–66. 119/225 were pregnant; 26/119 pregnant during hair sampling or within the last 6 mo Scalp hair (0–6 cm) ∑FAEE FAEE detection offers 90% sensitivity and specificity. A high rate of excessive alcohol use among parents (33%) is revealed with this method; the majority of samples having FAEE levels above a level that typically excludes strict abstinence. 27 Hutson et al./2009 10 samples Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) The method uses 50 mg of sample compared to 1 g required by previous methods. 28 Zelner et al./2010 31 neonates (28 samples) Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) 14/28 FAEE qualitatively identified; 9/28 nonanalyzable for the presence of contaminants and/or transitional stool interfering with chromatography. 5/28 were under analysis. A cumulative sum of greater or equal to 2.00 nmol/g of meconium is considered positive with 100% sensitivity and 98.4% specificity. 29 Agius et al./2010 266 T (children) Scalp hair (segmentation n.r.) EtG The method proved to be able to identify picogram amounts of EtG in blank hair for the purpose of understand the cutoff suitable to distinguish T from SD. 30 Pragst et al./2010 174 n.r.*1 Scalp hair (proximal segment 0–6 cm in case of longer samples) ∑FAEE ROC analysis for evaluation of data. The paper highlights the high specificity of this marker as direct marker. The combined use of FAEE and EtG increases the accuracy of the diagnosis. 31 Suesse et al./2010 644 n.r.*2 Scalp and body hair (proximal segment 0–6 cm) ∑FAEE ROC analysis for evaluation of data. The comparison with other markers (CDT, GGT, ALT, AST) evidences the much longer time window and increase in sensitivity and specificity with FAEE. 32 Morini et al./2010 99 (T + SD) *3 Scalp hair (segmentation 0–9 cm for mothers)*4 ∑FAEE The negativity of all hair samples demonstrated that FAEEs are not able to diagnose ethanol intake lower than 30 g/d. 33 Bakdash et al./2010 602 n.r. *5 Meconium Ethyl myristate, ethyl oleate, ethyl palmitate, ethyl stearate, ethyl linoleate FAEE and EtG can be detected in meconium; it was confirmed that the sum of esters is more suitable than each ester alone; the combined use of both the markers is suggested for better estimation of the degree of alcohol use. 34 Hutson et al./2011 39 samples Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) The method use 50 mg of sample compared to 1 g required by previous methods. 35 Gonzales-Illan et al./2011 5–6 T, 5–6 LD, 5–6 SD, 5–6 A Skin surface lipids ∑FAEE Skin surface lipids collected with Sebutapes from the foreheads were analyzed and the relative FAEE allowed an evaluation of alcohol consumption. An SD volunteer monitored during 2 mo revealed that the highest mFAEE/msebum was 7–9 d after the days of high alcohol consumption. 36 Natekar et al./2012 588 COC users Scalp hair CE, ∑FAEE In logistic regression, COC use was associated with a positive CE result (OR = 15.56, 95% CI = 5.95–40.67, P < 0.001), as were positive FAEE results (OR = 2.437, 95% CI = 1.21–4.87, P = 0.012). FAEE samples negative for excessive drinking (<0.5 ng/mg) were 95.18% of the time also negative for CE, indicating a very low rate of false positivity. Specificity increased when including social drinking/non-drinking population, indicating that 97.28% of FAEE values < 0.2 ng/mg were also negative for CE. This indicates that the rate of false positivity decreases even further for non-drinking individual. In addition, with a PPV of 0.66, if a positive CE is detected, then the sample is 66% likely to be positive for FAEE (≥0.5 ng/mg). 37 Agius et al./2012 3464 A *6 and 662 n.r. Scalp hair (segmentation 0–3 cm and 3–12 cm) EtG Statistical and stability studies provide evidence that the detection of EtG shall not be limited to 3-cm-long hair lenghts. The higher number of positivity in samples shorter than 3 cm supports incorporation of EtG from sweat after recent intake. 38 Albermann et al./2014 73 (T + SD) Scalp hair (proximal segment 0–3 cm) ∑FAEE The method was successfully validated according to the guidelines of the GTFCh, showing good linearity, precision and accuracy, recovery and stability. Comparison of FAEE and EtG (4–10 pg/mg previously detected with other techniques) in 73 authentic hair samples showed coincidences > 70%, but a plot of FAEE concentrations against EtG concentrations (R2 = 0.008) showed no linear correlation between the 2 markers. However, FAEE analysis can be performed to confirm EtG results even for abstinence tests. 39 Suesse et al./2015 157 n.r. (45 M, 112 F) Scalp hair (segmentation for both 0–3 cm and 0–6 cm) ∑FAEE The interpretation is complicated by inter-individual variability in formation and incorporation; the variable lenght of hair contributes to the uncertainty. The disagreement of the results depends mainly on the cosmetic treatment that can increase FAEE and decrease EtG. The redefinition of hair lenght for both markers is suggested. 40 Dumitrascu et al./2018 10 n.r. (5 F and 5 M) Scalp hair (segmentation 0–3 cm) ∑FAEE Various cosmetics were evaluated in order to understand their influence on FAEE: alcohol based hairspray increase FAEE levels in a highly variable manner. Quantification performed both with ethyl palmitate alone or the sum of the 4 esters produce the same results. Ref. Author/year Population Matrix Substances identified Results 13 Pragst et al./2000 3 C and 3 T (children) Scalp hair (segmentation n.r.) Ethyl palmitate, ethyl oleate, ethyl stearate Ethyl palmitate, ethyl oleate and ethyl stearate were qualitatively identified in hair of alcoholics as useful markers by the method proposed. 5 B, 5 SD, 5 T Volatile compounds (such as alcohols, ketones, ...) Ethanol itself was found in almost every hair samples, mostly due to absorption from surrounding air, hence it cannot be used as a marker of alcohol consumption. 21 C FAEE were detected in all alcoholics but concentrations differ strongly from case to case. 14 Pragst et al./2001 10 SD Scalp and body hair (segmentation n.r.) ∑FAEE Ethyl palmitate was always detected at concentration clearly below that of alcoholics. 10 T Only traces of ethyl palmitate were sometimes detected. 15 Auwarter et al./2001 19 A, 10 C, 13 SD, 5 T Scalp hair (1–12 segments: 1–2.5 cm in the proximal region) ∑FAEE Mean of ∑FAEE concentration performed on 0–6 cm. No correlation between the reported daily alcohol consumption and hair concentration. Routes of incorporation widely discussed: in most cases, segmental concentration increases from proximal to distal demonstrating incorporation mainly from sebum. Large individual differences were described, but FAEE concentrations are considered useful for the diagnosis of excessive alcohol use also in postmortem. 16 Hartwig et al./2003 n.r. C, 75 (B + SD + T) Scalp hair in full length (segmentation in some case) ∑FAEE Traces of esters found in 49 hair care products, but no influence on the concentration of FAEE; false-positive results can be obtained with daily treatment with hair lotion containing ethanol. 17 Hartwig et al./2003 18 C, 5 SD, 1 T Scalp hair in full length and body hair ∑FAEE FAEE are deposited in all kinds of human hair after alcohol consumption; their concentration in body hair varies between different sites in the same subject. Cutoff n.r. ∑FAEE always lower than 0.8 ng/mg for SD and T. 18 Yegles et al./2004 10 A, 4 SD, 3 T, 11 C Scalp hair (proximal segment 0–6 cm) ∑FAEE A cutoff value of ∑FAEE ≥1 ng/mg and/or a positive EtG result in hair may evidence excessive alcohol consumption. No significant correlation between EtG and FAEE was shown. Segmental analysis of some of the specimens did not reveal the same distribution for EtG compared to FAEE in hair. No chronological accordance compared to the self-reported alcohol consumption was observed for both parameters. 19 Pragst et al./2004 13 T, 16 SD, 10 C Skin and sebum ∑ FAEE SQ was used as a natural reference substance to which FAEEs were related for the variable amounts of lipids. The ratio FAEE/SQ was calculated. The evaluation of abstinence from this ratio is not possible. The comparison with the self-reported ethanol amounts consumed the week before and during the test gave no good correlation (R2 = 0.42). FAEE concentration in skin surface lipids can be used for medium-term retrospective detection of heavy drinking. 20 Wurst et al./2004 18 A (14 M, 4 F, age 24–55) Scalp hair (segmentation n.r.) ∑FAEE ROC curve for ∑FAEE indicated sensitivity =100% and specificity = 90% for a cutoff of 0.29 ng/mg. By using a cutoff of 0.4 ng/mg, ∑FAEE identified 94.4% correctly. ∑FAEE and ethyl palmitate were significantly associated as were ∑FAEE and PEth. A cutoff value of 0.4 ng/mg for ∑FAEE allows to distinguish teetotallers/moderate/social drinkers from alcoholics. Comparison with other markers disclosed a good qualitative agreement between ∑FAEE, CDT and Peth. 10 SD, 10 T 21 Auwarter et al./2004 13 T – 16 SD–12 C–9 n.r. (unclear alcohol anamnesis) Scalp hair (0–6 cm segmentation) ∑FAEE SQ enables a control of the lipid content of hair and a correction of ∑FAEE in cases with deviations from the usual lipid content in a similar way as creatinine in urine. However, the relative concentration ∑FAEE/SQ cannot completely replace the absolute one. 22 De Giovanni et al./2007 12 B, 10 SD, 10 T Scalp hair (segmentation when hair length higher than 5 cm) ∑FAEE A partial overlapping between the 3 groups; 83% of alcoholics showed FAEE higher than 0.5 ng/mg; variation between segments in the same sample was observed. Special attention to the interpretation of results is suggested for the possible external contamination. 23 Schulz et al./2008 20 drivers and a volunteer Serum Eugenol Eugenol detected in 5/20 samples with positive BAC, ranging from 12.1 to 172.3 ng/mL related to amount and type of beverages and time of collection. 24 Schulz et al./2008 50 drivers and a volunteer Serum Anethole Anethol detected in 9/50 samples with concentrations ranging from 5.4 to 17.6 ng/mL related to amount and type of beverages and time of collection. 25 Schulz et al./2009 100 drivers and a volunteer Serum Menthone, isomenthone, neomenthol, menthol Menthone and isomenthone detected in 8/100 serum samples with positive BAC; neomenthol detected in 35 cases; menthol detected in 59 cases. Concentrations were related to amount and type of beverages and time of collection. 26 Kulaga et al./2009 324 (225 F, 96 M, 3 n.r.) Age 18–66. 119/225 were pregnant; 26/119 pregnant during hair sampling or within the last 6 mo Scalp hair (0–6 cm) ∑FAEE FAEE detection offers 90% sensitivity and specificity. A high rate of excessive alcohol use among parents (33%) is revealed with this method; the majority of samples having FAEE levels above a level that typically excludes strict abstinence. 27 Hutson et al./2009 10 samples Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) The method uses 50 mg of sample compared to 1 g required by previous methods. 28 Zelner et al./2010 31 neonates (28 samples) Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) 14/28 FAEE qualitatively identified; 9/28 nonanalyzable for the presence of contaminants and/or transitional stool interfering with chromatography. 5/28 were under analysis. A cumulative sum of greater or equal to 2.00 nmol/g of meconium is considered positive with 100% sensitivity and 98.4% specificity. 29 Agius et al./2010 266 T (children) Scalp hair (segmentation n.r.) EtG The method proved to be able to identify picogram amounts of EtG in blank hair for the purpose of understand the cutoff suitable to distinguish T from SD. 30 Pragst et al./2010 174 n.r.*1 Scalp hair (proximal segment 0–6 cm in case of longer samples) ∑FAEE ROC analysis for evaluation of data. The paper highlights the high specificity of this marker as direct marker. The combined use of FAEE and EtG increases the accuracy of the diagnosis. 31 Suesse et al./2010 644 n.r.*2 Scalp and body hair (proximal segment 0–6 cm) ∑FAEE ROC analysis for evaluation of data. The comparison with other markers (CDT, GGT, ALT, AST) evidences the much longer time window and increase in sensitivity and specificity with FAEE. 32 Morini et al./2010 99 (T + SD) *3 Scalp hair (segmentation 0–9 cm for mothers)*4 ∑FAEE The negativity of all hair samples demonstrated that FAEEs are not able to diagnose ethanol intake lower than 30 g/d. 33 Bakdash et al./2010 602 n.r. *5 Meconium Ethyl myristate, ethyl oleate, ethyl palmitate, ethyl stearate, ethyl linoleate FAEE and EtG can be detected in meconium; it was confirmed that the sum of esters is more suitable than each ester alone; the combined use of both the markers is suggested for better estimation of the degree of alcohol use. 34 Hutson et al./2011 39 samples Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) The method use 50 mg of sample compared to 1 g required by previous methods. 35 Gonzales-Illan et al./2011 5–6 T, 5–6 LD, 5–6 SD, 5–6 A Skin surface lipids ∑FAEE Skin surface lipids collected with Sebutapes from the foreheads were analyzed and the relative FAEE allowed an evaluation of alcohol consumption. An SD volunteer monitored during 2 mo revealed that the highest mFAEE/msebum was 7–9 d after the days of high alcohol consumption. 36 Natekar et al./2012 588 COC users Scalp hair CE, ∑FAEE In logistic regression, COC use was associated with a positive CE result (OR = 15.56, 95% CI = 5.95–40.67, P < 0.001), as were positive FAEE results (OR = 2.437, 95% CI = 1.21–4.87, P = 0.012). FAEE samples negative for excessive drinking (<0.5 ng/mg) were 95.18% of the time also negative for CE, indicating a very low rate of false positivity. Specificity increased when including social drinking/non-drinking population, indicating that 97.28% of FAEE values < 0.2 ng/mg were also negative for CE. This indicates that the rate of false positivity decreases even further for non-drinking individual. In addition, with a PPV of 0.66, if a positive CE is detected, then the sample is 66% likely to be positive for FAEE (≥0.5 ng/mg). 37 Agius et al./2012 3464 A *6 and 662 n.r. Scalp hair (segmentation 0–3 cm and 3–12 cm) EtG Statistical and stability studies provide evidence that the detection of EtG shall not be limited to 3-cm-long hair lenghts. The higher number of positivity in samples shorter than 3 cm supports incorporation of EtG from sweat after recent intake. 38 Albermann et al./2014 73 (T + SD) Scalp hair (proximal segment 0–3 cm) ∑FAEE The method was successfully validated according to the guidelines of the GTFCh, showing good linearity, precision and accuracy, recovery and stability. Comparison of FAEE and EtG (4–10 pg/mg previously detected with other techniques) in 73 authentic hair samples showed coincidences > 70%, but a plot of FAEE concentrations against EtG concentrations (R2 = 0.008) showed no linear correlation between the 2 markers. However, FAEE analysis can be performed to confirm EtG results even for abstinence tests. 39 Suesse et al./2015 157 n.r. (45 M, 112 F) Scalp hair (segmentation for both 0–3 cm and 0–6 cm) ∑FAEE The interpretation is complicated by inter-individual variability in formation and incorporation; the variable lenght of hair contributes to the uncertainty. The disagreement of the results depends mainly on the cosmetic treatment that can increase FAEE and decrease EtG. The redefinition of hair lenght for both markers is suggested. 40 Dumitrascu et al./2018 10 n.r. (5 F and 5 M) Scalp hair (segmentation 0–3 cm) ∑FAEE Various cosmetics were evaluated in order to understand their influence on FAEE: alcohol based hairspray increase FAEE levels in a highly variable manner. Quantification performed both with ethyl palmitate alone or the sum of the 4 esters produce the same results. A Alcoholics in withdrawal treatment *1 Samples collected in context of driving ability, for workplace testing and child custody B Alcoholics with a known history of alcohol abuse *2 Samples mainly collected from parents, in child protection cases C Fatalities with a history of excessive alcohol consumption confirmed by pathologic findings *3 Samples collected by mothers with suspicious alcohol abuse and children (mother–infant dyads) LD Light drinkers *4 The amount of neonatal hair has been reported scarce SD Self-reported social drinkers *5 No pre-selection of the cases with respect to alcohol abuse T Teetotallers *6 Samples tested for abstinence to regain revoked driving license ∑FAEE Ethyl myristate, ethyl oleate, ethyl palmitate, ethyl stearate (referred as their sum) n.r. not reported. Open in new tab Table 1 Headspace SPME GC–MS technique for the identification of markers of alcohol abuse Ref. Author/year Population Matrix Substances identified Results 13 Pragst et al./2000 3 C and 3 T (children) Scalp hair (segmentation n.r.) Ethyl palmitate, ethyl oleate, ethyl stearate Ethyl palmitate, ethyl oleate and ethyl stearate were qualitatively identified in hair of alcoholics as useful markers by the method proposed. 5 B, 5 SD, 5 T Volatile compounds (such as alcohols, ketones, ...) Ethanol itself was found in almost every hair samples, mostly due to absorption from surrounding air, hence it cannot be used as a marker of alcohol consumption. 21 C FAEE were detected in all alcoholics but concentrations differ strongly from case to case. 14 Pragst et al./2001 10 SD Scalp and body hair (segmentation n.r.) ∑FAEE Ethyl palmitate was always detected at concentration clearly below that of alcoholics. 10 T Only traces of ethyl palmitate were sometimes detected. 15 Auwarter et al./2001 19 A, 10 C, 13 SD, 5 T Scalp hair (1–12 segments: 1–2.5 cm in the proximal region) ∑FAEE Mean of ∑FAEE concentration performed on 0–6 cm. No correlation between the reported daily alcohol consumption and hair concentration. Routes of incorporation widely discussed: in most cases, segmental concentration increases from proximal to distal demonstrating incorporation mainly from sebum. Large individual differences were described, but FAEE concentrations are considered useful for the diagnosis of excessive alcohol use also in postmortem. 16 Hartwig et al./2003 n.r. C, 75 (B + SD + T) Scalp hair in full length (segmentation in some case) ∑FAEE Traces of esters found in 49 hair care products, but no influence on the concentration of FAEE; false-positive results can be obtained with daily treatment with hair lotion containing ethanol. 17 Hartwig et al./2003 18 C, 5 SD, 1 T Scalp hair in full length and body hair ∑FAEE FAEE are deposited in all kinds of human hair after alcohol consumption; their concentration in body hair varies between different sites in the same subject. Cutoff n.r. ∑FAEE always lower than 0.8 ng/mg for SD and T. 18 Yegles et al./2004 10 A, 4 SD, 3 T, 11 C Scalp hair (proximal segment 0–6 cm) ∑FAEE A cutoff value of ∑FAEE ≥1 ng/mg and/or a positive EtG result in hair may evidence excessive alcohol consumption. No significant correlation between EtG and FAEE was shown. Segmental analysis of some of the specimens did not reveal the same distribution for EtG compared to FAEE in hair. No chronological accordance compared to the self-reported alcohol consumption was observed for both parameters. 19 Pragst et al./2004 13 T, 16 SD, 10 C Skin and sebum ∑ FAEE SQ was used as a natural reference substance to which FAEEs were related for the variable amounts of lipids. The ratio FAEE/SQ was calculated. The evaluation of abstinence from this ratio is not possible. The comparison with the self-reported ethanol amounts consumed the week before and during the test gave no good correlation (R2 = 0.42). FAEE concentration in skin surface lipids can be used for medium-term retrospective detection of heavy drinking. 20 Wurst et al./2004 18 A (14 M, 4 F, age 24–55) Scalp hair (segmentation n.r.) ∑FAEE ROC curve for ∑FAEE indicated sensitivity =100% and specificity = 90% for a cutoff of 0.29 ng/mg. By using a cutoff of 0.4 ng/mg, ∑FAEE identified 94.4% correctly. ∑FAEE and ethyl palmitate were significantly associated as were ∑FAEE and PEth. A cutoff value of 0.4 ng/mg for ∑FAEE allows to distinguish teetotallers/moderate/social drinkers from alcoholics. Comparison with other markers disclosed a good qualitative agreement between ∑FAEE, CDT and Peth. 10 SD, 10 T 21 Auwarter et al./2004 13 T – 16 SD–12 C–9 n.r. (unclear alcohol anamnesis) Scalp hair (0–6 cm segmentation) ∑FAEE SQ enables a control of the lipid content of hair and a correction of ∑FAEE in cases with deviations from the usual lipid content in a similar way as creatinine in urine. However, the relative concentration ∑FAEE/SQ cannot completely replace the absolute one. 22 De Giovanni et al./2007 12 B, 10 SD, 10 T Scalp hair (segmentation when hair length higher than 5 cm) ∑FAEE A partial overlapping between the 3 groups; 83% of alcoholics showed FAEE higher than 0.5 ng/mg; variation between segments in the same sample was observed. Special attention to the interpretation of results is suggested for the possible external contamination. 23 Schulz et al./2008 20 drivers and a volunteer Serum Eugenol Eugenol detected in 5/20 samples with positive BAC, ranging from 12.1 to 172.3 ng/mL related to amount and type of beverages and time of collection. 24 Schulz et al./2008 50 drivers and a volunteer Serum Anethole Anethol detected in 9/50 samples with concentrations ranging from 5.4 to 17.6 ng/mL related to amount and type of beverages and time of collection. 25 Schulz et al./2009 100 drivers and a volunteer Serum Menthone, isomenthone, neomenthol, menthol Menthone and isomenthone detected in 8/100 serum samples with positive BAC; neomenthol detected in 35 cases; menthol detected in 59 cases. Concentrations were related to amount and type of beverages and time of collection. 26 Kulaga et al./2009 324 (225 F, 96 M, 3 n.r.) Age 18–66. 119/225 were pregnant; 26/119 pregnant during hair sampling or within the last 6 mo Scalp hair (0–6 cm) ∑FAEE FAEE detection offers 90% sensitivity and specificity. A high rate of excessive alcohol use among parents (33%) is revealed with this method; the majority of samples having FAEE levels above a level that typically excludes strict abstinence. 27 Hutson et al./2009 10 samples Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) The method uses 50 mg of sample compared to 1 g required by previous methods. 28 Zelner et al./2010 31 neonates (28 samples) Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) 14/28 FAEE qualitatively identified; 9/28 nonanalyzable for the presence of contaminants and/or transitional stool interfering with chromatography. 5/28 were under analysis. A cumulative sum of greater or equal to 2.00 nmol/g of meconium is considered positive with 100% sensitivity and 98.4% specificity. 29 Agius et al./2010 266 T (children) Scalp hair (segmentation n.r.) EtG The method proved to be able to identify picogram amounts of EtG in blank hair for the purpose of understand the cutoff suitable to distinguish T from SD. 30 Pragst et al./2010 174 n.r.*1 Scalp hair (proximal segment 0–6 cm in case of longer samples) ∑FAEE ROC analysis for evaluation of data. The paper highlights the high specificity of this marker as direct marker. The combined use of FAEE and EtG increases the accuracy of the diagnosis. 31 Suesse et al./2010 644 n.r.*2 Scalp and body hair (proximal segment 0–6 cm) ∑FAEE ROC analysis for evaluation of data. The comparison with other markers (CDT, GGT, ALT, AST) evidences the much longer time window and increase in sensitivity and specificity with FAEE. 32 Morini et al./2010 99 (T + SD) *3 Scalp hair (segmentation 0–9 cm for mothers)*4 ∑FAEE The negativity of all hair samples demonstrated that FAEEs are not able to diagnose ethanol intake lower than 30 g/d. 33 Bakdash et al./2010 602 n.r. *5 Meconium Ethyl myristate, ethyl oleate, ethyl palmitate, ethyl stearate, ethyl linoleate FAEE and EtG can be detected in meconium; it was confirmed that the sum of esters is more suitable than each ester alone; the combined use of both the markers is suggested for better estimation of the degree of alcohol use. 34 Hutson et al./2011 39 samples Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) The method use 50 mg of sample compared to 1 g required by previous methods. 35 Gonzales-Illan et al./2011 5–6 T, 5–6 LD, 5–6 SD, 5–6 A Skin surface lipids ∑FAEE Skin surface lipids collected with Sebutapes from the foreheads were analyzed and the relative FAEE allowed an evaluation of alcohol consumption. An SD volunteer monitored during 2 mo revealed that the highest mFAEE/msebum was 7–9 d after the days of high alcohol consumption. 36 Natekar et al./2012 588 COC users Scalp hair CE, ∑FAEE In logistic regression, COC use was associated with a positive CE result (OR = 15.56, 95% CI = 5.95–40.67, P < 0.001), as were positive FAEE results (OR = 2.437, 95% CI = 1.21–4.87, P = 0.012). FAEE samples negative for excessive drinking (<0.5 ng/mg) were 95.18% of the time also negative for CE, indicating a very low rate of false positivity. Specificity increased when including social drinking/non-drinking population, indicating that 97.28% of FAEE values < 0.2 ng/mg were also negative for CE. This indicates that the rate of false positivity decreases even further for non-drinking individual. In addition, with a PPV of 0.66, if a positive CE is detected, then the sample is 66% likely to be positive for FAEE (≥0.5 ng/mg). 37 Agius et al./2012 3464 A *6 and 662 n.r. Scalp hair (segmentation 0–3 cm and 3–12 cm) EtG Statistical and stability studies provide evidence that the detection of EtG shall not be limited to 3-cm-long hair lenghts. The higher number of positivity in samples shorter than 3 cm supports incorporation of EtG from sweat after recent intake. 38 Albermann et al./2014 73 (T + SD) Scalp hair (proximal segment 0–3 cm) ∑FAEE The method was successfully validated according to the guidelines of the GTFCh, showing good linearity, precision and accuracy, recovery and stability. Comparison of FAEE and EtG (4–10 pg/mg previously detected with other techniques) in 73 authentic hair samples showed coincidences > 70%, but a plot of FAEE concentrations against EtG concentrations (R2 = 0.008) showed no linear correlation between the 2 markers. However, FAEE analysis can be performed to confirm EtG results even for abstinence tests. 39 Suesse et al./2015 157 n.r. (45 M, 112 F) Scalp hair (segmentation for both 0–3 cm and 0–6 cm) ∑FAEE The interpretation is complicated by inter-individual variability in formation and incorporation; the variable lenght of hair contributes to the uncertainty. The disagreement of the results depends mainly on the cosmetic treatment that can increase FAEE and decrease EtG. The redefinition of hair lenght for both markers is suggested. 40 Dumitrascu et al./2018 10 n.r. (5 F and 5 M) Scalp hair (segmentation 0–3 cm) ∑FAEE Various cosmetics were evaluated in order to understand their influence on FAEE: alcohol based hairspray increase FAEE levels in a highly variable manner. Quantification performed both with ethyl palmitate alone or the sum of the 4 esters produce the same results. Ref. Author/year Population Matrix Substances identified Results 13 Pragst et al./2000 3 C and 3 T (children) Scalp hair (segmentation n.r.) Ethyl palmitate, ethyl oleate, ethyl stearate Ethyl palmitate, ethyl oleate and ethyl stearate were qualitatively identified in hair of alcoholics as useful markers by the method proposed. 5 B, 5 SD, 5 T Volatile compounds (such as alcohols, ketones, ...) Ethanol itself was found in almost every hair samples, mostly due to absorption from surrounding air, hence it cannot be used as a marker of alcohol consumption. 21 C FAEE were detected in all alcoholics but concentrations differ strongly from case to case. 14 Pragst et al./2001 10 SD Scalp and body hair (segmentation n.r.) ∑FAEE Ethyl palmitate was always detected at concentration clearly below that of alcoholics. 10 T Only traces of ethyl palmitate were sometimes detected. 15 Auwarter et al./2001 19 A, 10 C, 13 SD, 5 T Scalp hair (1–12 segments: 1–2.5 cm in the proximal region) ∑FAEE Mean of ∑FAEE concentration performed on 0–6 cm. No correlation between the reported daily alcohol consumption and hair concentration. Routes of incorporation widely discussed: in most cases, segmental concentration increases from proximal to distal demonstrating incorporation mainly from sebum. Large individual differences were described, but FAEE concentrations are considered useful for the diagnosis of excessive alcohol use also in postmortem. 16 Hartwig et al./2003 n.r. C, 75 (B + SD + T) Scalp hair in full length (segmentation in some case) ∑FAEE Traces of esters found in 49 hair care products, but no influence on the concentration of FAEE; false-positive results can be obtained with daily treatment with hair lotion containing ethanol. 17 Hartwig et al./2003 18 C, 5 SD, 1 T Scalp hair in full length and body hair ∑FAEE FAEE are deposited in all kinds of human hair after alcohol consumption; their concentration in body hair varies between different sites in the same subject. Cutoff n.r. ∑FAEE always lower than 0.8 ng/mg for SD and T. 18 Yegles et al./2004 10 A, 4 SD, 3 T, 11 C Scalp hair (proximal segment 0–6 cm) ∑FAEE A cutoff value of ∑FAEE ≥1 ng/mg and/or a positive EtG result in hair may evidence excessive alcohol consumption. No significant correlation between EtG and FAEE was shown. Segmental analysis of some of the specimens did not reveal the same distribution for EtG compared to FAEE in hair. No chronological accordance compared to the self-reported alcohol consumption was observed for both parameters. 19 Pragst et al./2004 13 T, 16 SD, 10 C Skin and sebum ∑ FAEE SQ was used as a natural reference substance to which FAEEs were related for the variable amounts of lipids. The ratio FAEE/SQ was calculated. The evaluation of abstinence from this ratio is not possible. The comparison with the self-reported ethanol amounts consumed the week before and during the test gave no good correlation (R2 = 0.42). FAEE concentration in skin surface lipids can be used for medium-term retrospective detection of heavy drinking. 20 Wurst et al./2004 18 A (14 M, 4 F, age 24–55) Scalp hair (segmentation n.r.) ∑FAEE ROC curve for ∑FAEE indicated sensitivity =100% and specificity = 90% for a cutoff of 0.29 ng/mg. By using a cutoff of 0.4 ng/mg, ∑FAEE identified 94.4% correctly. ∑FAEE and ethyl palmitate were significantly associated as were ∑FAEE and PEth. A cutoff value of 0.4 ng/mg for ∑FAEE allows to distinguish teetotallers/moderate/social drinkers from alcoholics. Comparison with other markers disclosed a good qualitative agreement between ∑FAEE, CDT and Peth. 10 SD, 10 T 21 Auwarter et al./2004 13 T – 16 SD–12 C–9 n.r. (unclear alcohol anamnesis) Scalp hair (0–6 cm segmentation) ∑FAEE SQ enables a control of the lipid content of hair and a correction of ∑FAEE in cases with deviations from the usual lipid content in a similar way as creatinine in urine. However, the relative concentration ∑FAEE/SQ cannot completely replace the absolute one. 22 De Giovanni et al./2007 12 B, 10 SD, 10 T Scalp hair (segmentation when hair length higher than 5 cm) ∑FAEE A partial overlapping between the 3 groups; 83% of alcoholics showed FAEE higher than 0.5 ng/mg; variation between segments in the same sample was observed. Special attention to the interpretation of results is suggested for the possible external contamination. 23 Schulz et al./2008 20 drivers and a volunteer Serum Eugenol Eugenol detected in 5/20 samples with positive BAC, ranging from 12.1 to 172.3 ng/mL related to amount and type of beverages and time of collection. 24 Schulz et al./2008 50 drivers and a volunteer Serum Anethole Anethol detected in 9/50 samples with concentrations ranging from 5.4 to 17.6 ng/mL related to amount and type of beverages and time of collection. 25 Schulz et al./2009 100 drivers and a volunteer Serum Menthone, isomenthone, neomenthol, menthol Menthone and isomenthone detected in 8/100 serum samples with positive BAC; neomenthol detected in 35 cases; menthol detected in 59 cases. Concentrations were related to amount and type of beverages and time of collection. 26 Kulaga et al./2009 324 (225 F, 96 M, 3 n.r.) Age 18–66. 119/225 were pregnant; 26/119 pregnant during hair sampling or within the last 6 mo Scalp hair (0–6 cm) ∑FAEE FAEE detection offers 90% sensitivity and specificity. A high rate of excessive alcohol use among parents (33%) is revealed with this method; the majority of samples having FAEE levels above a level that typically excludes strict abstinence. 27 Hutson et al./2009 10 samples Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) The method uses 50 mg of sample compared to 1 g required by previous methods. 28 Zelner et al./2010 31 neonates (28 samples) Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) 14/28 FAEE qualitatively identified; 9/28 nonanalyzable for the presence of contaminants and/or transitional stool interfering with chromatography. 5/28 were under analysis. A cumulative sum of greater or equal to 2.00 nmol/g of meconium is considered positive with 100% sensitivity and 98.4% specificity. 29 Agius et al./2010 266 T (children) Scalp hair (segmentation n.r.) EtG The method proved to be able to identify picogram amounts of EtG in blank hair for the purpose of understand the cutoff suitable to distinguish T from SD. 30 Pragst et al./2010 174 n.r.*1 Scalp hair (proximal segment 0–6 cm in case of longer samples) ∑FAEE ROC analysis for evaluation of data. The paper highlights the high specificity of this marker as direct marker. The combined use of FAEE and EtG increases the accuracy of the diagnosis. 31 Suesse et al./2010 644 n.r.*2 Scalp and body hair (proximal segment 0–6 cm) ∑FAEE ROC analysis for evaluation of data. The comparison with other markers (CDT, GGT, ALT, AST) evidences the much longer time window and increase in sensitivity and specificity with FAEE. 32 Morini et al./2010 99 (T + SD) *3 Scalp hair (segmentation 0–9 cm for mothers)*4 ∑FAEE The negativity of all hair samples demonstrated that FAEEs are not able to diagnose ethanol intake lower than 30 g/d. 33 Bakdash et al./2010 602 n.r. *5 Meconium Ethyl myristate, ethyl oleate, ethyl palmitate, ethyl stearate, ethyl linoleate FAEE and EtG can be detected in meconium; it was confirmed that the sum of esters is more suitable than each ester alone; the combined use of both the markers is suggested for better estimation of the degree of alcohol use. 34 Hutson et al./2011 39 samples Meconium ∑FAEE (ethyl palmitate, linolate, oleate, stearate) The method use 50 mg of sample compared to 1 g required by previous methods. 35 Gonzales-Illan et al./2011 5–6 T, 5–6 LD, 5–6 SD, 5–6 A Skin surface lipids ∑FAEE Skin surface lipids collected with Sebutapes from the foreheads were analyzed and the relative FAEE allowed an evaluation of alcohol consumption. An SD volunteer monitored during 2 mo revealed that the highest mFAEE/msebum was 7–9 d after the days of high alcohol consumption. 36 Natekar et al./2012 588 COC users Scalp hair CE, ∑FAEE In logistic regression, COC use was associated with a positive CE result (OR = 15.56, 95% CI = 5.95–40.67, P < 0.001), as were positive FAEE results (OR = 2.437, 95% CI = 1.21–4.87, P = 0.012). FAEE samples negative for excessive drinking (<0.5 ng/mg) were 95.18% of the time also negative for CE, indicating a very low rate of false positivity. Specificity increased when including social drinking/non-drinking population, indicating that 97.28% of FAEE values < 0.2 ng/mg were also negative for CE. This indicates that the rate of false positivity decreases even further for non-drinking individual. In addition, with a PPV of 0.66, if a positive CE is detected, then the sample is 66% likely to be positive for FAEE (≥0.5 ng/mg). 37 Agius et al./2012 3464 A *6 and 662 n.r. Scalp hair (segmentation 0–3 cm and 3–12 cm) EtG Statistical and stability studies provide evidence that the detection of EtG shall not be limited to 3-cm-long hair lenghts. The higher number of positivity in samples shorter than 3 cm supports incorporation of EtG from sweat after recent intake. 38 Albermann et al./2014 73 (T + SD) Scalp hair (proximal segment 0–3 cm) ∑FAEE The method was successfully validated according to the guidelines of the GTFCh, showing good linearity, precision and accuracy, recovery and stability. Comparison of FAEE and EtG (4–10 pg/mg previously detected with other techniques) in 73 authentic hair samples showed coincidences > 70%, but a plot of FAEE concentrations against EtG concentrations (R2 = 0.008) showed no linear correlation between the 2 markers. However, FAEE analysis can be performed to confirm EtG results even for abstinence tests. 39 Suesse et al./2015 157 n.r. (45 M, 112 F) Scalp hair (segmentation for both 0–3 cm and 0–6 cm) ∑FAEE The interpretation is complicated by inter-individual variability in formation and incorporation; the variable lenght of hair contributes to the uncertainty. The disagreement of the results depends mainly on the cosmetic treatment that can increase FAEE and decrease EtG. The redefinition of hair lenght for both markers is suggested. 40 Dumitrascu et al./2018 10 n.r. (5 F and 5 M) Scalp hair (segmentation 0–3 cm) ∑FAEE Various cosmetics were evaluated in order to understand their influence on FAEE: alcohol based hairspray increase FAEE levels in a highly variable manner. Quantification performed both with ethyl palmitate alone or the sum of the 4 esters produce the same results. A Alcoholics in withdrawal treatment *1 Samples collected in context of driving ability, for workplace testing and child custody B Alcoholics with a known history of alcohol abuse *2 Samples mainly collected from parents, in child protection cases C Fatalities with a history of excessive alcohol consumption confirmed by pathologic findings *3 Samples collected by mothers with suspicious alcohol abuse and children (mother–infant dyads) LD Light drinkers *4 The amount of neonatal hair has been reported scarce SD Self-reported social drinkers *5 No pre-selection of the cases with respect to alcohol abuse T Teetotallers *6 Samples tested for abstinence to regain revoked driving license ∑FAEE Ethyl myristate, ethyl oleate, ethyl palmitate, ethyl stearate (referred as their sum) n.r. not reported. Open in new tab Table 2 Solid-phase microextraction technique for the identification of VOCs for the human odor profile Ref. Author/year Population Matrix Results 41 Lorenzo et al./2003 n.r. Human remains Trimethylamine, 1-pentanol, hexanal, butanoic acid, pentanoic acid, heptanal, benzaldehyde, 2-pentyl furan, dimethyl disulfide, hexanoic acid, heptanoic acid, nonanoic acid and octanoic acid were identified in human decomposition fluids. 42 Curran et al./2005 8 subjects (4 M and 4 F) Axillary sweat The majority of compounds were aldehydes, alcohols, alkanes and esters in different patterns in males and females. Respect to previous literature: 6-methyl-5-hepten-2-one is an axillary sweat human compound; 2-nonenal is not related to individual ≥40 y old. 43 Curran et al./2007 60 subjects (30 M, 30 F) Hands Method developed to extract hand odor compounds; 63 VOCs were identified and 50 molecules had low frequence. Spearman correlation coefficient was able to distinguish individuals based on the VOCs identified. 44 Hoffman et al./2009 14 tissue samples Human remains (blood clot, blood clot-placenta, blood, muscle, testicle, skin, body fat, adipocere, bone, teeth) 33 VOCs were investigated. 7 chemical classes (acid esters, alcohols, aldehydes, halogens, aromatic hydrocarbons, ketones, sulfides). p-Xylene was present in all the samples except teeth. The 2 major classes observed were alcohols and aldehydes. 45 Hudson et al./2009 6 subjects Sorbent materials and storage containers Glass had been determined to be the optimal storage material; scent profile change during time and the sorbent material should not be exposed to UVA/UVB lights for the possible oxidation reactions resulting in the formation of VOC such as aldehydes and methyl esters with consequent alteration of the profile. 46 Curran et al./2010 10 subjects (5 M, 5 F) Hands A total of 24 compounds were considered part of a “primary odor profile”; the predominance of the functional groups has shifted with alkanes and aldehydes comprising 50% of the compounds considered. 2-Nonenal was detected in 2/10 subjects from 17 to 28 y old. 47 Prada et al./2011 6 subjects (3 M, 3 F) Textiles STU-100 used as non-contact odor collection. Comparison of contact and non-contact human scent collection procedures across various textiles. Contact sampling with cotton material demonstrated the greatest reproducibility with high discrimination. 48 Kusano et al./2011 1 M Blood, breath, urine, buccal cells A direct comparison between VOC chromatograms of blood, breath, buccal swabs and urine taken from the same individual demonstrated differences in VOC profile in respect to each matrix; however, the scent profiling ability of the different biological specimens was noted. 49 DeGreeff et al./2011 21 deceased bodies Human remains Sample collection was performed by STU-100. 13/50 compounds identified were common among human remains collected from 2 different locations (morgue and cremation service). 50 Brown et al./2013 20 subjects Hand, hair, fingernails, saliva VOC collection is performed according to Curran et al. (2007) (43). 42 VOCs were detected (acids, alcohols, aldehydes, esters, hydrocarbons, ketones, heterocyclic compounds). Hand odor samples produced 24 compounds, fingernail 18, hair 23. High cross-correlation between hair and nails, no correlation between hand and saliva. 51 Kusano et al./2013 31 healthy individuals Hand, breath, blood, urine, saliva VOC collection performed according to Curran et al. (2007) (43).Quali-quantitative VOC chromatograms comparison from different biological specimens (hand odor, buccal swabs, breath urine and blood) of the same individual showed differences that can be used for matching purposes. Results from the 6-mo study of the VOC profiles of 2 subjects support the individual odor hypothesis that human scent is stable over time and distinguishable between individuals. 52 Choi et al./2014 7 subjects Sweat (after induced exercises) from face and upper part of body It is not clear the collection method used. Many factors (illness, physical fitness, age…) alter an individual’s VOC profile due to their effects on apocrine glands. A total of 574 components (alcohols, aldehydes, aliphatics/aromatics, carboxylic acids, esters, ketones, amides/amines, thio/thioesters, oxide, sulfides, nitro) in human sweat were identified. 1-Tridecanol, 1,3-bis(1,1-dimethyl ethyl)-benzene, 4,4′-(1-methylethylidene) bis-phenol and 7-acetyl-6-ethyl-1,1,4,4,-tetramethyltetraline were common components in all donor’s sweat volatile samples. Age-related specific compounds were also detected. 53 Forbes et al./2014 2 F: 25–35 y old Blood VOC profile of fresh (0–48 h) and degraded blood (1–6 wk) showed differences in chemical patterns and concentration related to time and storage conditions. 54 Rust et al./2016 A blood donor Blood Blood samples on non-porous (aluminum) and porous surfaces (cotton) aged over time (periodically examined from day 0 to 12 mo) showed the VOC profile changing based on the surface type and age of the sample. 55 Caraballo et al./2016 20 subjects (10 F, 10 M) Hands STU-100 was positively used to transfer VOC from an object/person onto a collection material. Over 20 VOCs consisting of 5 different functional groups were identified. 56 Chilcote et al./2018 1 F Blood wood and concrete surfaces Samples surfaces weathered in an outdoor environment over a 3-mo period detected up to 59 d with the majority of the odor profile consisting of nonanal, heptanal and 2-pentyl furan. Ref. Author/year Population Matrix Results 41 Lorenzo et al./2003 n.r. Human remains Trimethylamine, 1-pentanol, hexanal, butanoic acid, pentanoic acid, heptanal, benzaldehyde, 2-pentyl furan, dimethyl disulfide, hexanoic acid, heptanoic acid, nonanoic acid and octanoic acid were identified in human decomposition fluids. 42 Curran et al./2005 8 subjects (4 M and 4 F) Axillary sweat The majority of compounds were aldehydes, alcohols, alkanes and esters in different patterns in males and females. Respect to previous literature: 6-methyl-5-hepten-2-one is an axillary sweat human compound; 2-nonenal is not related to individual ≥40 y old. 43 Curran et al./2007 60 subjects (30 M, 30 F) Hands Method developed to extract hand odor compounds; 63 VOCs were identified and 50 molecules had low frequence. Spearman correlation coefficient was able to distinguish individuals based on the VOCs identified. 44 Hoffman et al./2009 14 tissue samples Human remains (blood clot, blood clot-placenta, blood, muscle, testicle, skin, body fat, adipocere, bone, teeth) 33 VOCs were investigated. 7 chemical classes (acid esters, alcohols, aldehydes, halogens, aromatic hydrocarbons, ketones, sulfides). p-Xylene was present in all the samples except teeth. The 2 major classes observed were alcohols and aldehydes. 45 Hudson et al./2009 6 subjects Sorbent materials and storage containers Glass had been determined to be the optimal storage material; scent profile change during time and the sorbent material should not be exposed to UVA/UVB lights for the possible oxidation reactions resulting in the formation of VOC such as aldehydes and methyl esters with consequent alteration of the profile. 46 Curran et al./2010 10 subjects (5 M, 5 F) Hands A total of 24 compounds were considered part of a “primary odor profile”; the predominance of the functional groups has shifted with alkanes and aldehydes comprising 50% of the compounds considered. 2-Nonenal was detected in 2/10 subjects from 17 to 28 y old. 47 Prada et al./2011 6 subjects (3 M, 3 F) Textiles STU-100 used as non-contact odor collection. Comparison of contact and non-contact human scent collection procedures across various textiles. Contact sampling with cotton material demonstrated the greatest reproducibility with high discrimination. 48 Kusano et al./2011 1 M Blood, breath, urine, buccal cells A direct comparison between VOC chromatograms of blood, breath, buccal swabs and urine taken from the same individual demonstrated differences in VOC profile in respect to each matrix; however, the scent profiling ability of the different biological specimens was noted. 49 DeGreeff et al./2011 21 deceased bodies Human remains Sample collection was performed by STU-100. 13/50 compounds identified were common among human remains collected from 2 different locations (morgue and cremation service). 50 Brown et al./2013 20 subjects Hand, hair, fingernails, saliva VOC collection is performed according to Curran et al. (2007) (43). 42 VOCs were detected (acids, alcohols, aldehydes, esters, hydrocarbons, ketones, heterocyclic compounds). Hand odor samples produced 24 compounds, fingernail 18, hair 23. High cross-correlation between hair and nails, no correlation between hand and saliva. 51 Kusano et al./2013 31 healthy individuals Hand, breath, blood, urine, saliva VOC collection performed according to Curran et al. (2007) (43).Quali-quantitative VOC chromatograms comparison from different biological specimens (hand odor, buccal swabs, breath urine and blood) of the same individual showed differences that can be used for matching purposes. Results from the 6-mo study of the VOC profiles of 2 subjects support the individual odor hypothesis that human scent is stable over time and distinguishable between individuals. 52 Choi et al./2014 7 subjects Sweat (after induced exercises) from face and upper part of body It is not clear the collection method used. Many factors (illness, physical fitness, age…) alter an individual’s VOC profile due to their effects on apocrine glands. A total of 574 components (alcohols, aldehydes, aliphatics/aromatics, carboxylic acids, esters, ketones, amides/amines, thio/thioesters, oxide, sulfides, nitro) in human sweat were identified. 1-Tridecanol, 1,3-bis(1,1-dimethyl ethyl)-benzene, 4,4′-(1-methylethylidene) bis-phenol and 7-acetyl-6-ethyl-1,1,4,4,-tetramethyltetraline were common components in all donor’s sweat volatile samples. Age-related specific compounds were also detected. 53 Forbes et al./2014 2 F: 25–35 y old Blood VOC profile of fresh (0–48 h) and degraded blood (1–6 wk) showed differences in chemical patterns and concentration related to time and storage conditions. 54 Rust et al./2016 A blood donor Blood Blood samples on non-porous (aluminum) and porous surfaces (cotton) aged over time (periodically examined from day 0 to 12 mo) showed the VOC profile changing based on the surface type and age of the sample. 55 Caraballo et al./2016 20 subjects (10 F, 10 M) Hands STU-100 was positively used to transfer VOC from an object/person onto a collection material. Over 20 VOCs consisting of 5 different functional groups were identified. 56 Chilcote et al./2018 1 F Blood wood and concrete surfaces Samples surfaces weathered in an outdoor environment over a 3-mo period detected up to 59 d with the majority of the odor profile consisting of nonanal, heptanal and 2-pentyl furan. n.r., not reported. Open in new tab Table 2 Solid-phase microextraction technique for the identification of VOCs for the human odor profile Ref. Author/year Population Matrix Results 41 Lorenzo et al./2003 n.r. Human remains Trimethylamine, 1-pentanol, hexanal, butanoic acid, pentanoic acid, heptanal, benzaldehyde, 2-pentyl furan, dimethyl disulfide, hexanoic acid, heptanoic acid, nonanoic acid and octanoic acid were identified in human decomposition fluids. 42 Curran et al./2005 8 subjects (4 M and 4 F) Axillary sweat The majority of compounds were aldehydes, alcohols, alkanes and esters in different patterns in males and females. Respect to previous literature: 6-methyl-5-hepten-2-one is an axillary sweat human compound; 2-nonenal is not related to individual ≥40 y old. 43 Curran et al./2007 60 subjects (30 M, 30 F) Hands Method developed to extract hand odor compounds; 63 VOCs were identified and 50 molecules had low frequence. Spearman correlation coefficient was able to distinguish individuals based on the VOCs identified. 44 Hoffman et al./2009 14 tissue samples Human remains (blood clot, blood clot-placenta, blood, muscle, testicle, skin, body fat, adipocere, bone, teeth) 33 VOCs were investigated. 7 chemical classes (acid esters, alcohols, aldehydes, halogens, aromatic hydrocarbons, ketones, sulfides). p-Xylene was present in all the samples except teeth. The 2 major classes observed were alcohols and aldehydes. 45 Hudson et al./2009 6 subjects Sorbent materials and storage containers Glass had been determined to be the optimal storage material; scent profile change during time and the sorbent material should not be exposed to UVA/UVB lights for the possible oxidation reactions resulting in the formation of VOC such as aldehydes and methyl esters with consequent alteration of the profile. 46 Curran et al./2010 10 subjects (5 M, 5 F) Hands A total of 24 compounds were considered part of a “primary odor profile”; the predominance of the functional groups has shifted with alkanes and aldehydes comprising 50% of the compounds considered. 2-Nonenal was detected in 2/10 subjects from 17 to 28 y old. 47 Prada et al./2011 6 subjects (3 M, 3 F) Textiles STU-100 used as non-contact odor collection. Comparison of contact and non-contact human scent collection procedures across various textiles. Contact sampling with cotton material demonstrated the greatest reproducibility with high discrimination. 48 Kusano et al./2011 1 M Blood, breath, urine, buccal cells A direct comparison between VOC chromatograms of blood, breath, buccal swabs and urine taken from the same individual demonstrated differences in VOC profile in respect to each matrix; however, the scent profiling ability of the different biological specimens was noted. 49 DeGreeff et al./2011 21 deceased bodies Human remains Sample collection was performed by STU-100. 13/50 compounds identified were common among human remains collected from 2 different locations (morgue and cremation service). 50 Brown et al./2013 20 subjects Hand, hair, fingernails, saliva VOC collection is performed according to Curran et al. (2007) (43). 42 VOCs were detected (acids, alcohols, aldehydes, esters, hydrocarbons, ketones, heterocyclic compounds). Hand odor samples produced 24 compounds, fingernail 18, hair 23. High cross-correlation between hair and nails, no correlation between hand and saliva. 51 Kusano et al./2013 31 healthy individuals Hand, breath, blood, urine, saliva VOC collection performed according to Curran et al. (2007) (43).Quali-quantitative VOC chromatograms comparison from different biological specimens (hand odor, buccal swabs, breath urine and blood) of the same individual showed differences that can be used for matching purposes. Results from the 6-mo study of the VOC profiles of 2 subjects support the individual odor hypothesis that human scent is stable over time and distinguishable between individuals. 52 Choi et al./2014 7 subjects Sweat (after induced exercises) from face and upper part of body It is not clear the collection method used. Many factors (illness, physical fitness, age…) alter an individual’s VOC profile due to their effects on apocrine glands. A total of 574 components (alcohols, aldehydes, aliphatics/aromatics, carboxylic acids, esters, ketones, amides/amines, thio/thioesters, oxide, sulfides, nitro) in human sweat were identified. 1-Tridecanol, 1,3-bis(1,1-dimethyl ethyl)-benzene, 4,4′-(1-methylethylidene) bis-phenol and 7-acetyl-6-ethyl-1,1,4,4,-tetramethyltetraline were common components in all donor’s sweat volatile samples. Age-related specific compounds were also detected. 53 Forbes et al./2014 2 F: 25–35 y old Blood VOC profile of fresh (0–48 h) and degraded blood (1–6 wk) showed differences in chemical patterns and concentration related to time and storage conditions. 54 Rust et al./2016 A blood donor Blood Blood samples on non-porous (aluminum) and porous surfaces (cotton) aged over time (periodically examined from day 0 to 12 mo) showed the VOC profile changing based on the surface type and age of the sample. 55 Caraballo et al./2016 20 subjects (10 F, 10 M) Hands STU-100 was positively used to transfer VOC from an object/person onto a collection material. Over 20 VOCs consisting of 5 different functional groups were identified. 56 Chilcote et al./2018 1 F Blood wood and concrete surfaces Samples surfaces weathered in an outdoor environment over a 3-mo period detected up to 59 d with the majority of the odor profile consisting of nonanal, heptanal and 2-pentyl furan. Ref. Author/year Population Matrix Results 41 Lorenzo et al./2003 n.r. Human remains Trimethylamine, 1-pentanol, hexanal, butanoic acid, pentanoic acid, heptanal, benzaldehyde, 2-pentyl furan, dimethyl disulfide, hexanoic acid, heptanoic acid, nonanoic acid and octanoic acid were identified in human decomposition fluids. 42 Curran et al./2005 8 subjects (4 M and 4 F) Axillary sweat The majority of compounds were aldehydes, alcohols, alkanes and esters in different patterns in males and females. Respect to previous literature: 6-methyl-5-hepten-2-one is an axillary sweat human compound; 2-nonenal is not related to individual ≥40 y old. 43 Curran et al./2007 60 subjects (30 M, 30 F) Hands Method developed to extract hand odor compounds; 63 VOCs were identified and 50 molecules had low frequence. Spearman correlation coefficient was able to distinguish individuals based on the VOCs identified. 44 Hoffman et al./2009 14 tissue samples Human remains (blood clot, blood clot-placenta, blood, muscle, testicle, skin, body fat, adipocere, bone, teeth) 33 VOCs were investigated. 7 chemical classes (acid esters, alcohols, aldehydes, halogens, aromatic hydrocarbons, ketones, sulfides). p-Xylene was present in all the samples except teeth. The 2 major classes observed were alcohols and aldehydes. 45 Hudson et al./2009 6 subjects Sorbent materials and storage containers Glass had been determined to be the optimal storage material; scent profile change during time and the sorbent material should not be exposed to UVA/UVB lights for the possible oxidation reactions resulting in the formation of VOC such as aldehydes and methyl esters with consequent alteration of the profile. 46 Curran et al./2010 10 subjects (5 M, 5 F) Hands A total of 24 compounds were considered part of a “primary odor profile”; the predominance of the functional groups has shifted with alkanes and aldehydes comprising 50% of the compounds considered. 2-Nonenal was detected in 2/10 subjects from 17 to 28 y old. 47 Prada et al./2011 6 subjects (3 M, 3 F) Textiles STU-100 used as non-contact odor collection. Comparison of contact and non-contact human scent collection procedures across various textiles. Contact sampling with cotton material demonstrated the greatest reproducibility with high discrimination. 48 Kusano et al./2011 1 M Blood, breath, urine, buccal cells A direct comparison between VOC chromatograms of blood, breath, buccal swabs and urine taken from the same individual demonstrated differences in VOC profile in respect to each matrix; however, the scent profiling ability of the different biological specimens was noted. 49 DeGreeff et al./2011 21 deceased bodies Human remains Sample collection was performed by STU-100. 13/50 compounds identified were common among human remains collected from 2 different locations (morgue and cremation service). 50 Brown et al./2013 20 subjects Hand, hair, fingernails, saliva VOC collection is performed according to Curran et al. (2007) (43). 42 VOCs were detected (acids, alcohols, aldehydes, esters, hydrocarbons, ketones, heterocyclic compounds). Hand odor samples produced 24 compounds, fingernail 18, hair 23. High cross-correlation between hair and nails, no correlation between hand and saliva. 51 Kusano et al./2013 31 healthy individuals Hand, breath, blood, urine, saliva VOC collection performed according to Curran et al. (2007) (43).Quali-quantitative VOC chromatograms comparison from different biological specimens (hand odor, buccal swabs, breath urine and blood) of the same individual showed differences that can be used for matching purposes. Results from the 6-mo study of the VOC profiles of 2 subjects support the individual odor hypothesis that human scent is stable over time and distinguishable between individuals. 52 Choi et al./2014 7 subjects Sweat (after induced exercises) from face and upper part of body It is not clear the collection method used. Many factors (illness, physical fitness, age…) alter an individual’s VOC profile due to their effects on apocrine glands. A total of 574 components (alcohols, aldehydes, aliphatics/aromatics, carboxylic acids, esters, ketones, amides/amines, thio/thioesters, oxide, sulfides, nitro) in human sweat were identified. 1-Tridecanol, 1,3-bis(1,1-dimethyl ethyl)-benzene, 4,4′-(1-methylethylidene) bis-phenol and 7-acetyl-6-ethyl-1,1,4,4,-tetramethyltetraline were common components in all donor’s sweat volatile samples. Age-related specific compounds were also detected. 53 Forbes et al./2014 2 F: 25–35 y old Blood VOC profile of fresh (0–48 h) and degraded blood (1–6 wk) showed differences in chemical patterns and concentration related to time and storage conditions. 54 Rust et al./2016 A blood donor Blood Blood samples on non-porous (aluminum) and porous surfaces (cotton) aged over time (periodically examined from day 0 to 12 mo) showed the VOC profile changing based on the surface type and age of the sample. 55 Caraballo et al./2016 20 subjects (10 F, 10 M) Hands STU-100 was positively used to transfer VOC from an object/person onto a collection material. Over 20 VOCs consisting of 5 different functional groups were identified. 56 Chilcote et al./2018 1 F Blood wood and concrete surfaces Samples surfaces weathered in an outdoor environment over a 3-mo period detected up to 59 d with the majority of the odor profile consisting of nonanal, heptanal and 2-pentyl furan. n.r., not reported. Open in new tab Discussion Headspace solid-phase microextraction, GC–MS technique for the identification of markers of alcohol abuse Our review shows that the HS-SPME technique applied to the detection of objective markers of alcohol abuse was evaluated for the first time by Pragst et al. (13); to demonstrate its usefulness in the analysis of ethyl esters of fatty acids in the hair of alcoholics, they developed a method for extraction and GC–MS identification; successively, they improved the technique, demonstrating that the sum of palmitic, myristic, stearic and oleic ethyl esters (∑FAEE, fatty acid ethyl esters) can be used to discriminate among people with alcoholism, those who drink socially and those who abstain from drinking (Table 1). This review reports SPME in diagnosing chronically excessive alcohol abuse by exploring the presence of fatty acid ethyl esters (FAEEs) and ethyl glucuronide (EtG), mainly on hair samples, sometimes collected during driving ability examination and child custody cases. EtG is a non-volatile, water-soluble metabolite formed in the liver via conjugation of ethanol with activated glucuronate; the semivolatile FAEEs are products of non-oxidative ethanol metabolism, mainly originating from the esterification of fatty acids with ethanol. These compounds can be detected in biological samples, and they are durably deposited in the hair, which enables taking advantage of the long diagnostic time window of this sample material, making them suitable long-term markers of chronically elevated alcohol consumption. Twenty-eight of the 133 papers were selected, including seven papers identified in the references. Of those 28 papers, 25 were about SPME GC–MS technique for the identification of biological markers of chronic alcohol abuse; three of the 28 papers examined the same technique to detect beverage flavoring compounds as markers for intake of special alcoholic beverages for verification of drinking claims. Since 2000, the developed technique (always HS-SPME GC–MS using a 65-μm PDMS/divinylbenzene [DVB] fiber) did not show significant changes over time when analyzing FAEEs in hair (13–18, 20–22, 26, 30–32, 36, 38–40); some diversification has been described for different matrices (skin, meconium). EtG needs derivatization and requires more sensitive techniques; hence, its quantification by HS-SPME is performed by a different kind of fiber (a 75-μm CAR/PDMS fiber), and gas chromatography tandem mass spectrometry (GC–MS-MS) is used for its detection (29, 37). Regarding chronic alcohol abuse, most of the papers (23/25) investigated FAEEs (mainly as the sum of four ethyl esters); one of the papers also detected cocaethylene (CE). Additionally, two of the 25 studies performed an investigation on EtG (29, 37). The possible use of SPME to detect volatile compounds, such as ethanol itself, was also discussed (13). The study population usually involved people who abstain from drinking (sometimes children), those who drink socially, and those with alcoholism; in eight of the 25 studies, dead people were also considered (13–19, 21). FAEEs were mainly detected on the scalp hair (17/23), while in three of the 17 papers, different sites of the body were also explored (14, 17, 31). The analysis of one or more samples collected from various body sites gave different concentrations in the same subject (17); nevertheless, body hair can be collected when head hair is not available, as suggested by the Society of Hair Testing (SoHT) (2). Moreover, body hair can be used to confirm scalp hair analysis (17) because FAEEs in scalp hair may also be present for the use of ethanol-containing hair care products (e.g., hairspray or hair lotions), which may lead to false-positive FAEE detection. The cosmetic interferences were explored by some researchers (16, 39, 40), demonstrating that the regular use of alcohol-containing cosmetic products can increase the concentration of FAEEs detected in the hair, leading to false-positive results. The papers of the present review discussing segmentation (10/23) highlighted that FAEE concentration was different with respect to the segments analyzed (15, 16, 18, 22, 30–32, 38–40). The variable length of the proximal segment examined between 3 cm (38–40) and 6 cm (18, 21, 26, 30, 31, 39) additionally contributes to the uncertainty of interpretation caused by biological variability and hair cosmetics. Moreover, Morini et al. (32) examined 0- to 9-cm proximal segments from mothers. De Giovanni et al. (22) performed segmentation only when the length was higher than 5 cm; in some cases, full-length scalp hair was examined (16, 17), while in four cases, this information was not reported (13, 14, 20, 36). These results encouraged SoHT (2) to suggest that segmentation of the hair shaft may provide additional information with respect to the interpretation of biomarkers, and the results should be interpreted with caution when samples are less than 3 cm or greater than 6 cm. Routes of incorporation of FAEEs are discussed by Auwarter et al. (15) and Suesse et al. (39); they underlined that incorporation of FAEEs into hair arises mainly through sebum. For this reason, inter-individual differences in the activity of the sebum glands may influence FAEE concentration (15). To minimize inter-individual differences, the ratio of FAEEs/squalene (SQ) in sebum was examined (19, 21). In fact, sebum contains approximately 10–20% SQ; thus, it can be used as a natural reference compound. Abstinence was considered by 12 of the 25 papers both when samples examined were described or in the conclusion sections (15, 17, 19, 21, 26, 28, 30, 31, 35, 37–39); comparisons with self-reported data are available for eight of the 25 studies (15, 18–21, 28, 31, 37), but no correlation was found. The correlation between FAEE and EtG markers in scalp hair has been evaluated (18, 30, 38, 39), although in these studies, EtG was not quantified by the use of SPME, and no correspondence was described (18, 38). However, the combined use of both biological markers is reported to increase accuracy in the diagnosis of abstinence, social or excessive drinking when detected in meconium (33). In recent years, FAEEs and EtG have also been used as biomarkers for alcohol abuse in pregnancy, often using meconium as a reliable biological matrix for establishing gestational ethanol exposure. Whether EtG in maternal hair measured during the three trimesters of pregnancy correlated with EtG and FAEEs in neonatal meconium was also investigated (33). The results confirm that FAEEs and EtG in meconium are the best biomarkers to assess in utero exposure to maternal alcohol misuse. The retrospective detection of alcohol consumption during pregnancy can help with the diagnosis of fetal alcohol syndrome (26, 32). According to Pragst et al. (146), FAEEs and/or EtG in the hair of the mothers have been determined. The validation of the developed analytical method was performed by almost all authors, except when the method was proposed by Pragst et al. (13) and when the aim of the study was the evaluation of various cosmetics to explain their influence on FAEE production (40). Quantitative detection was always performed except in three papers (13, 16, 28), in particular when the main purpose of the study was to test the presence of FAEEs in care products (16). Most of the papers evaluated the cutoff for the interpretation of results to distinguish alcoholics with respect to social drinkers. Only six of the 25 papers (20, 26, 28, 30, 31, 36) performed statistical evaluation of the method (receiver operating characteristic [ROC] evaluation or at least sensitivity and specificity of the marker). Solid-phase microextraction technique for the identification of Volatile Organic Compounds for the human odor profile (Table 2) The detection of human odor is of particular interest in the forensic field for the localization and identification of living people as well as dead bodies, usually performed by means of trained dogs. Dogs are said to recognize scents from different parts of the body, although this type of forensic evidence is still questioned in court (147). Several studies have been carried out on the development of SPME as an analytical strategy to characterize human odor, mainly to give relevance to the information obtained from dogs. Our systematic review selected a total of 16 of the 133 papers, and the first SPME application in this context dates back to 2003 (41), regarding human remains. Later, SPME applied to human remains was implemented by Hoffmann et al. (44) using blood clot, placenta, blood, muscle, testicle, skin, body fat, adipocere, bone and teeth collected from deceased people and by De Greef et al. (49), who sampled 21 deceased bodies without any contact using a device called “Scent Transfer Unit” (STU-100). Various biological matrices from living people have been sampled, including sweat from armpits (42), from the face and from the upper part of the body (52); hands alone (43, 46, 55) or together other body areas (50, 51) were also investigated. Other authors collected blood (53, 54, 56) and blood and urine together (48, 51) for odor profile identification. Browns et al. (50) showed that hand odor samples produce a higher number of compounds than hair, saliva or fingernails. Kusano et al. (51) reported that chromatogram comparison from different biological specimens (hand odor, buccal swabs, breath, urine and blood) of the same individual shows differences in the presence of VOCs; this characteristic can then be used for matching purposes. The results from the 6-mo study of the VOC profiles of two subjects support the individual odor hypothesis that the human scent is stable over time and distinguishable among individuals (51). Focusing on the odor of living humans, different groups of chemical compounds have been identified: acids, alcohols, aldehydes, esters, hydrocarbons, ketones, heterocyclic compounds and even sulfur-containing compounds in armpit samples (52). Many factors have been considered to influence the qualitative and quantitative composition of human scents, such as exogenous (soap or deodorant) and endogenous factors (52), sex, method of collection (47, 48, 54), sorbent material and storage containers (45). With respect to the storage material, glass was determined to be optimal (45), although the scent profile changed over time. Sorbent matter exposed to UVA/UVB lights (45) produces oxidation reactions resulting in the formation of VOCs, such as aldehydes and methyl esters, with consequent alteration of the profile. The stability of chemical compounds was observed to be over 6 months (51) with variations between fresh and degraded blood (53). Some authors asked the examined subjects to perform some exercises before sampling to increase the production of sweat (52). Many different components were identified in all the matrices investigated, and a total of 574 VOCs were recognized in human sweat (52), although many factors (such as illness, physical fitness, age) may alter an individual’s VOC profile due to their effects on apocrine glands. 1-Tridecanol, 1,3-bis(1,1-dimethylethyl)-benzene, 4,4′-(1-methylethylidene) bisphenol and 7-acetyl-6-ethyl-1,1,4,4-tetramethyltetraline were recognized as common components in all donors’ sweat volatile samples (52). It has also been noted in comparison with previous literature that 6-methyl-5-hepten-2-one was observed in axillary sweat and that 2-nonenal is not related to individuals older than 40 years of age (42, 52). Two types of sampling methods have been described to collect “human odor.” Most studies used sampling by direct contact consisting of placing an adsorbent phase directly on the skin of the subject to collect compounds. The sampling material, whatever the composition, has to be pre-treated to eliminate contaminants. Curran et al. (42) showed that the analysis of blank gauzes may indicate the presence of several key odor compounds, including decanal and nonanal, because of their initial presence in the headspace of the absorbent material. Two studies performed indirect sampling without contact, using air suction from human remains (49) and from skin of the hands (55) by the STU-100. The device is a vacuum pump connected to a piece of gauze that allows the collection of VOC compounds. The same device was also employed for the non-contact odor collection from textiles (47). Prada et al. (47) compared contact and non-contact human scent sampling across various textiles and showed that cotton material sampling has the greatest reproducibility with high discrimination. Caraballo et al. (55) identified 20 VOCs consisting of five different functional groups from the hands of 20 subjects. Degreeff et al. (49) identified 13 of the 50 compounds common among human remains collected from two locations (morgue and cremation service). Post-mortem VOC analysis identified a wide range of compounds, including alcohols, aldehydes, aromatics, carboxylic acids, esters, ethers, hydrocarbons, ketones, nitrogen-containing compounds and sulfur-containing compounds, where the two major classes described were alcohols and aldehydes. Individual post-mortem VOCs also demonstrated a dynamic profile even after days, weeks or months of detection (41, 44, 49). The decomposition odor is important to understand in the context of missing persons or victims of homicide who may have been missing for a long time since death. VOC markers for the evaluation of post-mortem interval (PMI) (84) were also detected by investigating cadaveric gases. Regarding the analytical characteristics of the technique employed in the studies reviewed, it can be highlighted that SPME was always used in the HS mode, although each researcher employed different kinds of fibers, that is, PDMS/DVB, PA or, more frequently, DVB/CAR on PDMS. GC–MS was the technique of choice in all the papers; however, second-dimensional GC coupled with time-of-flight MS (GCxGC TOFMS) was employed by Choi et al. (52) on sweat matrices, Rust et al. (54) on blood and Chilcote et al. (56) on blood from wood and concrete surfaces. In seven of the 16 studies, pre-treatment of samples was performed (42, 43, 45, 46, 48, 50, 51) sometimes using supercritical fluid extraction (42, 43, 45, 46). The validation of the method was never performed, probably due to the difficulty of detecting and quantitating so many compounds, considering the most important comparison of qualitative profiles. Only Kusano et al. (48) determined the limit of detection (LOD) and the limit of quantitation (LOQ) for each detected compound commonly found with respect to each matrix. Nevertheless, many studies performed correlations and/or statistical evaluation, and Spearman’s rank correlation was sometimes used to discriminate samples (47, 50, 51). Statistical evaluation was performed using principal component analysis (PCA) (47, 49–51, 56), hierarchical cluster analysis (HCA) (56) and three-dimensional covariance mapping (45). Solid-phase microextraction in forensic toxicology investigation (Tables 3–5) Table 3 Solid-phase microextraction application in the forensic toxicology investigation concerning death Ref. Author/year Population Matrix Substances identified Results 57 Nagasawa et al./1996 3 M and 2 F (2 brain injuries; 3 MA poisoning) Blood MA and AP 3 MA poisoning: range 3.23–5.31 μmol/100 g. MA and AP were detected in 4 out of the 5 cases. 58 Namera et al./1997 A 50-y-old M (acute intoxication) Blood Malathion Malathion detected in blood: left heart 41 μg/g; right heart 5.4 μg/g. 59 Brewer et al./1997 2 victims (traffic fatalities) Urine, gastric content, blood VOC (methylene chloride, light hydrocarbons) Case 1: urine concentration of methylene chloride at the time of the accident (23 mg/L) was considered toxic because greater than expected from occupational exposure; blood was not available. Case 2: charcoal lighter fluid was detected in gastric content but not in blood sample. Circumstantial data support a suicide as manner of death due to traffic accident. 60 Watanabe et al./1998 One victim (local anaestetic poisoning) Blood 5 local anesthetics (prilocaine, lidocaine, mepivacaine, bupivacaine, dibucaine) Mepivacaine and lidocaine were detected in the left and right heart blood samples at concentrations of 18.6 and 15.8 mg/g for mepivacaine and 0.14 and 0.17 mg/g for lidocaine, respectively. Mepivacaine concentration in blood was higher than therapeutic levels. 61 Namera et al./1998 1 victim (acute intoxication) Blood Tetracyclic antidepressants (setiptiline, mianserin, maprotiline) Setiptiline was found in the left and right heart blood samples at concentrations of 1.77 and 0.78 μg/g, respectively. It was about 200 times higher than therapeutic level (1.0–5.0 ng/g). 62 Sporkert et al./2000 49 victims (drug fatalities) Hair (also segmental analysis) Lidocaine 32/49 cases were positive to lidocaine with concentrations between 0.4 and 675 ng/mg (18/32 were > 10 ng/mg; 4/32 were > 150 ng/mg). 63 Dehon et al./2000 A 45-y-old woman (acute intoxication) Blood, urine, tissue homogenates Tetrachloroethylene (C2Cl4) and TCE C2Cl4 and TCE were found in blood and urine of the woman who died 7 d after admission to the hospital. Tissue concentration of TCE was higher in liposoluble tissues (brain); C2Cl4 was found in all post-mortem samples. 64 Musshoff et al./2000 A 75-y-old man and a 82-y-old woman (double homicide by smothering) Blood, urine, bile, tissues Halothane First toxicological analysis was performed 3-mo post-mortem with GC and after 15 mo with HS-SPME–GC–MS. Higher concentration in the brain and the lowest in liver and bile. Both the techniques gave similar analytical results. Halothane concentrations were consistent with intoxication. Manner of death was homicide by forced smothering of halothane. 65 Namera et al./2000 A 25-y-old woman (suicide by intoxication) Blood AP, MA, fenfluramine The developed HS–SPME method was optimized for the simultaneous determination of fenfluramine and amphetamines in human blood using the on-column derivatization method. Fenfluramine was detected in the blood sample after 4-h hospitalization at fatal concentration (7.7 μg/g). The death was due to suicide. 66 Sporkert et al./2000 26 victims (drug intoxication) 3-cm proximal hair Methadone, EDDP, EMDP Methadone was detected in 19 cases, while EDDP was found in 17 cases. EMPD was detected in 2/7 methadone positivities. 67 Liu et al./2001 A 43-y-old cadaver (n.r.) Hair AP, MA The method was applied to 5 strands of hair from a crime-suspected person and the results were compared with a usual method. MA concentration range from proximal (about 42 pmol/cm) to distal (about 3 pmol/cm). 68 Tranthim-Fryer et al./2001 3 M, 1 F (VOC-related deaths) Blood tissues VOC (butane, halotane, toluene and petrol residues) Only qualitative results. Case 1: a 17-y-old male died after allegedly inhaling butane that was found in all the samples. Case 2: a 13-y-old female died due to hanging; it was believed that she sniffed solvents prior to death: toluene was found in postmortem samples. Case 3: A 22-y-old male died in a crash event; it was believed that he sniffed petrol prior to crash: petrol residues were found in blood. Case 4: A 17-y-old male, known drug user, died suddenly under suspicious circumstances: halotane was detected in biological samples. 69 Mosaddegh et al./2001 16 victims (n.r.) Urine, plasma and blood Benzodiazepines, tricyclic antidepressants, local anesthetics Simultaneous qualitative identification of many different drugs with a simple and fast procedure. 70 Okajima et al./2001 2 victims (acute intoxication) Blood AP, MA Case 1: MA and AP concentrations in the mixed left and right heart blood were 165 and 36.9 ng/g, respectively. Case 2: MA and AP concentrations were 1.79 and 0.119 μg/g in the left heart blood and 1.27 and 0.074 μg/g in the right heart blood, respectively. In both cases, the concentrations of MA in blood were higher than therapeutic levels. 71 Namera et al./2002 1 victim (n.r.) Urine AP, MA, phenethylamine MA, AP and phenethylamine detected in the urine from the victim of a medicolegal case. No quantitative data are reported. 72 De Martinis et al./2002 A 39-y-old woman (acute intoxication) Blood, urine, vitreous humor Ethanol and other VOCs The autopsy showed ischemic and hypertensive cardiomyopathy. Ethanol concentration was found to be 3.1, 3.9 and 5.5 g/L in blood, urine and vitreous humor, respectively. 73 Toennes et al./2002 A 55-y-old man (acute intoxication) Blood, gastric content Allyl alcohol and its metabolite acrolein Allyl alcohol detected in high amount in the gastric content indicated the oral ingestion. Lethal allyl alcohol concentration was found in blood, bile and urine (309, 963 and 26 mg/L, respectively). Acrolein found at concentrations: 7.2, 13.8 and 5.4 mg/L in blood, bile and urine, respectively. The cause of death was acrolein cardiotoxicity. 74 de Toledo et al./2003 1 victim (n.r.) Scalp hair COC, BE, CE After the development and the validation, the method applied to the real hair sample showed COC (47.4 ng/mg), butylbenzoylecgonine (4.1 ng/mg) and CE (1.3 ng/mg). 75 Tsoukali et al./2004 A 21-y-old woman (suicide) Liver, gastric content, kidneys, blood Methyl-parathion A case of intravenous methyl-parathion injection is reported. Methyl-parathion was determined blood at a concentration of 24 ug/mL. Ingestion was excluded due to the lack of the pesticide in the stomach content. The negative results in other samples were attributed to the instantaneous death due to suicide. 76 Frison et al./2006 Fire victim Blood CN Carbon monoxide saturation in blood was 30% and cyanide concentration was 2 μg/mL. 77 Musshoff et al./2006 A 26-y-old M; a 41-y-old M; a 14-y-old M; a 30-y-old male; a 30-y-old M (acute intoxication). Blood, lung, adipose tissue, brain, kidney, liver, clothes Chloroform, toluene, xylene, ethanol, isopropanol, hydrocarbons (C6-C13), propane, butane VSA lethal intoxication vs VSA autoerotic fatal accident. 4/5 showed circumstantial data for AAD; 1/5 was classified as an accident. Case 1: chloroform was detected in all the samples. Case 2: toluene and xylene were identified in all the specimens. Case 3: a witness reported butane inhalation with negative toxicological results. Case 4: ethanol, methyl pentane and methylhexane in the blood; hydrocarbons in other samples and cloth. BAC 2.46 g/L, propane and butane positivities in case 5. 78 Walker et al./2006 3 victims (acute intoxication) Blood, urine, adipose, brain and lung tissues LPG, ethanol Case 1: 3-hydroxy-2-butanone, ethanol (0.69 g/L) and traces of 2,3-butanedione, n-butanol and 2-butanol in the blood. Case 2: large amounts of 2-butanone and smaller quantities of 2-butanol, 3-hydroxy-2-butanone and 2,3-butanediol in blood, lung and brain. Case 3: large amount of n-butanol and butyric acid in blood. 79 Gottzein et al./2010 A fire victim Blood and viscera (brain, skin, lung, muscle) VOC (toluene, benzene, isooctane, xylene) Toluene was found in all viscera but muscle; benzene found in the lung, brain and muscle; isooctane found in lungs; xylene in skin and blood. 80 Argo et al./2010 A 15-y-old boy (suicide) Blood, gastric content, liver, kidney, lung Toluene and xylene Gastric content: toluene, xylene; blood: toluene 0.06 ppm, xylene 0.457 ppm; liver: toluene 41.5 ppm, xylene 33 ppm; kidney: toluene 0.939 ppm, xylene 9.09 ppm. Nothing found in lung. 81 Mastrogianni et al./2012 A 55-y-old F (acute intoxication) Blood Venlafaxine Venlafaxine concentrations were as follows: 13.7 μg/mL in blood, 729 μg/mL in the gastric content and > 9 mg/mL in the esophagus. These concentrations are much above the therapeutic range; hence, the cause of death was ruled as acute venlafaxine intoxication. 82 Gottzein et al./2013 A 40-y-old F (suicide) Femoral and heart blood, gastric content Captan Captan was detected in gastric content, but not found in blood, due to its rapid hydrolysis to THPI. THPI detected by other techniques: heart blood 0.35 μg/mL, bile 0.30 μg/mL, liver 0.24 μg/g, femoral blood 0.22 μg/mL, kidneys 0.14 μg/mL and brain 0.06 μg/mL. 83 Zheng et al./2016 2 victims (homicide, suicide) Blood and gastric content ACH–FCH Cyanide and acetone were detected in the homicide case. FCH intoxication was demonstrated in the suicide. 84 Perrault et al./2017 7 bodies (n.r.) Intracadaveric gases 29 VOC markers To detect circumstances of death and evaluate PMI, VOC profile for each sample had been performed; a standard method was developed. Differences between VOC profiles from different samples were demonstrated. 85 Waters et al./2017 30 fire victims (20 M and 10 F) Blood 24 VHC (accelerants) To interpret the fire-related incidents by using VHC pattern. To evaluate the presence of accelerants, kerosene and gasoline should be distinghished; the absence of some hydrocarbons may indicate a normal construction fire. Ref. Author/year Population Matrix Substances identified Results 57 Nagasawa et al./1996 3 M and 2 F (2 brain injuries; 3 MA poisoning) Blood MA and AP 3 MA poisoning: range 3.23–5.31 μmol/100 g. MA and AP were detected in 4 out of the 5 cases. 58 Namera et al./1997 A 50-y-old M (acute intoxication) Blood Malathion Malathion detected in blood: left heart 41 μg/g; right heart 5.4 μg/g. 59 Brewer et al./1997 2 victims (traffic fatalities) Urine, gastric content, blood VOC (methylene chloride, light hydrocarbons) Case 1: urine concentration of methylene chloride at the time of the accident (23 mg/L) was considered toxic because greater than expected from occupational exposure; blood was not available. Case 2: charcoal lighter fluid was detected in gastric content but not in blood sample. Circumstantial data support a suicide as manner of death due to traffic accident. 60 Watanabe et al./1998 One victim (local anaestetic poisoning) Blood 5 local anesthetics (prilocaine, lidocaine, mepivacaine, bupivacaine, dibucaine) Mepivacaine and lidocaine were detected in the left and right heart blood samples at concentrations of 18.6 and 15.8 mg/g for mepivacaine and 0.14 and 0.17 mg/g for lidocaine, respectively. Mepivacaine concentration in blood was higher than therapeutic levels. 61 Namera et al./1998 1 victim (acute intoxication) Blood Tetracyclic antidepressants (setiptiline, mianserin, maprotiline) Setiptiline was found in the left and right heart blood samples at concentrations of 1.77 and 0.78 μg/g, respectively. It was about 200 times higher than therapeutic level (1.0–5.0 ng/g). 62 Sporkert et al./2000 49 victims (drug fatalities) Hair (also segmental analysis) Lidocaine 32/49 cases were positive to lidocaine with concentrations between 0.4 and 675 ng/mg (18/32 were > 10 ng/mg; 4/32 were > 150 ng/mg). 63 Dehon et al./2000 A 45-y-old woman (acute intoxication) Blood, urine, tissue homogenates Tetrachloroethylene (C2Cl4) and TCE C2Cl4 and TCE were found in blood and urine of the woman who died 7 d after admission to the hospital. Tissue concentration of TCE was higher in liposoluble tissues (brain); C2Cl4 was found in all post-mortem samples. 64 Musshoff et al./2000 A 75-y-old man and a 82-y-old woman (double homicide by smothering) Blood, urine, bile, tissues Halothane First toxicological analysis was performed 3-mo post-mortem with GC and after 15 mo with HS-SPME–GC–MS. Higher concentration in the brain and the lowest in liver and bile. Both the techniques gave similar analytical results. Halothane concentrations were consistent with intoxication. Manner of death was homicide by forced smothering of halothane. 65 Namera et al./2000 A 25-y-old woman (suicide by intoxication) Blood AP, MA, fenfluramine The developed HS–SPME method was optimized for the simultaneous determination of fenfluramine and amphetamines in human blood using the on-column derivatization method. Fenfluramine was detected in the blood sample after 4-h hospitalization at fatal concentration (7.7 μg/g). The death was due to suicide. 66 Sporkert et al./2000 26 victims (drug intoxication) 3-cm proximal hair Methadone, EDDP, EMDP Methadone was detected in 19 cases, while EDDP was found in 17 cases. EMPD was detected in 2/7 methadone positivities. 67 Liu et al./2001 A 43-y-old cadaver (n.r.) Hair AP, MA The method was applied to 5 strands of hair from a crime-suspected person and the results were compared with a usual method. MA concentration range from proximal (about 42 pmol/cm) to distal (about 3 pmol/cm). 68 Tranthim-Fryer et al./2001 3 M, 1 F (VOC-related deaths) Blood tissues VOC (butane, halotane, toluene and petrol residues) Only qualitative results. Case 1: a 17-y-old male died after allegedly inhaling butane that was found in all the samples. Case 2: a 13-y-old female died due to hanging; it was believed that she sniffed solvents prior to death: toluene was found in postmortem samples. Case 3: A 22-y-old male died in a crash event; it was believed that he sniffed petrol prior to crash: petrol residues were found in blood. Case 4: A 17-y-old male, known drug user, died suddenly under suspicious circumstances: halotane was detected in biological samples. 69 Mosaddegh et al./2001 16 victims (n.r.) Urine, plasma and blood Benzodiazepines, tricyclic antidepressants, local anesthetics Simultaneous qualitative identification of many different drugs with a simple and fast procedure. 70 Okajima et al./2001 2 victims (acute intoxication) Blood AP, MA Case 1: MA and AP concentrations in the mixed left and right heart blood were 165 and 36.9 ng/g, respectively. Case 2: MA and AP concentrations were 1.79 and 0.119 μg/g in the left heart blood and 1.27 and 0.074 μg/g in the right heart blood, respectively. In both cases, the concentrations of MA in blood were higher than therapeutic levels. 71 Namera et al./2002 1 victim (n.r.) Urine AP, MA, phenethylamine MA, AP and phenethylamine detected in the urine from the victim of a medicolegal case. No quantitative data are reported. 72 De Martinis et al./2002 A 39-y-old woman (acute intoxication) Blood, urine, vitreous humor Ethanol and other VOCs The autopsy showed ischemic and hypertensive cardiomyopathy. Ethanol concentration was found to be 3.1, 3.9 and 5.5 g/L in blood, urine and vitreous humor, respectively. 73 Toennes et al./2002 A 55-y-old man (acute intoxication) Blood, gastric content Allyl alcohol and its metabolite acrolein Allyl alcohol detected in high amount in the gastric content indicated the oral ingestion. Lethal allyl alcohol concentration was found in blood, bile and urine (309, 963 and 26 mg/L, respectively). Acrolein found at concentrations: 7.2, 13.8 and 5.4 mg/L in blood, bile and urine, respectively. The cause of death was acrolein cardiotoxicity. 74 de Toledo et al./2003 1 victim (n.r.) Scalp hair COC, BE, CE After the development and the validation, the method applied to the real hair sample showed COC (47.4 ng/mg), butylbenzoylecgonine (4.1 ng/mg) and CE (1.3 ng/mg). 75 Tsoukali et al./2004 A 21-y-old woman (suicide) Liver, gastric content, kidneys, blood Methyl-parathion A case of intravenous methyl-parathion injection is reported. Methyl-parathion was determined blood at a concentration of 24 ug/mL. Ingestion was excluded due to the lack of the pesticide in the stomach content. The negative results in other samples were attributed to the instantaneous death due to suicide. 76 Frison et al./2006 Fire victim Blood CN Carbon monoxide saturation in blood was 30% and cyanide concentration was 2 μg/mL. 77 Musshoff et al./2006 A 26-y-old M; a 41-y-old M; a 14-y-old M; a 30-y-old male; a 30-y-old M (acute intoxication). Blood, lung, adipose tissue, brain, kidney, liver, clothes Chloroform, toluene, xylene, ethanol, isopropanol, hydrocarbons (C6-C13), propane, butane VSA lethal intoxication vs VSA autoerotic fatal accident. 4/5 showed circumstantial data for AAD; 1/5 was classified as an accident. Case 1: chloroform was detected in all the samples. Case 2: toluene and xylene were identified in all the specimens. Case 3: a witness reported butane inhalation with negative toxicological results. Case 4: ethanol, methyl pentane and methylhexane in the blood; hydrocarbons in other samples and cloth. BAC 2.46 g/L, propane and butane positivities in case 5. 78 Walker et al./2006 3 victims (acute intoxication) Blood, urine, adipose, brain and lung tissues LPG, ethanol Case 1: 3-hydroxy-2-butanone, ethanol (0.69 g/L) and traces of 2,3-butanedione, n-butanol and 2-butanol in the blood. Case 2: large amounts of 2-butanone and smaller quantities of 2-butanol, 3-hydroxy-2-butanone and 2,3-butanediol in blood, lung and brain. Case 3: large amount of n-butanol and butyric acid in blood. 79 Gottzein et al./2010 A fire victim Blood and viscera (brain, skin, lung, muscle) VOC (toluene, benzene, isooctane, xylene) Toluene was found in all viscera but muscle; benzene found in the lung, brain and muscle; isooctane found in lungs; xylene in skin and blood. 80 Argo et al./2010 A 15-y-old boy (suicide) Blood, gastric content, liver, kidney, lung Toluene and xylene Gastric content: toluene, xylene; blood: toluene 0.06 ppm, xylene 0.457 ppm; liver: toluene 41.5 ppm, xylene 33 ppm; kidney: toluene 0.939 ppm, xylene 9.09 ppm. Nothing found in lung. 81 Mastrogianni et al./2012 A 55-y-old F (acute intoxication) Blood Venlafaxine Venlafaxine concentrations were as follows: 13.7 μg/mL in blood, 729 μg/mL in the gastric content and > 9 mg/mL in the esophagus. These concentrations are much above the therapeutic range; hence, the cause of death was ruled as acute venlafaxine intoxication. 82 Gottzein et al./2013 A 40-y-old F (suicide) Femoral and heart blood, gastric content Captan Captan was detected in gastric content, but not found in blood, due to its rapid hydrolysis to THPI. THPI detected by other techniques: heart blood 0.35 μg/mL, bile 0.30 μg/mL, liver 0.24 μg/g, femoral blood 0.22 μg/mL, kidneys 0.14 μg/mL and brain 0.06 μg/mL. 83 Zheng et al./2016 2 victims (homicide, suicide) Blood and gastric content ACH–FCH Cyanide and acetone were detected in the homicide case. FCH intoxication was demonstrated in the suicide. 84 Perrault et al./2017 7 bodies (n.r.) Intracadaveric gases 29 VOC markers To detect circumstances of death and evaluate PMI, VOC profile for each sample had been performed; a standard method was developed. Differences between VOC profiles from different samples were demonstrated. 85 Waters et al./2017 30 fire victims (20 M and 10 F) Blood 24 VHC (accelerants) To interpret the fire-related incidents by using VHC pattern. To evaluate the presence of accelerants, kerosene and gasoline should be distinghished; the absence of some hydrocarbons may indicate a normal construction fire. n.r., not reported. Open in new tab Table 3 Solid-phase microextraction application in the forensic toxicology investigation concerning death Ref. Author/year Population Matrix Substances identified Results 57 Nagasawa et al./1996 3 M and 2 F (2 brain injuries; 3 MA poisoning) Blood MA and AP 3 MA poisoning: range 3.23–5.31 μmol/100 g. MA and AP were detected in 4 out of the 5 cases. 58 Namera et al./1997 A 50-y-old M (acute intoxication) Blood Malathion Malathion detected in blood: left heart 41 μg/g; right heart 5.4 μg/g. 59 Brewer et al./1997 2 victims (traffic fatalities) Urine, gastric content, blood VOC (methylene chloride, light hydrocarbons) Case 1: urine concentration of methylene chloride at the time of the accident (23 mg/L) was considered toxic because greater than expected from occupational exposure; blood was not available. Case 2: charcoal lighter fluid was detected in gastric content but not in blood sample. Circumstantial data support a suicide as manner of death due to traffic accident. 60 Watanabe et al./1998 One victim (local anaestetic poisoning) Blood 5 local anesthetics (prilocaine, lidocaine, mepivacaine, bupivacaine, dibucaine) Mepivacaine and lidocaine were detected in the left and right heart blood samples at concentrations of 18.6 and 15.8 mg/g for mepivacaine and 0.14 and 0.17 mg/g for lidocaine, respectively. Mepivacaine concentration in blood was higher than therapeutic levels. 61 Namera et al./1998 1 victim (acute intoxication) Blood Tetracyclic antidepressants (setiptiline, mianserin, maprotiline) Setiptiline was found in the left and right heart blood samples at concentrations of 1.77 and 0.78 μg/g, respectively. It was about 200 times higher than therapeutic level (1.0–5.0 ng/g). 62 Sporkert et al./2000 49 victims (drug fatalities) Hair (also segmental analysis) Lidocaine 32/49 cases were positive to lidocaine with concentrations between 0.4 and 675 ng/mg (18/32 were > 10 ng/mg; 4/32 were > 150 ng/mg). 63 Dehon et al./2000 A 45-y-old woman (acute intoxication) Blood, urine, tissue homogenates Tetrachloroethylene (C2Cl4) and TCE C2Cl4 and TCE were found in blood and urine of the woman who died 7 d after admission to the hospital. Tissue concentration of TCE was higher in liposoluble tissues (brain); C2Cl4 was found in all post-mortem samples. 64 Musshoff et al./2000 A 75-y-old man and a 82-y-old woman (double homicide by smothering) Blood, urine, bile, tissues Halothane First toxicological analysis was performed 3-mo post-mortem with GC and after 15 mo with HS-SPME–GC–MS. Higher concentration in the brain and the lowest in liver and bile. Both the techniques gave similar analytical results. Halothane concentrations were consistent with intoxication. Manner of death was homicide by forced smothering of halothane. 65 Namera et al./2000 A 25-y-old woman (suicide by intoxication) Blood AP, MA, fenfluramine The developed HS–SPME method was optimized for the simultaneous determination of fenfluramine and amphetamines in human blood using the on-column derivatization method. Fenfluramine was detected in the blood sample after 4-h hospitalization at fatal concentration (7.7 μg/g). The death was due to suicide. 66 Sporkert et al./2000 26 victims (drug intoxication) 3-cm proximal hair Methadone, EDDP, EMDP Methadone was detected in 19 cases, while EDDP was found in 17 cases. EMPD was detected in 2/7 methadone positivities. 67 Liu et al./2001 A 43-y-old cadaver (n.r.) Hair AP, MA The method was applied to 5 strands of hair from a crime-suspected person and the results were compared with a usual method. MA concentration range from proximal (about 42 pmol/cm) to distal (about 3 pmol/cm). 68 Tranthim-Fryer et al./2001 3 M, 1 F (VOC-related deaths) Blood tissues VOC (butane, halotane, toluene and petrol residues) Only qualitative results. Case 1: a 17-y-old male died after allegedly inhaling butane that was found in all the samples. Case 2: a 13-y-old female died due to hanging; it was believed that she sniffed solvents prior to death: toluene was found in postmortem samples. Case 3: A 22-y-old male died in a crash event; it was believed that he sniffed petrol prior to crash: petrol residues were found in blood. Case 4: A 17-y-old male, known drug user, died suddenly under suspicious circumstances: halotane was detected in biological samples. 69 Mosaddegh et al./2001 16 victims (n.r.) Urine, plasma and blood Benzodiazepines, tricyclic antidepressants, local anesthetics Simultaneous qualitative identification of many different drugs with a simple and fast procedure. 70 Okajima et al./2001 2 victims (acute intoxication) Blood AP, MA Case 1: MA and AP concentrations in the mixed left and right heart blood were 165 and 36.9 ng/g, respectively. Case 2: MA and AP concentrations were 1.79 and 0.119 μg/g in the left heart blood and 1.27 and 0.074 μg/g in the right heart blood, respectively. In both cases, the concentrations of MA in blood were higher than therapeutic levels. 71 Namera et al./2002 1 victim (n.r.) Urine AP, MA, phenethylamine MA, AP and phenethylamine detected in the urine from the victim of a medicolegal case. No quantitative data are reported. 72 De Martinis et al./2002 A 39-y-old woman (acute intoxication) Blood, urine, vitreous humor Ethanol and other VOCs The autopsy showed ischemic and hypertensive cardiomyopathy. Ethanol concentration was found to be 3.1, 3.9 and 5.5 g/L in blood, urine and vitreous humor, respectively. 73 Toennes et al./2002 A 55-y-old man (acute intoxication) Blood, gastric content Allyl alcohol and its metabolite acrolein Allyl alcohol detected in high amount in the gastric content indicated the oral ingestion. Lethal allyl alcohol concentration was found in blood, bile and urine (309, 963 and 26 mg/L, respectively). Acrolein found at concentrations: 7.2, 13.8 and 5.4 mg/L in blood, bile and urine, respectively. The cause of death was acrolein cardiotoxicity. 74 de Toledo et al./2003 1 victim (n.r.) Scalp hair COC, BE, CE After the development and the validation, the method applied to the real hair sample showed COC (47.4 ng/mg), butylbenzoylecgonine (4.1 ng/mg) and CE (1.3 ng/mg). 75 Tsoukali et al./2004 A 21-y-old woman (suicide) Liver, gastric content, kidneys, blood Methyl-parathion A case of intravenous methyl-parathion injection is reported. Methyl-parathion was determined blood at a concentration of 24 ug/mL. Ingestion was excluded due to the lack of the pesticide in the stomach content. The negative results in other samples were attributed to the instantaneous death due to suicide. 76 Frison et al./2006 Fire victim Blood CN Carbon monoxide saturation in blood was 30% and cyanide concentration was 2 μg/mL. 77 Musshoff et al./2006 A 26-y-old M; a 41-y-old M; a 14-y-old M; a 30-y-old male; a 30-y-old M (acute intoxication). Blood, lung, adipose tissue, brain, kidney, liver, clothes Chloroform, toluene, xylene, ethanol, isopropanol, hydrocarbons (C6-C13), propane, butane VSA lethal intoxication vs VSA autoerotic fatal accident. 4/5 showed circumstantial data for AAD; 1/5 was classified as an accident. Case 1: chloroform was detected in all the samples. Case 2: toluene and xylene were identified in all the specimens. Case 3: a witness reported butane inhalation with negative toxicological results. Case 4: ethanol, methyl pentane and methylhexane in the blood; hydrocarbons in other samples and cloth. BAC 2.46 g/L, propane and butane positivities in case 5. 78 Walker et al./2006 3 victims (acute intoxication) Blood, urine, adipose, brain and lung tissues LPG, ethanol Case 1: 3-hydroxy-2-butanone, ethanol (0.69 g/L) and traces of 2,3-butanedione, n-butanol and 2-butanol in the blood. Case 2: large amounts of 2-butanone and smaller quantities of 2-butanol, 3-hydroxy-2-butanone and 2,3-butanediol in blood, lung and brain. Case 3: large amount of n-butanol and butyric acid in blood. 79 Gottzein et al./2010 A fire victim Blood and viscera (brain, skin, lung, muscle) VOC (toluene, benzene, isooctane, xylene) Toluene was found in all viscera but muscle; benzene found in the lung, brain and muscle; isooctane found in lungs; xylene in skin and blood. 80 Argo et al./2010 A 15-y-old boy (suicide) Blood, gastric content, liver, kidney, lung Toluene and xylene Gastric content: toluene, xylene; blood: toluene 0.06 ppm, xylene 0.457 ppm; liver: toluene 41.5 ppm, xylene 33 ppm; kidney: toluene 0.939 ppm, xylene 9.09 ppm. Nothing found in lung. 81 Mastrogianni et al./2012 A 55-y-old F (acute intoxication) Blood Venlafaxine Venlafaxine concentrations were as follows: 13.7 μg/mL in blood, 729 μg/mL in the gastric content and > 9 mg/mL in the esophagus. These concentrations are much above the therapeutic range; hence, the cause of death was ruled as acute venlafaxine intoxication. 82 Gottzein et al./2013 A 40-y-old F (suicide) Femoral and heart blood, gastric content Captan Captan was detected in gastric content, but not found in blood, due to its rapid hydrolysis to THPI. THPI detected by other techniques: heart blood 0.35 μg/mL, bile 0.30 μg/mL, liver 0.24 μg/g, femoral blood 0.22 μg/mL, kidneys 0.14 μg/mL and brain 0.06 μg/mL. 83 Zheng et al./2016 2 victims (homicide, suicide) Blood and gastric content ACH–FCH Cyanide and acetone were detected in the homicide case. FCH intoxication was demonstrated in the suicide. 84 Perrault et al./2017 7 bodies (n.r.) Intracadaveric gases 29 VOC markers To detect circumstances of death and evaluate PMI, VOC profile for each sample had been performed; a standard method was developed. Differences between VOC profiles from different samples were demonstrated. 85 Waters et al./2017 30 fire victims (20 M and 10 F) Blood 24 VHC (accelerants) To interpret the fire-related incidents by using VHC pattern. To evaluate the presence of accelerants, kerosene and gasoline should be distinghished; the absence of some hydrocarbons may indicate a normal construction fire. Ref. Author/year Population Matrix Substances identified Results 57 Nagasawa et al./1996 3 M and 2 F (2 brain injuries; 3 MA poisoning) Blood MA and AP 3 MA poisoning: range 3.23–5.31 μmol/100 g. MA and AP were detected in 4 out of the 5 cases. 58 Namera et al./1997 A 50-y-old M (acute intoxication) Blood Malathion Malathion detected in blood: left heart 41 μg/g; right heart 5.4 μg/g. 59 Brewer et al./1997 2 victims (traffic fatalities) Urine, gastric content, blood VOC (methylene chloride, light hydrocarbons) Case 1: urine concentration of methylene chloride at the time of the accident (23 mg/L) was considered toxic because greater than expected from occupational exposure; blood was not available. Case 2: charcoal lighter fluid was detected in gastric content but not in blood sample. Circumstantial data support a suicide as manner of death due to traffic accident. 60 Watanabe et al./1998 One victim (local anaestetic poisoning) Blood 5 local anesthetics (prilocaine, lidocaine, mepivacaine, bupivacaine, dibucaine) Mepivacaine and lidocaine were detected in the left and right heart blood samples at concentrations of 18.6 and 15.8 mg/g for mepivacaine and 0.14 and 0.17 mg/g for lidocaine, respectively. Mepivacaine concentration in blood was higher than therapeutic levels. 61 Namera et al./1998 1 victim (acute intoxication) Blood Tetracyclic antidepressants (setiptiline, mianserin, maprotiline) Setiptiline was found in the left and right heart blood samples at concentrations of 1.77 and 0.78 μg/g, respectively. It was about 200 times higher than therapeutic level (1.0–5.0 ng/g). 62 Sporkert et al./2000 49 victims (drug fatalities) Hair (also segmental analysis) Lidocaine 32/49 cases were positive to lidocaine with concentrations between 0.4 and 675 ng/mg (18/32 were > 10 ng/mg; 4/32 were > 150 ng/mg). 63 Dehon et al./2000 A 45-y-old woman (acute intoxication) Blood, urine, tissue homogenates Tetrachloroethylene (C2Cl4) and TCE C2Cl4 and TCE were found in blood and urine of the woman who died 7 d after admission to the hospital. Tissue concentration of TCE was higher in liposoluble tissues (brain); C2Cl4 was found in all post-mortem samples. 64 Musshoff et al./2000 A 75-y-old man and a 82-y-old woman (double homicide by smothering) Blood, urine, bile, tissues Halothane First toxicological analysis was performed 3-mo post-mortem with GC and after 15 mo with HS-SPME–GC–MS. Higher concentration in the brain and the lowest in liver and bile. Both the techniques gave similar analytical results. Halothane concentrations were consistent with intoxication. Manner of death was homicide by forced smothering of halothane. 65 Namera et al./2000 A 25-y-old woman (suicide by intoxication) Blood AP, MA, fenfluramine The developed HS–SPME method was optimized for the simultaneous determination of fenfluramine and amphetamines in human blood using the on-column derivatization method. Fenfluramine was detected in the blood sample after 4-h hospitalization at fatal concentration (7.7 μg/g). The death was due to suicide. 66 Sporkert et al./2000 26 victims (drug intoxication) 3-cm proximal hair Methadone, EDDP, EMDP Methadone was detected in 19 cases, while EDDP was found in 17 cases. EMPD was detected in 2/7 methadone positivities. 67 Liu et al./2001 A 43-y-old cadaver (n.r.) Hair AP, MA The method was applied to 5 strands of hair from a crime-suspected person and the results were compared with a usual method. MA concentration range from proximal (about 42 pmol/cm) to distal (about 3 pmol/cm). 68 Tranthim-Fryer et al./2001 3 M, 1 F (VOC-related deaths) Blood tissues VOC (butane, halotane, toluene and petrol residues) Only qualitative results. Case 1: a 17-y-old male died after allegedly inhaling butane that was found in all the samples. Case 2: a 13-y-old female died due to hanging; it was believed that she sniffed solvents prior to death: toluene was found in postmortem samples. Case 3: A 22-y-old male died in a crash event; it was believed that he sniffed petrol prior to crash: petrol residues were found in blood. Case 4: A 17-y-old male, known drug user, died suddenly under suspicious circumstances: halotane was detected in biological samples. 69 Mosaddegh et al./2001 16 victims (n.r.) Urine, plasma and blood Benzodiazepines, tricyclic antidepressants, local anesthetics Simultaneous qualitative identification of many different drugs with a simple and fast procedure. 70 Okajima et al./2001 2 victims (acute intoxication) Blood AP, MA Case 1: MA and AP concentrations in the mixed left and right heart blood were 165 and 36.9 ng/g, respectively. Case 2: MA and AP concentrations were 1.79 and 0.119 μg/g in the left heart blood and 1.27 and 0.074 μg/g in the right heart blood, respectively. In both cases, the concentrations of MA in blood were higher than therapeutic levels. 71 Namera et al./2002 1 victim (n.r.) Urine AP, MA, phenethylamine MA, AP and phenethylamine detected in the urine from the victim of a medicolegal case. No quantitative data are reported. 72 De Martinis et al./2002 A 39-y-old woman (acute intoxication) Blood, urine, vitreous humor Ethanol and other VOCs The autopsy showed ischemic and hypertensive cardiomyopathy. Ethanol concentration was found to be 3.1, 3.9 and 5.5 g/L in blood, urine and vitreous humor, respectively. 73 Toennes et al./2002 A 55-y-old man (acute intoxication) Blood, gastric content Allyl alcohol and its metabolite acrolein Allyl alcohol detected in high amount in the gastric content indicated the oral ingestion. Lethal allyl alcohol concentration was found in blood, bile and urine (309, 963 and 26 mg/L, respectively). Acrolein found at concentrations: 7.2, 13.8 and 5.4 mg/L in blood, bile and urine, respectively. The cause of death was acrolein cardiotoxicity. 74 de Toledo et al./2003 1 victim (n.r.) Scalp hair COC, BE, CE After the development and the validation, the method applied to the real hair sample showed COC (47.4 ng/mg), butylbenzoylecgonine (4.1 ng/mg) and CE (1.3 ng/mg). 75 Tsoukali et al./2004 A 21-y-old woman (suicide) Liver, gastric content, kidneys, blood Methyl-parathion A case of intravenous methyl-parathion injection is reported. Methyl-parathion was determined blood at a concentration of 24 ug/mL. Ingestion was excluded due to the lack of the pesticide in the stomach content. The negative results in other samples were attributed to the instantaneous death due to suicide. 76 Frison et al./2006 Fire victim Blood CN Carbon monoxide saturation in blood was 30% and cyanide concentration was 2 μg/mL. 77 Musshoff et al./2006 A 26-y-old M; a 41-y-old M; a 14-y-old M; a 30-y-old male; a 30-y-old M (acute intoxication). Blood, lung, adipose tissue, brain, kidney, liver, clothes Chloroform, toluene, xylene, ethanol, isopropanol, hydrocarbons (C6-C13), propane, butane VSA lethal intoxication vs VSA autoerotic fatal accident. 4/5 showed circumstantial data for AAD; 1/5 was classified as an accident. Case 1: chloroform was detected in all the samples. Case 2: toluene and xylene were identified in all the specimens. Case 3: a witness reported butane inhalation with negative toxicological results. Case 4: ethanol, methyl pentane and methylhexane in the blood; hydrocarbons in other samples and cloth. BAC 2.46 g/L, propane and butane positivities in case 5. 78 Walker et al./2006 3 victims (acute intoxication) Blood, urine, adipose, brain and lung tissues LPG, ethanol Case 1: 3-hydroxy-2-butanone, ethanol (0.69 g/L) and traces of 2,3-butanedione, n-butanol and 2-butanol in the blood. Case 2: large amounts of 2-butanone and smaller quantities of 2-butanol, 3-hydroxy-2-butanone and 2,3-butanediol in blood, lung and brain. Case 3: large amount of n-butanol and butyric acid in blood. 79 Gottzein et al./2010 A fire victim Blood and viscera (brain, skin, lung, muscle) VOC (toluene, benzene, isooctane, xylene) Toluene was found in all viscera but muscle; benzene found in the lung, brain and muscle; isooctane found in lungs; xylene in skin and blood. 80 Argo et al./2010 A 15-y-old boy (suicide) Blood, gastric content, liver, kidney, lung Toluene and xylene Gastric content: toluene, xylene; blood: toluene 0.06 ppm, xylene 0.457 ppm; liver: toluene 41.5 ppm, xylene 33 ppm; kidney: toluene 0.939 ppm, xylene 9.09 ppm. Nothing found in lung. 81 Mastrogianni et al./2012 A 55-y-old F (acute intoxication) Blood Venlafaxine Venlafaxine concentrations were as follows: 13.7 μg/mL in blood, 729 μg/mL in the gastric content and > 9 mg/mL in the esophagus. These concentrations are much above the therapeutic range; hence, the cause of death was ruled as acute venlafaxine intoxication. 82 Gottzein et al./2013 A 40-y-old F (suicide) Femoral and heart blood, gastric content Captan Captan was detected in gastric content, but not found in blood, due to its rapid hydrolysis to THPI. THPI detected by other techniques: heart blood 0.35 μg/mL, bile 0.30 μg/mL, liver 0.24 μg/g, femoral blood 0.22 μg/mL, kidneys 0.14 μg/mL and brain 0.06 μg/mL. 83 Zheng et al./2016 2 victims (homicide, suicide) Blood and gastric content ACH–FCH Cyanide and acetone were detected in the homicide case. FCH intoxication was demonstrated in the suicide. 84 Perrault et al./2017 7 bodies (n.r.) Intracadaveric gases 29 VOC markers To detect circumstances of death and evaluate PMI, VOC profile for each sample had been performed; a standard method was developed. Differences between VOC profiles from different samples were demonstrated. 85 Waters et al./2017 30 fire victims (20 M and 10 F) Blood 24 VHC (accelerants) To interpret the fire-related incidents by using VHC pattern. To evaluate the presence of accelerants, kerosene and gasoline should be distinghished; the absence of some hydrocarbons may indicate a normal construction fire. n.r., not reported. Open in new tab Table 4 Solid-phase microextraction in forensic toxicology on living people Ref. Author/year Population Matrix Substances identified Results 67 Liu et al./2001 Stimulant abusers Scalp hair AP, MA In order to measure MA distribution, hair specimens were cut into 1-cm lengths from the root, and 5 pieces of each of these lengths were, respectively, collected in a vial. 71 Namera et al./2002 8 patients with MA intoxication Urine AP, MA, phenethylamine MA and AP detected in urine collected from patients involved in clinical cases. No quantitative data are reported. 86 Namera et al./1999 A 40-y-old M suspected to attempt suicide Serum The herbicide Kusanon A containing propanil, carbaryl, xylene and surfactant. Propanil (the main ingredient in the herbicide) was detected in the 8 samples collected during hospitalization at the concentration range from 26.7 (on admission 6 h after drinking) to 1.1 μg/mL (60 h after admission). 87 Liu et al./1999 A 20-y-old M intoxicated by ingestion of a pesticide Blood 3 isomers of DCB At admission (4 h after drinking): o-DCB 20.7 μg/g; m-DCB 0.22 μg/g; p-DCB 4.6 μg/g. By 36 h: o-DCB 0.34 μg/g; m-DCB 0.02 μg/g; p-DCB 0.77 μg/g 88 Namera et al./1999 A 83-y-old hospitalized woman suspected to attempt suicide Serum The herbicide Padan 4R containing 4% Cartap HCl (nereistoxin analogue) Patient hospitalized about 3 h after ingestion. Nereistoxin and its metabolites were detected in the serum at concentration of 2.69 μg/mL (0.5 h after admission) and 0.09 μg/mL (after 24 h). 89 Kim et al./2000 Glue-sniffers Blood, urine Toluene Urine: 22/25 positive to toluene; blood: 47/49 positive to toluene. The range of toluene concentrations as positive meaning were: 0.1–14.0 μg/mL and 0.1–17.6 μg/mL in urine and blood, respectively. 90 Blair et al./2001 4 suspected abusers and an accidental GHB overdose Urine GHB GHB urine concentrations were 130, 4730, 406, 951 and 5400 μg/mL, respectively. The results were compared with a conventional method and a good agreement was found. 91 Frison et al./2003 A 61-y-old woman (DFSA) Scalp hair (1.5 cm long of proximal and distal ends) and pubic hair Thiopental, pentobarbital Results of 3 proximal hair samples: pentobarbital 0.40, 0.20 and 0.20 ng/mg and thiopental 0.30, 0.20 and 0.15 ng/mg. Distal samples were negative. Pubic hair: pentobarbital 0.40 ng/mg and thiopental 0.25 ng/mg. 92 Barroso et al./2005 3 intoxicated persons Blood Strychnine Strychnine concentration in the 3 cases were as follows: 2.39, 1.27 and 1.03 μg/mL. 93 Wang et al./2005 12 WDT specimens Urine AP, MA Quantitative determination of the enantiomeric pairs: MA L-form 219–5836 ng/mL; D-form 1821–13845 ng/mL. AP: L-form 36–403 ng/mL; D-form 580–3324 ng/mL. 94 Zeng et al./2005 3 tetramine toxicosis patients Urine Tetramine Tetramine was detected in urine samples of the 3 patients examined with the time concentration profile (time peak at 90 h). 95 Nadulski et al./2007 >250 samples Scalp hair (proximal segment 0–6 cm) THC, CBD, CBN THC in 77 samples (0.012–4.2 ng/mg); CBD (0.013–12.1 ng/mg); CBN (0.016–0.85 ng/mg). Six samples analyzed in 5–14 segments. 96 Aleksa et al./2012 16 samples from the hospital of sick children Scalp hair METH, COC, CE, BE, nor-COC, meperidine, AP, MA, MDMA, MDA, COD, 6-MAM, MOR, hydromorphone, oxymorphone, oxycodone, hydrocodone 11/16 were found to be poly-drug abusers; 5/16 only 1 drug was detected. 97 Pragst et al./2013 Children with parents suspected of illicit drug abuse: 149 children (1 to 14 y old), 124 parents Scalp hair THC, CBD, CBN 24/149 children were positive to THC (range = 0.08–0.72 ng/mg), 22/24 positive to CBN (range = 0.02–0.23 ng/mg), 9/24 positive to CBD (range = 0.02–0.13 ng/mg). 14/124 parents: 7/14 not tested; 5/7 positive to THC (0.028–1.87 ng/mg); 2/7 not detected. 98 Anzillotti et al./2014 70 drivers Oral fluid THC, CBD, CBN THC in 42/70 samples (2–690 ng/mL), CBD in 21/42, CBN in 14/42. 99 Tzatzarakis et al./2014 72 children living in urban (15) and rural (57) regions (4 to 12 y old) Scalp hair DDT, PCB congeners PCB28 and PCB52 detection rates were higher in the investigated samples for rural area. 94.4% were positive for at least one of the examined DDTs especially for rural area. Quantitative data reported in the paper. 100 Rainey et al./2014 3 smokers Scalp hair Nicotine, cotinine Nicotine concentration in hair of a female tobacco user (4 packs of cigarettes/weeks) was 16 ng/mg. Two smokers had concentration of nicotine of 22 and 28 ng/mg. 101 Moosman et al./2015 Children with parents suspected of illicit drug abuse: 41 children (7 mo–12 y), 4 teenagers (13–17 y), 34 parents (18–59 y) Scalp hair (proximal segments 0–6 cm) THC, CBD, CBN Comparison of SPME with LC/MS/MS. 32/41 children positive to THC (11–4330 pg/mg); 32/34 adults positive to THC (11–1360 pg/mg); 4/4 teenagers positive to THC (100–4330 pg/mg). 102 Gentili et al./2016 60 drivers Sweat AP, MA, MDA, MDMA, MDEA, MBDB, ketamine, METH, COC, CE, THC 60/66 samples showed 1 or more drugs; the sweat pads are suggested for the non-invasive monitoring of exposure of drugs of abuse in drivers. 103 Muller et al./2017 An injured woman Burned clothes remains White phosphorous Identification of white phosphorous as cause of burning. 104 Sporkert et al./2000 53 dead people; 10 n.r.; 4 living people Scalp Hair Cannabinoids, clomethiazole and many lipophilic basic drugs (nicotine, amphetamine derivatives, local anesthetics, phencyclidine, ketamine, methadone, diphenhydramine, tramadol, tricyclic antidepressants and phenothiazines) Dead people: 32/53 were positive to lidocaine (0.4–300 ng/mg); 2/53 positive to diphenadrimine (2–3.7 ng/mg); 2/53 positive to methadone (9.9–20.1 ng/mg). Living people: 2/4 tramadol (0.78–1.14 ng/mg); Clometiazole 2/4 (15.3–115 ng/mg). N.R. 7/10 tricyclic antidepressant; 3/10 nicotine. 105 Musshoff et al./2002 25 drug abusers (18 M, 7 F, age 29 ± 11 y): deceased persons and persons examined for driving liability Scalp hair THC, CBD, CBN The developed method was proved to be useful to detect cannabis in hair of abusers at the following concentration ranges: THC 0.29–2.20 ng/mg, CBN 0.55–4.54 ng/mg and CBD 0.53–18.36 ng/mg. Ref. Author/year Population Matrix Substances identified Results 67 Liu et al./2001 Stimulant abusers Scalp hair AP, MA In order to measure MA distribution, hair specimens were cut into 1-cm lengths from the root, and 5 pieces of each of these lengths were, respectively, collected in a vial. 71 Namera et al./2002 8 patients with MA intoxication Urine AP, MA, phenethylamine MA and AP detected in urine collected from patients involved in clinical cases. No quantitative data are reported. 86 Namera et al./1999 A 40-y-old M suspected to attempt suicide Serum The herbicide Kusanon A containing propanil, carbaryl, xylene and surfactant. Propanil (the main ingredient in the herbicide) was detected in the 8 samples collected during hospitalization at the concentration range from 26.7 (on admission 6 h after drinking) to 1.1 μg/mL (60 h after admission). 87 Liu et al./1999 A 20-y-old M intoxicated by ingestion of a pesticide Blood 3 isomers of DCB At admission (4 h after drinking): o-DCB 20.7 μg/g; m-DCB 0.22 μg/g; p-DCB 4.6 μg/g. By 36 h: o-DCB 0.34 μg/g; m-DCB 0.02 μg/g; p-DCB 0.77 μg/g 88 Namera et al./1999 A 83-y-old hospitalized woman suspected to attempt suicide Serum The herbicide Padan 4R containing 4% Cartap HCl (nereistoxin analogue) Patient hospitalized about 3 h after ingestion. Nereistoxin and its metabolites were detected in the serum at concentration of 2.69 μg/mL (0.5 h after admission) and 0.09 μg/mL (after 24 h). 89 Kim et al./2000 Glue-sniffers Blood, urine Toluene Urine: 22/25 positive to toluene; blood: 47/49 positive to toluene. The range of toluene concentrations as positive meaning were: 0.1–14.0 μg/mL and 0.1–17.6 μg/mL in urine and blood, respectively. 90 Blair et al./2001 4 suspected abusers and an accidental GHB overdose Urine GHB GHB urine concentrations were 130, 4730, 406, 951 and 5400 μg/mL, respectively. The results were compared with a conventional method and a good agreement was found. 91 Frison et al./2003 A 61-y-old woman (DFSA) Scalp hair (1.5 cm long of proximal and distal ends) and pubic hair Thiopental, pentobarbital Results of 3 proximal hair samples: pentobarbital 0.40, 0.20 and 0.20 ng/mg and thiopental 0.30, 0.20 and 0.15 ng/mg. Distal samples were negative. Pubic hair: pentobarbital 0.40 ng/mg and thiopental 0.25 ng/mg. 92 Barroso et al./2005 3 intoxicated persons Blood Strychnine Strychnine concentration in the 3 cases were as follows: 2.39, 1.27 and 1.03 μg/mL. 93 Wang et al./2005 12 WDT specimens Urine AP, MA Quantitative determination of the enantiomeric pairs: MA L-form 219–5836 ng/mL; D-form 1821–13845 ng/mL. AP: L-form 36–403 ng/mL; D-form 580–3324 ng/mL. 94 Zeng et al./2005 3 tetramine toxicosis patients Urine Tetramine Tetramine was detected in urine samples of the 3 patients examined with the time concentration profile (time peak at 90 h). 95 Nadulski et al./2007 >250 samples Scalp hair (proximal segment 0–6 cm) THC, CBD, CBN THC in 77 samples (0.012–4.2 ng/mg); CBD (0.013–12.1 ng/mg); CBN (0.016–0.85 ng/mg). Six samples analyzed in 5–14 segments. 96 Aleksa et al./2012 16 samples from the hospital of sick children Scalp hair METH, COC, CE, BE, nor-COC, meperidine, AP, MA, MDMA, MDA, COD, 6-MAM, MOR, hydromorphone, oxymorphone, oxycodone, hydrocodone 11/16 were found to be poly-drug abusers; 5/16 only 1 drug was detected. 97 Pragst et al./2013 Children with parents suspected of illicit drug abuse: 149 children (1 to 14 y old), 124 parents Scalp hair THC, CBD, CBN 24/149 children were positive to THC (range = 0.08–0.72 ng/mg), 22/24 positive to CBN (range = 0.02–0.23 ng/mg), 9/24 positive to CBD (range = 0.02–0.13 ng/mg). 14/124 parents: 7/14 not tested; 5/7 positive to THC (0.028–1.87 ng/mg); 2/7 not detected. 98 Anzillotti et al./2014 70 drivers Oral fluid THC, CBD, CBN THC in 42/70 samples (2–690 ng/mL), CBD in 21/42, CBN in 14/42. 99 Tzatzarakis et al./2014 72 children living in urban (15) and rural (57) regions (4 to 12 y old) Scalp hair DDT, PCB congeners PCB28 and PCB52 detection rates were higher in the investigated samples for rural area. 94.4% were positive for at least one of the examined DDTs especially for rural area. Quantitative data reported in the paper. 100 Rainey et al./2014 3 smokers Scalp hair Nicotine, cotinine Nicotine concentration in hair of a female tobacco user (4 packs of cigarettes/weeks) was 16 ng/mg. Two smokers had concentration of nicotine of 22 and 28 ng/mg. 101 Moosman et al./2015 Children with parents suspected of illicit drug abuse: 41 children (7 mo–12 y), 4 teenagers (13–17 y), 34 parents (18–59 y) Scalp hair (proximal segments 0–6 cm) THC, CBD, CBN Comparison of SPME with LC/MS/MS. 32/41 children positive to THC (11–4330 pg/mg); 32/34 adults positive to THC (11–1360 pg/mg); 4/4 teenagers positive to THC (100–4330 pg/mg). 102 Gentili et al./2016 60 drivers Sweat AP, MA, MDA, MDMA, MDEA, MBDB, ketamine, METH, COC, CE, THC 60/66 samples showed 1 or more drugs; the sweat pads are suggested for the non-invasive monitoring of exposure of drugs of abuse in drivers. 103 Muller et al./2017 An injured woman Burned clothes remains White phosphorous Identification of white phosphorous as cause of burning. 104 Sporkert et al./2000 53 dead people; 10 n.r.; 4 living people Scalp Hair Cannabinoids, clomethiazole and many lipophilic basic drugs (nicotine, amphetamine derivatives, local anesthetics, phencyclidine, ketamine, methadone, diphenhydramine, tramadol, tricyclic antidepressants and phenothiazines) Dead people: 32/53 were positive to lidocaine (0.4–300 ng/mg); 2/53 positive to diphenadrimine (2–3.7 ng/mg); 2/53 positive to methadone (9.9–20.1 ng/mg). Living people: 2/4 tramadol (0.78–1.14 ng/mg); Clometiazole 2/4 (15.3–115 ng/mg). N.R. 7/10 tricyclic antidepressant; 3/10 nicotine. 105 Musshoff et al./2002 25 drug abusers (18 M, 7 F, age 29 ± 11 y): deceased persons and persons examined for driving liability Scalp hair THC, CBD, CBN The developed method was proved to be useful to detect cannabis in hair of abusers at the following concentration ranges: THC 0.29–2.20 ng/mg, CBN 0.55–4.54 ng/mg and CBD 0.53–18.36 ng/mg. n.r., not reported. Open in new tab Table 4 Solid-phase microextraction in forensic toxicology on living people Ref. Author/year Population Matrix Substances identified Results 67 Liu et al./2001 Stimulant abusers Scalp hair AP, MA In order to measure MA distribution, hair specimens were cut into 1-cm lengths from the root, and 5 pieces of each of these lengths were, respectively, collected in a vial. 71 Namera et al./2002 8 patients with MA intoxication Urine AP, MA, phenethylamine MA and AP detected in urine collected from patients involved in clinical cases. No quantitative data are reported. 86 Namera et al./1999 A 40-y-old M suspected to attempt suicide Serum The herbicide Kusanon A containing propanil, carbaryl, xylene and surfactant. Propanil (the main ingredient in the herbicide) was detected in the 8 samples collected during hospitalization at the concentration range from 26.7 (on admission 6 h after drinking) to 1.1 μg/mL (60 h after admission). 87 Liu et al./1999 A 20-y-old M intoxicated by ingestion of a pesticide Blood 3 isomers of DCB At admission (4 h after drinking): o-DCB 20.7 μg/g; m-DCB 0.22 μg/g; p-DCB 4.6 μg/g. By 36 h: o-DCB 0.34 μg/g; m-DCB 0.02 μg/g; p-DCB 0.77 μg/g 88 Namera et al./1999 A 83-y-old hospitalized woman suspected to attempt suicide Serum The herbicide Padan 4R containing 4% Cartap HCl (nereistoxin analogue) Patient hospitalized about 3 h after ingestion. Nereistoxin and its metabolites were detected in the serum at concentration of 2.69 μg/mL (0.5 h after admission) and 0.09 μg/mL (after 24 h). 89 Kim et al./2000 Glue-sniffers Blood, urine Toluene Urine: 22/25 positive to toluene; blood: 47/49 positive to toluene. The range of toluene concentrations as positive meaning were: 0.1–14.0 μg/mL and 0.1–17.6 μg/mL in urine and blood, respectively. 90 Blair et al./2001 4 suspected abusers and an accidental GHB overdose Urine GHB GHB urine concentrations were 130, 4730, 406, 951 and 5400 μg/mL, respectively. The results were compared with a conventional method and a good agreement was found. 91 Frison et al./2003 A 61-y-old woman (DFSA) Scalp hair (1.5 cm long of proximal and distal ends) and pubic hair Thiopental, pentobarbital Results of 3 proximal hair samples: pentobarbital 0.40, 0.20 and 0.20 ng/mg and thiopental 0.30, 0.20 and 0.15 ng/mg. Distal samples were negative. Pubic hair: pentobarbital 0.40 ng/mg and thiopental 0.25 ng/mg. 92 Barroso et al./2005 3 intoxicated persons Blood Strychnine Strychnine concentration in the 3 cases were as follows: 2.39, 1.27 and 1.03 μg/mL. 93 Wang et al./2005 12 WDT specimens Urine AP, MA Quantitative determination of the enantiomeric pairs: MA L-form 219–5836 ng/mL; D-form 1821–13845 ng/mL. AP: L-form 36–403 ng/mL; D-form 580–3324 ng/mL. 94 Zeng et al./2005 3 tetramine toxicosis patients Urine Tetramine Tetramine was detected in urine samples of the 3 patients examined with the time concentration profile (time peak at 90 h). 95 Nadulski et al./2007 >250 samples Scalp hair (proximal segment 0–6 cm) THC, CBD, CBN THC in 77 samples (0.012–4.2 ng/mg); CBD (0.013–12.1 ng/mg); CBN (0.016–0.85 ng/mg). Six samples analyzed in 5–14 segments. 96 Aleksa et al./2012 16 samples from the hospital of sick children Scalp hair METH, COC, CE, BE, nor-COC, meperidine, AP, MA, MDMA, MDA, COD, 6-MAM, MOR, hydromorphone, oxymorphone, oxycodone, hydrocodone 11/16 were found to be poly-drug abusers; 5/16 only 1 drug was detected. 97 Pragst et al./2013 Children with parents suspected of illicit drug abuse: 149 children (1 to 14 y old), 124 parents Scalp hair THC, CBD, CBN 24/149 children were positive to THC (range = 0.08–0.72 ng/mg), 22/24 positive to CBN (range = 0.02–0.23 ng/mg), 9/24 positive to CBD (range = 0.02–0.13 ng/mg). 14/124 parents: 7/14 not tested; 5/7 positive to THC (0.028–1.87 ng/mg); 2/7 not detected. 98 Anzillotti et al./2014 70 drivers Oral fluid THC, CBD, CBN THC in 42/70 samples (2–690 ng/mL), CBD in 21/42, CBN in 14/42. 99 Tzatzarakis et al./2014 72 children living in urban (15) and rural (57) regions (4 to 12 y old) Scalp hair DDT, PCB congeners PCB28 and PCB52 detection rates were higher in the investigated samples for rural area. 94.4% were positive for at least one of the examined DDTs especially for rural area. Quantitative data reported in the paper. 100 Rainey et al./2014 3 smokers Scalp hair Nicotine, cotinine Nicotine concentration in hair of a female tobacco user (4 packs of cigarettes/weeks) was 16 ng/mg. Two smokers had concentration of nicotine of 22 and 28 ng/mg. 101 Moosman et al./2015 Children with parents suspected of illicit drug abuse: 41 children (7 mo–12 y), 4 teenagers (13–17 y), 34 parents (18–59 y) Scalp hair (proximal segments 0–6 cm) THC, CBD, CBN Comparison of SPME with LC/MS/MS. 32/41 children positive to THC (11–4330 pg/mg); 32/34 adults positive to THC (11–1360 pg/mg); 4/4 teenagers positive to THC (100–4330 pg/mg). 102 Gentili et al./2016 60 drivers Sweat AP, MA, MDA, MDMA, MDEA, MBDB, ketamine, METH, COC, CE, THC 60/66 samples showed 1 or more drugs; the sweat pads are suggested for the non-invasive monitoring of exposure of drugs of abuse in drivers. 103 Muller et al./2017 An injured woman Burned clothes remains White phosphorous Identification of white phosphorous as cause of burning. 104 Sporkert et al./2000 53 dead people; 10 n.r.; 4 living people Scalp Hair Cannabinoids, clomethiazole and many lipophilic basic drugs (nicotine, amphetamine derivatives, local anesthetics, phencyclidine, ketamine, methadone, diphenhydramine, tramadol, tricyclic antidepressants and phenothiazines) Dead people: 32/53 were positive to lidocaine (0.4–300 ng/mg); 2/53 positive to diphenadrimine (2–3.7 ng/mg); 2/53 positive to methadone (9.9–20.1 ng/mg). Living people: 2/4 tramadol (0.78–1.14 ng/mg); Clometiazole 2/4 (15.3–115 ng/mg). N.R. 7/10 tricyclic antidepressant; 3/10 nicotine. 105 Musshoff et al./2002 25 drug abusers (18 M, 7 F, age 29 ± 11 y): deceased persons and persons examined for driving liability Scalp hair THC, CBD, CBN The developed method was proved to be useful to detect cannabis in hair of abusers at the following concentration ranges: THC 0.29–2.20 ng/mg, CBN 0.55–4.54 ng/mg and CBD 0.53–18.36 ng/mg. Ref. Author/year Population Matrix Substances identified Results 67 Liu et al./2001 Stimulant abusers Scalp hair AP, MA In order to measure MA distribution, hair specimens were cut into 1-cm lengths from the root, and 5 pieces of each of these lengths were, respectively, collected in a vial. 71 Namera et al./2002 8 patients with MA intoxication Urine AP, MA, phenethylamine MA and AP detected in urine collected from patients involved in clinical cases. No quantitative data are reported. 86 Namera et al./1999 A 40-y-old M suspected to attempt suicide Serum The herbicide Kusanon A containing propanil, carbaryl, xylene and surfactant. Propanil (the main ingredient in the herbicide) was detected in the 8 samples collected during hospitalization at the concentration range from 26.7 (on admission 6 h after drinking) to 1.1 μg/mL (60 h after admission). 87 Liu et al./1999 A 20-y-old M intoxicated by ingestion of a pesticide Blood 3 isomers of DCB At admission (4 h after drinking): o-DCB 20.7 μg/g; m-DCB 0.22 μg/g; p-DCB 4.6 μg/g. By 36 h: o-DCB 0.34 μg/g; m-DCB 0.02 μg/g; p-DCB 0.77 μg/g 88 Namera et al./1999 A 83-y-old hospitalized woman suspected to attempt suicide Serum The herbicide Padan 4R containing 4% Cartap HCl (nereistoxin analogue) Patient hospitalized about 3 h after ingestion. Nereistoxin and its metabolites were detected in the serum at concentration of 2.69 μg/mL (0.5 h after admission) and 0.09 μg/mL (after 24 h). 89 Kim et al./2000 Glue-sniffers Blood, urine Toluene Urine: 22/25 positive to toluene; blood: 47/49 positive to toluene. The range of toluene concentrations as positive meaning were: 0.1–14.0 μg/mL and 0.1–17.6 μg/mL in urine and blood, respectively. 90 Blair et al./2001 4 suspected abusers and an accidental GHB overdose Urine GHB GHB urine concentrations were 130, 4730, 406, 951 and 5400 μg/mL, respectively. The results were compared with a conventional method and a good agreement was found. 91 Frison et al./2003 A 61-y-old woman (DFSA) Scalp hair (1.5 cm long of proximal and distal ends) and pubic hair Thiopental, pentobarbital Results of 3 proximal hair samples: pentobarbital 0.40, 0.20 and 0.20 ng/mg and thiopental 0.30, 0.20 and 0.15 ng/mg. Distal samples were negative. Pubic hair: pentobarbital 0.40 ng/mg and thiopental 0.25 ng/mg. 92 Barroso et al./2005 3 intoxicated persons Blood Strychnine Strychnine concentration in the 3 cases were as follows: 2.39, 1.27 and 1.03 μg/mL. 93 Wang et al./2005 12 WDT specimens Urine AP, MA Quantitative determination of the enantiomeric pairs: MA L-form 219–5836 ng/mL; D-form 1821–13845 ng/mL. AP: L-form 36–403 ng/mL; D-form 580–3324 ng/mL. 94 Zeng et al./2005 3 tetramine toxicosis patients Urine Tetramine Tetramine was detected in urine samples of the 3 patients examined with the time concentration profile (time peak at 90 h). 95 Nadulski et al./2007 >250 samples Scalp hair (proximal segment 0–6 cm) THC, CBD, CBN THC in 77 samples (0.012–4.2 ng/mg); CBD (0.013–12.1 ng/mg); CBN (0.016–0.85 ng/mg). Six samples analyzed in 5–14 segments. 96 Aleksa et al./2012 16 samples from the hospital of sick children Scalp hair METH, COC, CE, BE, nor-COC, meperidine, AP, MA, MDMA, MDA, COD, 6-MAM, MOR, hydromorphone, oxymorphone, oxycodone, hydrocodone 11/16 were found to be poly-drug abusers; 5/16 only 1 drug was detected. 97 Pragst et al./2013 Children with parents suspected of illicit drug abuse: 149 children (1 to 14 y old), 124 parents Scalp hair THC, CBD, CBN 24/149 children were positive to THC (range = 0.08–0.72 ng/mg), 22/24 positive to CBN (range = 0.02–0.23 ng/mg), 9/24 positive to CBD (range = 0.02–0.13 ng/mg). 14/124 parents: 7/14 not tested; 5/7 positive to THC (0.028–1.87 ng/mg); 2/7 not detected. 98 Anzillotti et al./2014 70 drivers Oral fluid THC, CBD, CBN THC in 42/70 samples (2–690 ng/mL), CBD in 21/42, CBN in 14/42. 99 Tzatzarakis et al./2014 72 children living in urban (15) and rural (57) regions (4 to 12 y old) Scalp hair DDT, PCB congeners PCB28 and PCB52 detection rates were higher in the investigated samples for rural area. 94.4% were positive for at least one of the examined DDTs especially for rural area. Quantitative data reported in the paper. 100 Rainey et al./2014 3 smokers Scalp hair Nicotine, cotinine Nicotine concentration in hair of a female tobacco user (4 packs of cigarettes/weeks) was 16 ng/mg. Two smokers had concentration of nicotine of 22 and 28 ng/mg. 101 Moosman et al./2015 Children with parents suspected of illicit drug abuse: 41 children (7 mo–12 y), 4 teenagers (13–17 y), 34 parents (18–59 y) Scalp hair (proximal segments 0–6 cm) THC, CBD, CBN Comparison of SPME with LC/MS/MS. 32/41 children positive to THC (11–4330 pg/mg); 32/34 adults positive to THC (11–1360 pg/mg); 4/4 teenagers positive to THC (100–4330 pg/mg). 102 Gentili et al./2016 60 drivers Sweat AP, MA, MDA, MDMA, MDEA, MBDB, ketamine, METH, COC, CE, THC 60/66 samples showed 1 or more drugs; the sweat pads are suggested for the non-invasive monitoring of exposure of drugs of abuse in drivers. 103 Muller et al./2017 An injured woman Burned clothes remains White phosphorous Identification of white phosphorous as cause of burning. 104 Sporkert et al./2000 53 dead people; 10 n.r.; 4 living people Scalp Hair Cannabinoids, clomethiazole and many lipophilic basic drugs (nicotine, amphetamine derivatives, local anesthetics, phencyclidine, ketamine, methadone, diphenhydramine, tramadol, tricyclic antidepressants and phenothiazines) Dead people: 32/53 were positive to lidocaine (0.4–300 ng/mg); 2/53 positive to diphenadrimine (2–3.7 ng/mg); 2/53 positive to methadone (9.9–20.1 ng/mg). Living people: 2/4 tramadol (0.78–1.14 ng/mg); Clometiazole 2/4 (15.3–115 ng/mg). N.R. 7/10 tricyclic antidepressant; 3/10 nicotine. 105 Musshoff et al./2002 25 drug abusers (18 M, 7 F, age 29 ± 11 y): deceased persons and persons examined for driving liability Scalp hair THC, CBD, CBN The developed method was proved to be useful to detect cannabis in hair of abusers at the following concentration ranges: THC 0.29–2.20 ng/mg, CBN 0.55–4.54 ng/mg and CBD 0.53–18.36 ng/mg. n.r., not reported. Open in new tab Table 5 Analytical studies of the SPME technique for the detection of xenobiotics in biological samples Ref. Author/year Population Matrix Substances identified Results 106 Yashiki et al./1995 Suspected drug abusers Urine MA, AP No quantitative data. The method was found to be suitable for analysis of AP and MA in urine. 107 Lord et al./1997 4 clinical samples Urine MA, AP External calibration: AP range 0.2–4.9 μg/mL and MA 0–52 μg/mL. Standard addition: AP range 0.41–8.6 μg/mL and MA 0.05–39 μg/mL. The methods were then applied to several narcotic analgesics. 108 Myung et al./1998 2 subjects who were administered the stimulants Urine AP, MA, dimethamphetamine Case 1: dimethamphetamine and MA detected at 0.51 ± 0.01 and 0.15 ± 0.03 μg/mL, respectively. Case 2: AP and MA detected at 0.35 ± 0.02 and 1.08 ± 0.07 μg/mL, respectively. 109 Ugland et al./1999 1 real sample Urine AP, MA, MDMA, MDA, MDEA The method was developed with urine spiked samples; a real specimen showed the presence of MDA and MDMA. No quantitative data reported. 110 Gentili et al./2002 27 young people attending a disco (18 M, 9 F, mean age = 22 ± 5 y) 20 hair and 7 OF samples Amphetamine-like drugs and ketamine 3/20 hair samples were positive for ketamine (30–40 ng/mg); 1/3 was also positive for MDMA (2.3 ng/mg). 6/7 saliva samples were positive for ketamine, 1/6 also positive for MA, MDA and MDMA. 111 Musshoff et al./2002 Drug abusers 20 hair samples Amphetamines and synthetic designer drugs AP 0.2–21.8 ng/mg; MA 0.1–9.6 ng/mg; MDA 1.3–26.4 ng/mg; MDMA 0.5–12.4 ng/mg; MDEA 0.1–9.9 ng/mg; MBDB 0.2–0.9 ng/mg. 112 Musshoff et al./2002 Drug abusers Hair Amphetamines and synthetic designer drugs No quantitative data of real samples. The method was found to be suitable for analysis of hair samples from drug abusers. 113 Nishida et al./2006 A 37-y-old M Hair (also segmental analysis) MA, AP MA and AP were detected in 1-cm-long hair segments. 114 Junkuy et al./2014 502 yaba users (75% M and 25% F; 18–25 y old) Urine, hair (3 cm proximal) MA Urinalysis significantly better detect within 7 d of testing; hair analysis had better results after 14 d up to 3 mo. No urine positive after 30 d. 115 Strano Rossi et al./1999 20 subjects in methadone treatment (hashish users) Hair THC, CBD, CBN THC positive in 11/20 in the range 0.1–0.7 ng/mg CBD (0.7–14.1 ng/mg) and CBN (0.4/0.7 ng/mg). 116 Fucci et al./2001 24 heroin abusers OF THC, CBD, CBN Qualitative results: THC positive in 8/24 samples; CBD positive in 12/24; CBN positive in 13/24 specimens. 117 Musshoff et al./2003 20 drug abusers Hair THC, CBD, CBN 20/20 THC concentration range 0.25–0.73 ng/mg; CBD 0.81–19.02 ng/mg; CBN 0.12–1.48 ng/mg. 118 Dizioli Rodrigues et al./2007 8 cannabis users both sex, older than 21 y old. Hair THC, CBD, CBN 3/8 THC (0.13–0.31 ng/mg); CBD 0.12–0.31 ng/mg; CBN 0.16–0.48 ng/mg. 119 Emidio et al./2010 10 cannabis users (M) Hair THC, CBD, CBN The average concentrations of CBD, THC and CBN were as follows: 0.014 ng/mg, 0.056 ng/mg and 0.096 ng/mg, respectively. 120 Hall et al./1999 24 clinical samples Urine BE 22/24 qualitative detection of BE. 2/24 BE was 3.3 μg/mL and 7.1 μg/mL, respectively. 121 Follador et al./2004 11 COC users both sex, > 21 y old 21 sweat samples COC, CE Qualitative detection. 9/21 sweat samples positive for COC, 1/9 positivity for CE. 122 Bermejo et al./2006 15 COC users 20–40 y old (12 M, 3 F) Scalp hair (segmentation when lenght>10 cm) COC, CE All samples were positive for COC (0.43–8.98 ng/mg); CE often detected (0.42–2.23 ng/mg). 123 Alvarez et al./2007 10 COC and alcohol abusers Plasma COC, CE COC always detected (50.17–635.4 ng/mL); CE always found (25.39–59.61 ng/mL). 124 Fucci et al./2014 Cocaine abusers 50 scalp hair Levamisole, COC, tropococaine, norcocaine, CE COC (0.5–18 ng/mg) always detected; levamisole (0.2–0.8 ng/mg) found in 38% of examined samples seems to correlate with COC concentration. 125 Gambelunghe et al./2017 90 COC addicts (20 F, 70 M) Scalp hair COC, nor-COC, CE, tropococaine COC (1.63–39.29 ng/mg) always detected; norcocaine found in 58.9% of cases, CE in 20 samples, tropococaine randomly detected. 126 Lucas et al./2000 8 patients of methadone maintenance program (25–35 y old) Hair METH, EDDP METH ranged from 2.45 to 78.1 ng/mg; EDDP was in the range 0–7.76 ng/mg. The correlation between methadone dose and concentration in hair was statistically significant (p < 0.02). 127 Bermejo et al./2000 10 methadone-treated patients Plasma METH, EDDP METH concentration was in the range 0.16–1.92 μg/mL; EDDP range 0.05–0.47 μg/mL. 128 dos Santos Lucas et al./2000 4 patients in a methadone maintenance program OF METH, EDDP METH concentration was in the range 1.05–2.57 μg/mL; EDDP range 0.06–0.12 μg/mL. 129 Musshoff et al./2005 41 patients in maintenance program Hair METH, EDDP METH concentration: 0.25–13.29 ng/mg; EDDP concentration 0.05–2.17 ng/mg. A statistically significant correlation between dose and concentrations of METH and EDDP in hair found only in the proximal segments. 130 Fucci et al./2008 59 heroin addicts in methadone treatment OF METH, EDDP A stability study of methadone concentration in saliva during 12 mo showed METH stable at 4°C during 2 mo; EDDP was not stable during time. 131 Moller et al./2010 12 clinical samples Hair COD, MOR, 6-MAM 2/12 MOR positivities (0.503, 0.484 ng/mg) and MAM positivities (1.333, 1.301 ng/mg); 9/12 COD positivities (0.212–2.896 ng/mg). 132 Beck et al./2011 3 patients in maintenance program sampled by SPME Exhaled breath METH The experiment with the SPME based on the trapping of volatile analytes from air showed promising results, but the trapped amount was very low: methadone trapped on the fiber was < 1 pg/min. 133 Lee et al./1997 A 74-kg 31-y-old M volunteer Blood Tricyclic antidepressants After 4-h oral administration of 50 mg of imipramine, the concentration was 23.9 ng/mL 134 Guan et al./1999 A 68-kg 36-y-old M volunteer Urine Flunitrazepam, oxazolam Therapeutic oral doses of flunitrazepam (1.2 mg) and 4 d later oxazolam (2.5 mg). Urine was collected at 8.5, 11.5, 16.5 and 19 h for flunitrazepam showing a concentration of 71, 101, 44 and 13 ng/mL; Urine was collected at 8, 10, 16 and 23 h for oxazolam showing a concentration of 31, 50, 45 and 51 ng/mL. 135 Frison et al./2000 A patient under treatment with GHB Plasma, urine GHB Chromatographic evidence of GHB detection. 136 Frison et al./2001 A patient in ICU under treatment Plasma Midazolam Midazolam was qualitatively identified in the real sample. 137 Iwai et al./2004 2 volunteers: a 41-y-old M, a 30-y-old F Blood, urine Pentobarbital Oral administration of 50 mg Pentobarbital with collection at 8 and 24 h. Blood: male 0.502 and 0.292 μg/mL, female 0.794 and 0.454 μg/mL. Urine: 0.145 μg/mL and 0.15 μg/mL for male; 0.422 and 0.192 μg/mL for female. 138 Raikos et al./2009 9 patients with coronary bypass Urine Fentanyl, midazolam, lidocaine Dose: lidocaine 1 mg/kg; midazolam 0.025 mg/kg; fentanyl 0.01 mg/kg. Lidocaine 9/9 positive (0.73–4.88 μg/mL); midazolam 2/4 positive (0.34 and 0.37 μg/mL); fentanyl 3/7 positive (0.03–0.16 μg/mL). 139 De Giovanni et al./2002 25 heroin addicts in methadone treatment and 14 young people at the exit of a discotheque OF AP, MA, MDA, MDMA, METH, EDDP, COC, CE, THC, CBD, CBN SPME used to confirm immunochemical screening results. Heroin addicts: 21/25 positive to methadone and metabolite; 9/21 positive to other drugs (THC, CBD, CBN, COC, MDA, MDMA). Young people: 9/14 positive to at least 1 drug; 5/9 positive for 1 substance and 4/9 were poly-drug users. AP, MA and CE never detected. 140 Fucci et al./2003 53 drug abusers OF METH, EDDP, COC, CE, AP, MA, MDMA, MDEA, MBDB, CBD, CBN, THC The comparison between HS-SPME and DI-SPME disclosed advantages and disadvantages for each modality. 35/53 were positive to methadone (10–3800 ng/mL); 8/53 disclosed methadone < 10 ng/mL; 14/53 were positive to cannabinoids; 4/53 were positive to COC; 6/53 positive for MA (near to the LOD value). 141 Lachenmeier et al./2003 n.r. Hair METH, EDDP, COC, AP, MA, MDMA, MDA, MDEA, MBDB, BDB, CBD, CBN, THC Authentic samples from non-drug and multi-drug users were analyzed. In the case of a cannabis user, CBD 0.50 ng/mg, THC 0.95 ng/mg and CBN 1.93 ng/mg were detected. 142 Gentili et al./2004 183 young people (138 M, 45 F, 14–40 y old) Hair METH, COC, AP, MA, MDMA, MDA, MDEA, MBDB, KET 43/183 were drug free; 78/183 disclosed the use of 1 compound (73/78 COC); 62/183 were poly-drug abusers. 143 Strano Rossi et al./2005 2 people Urine 59 compounds (stimulants and narcotics) The method was developed for 59 compounds; it was applied to 2 real samples: case 1 methadone 200 ng/mL; case 2 ephedrine 20.2 μg/mL. 144 Merola et al./2010 32 young people in a dancing venue Hair KET, METH, COC, CE, THC, AP, MA, MDA, MDMA, MDEA, MBDB 27/32 tested positive for 1 or more substances. 15/32 were positive to COC that was quantified; 9/32 qualitative COC detection. 15/32 THC positivities were also quantified. 145 Gentili et al./2016 83 club goers (young people) OF METH, COC, CE, AP, MA, MDMA, MDA, COD, 6-MAM, MOR, KET, MDEA, MBDB, THC 75/83 samples were positive for 1 or more substances; 35/83 were poly-drug users, 21/83 were THC positive, 7/83 were MDMA positive; 8/83 COC detected, 4/83 KETdetected. Ref. Author/year Population Matrix Substances identified Results 106 Yashiki et al./1995 Suspected drug abusers Urine MA, AP No quantitative data. The method was found to be suitable for analysis of AP and MA in urine. 107 Lord et al./1997 4 clinical samples Urine MA, AP External calibration: AP range 0.2–4.9 μg/mL and MA 0–52 μg/mL. Standard addition: AP range 0.41–8.6 μg/mL and MA 0.05–39 μg/mL. The methods were then applied to several narcotic analgesics. 108 Myung et al./1998 2 subjects who were administered the stimulants Urine AP, MA, dimethamphetamine Case 1: dimethamphetamine and MA detected at 0.51 ± 0.01 and 0.15 ± 0.03 μg/mL, respectively. Case 2: AP and MA detected at 0.35 ± 0.02 and 1.08 ± 0.07 μg/mL, respectively. 109 Ugland et al./1999 1 real sample Urine AP, MA, MDMA, MDA, MDEA The method was developed with urine spiked samples; a real specimen showed the presence of MDA and MDMA. No quantitative data reported. 110 Gentili et al./2002 27 young people attending a disco (18 M, 9 F, mean age = 22 ± 5 y) 20 hair and 7 OF samples Amphetamine-like drugs and ketamine 3/20 hair samples were positive for ketamine (30–40 ng/mg); 1/3 was also positive for MDMA (2.3 ng/mg). 6/7 saliva samples were positive for ketamine, 1/6 also positive for MA, MDA and MDMA. 111 Musshoff et al./2002 Drug abusers 20 hair samples Amphetamines and synthetic designer drugs AP 0.2–21.8 ng/mg; MA 0.1–9.6 ng/mg; MDA 1.3–26.4 ng/mg; MDMA 0.5–12.4 ng/mg; MDEA 0.1–9.9 ng/mg; MBDB 0.2–0.9 ng/mg. 112 Musshoff et al./2002 Drug abusers Hair Amphetamines and synthetic designer drugs No quantitative data of real samples. The method was found to be suitable for analysis of hair samples from drug abusers. 113 Nishida et al./2006 A 37-y-old M Hair (also segmental analysis) MA, AP MA and AP were detected in 1-cm-long hair segments. 114 Junkuy et al./2014 502 yaba users (75% M and 25% F; 18–25 y old) Urine, hair (3 cm proximal) MA Urinalysis significantly better detect within 7 d of testing; hair analysis had better results after 14 d up to 3 mo. No urine positive after 30 d. 115 Strano Rossi et al./1999 20 subjects in methadone treatment (hashish users) Hair THC, CBD, CBN THC positive in 11/20 in the range 0.1–0.7 ng/mg CBD (0.7–14.1 ng/mg) and CBN (0.4/0.7 ng/mg). 116 Fucci et al./2001 24 heroin abusers OF THC, CBD, CBN Qualitative results: THC positive in 8/24 samples; CBD positive in 12/24; CBN positive in 13/24 specimens. 117 Musshoff et al./2003 20 drug abusers Hair THC, CBD, CBN 20/20 THC concentration range 0.25–0.73 ng/mg; CBD 0.81–19.02 ng/mg; CBN 0.12–1.48 ng/mg. 118 Dizioli Rodrigues et al./2007 8 cannabis users both sex, older than 21 y old. Hair THC, CBD, CBN 3/8 THC (0.13–0.31 ng/mg); CBD 0.12–0.31 ng/mg; CBN 0.16–0.48 ng/mg. 119 Emidio et al./2010 10 cannabis users (M) Hair THC, CBD, CBN The average concentrations of CBD, THC and CBN were as follows: 0.014 ng/mg, 0.056 ng/mg and 0.096 ng/mg, respectively. 120 Hall et al./1999 24 clinical samples Urine BE 22/24 qualitative detection of BE. 2/24 BE was 3.3 μg/mL and 7.1 μg/mL, respectively. 121 Follador et al./2004 11 COC users both sex, > 21 y old 21 sweat samples COC, CE Qualitative detection. 9/21 sweat samples positive for COC, 1/9 positivity for CE. 122 Bermejo et al./2006 15 COC users 20–40 y old (12 M, 3 F) Scalp hair (segmentation when lenght>10 cm) COC, CE All samples were positive for COC (0.43–8.98 ng/mg); CE often detected (0.42–2.23 ng/mg). 123 Alvarez et al./2007 10 COC and alcohol abusers Plasma COC, CE COC always detected (50.17–635.4 ng/mL); CE always found (25.39–59.61 ng/mL). 124 Fucci et al./2014 Cocaine abusers 50 scalp hair Levamisole, COC, tropococaine, norcocaine, CE COC (0.5–18 ng/mg) always detected; levamisole (0.2–0.8 ng/mg) found in 38% of examined samples seems to correlate with COC concentration. 125 Gambelunghe et al./2017 90 COC addicts (20 F, 70 M) Scalp hair COC, nor-COC, CE, tropococaine COC (1.63–39.29 ng/mg) always detected; norcocaine found in 58.9% of cases, CE in 20 samples, tropococaine randomly detected. 126 Lucas et al./2000 8 patients of methadone maintenance program (25–35 y old) Hair METH, EDDP METH ranged from 2.45 to 78.1 ng/mg; EDDP was in the range 0–7.76 ng/mg. The correlation between methadone dose and concentration in hair was statistically significant (p < 0.02). 127 Bermejo et al./2000 10 methadone-treated patients Plasma METH, EDDP METH concentration was in the range 0.16–1.92 μg/mL; EDDP range 0.05–0.47 μg/mL. 128 dos Santos Lucas et al./2000 4 patients in a methadone maintenance program OF METH, EDDP METH concentration was in the range 1.05–2.57 μg/mL; EDDP range 0.06–0.12 μg/mL. 129 Musshoff et al./2005 41 patients in maintenance program Hair METH, EDDP METH concentration: 0.25–13.29 ng/mg; EDDP concentration 0.05–2.17 ng/mg. A statistically significant correlation between dose and concentrations of METH and EDDP in hair found only in the proximal segments. 130 Fucci et al./2008 59 heroin addicts in methadone treatment OF METH, EDDP A stability study of methadone concentration in saliva during 12 mo showed METH stable at 4°C during 2 mo; EDDP was not stable during time. 131 Moller et al./2010 12 clinical samples Hair COD, MOR, 6-MAM 2/12 MOR positivities (0.503, 0.484 ng/mg) and MAM positivities (1.333, 1.301 ng/mg); 9/12 COD positivities (0.212–2.896 ng/mg). 132 Beck et al./2011 3 patients in maintenance program sampled by SPME Exhaled breath METH The experiment with the SPME based on the trapping of volatile analytes from air showed promising results, but the trapped amount was very low: methadone trapped on the fiber was < 1 pg/min. 133 Lee et al./1997 A 74-kg 31-y-old M volunteer Blood Tricyclic antidepressants After 4-h oral administration of 50 mg of imipramine, the concentration was 23.9 ng/mL 134 Guan et al./1999 A 68-kg 36-y-old M volunteer Urine Flunitrazepam, oxazolam Therapeutic oral doses of flunitrazepam (1.2 mg) and 4 d later oxazolam (2.5 mg). Urine was collected at 8.5, 11.5, 16.5 and 19 h for flunitrazepam showing a concentration of 71, 101, 44 and 13 ng/mL; Urine was collected at 8, 10, 16 and 23 h for oxazolam showing a concentration of 31, 50, 45 and 51 ng/mL. 135 Frison et al./2000 A patient under treatment with GHB Plasma, urine GHB Chromatographic evidence of GHB detection. 136 Frison et al./2001 A patient in ICU under treatment Plasma Midazolam Midazolam was qualitatively identified in the real sample. 137 Iwai et al./2004 2 volunteers: a 41-y-old M, a 30-y-old F Blood, urine Pentobarbital Oral administration of 50 mg Pentobarbital with collection at 8 and 24 h. Blood: male 0.502 and 0.292 μg/mL, female 0.794 and 0.454 μg/mL. Urine: 0.145 μg/mL and 0.15 μg/mL for male; 0.422 and 0.192 μg/mL for female. 138 Raikos et al./2009 9 patients with coronary bypass Urine Fentanyl, midazolam, lidocaine Dose: lidocaine 1 mg/kg; midazolam 0.025 mg/kg; fentanyl 0.01 mg/kg. Lidocaine 9/9 positive (0.73–4.88 μg/mL); midazolam 2/4 positive (0.34 and 0.37 μg/mL); fentanyl 3/7 positive (0.03–0.16 μg/mL). 139 De Giovanni et al./2002 25 heroin addicts in methadone treatment and 14 young people at the exit of a discotheque OF AP, MA, MDA, MDMA, METH, EDDP, COC, CE, THC, CBD, CBN SPME used to confirm immunochemical screening results. Heroin addicts: 21/25 positive to methadone and metabolite; 9/21 positive to other drugs (THC, CBD, CBN, COC, MDA, MDMA). Young people: 9/14 positive to at least 1 drug; 5/9 positive for 1 substance and 4/9 were poly-drug users. AP, MA and CE never detected. 140 Fucci et al./2003 53 drug abusers OF METH, EDDP, COC, CE, AP, MA, MDMA, MDEA, MBDB, CBD, CBN, THC The comparison between HS-SPME and DI-SPME disclosed advantages and disadvantages for each modality. 35/53 were positive to methadone (10–3800 ng/mL); 8/53 disclosed methadone < 10 ng/mL; 14/53 were positive to cannabinoids; 4/53 were positive to COC; 6/53 positive for MA (near to the LOD value). 141 Lachenmeier et al./2003 n.r. Hair METH, EDDP, COC, AP, MA, MDMA, MDA, MDEA, MBDB, BDB, CBD, CBN, THC Authentic samples from non-drug and multi-drug users were analyzed. In the case of a cannabis user, CBD 0.50 ng/mg, THC 0.95 ng/mg and CBN 1.93 ng/mg were detected. 142 Gentili et al./2004 183 young people (138 M, 45 F, 14–40 y old) Hair METH, COC, AP, MA, MDMA, MDA, MDEA, MBDB, KET 43/183 were drug free; 78/183 disclosed the use of 1 compound (73/78 COC); 62/183 were poly-drug abusers. 143 Strano Rossi et al./2005 2 people Urine 59 compounds (stimulants and narcotics) The method was developed for 59 compounds; it was applied to 2 real samples: case 1 methadone 200 ng/mL; case 2 ephedrine 20.2 μg/mL. 144 Merola et al./2010 32 young people in a dancing venue Hair KET, METH, COC, CE, THC, AP, MA, MDA, MDMA, MDEA, MBDB 27/32 tested positive for 1 or more substances. 15/32 were positive to COC that was quantified; 9/32 qualitative COC detection. 15/32 THC positivities were also quantified. 145 Gentili et al./2016 83 club goers (young people) OF METH, COC, CE, AP, MA, MDMA, MDA, COD, 6-MAM, MOR, KET, MDEA, MBDB, THC 75/83 samples were positive for 1 or more substances; 35/83 were poly-drug users, 21/83 were THC positive, 7/83 were MDMA positive; 8/83 COC detected, 4/83 KETdetected. n.r., not reported. Open in new tab Table 5 Analytical studies of the SPME technique for the detection of xenobiotics in biological samples Ref. Author/year Population Matrix Substances identified Results 106 Yashiki et al./1995 Suspected drug abusers Urine MA, AP No quantitative data. The method was found to be suitable for analysis of AP and MA in urine. 107 Lord et al./1997 4 clinical samples Urine MA, AP External calibration: AP range 0.2–4.9 μg/mL and MA 0–52 μg/mL. Standard addition: AP range 0.41–8.6 μg/mL and MA 0.05–39 μg/mL. The methods were then applied to several narcotic analgesics. 108 Myung et al./1998 2 subjects who were administered the stimulants Urine AP, MA, dimethamphetamine Case 1: dimethamphetamine and MA detected at 0.51 ± 0.01 and 0.15 ± 0.03 μg/mL, respectively. Case 2: AP and MA detected at 0.35 ± 0.02 and 1.08 ± 0.07 μg/mL, respectively. 109 Ugland et al./1999 1 real sample Urine AP, MA, MDMA, MDA, MDEA The method was developed with urine spiked samples; a real specimen showed the presence of MDA and MDMA. No quantitative data reported. 110 Gentili et al./2002 27 young people attending a disco (18 M, 9 F, mean age = 22 ± 5 y) 20 hair and 7 OF samples Amphetamine-like drugs and ketamine 3/20 hair samples were positive for ketamine (30–40 ng/mg); 1/3 was also positive for MDMA (2.3 ng/mg). 6/7 saliva samples were positive for ketamine, 1/6 also positive for MA, MDA and MDMA. 111 Musshoff et al./2002 Drug abusers 20 hair samples Amphetamines and synthetic designer drugs AP 0.2–21.8 ng/mg; MA 0.1–9.6 ng/mg; MDA 1.3–26.4 ng/mg; MDMA 0.5–12.4 ng/mg; MDEA 0.1–9.9 ng/mg; MBDB 0.2–0.9 ng/mg. 112 Musshoff et al./2002 Drug abusers Hair Amphetamines and synthetic designer drugs No quantitative data of real samples. The method was found to be suitable for analysis of hair samples from drug abusers. 113 Nishida et al./2006 A 37-y-old M Hair (also segmental analysis) MA, AP MA and AP were detected in 1-cm-long hair segments. 114 Junkuy et al./2014 502 yaba users (75% M and 25% F; 18–25 y old) Urine, hair (3 cm proximal) MA Urinalysis significantly better detect within 7 d of testing; hair analysis had better results after 14 d up to 3 mo. No urine positive after 30 d. 115 Strano Rossi et al./1999 20 subjects in methadone treatment (hashish users) Hair THC, CBD, CBN THC positive in 11/20 in the range 0.1–0.7 ng/mg CBD (0.7–14.1 ng/mg) and CBN (0.4/0.7 ng/mg). 116 Fucci et al./2001 24 heroin abusers OF THC, CBD, CBN Qualitative results: THC positive in 8/24 samples; CBD positive in 12/24; CBN positive in 13/24 specimens. 117 Musshoff et al./2003 20 drug abusers Hair THC, CBD, CBN 20/20 THC concentration range 0.25–0.73 ng/mg; CBD 0.81–19.02 ng/mg; CBN 0.12–1.48 ng/mg. 118 Dizioli Rodrigues et al./2007 8 cannabis users both sex, older than 21 y old. Hair THC, CBD, CBN 3/8 THC (0.13–0.31 ng/mg); CBD 0.12–0.31 ng/mg; CBN 0.16–0.48 ng/mg. 119 Emidio et al./2010 10 cannabis users (M) Hair THC, CBD, CBN The average concentrations of CBD, THC and CBN were as follows: 0.014 ng/mg, 0.056 ng/mg and 0.096 ng/mg, respectively. 120 Hall et al./1999 24 clinical samples Urine BE 22/24 qualitative detection of BE. 2/24 BE was 3.3 μg/mL and 7.1 μg/mL, respectively. 121 Follador et al./2004 11 COC users both sex, > 21 y old 21 sweat samples COC, CE Qualitative detection. 9/21 sweat samples positive for COC, 1/9 positivity for CE. 122 Bermejo et al./2006 15 COC users 20–40 y old (12 M, 3 F) Scalp hair (segmentation when lenght>10 cm) COC, CE All samples were positive for COC (0.43–8.98 ng/mg); CE often detected (0.42–2.23 ng/mg). 123 Alvarez et al./2007 10 COC and alcohol abusers Plasma COC, CE COC always detected (50.17–635.4 ng/mL); CE always found (25.39–59.61 ng/mL). 124 Fucci et al./2014 Cocaine abusers 50 scalp hair Levamisole, COC, tropococaine, norcocaine, CE COC (0.5–18 ng/mg) always detected; levamisole (0.2–0.8 ng/mg) found in 38% of examined samples seems to correlate with COC concentration. 125 Gambelunghe et al./2017 90 COC addicts (20 F, 70 M) Scalp hair COC, nor-COC, CE, tropococaine COC (1.63–39.29 ng/mg) always detected; norcocaine found in 58.9% of cases, CE in 20 samples, tropococaine randomly detected. 126 Lucas et al./2000 8 patients of methadone maintenance program (25–35 y old) Hair METH, EDDP METH ranged from 2.45 to 78.1 ng/mg; EDDP was in the range 0–7.76 ng/mg. The correlation between methadone dose and concentration in hair was statistically significant (p < 0.02). 127 Bermejo et al./2000 10 methadone-treated patients Plasma METH, EDDP METH concentration was in the range 0.16–1.92 μg/mL; EDDP range 0.05–0.47 μg/mL. 128 dos Santos Lucas et al./2000 4 patients in a methadone maintenance program OF METH, EDDP METH concentration was in the range 1.05–2.57 μg/mL; EDDP range 0.06–0.12 μg/mL. 129 Musshoff et al./2005 41 patients in maintenance program Hair METH, EDDP METH concentration: 0.25–13.29 ng/mg; EDDP concentration 0.05–2.17 ng/mg. A statistically significant correlation between dose and concentrations of METH and EDDP in hair found only in the proximal segments. 130 Fucci et al./2008 59 heroin addicts in methadone treatment OF METH, EDDP A stability study of methadone concentration in saliva during 12 mo showed METH stable at 4°C during 2 mo; EDDP was not stable during time. 131 Moller et al./2010 12 clinical samples Hair COD, MOR, 6-MAM 2/12 MOR positivities (0.503, 0.484 ng/mg) and MAM positivities (1.333, 1.301 ng/mg); 9/12 COD positivities (0.212–2.896 ng/mg). 132 Beck et al./2011 3 patients in maintenance program sampled by SPME Exhaled breath METH The experiment with the SPME based on the trapping of volatile analytes from air showed promising results, but the trapped amount was very low: methadone trapped on the fiber was < 1 pg/min. 133 Lee et al./1997 A 74-kg 31-y-old M volunteer Blood Tricyclic antidepressants After 4-h oral administration of 50 mg of imipramine, the concentration was 23.9 ng/mL 134 Guan et al./1999 A 68-kg 36-y-old M volunteer Urine Flunitrazepam, oxazolam Therapeutic oral doses of flunitrazepam (1.2 mg) and 4 d later oxazolam (2.5 mg). Urine was collected at 8.5, 11.5, 16.5 and 19 h for flunitrazepam showing a concentration of 71, 101, 44 and 13 ng/mL; Urine was collected at 8, 10, 16 and 23 h for oxazolam showing a concentration of 31, 50, 45 and 51 ng/mL. 135 Frison et al./2000 A patient under treatment with GHB Plasma, urine GHB Chromatographic evidence of GHB detection. 136 Frison et al./2001 A patient in ICU under treatment Plasma Midazolam Midazolam was qualitatively identified in the real sample. 137 Iwai et al./2004 2 volunteers: a 41-y-old M, a 30-y-old F Blood, urine Pentobarbital Oral administration of 50 mg Pentobarbital with collection at 8 and 24 h. Blood: male 0.502 and 0.292 μg/mL, female 0.794 and 0.454 μg/mL. Urine: 0.145 μg/mL and 0.15 μg/mL for male; 0.422 and 0.192 μg/mL for female. 138 Raikos et al./2009 9 patients with coronary bypass Urine Fentanyl, midazolam, lidocaine Dose: lidocaine 1 mg/kg; midazolam 0.025 mg/kg; fentanyl 0.01 mg/kg. Lidocaine 9/9 positive (0.73–4.88 μg/mL); midazolam 2/4 positive (0.34 and 0.37 μg/mL); fentanyl 3/7 positive (0.03–0.16 μg/mL). 139 De Giovanni et al./2002 25 heroin addicts in methadone treatment and 14 young people at the exit of a discotheque OF AP, MA, MDA, MDMA, METH, EDDP, COC, CE, THC, CBD, CBN SPME used to confirm immunochemical screening results. Heroin addicts: 21/25 positive to methadone and metabolite; 9/21 positive to other drugs (THC, CBD, CBN, COC, MDA, MDMA). Young people: 9/14 positive to at least 1 drug; 5/9 positive for 1 substance and 4/9 were poly-drug users. AP, MA and CE never detected. 140 Fucci et al./2003 53 drug abusers OF METH, EDDP, COC, CE, AP, MA, MDMA, MDEA, MBDB, CBD, CBN, THC The comparison between HS-SPME and DI-SPME disclosed advantages and disadvantages for each modality. 35/53 were positive to methadone (10–3800 ng/mL); 8/53 disclosed methadone < 10 ng/mL; 14/53 were positive to cannabinoids; 4/53 were positive to COC; 6/53 positive for MA (near to the LOD value). 141 Lachenmeier et al./2003 n.r. Hair METH, EDDP, COC, AP, MA, MDMA, MDA, MDEA, MBDB, BDB, CBD, CBN, THC Authentic samples from non-drug and multi-drug users were analyzed. In the case of a cannabis user, CBD 0.50 ng/mg, THC 0.95 ng/mg and CBN 1.93 ng/mg were detected. 142 Gentili et al./2004 183 young people (138 M, 45 F, 14–40 y old) Hair METH, COC, AP, MA, MDMA, MDA, MDEA, MBDB, KET 43/183 were drug free; 78/183 disclosed the use of 1 compound (73/78 COC); 62/183 were poly-drug abusers. 143 Strano Rossi et al./2005 2 people Urine 59 compounds (stimulants and narcotics) The method was developed for 59 compounds; it was applied to 2 real samples: case 1 methadone 200 ng/mL; case 2 ephedrine 20.2 μg/mL. 144 Merola et al./2010 32 young people in a dancing venue Hair KET, METH, COC, CE, THC, AP, MA, MDA, MDMA, MDEA, MBDB 27/32 tested positive for 1 or more substances. 15/32 were positive to COC that was quantified; 9/32 qualitative COC detection. 15/32 THC positivities were also quantified. 145 Gentili et al./2016 83 club goers (young people) OF METH, COC, CE, AP, MA, MDMA, MDA, COD, 6-MAM, MOR, KET, MDEA, MBDB, THC 75/83 samples were positive for 1 or more substances; 35/83 were poly-drug users, 21/83 were THC positive, 7/83 were MDMA positive; 8/83 COC detected, 4/83 KETdetected. Ref. Author/year Population Matrix Substances identified Results 106 Yashiki et al./1995 Suspected drug abusers Urine MA, AP No quantitative data. The method was found to be suitable for analysis of AP and MA in urine. 107 Lord et al./1997 4 clinical samples Urine MA, AP External calibration: AP range 0.2–4.9 μg/mL and MA 0–52 μg/mL. Standard addition: AP range 0.41–8.6 μg/mL and MA 0.05–39 μg/mL. The methods were then applied to several narcotic analgesics. 108 Myung et al./1998 2 subjects who were administered the stimulants Urine AP, MA, dimethamphetamine Case 1: dimethamphetamine and MA detected at 0.51 ± 0.01 and 0.15 ± 0.03 μg/mL, respectively. Case 2: AP and MA detected at 0.35 ± 0.02 and 1.08 ± 0.07 μg/mL, respectively. 109 Ugland et al./1999 1 real sample Urine AP, MA, MDMA, MDA, MDEA The method was developed with urine spiked samples; a real specimen showed the presence of MDA and MDMA. No quantitative data reported. 110 Gentili et al./2002 27 young people attending a disco (18 M, 9 F, mean age = 22 ± 5 y) 20 hair and 7 OF samples Amphetamine-like drugs and ketamine 3/20 hair samples were positive for ketamine (30–40 ng/mg); 1/3 was also positive for MDMA (2.3 ng/mg). 6/7 saliva samples were positive for ketamine, 1/6 also positive for MA, MDA and MDMA. 111 Musshoff et al./2002 Drug abusers 20 hair samples Amphetamines and synthetic designer drugs AP 0.2–21.8 ng/mg; MA 0.1–9.6 ng/mg; MDA 1.3–26.4 ng/mg; MDMA 0.5–12.4 ng/mg; MDEA 0.1–9.9 ng/mg; MBDB 0.2–0.9 ng/mg. 112 Musshoff et al./2002 Drug abusers Hair Amphetamines and synthetic designer drugs No quantitative data of real samples. The method was found to be suitable for analysis of hair samples from drug abusers. 113 Nishida et al./2006 A 37-y-old M Hair (also segmental analysis) MA, AP MA and AP were detected in 1-cm-long hair segments. 114 Junkuy et al./2014 502 yaba users (75% M and 25% F; 18–25 y old) Urine, hair (3 cm proximal) MA Urinalysis significantly better detect within 7 d of testing; hair analysis had better results after 14 d up to 3 mo. No urine positive after 30 d. 115 Strano Rossi et al./1999 20 subjects in methadone treatment (hashish users) Hair THC, CBD, CBN THC positive in 11/20 in the range 0.1–0.7 ng/mg CBD (0.7–14.1 ng/mg) and CBN (0.4/0.7 ng/mg). 116 Fucci et al./2001 24 heroin abusers OF THC, CBD, CBN Qualitative results: THC positive in 8/24 samples; CBD positive in 12/24; CBN positive in 13/24 specimens. 117 Musshoff et al./2003 20 drug abusers Hair THC, CBD, CBN 20/20 THC concentration range 0.25–0.73 ng/mg; CBD 0.81–19.02 ng/mg; CBN 0.12–1.48 ng/mg. 118 Dizioli Rodrigues et al./2007 8 cannabis users both sex, older than 21 y old. Hair THC, CBD, CBN 3/8 THC (0.13–0.31 ng/mg); CBD 0.12–0.31 ng/mg; CBN 0.16–0.48 ng/mg. 119 Emidio et al./2010 10 cannabis users (M) Hair THC, CBD, CBN The average concentrations of CBD, THC and CBN were as follows: 0.014 ng/mg, 0.056 ng/mg and 0.096 ng/mg, respectively. 120 Hall et al./1999 24 clinical samples Urine BE 22/24 qualitative detection of BE. 2/24 BE was 3.3 μg/mL and 7.1 μg/mL, respectively. 121 Follador et al./2004 11 COC users both sex, > 21 y old 21 sweat samples COC, CE Qualitative detection. 9/21 sweat samples positive for COC, 1/9 positivity for CE. 122 Bermejo et al./2006 15 COC users 20–40 y old (12 M, 3 F) Scalp hair (segmentation when lenght>10 cm) COC, CE All samples were positive for COC (0.43–8.98 ng/mg); CE often detected (0.42–2.23 ng/mg). 123 Alvarez et al./2007 10 COC and alcohol abusers Plasma COC, CE COC always detected (50.17–635.4 ng/mL); CE always found (25.39–59.61 ng/mL). 124 Fucci et al./2014 Cocaine abusers 50 scalp hair Levamisole, COC, tropococaine, norcocaine, CE COC (0.5–18 ng/mg) always detected; levamisole (0.2–0.8 ng/mg) found in 38% of examined samples seems to correlate with COC concentration. 125 Gambelunghe et al./2017 90 COC addicts (20 F, 70 M) Scalp hair COC, nor-COC, CE, tropococaine COC (1.63–39.29 ng/mg) always detected; norcocaine found in 58.9% of cases, CE in 20 samples, tropococaine randomly detected. 126 Lucas et al./2000 8 patients of methadone maintenance program (25–35 y old) Hair METH, EDDP METH ranged from 2.45 to 78.1 ng/mg; EDDP was in the range 0–7.76 ng/mg. The correlation between methadone dose and concentration in hair was statistically significant (p < 0.02). 127 Bermejo et al./2000 10 methadone-treated patients Plasma METH, EDDP METH concentration was in the range 0.16–1.92 μg/mL; EDDP range 0.05–0.47 μg/mL. 128 dos Santos Lucas et al./2000 4 patients in a methadone maintenance program OF METH, EDDP METH concentration was in the range 1.05–2.57 μg/mL; EDDP range 0.06–0.12 μg/mL. 129 Musshoff et al./2005 41 patients in maintenance program Hair METH, EDDP METH concentration: 0.25–13.29 ng/mg; EDDP concentration 0.05–2.17 ng/mg. A statistically significant correlation between dose and concentrations of METH and EDDP in hair found only in the proximal segments. 130 Fucci et al./2008 59 heroin addicts in methadone treatment OF METH, EDDP A stability study of methadone concentration in saliva during 12 mo showed METH stable at 4°C during 2 mo; EDDP was not stable during time. 131 Moller et al./2010 12 clinical samples Hair COD, MOR, 6-MAM 2/12 MOR positivities (0.503, 0.484 ng/mg) and MAM positivities (1.333, 1.301 ng/mg); 9/12 COD positivities (0.212–2.896 ng/mg). 132 Beck et al./2011 3 patients in maintenance program sampled by SPME Exhaled breath METH The experiment with the SPME based on the trapping of volatile analytes from air showed promising results, but the trapped amount was very low: methadone trapped on the fiber was < 1 pg/min. 133 Lee et al./1997 A 74-kg 31-y-old M volunteer Blood Tricyclic antidepressants After 4-h oral administration of 50 mg of imipramine, the concentration was 23.9 ng/mL 134 Guan et al./1999 A 68-kg 36-y-old M volunteer Urine Flunitrazepam, oxazolam Therapeutic oral doses of flunitrazepam (1.2 mg) and 4 d later oxazolam (2.5 mg). Urine was collected at 8.5, 11.5, 16.5 and 19 h for flunitrazepam showing a concentration of 71, 101, 44 and 13 ng/mL; Urine was collected at 8, 10, 16 and 23 h for oxazolam showing a concentration of 31, 50, 45 and 51 ng/mL. 135 Frison et al./2000 A patient under treatment with GHB Plasma, urine GHB Chromatographic evidence of GHB detection. 136 Frison et al./2001 A patient in ICU under treatment Plasma Midazolam Midazolam was qualitatively identified in the real sample. 137 Iwai et al./2004 2 volunteers: a 41-y-old M, a 30-y-old F Blood, urine Pentobarbital Oral administration of 50 mg Pentobarbital with collection at 8 and 24 h. Blood: male 0.502 and 0.292 μg/mL, female 0.794 and 0.454 μg/mL. Urine: 0.145 μg/mL and 0.15 μg/mL for male; 0.422 and 0.192 μg/mL for female. 138 Raikos et al./2009 9 patients with coronary bypass Urine Fentanyl, midazolam, lidocaine Dose: lidocaine 1 mg/kg; midazolam 0.025 mg/kg; fentanyl 0.01 mg/kg. Lidocaine 9/9 positive (0.73–4.88 μg/mL); midazolam 2/4 positive (0.34 and 0.37 μg/mL); fentanyl 3/7 positive (0.03–0.16 μg/mL). 139 De Giovanni et al./2002 25 heroin addicts in methadone treatment and 14 young people at the exit of a discotheque OF AP, MA, MDA, MDMA, METH, EDDP, COC, CE, THC, CBD, CBN SPME used to confirm immunochemical screening results. Heroin addicts: 21/25 positive to methadone and metabolite; 9/21 positive to other drugs (THC, CBD, CBN, COC, MDA, MDMA). Young people: 9/14 positive to at least 1 drug; 5/9 positive for 1 substance and 4/9 were poly-drug users. AP, MA and CE never detected. 140 Fucci et al./2003 53 drug abusers OF METH, EDDP, COC, CE, AP, MA, MDMA, MDEA, MBDB, CBD, CBN, THC The comparison between HS-SPME and DI-SPME disclosed advantages and disadvantages for each modality. 35/53 were positive to methadone (10–3800 ng/mL); 8/53 disclosed methadone < 10 ng/mL; 14/53 were positive to cannabinoids; 4/53 were positive to COC; 6/53 positive for MA (near to the LOD value). 141 Lachenmeier et al./2003 n.r. Hair METH, EDDP, COC, AP, MA, MDMA, MDA, MDEA, MBDB, BDB, CBD, CBN, THC Authentic samples from non-drug and multi-drug users were analyzed. In the case of a cannabis user, CBD 0.50 ng/mg, THC 0.95 ng/mg and CBN 1.93 ng/mg were detected. 142 Gentili et al./2004 183 young people (138 M, 45 F, 14–40 y old) Hair METH, COC, AP, MA, MDMA, MDA, MDEA, MBDB, KET 43/183 were drug free; 78/183 disclosed the use of 1 compound (73/78 COC); 62/183 were poly-drug abusers. 143 Strano Rossi et al./2005 2 people Urine 59 compounds (stimulants and narcotics) The method was developed for 59 compounds; it was applied to 2 real samples: case 1 methadone 200 ng/mL; case 2 ephedrine 20.2 μg/mL. 144 Merola et al./2010 32 young people in a dancing venue Hair KET, METH, COC, CE, THC, AP, MA, MDA, MDMA, MDEA, MBDB 27/32 tested positive for 1 or more substances. 15/32 were positive to COC that was quantified; 9/32 qualitative COC detection. 15/32 THC positivities were also quantified. 145 Gentili et al./2016 83 club goers (young people) OF METH, COC, CE, AP, MA, MDMA, MDA, COD, 6-MAM, MOR, KET, MDEA, MBDB, THC 75/83 samples were positive for 1 or more substances; 35/83 were poly-drug users, 21/83 were THC positive, 7/83 were MDMA positive; 8/83 COC detected, 4/83 KETdetected. n.r., not reported. Open in new tab A review of the application of SPME analysis of forensic specimens included diagnosis of acute intoxication with illicit and therapeutic drugs as well as determination of volatile toxic molecules and environmental xenobiotics. SPME has also been used to identify white phosphorous as a cause of burning of an accidentally injured woman (103). According to Tables 3–5, a total of 89 of the 133 articles were selected. The wide variability of the results allows only tentative assessments of SPME with respect to the xenobiotics investigated and to some specific forensic contexts. Xenobiotic detection Amphetamines and congeners Since 1995, a total of 25 of the 89 papers have been published about the simultaneous detection of AP-like compounds by SPME, both detecting only some of the AP congeners or identifying multiple illicit drugs, including this class of compounds. The applicability of the methods was demonstrated in some actual specimens mainly obtained from people who abuse drugs or young people approached out of dance clubs. The technique was applied to urine (71, 93, 106–109, 114, 143), hair samples (67, 96, 104, 110–114, 141, 142, 144), oral fluid (110, 139, 140, 145) and sweat (102). Most of the papers considered only AP and/or methamphetamine (MA) detection (57, 67, 70, 93, 106, 107, 113, 114). Some papers also investigated 3,4-methylendioxymethamphetamine (MDMA), 3,4-methylendioxyamphetamine (MDA), 3,4-methylendioxyethylmphetamine (MDEA) and/or other congeners (65, 71, 108, 109). Additionally, 14 of the 25 papers examined the possibility of simultaneous detection of APs together with other illicit compounds (96, 102, 104, 109–112, 139–145). SPME was mainly carried out in the headspace mode. Myung et al. (108) and De Giovanni et al. (139) used direct immersion of the fiber. The fiber chosen was always PDMS with different film thicknesses, except Junkuy et al. (114), who identified MA in urine and hair of 502 yaba users by means of a PDMS/DVB fiber. Lachenmeier et al. (141)—for the simultaneous detection of various illicit drugs in hair—and Musshoff et al. (112)—for the detection of AP-like and synthetic designer drugs from hair of people who abuse drugs—employed HS-SPDE. As derivatization can improve the performance of the method, derivatization is often used with different kinds of reagents applying in-sample derivatization, on-fiber or in the injector of the GC. In-matrix derivatization was preferably performed (67, 70, 71, 93, 96, 109, 113, 145), but on-fiber derivatization (111, 112, 141, 144) and in the injector of the GC (57, 65) were also used. For enantioselective determination, the APs were in-sample derivatized with trifluoroacetyl-prolyl chloride as the chiral derivatizing reagent directly added to urine with an LOD of 30 ng/mL for both pairs of enantiomers with the purpose of workplace drug testing (93). In the studies reviewed, validation was always performed, and the IS used was mainly the corresponding deuterated (67, 71, 106, 111–113, 139–141) or consisting of different compounds (109, 110, 114, 142–145). In two studies, no IS was used (107, 108). Cannabinoids The literature reviewed disclosed 18 of the 89 applications of SPME to the detection of cannabinoids, eight of which examined the psychoactive cannabis substances together with other illegal drugs. Most papers simultaneously analyzed tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN) in hair samples (95, 97, 101, 105, 115, 117–119), as well as other drugs (104, 141, 144). In oral fluid, these compounds were detected alone (98, 116) and together with other drugs (139, 140, 145). In sweat (102) and urine (143), THC was detected together with other drugs. Two studies were performed on the hair of a large population, including children with parents abusing illicit drugs (97, 101). The SPME technique was employed either by headspace mode (95, 97, 101, 102, 104, 105, 118, 119, 144, 145) or by direct immersion (98, 115, 116, 139, 140); both methodologies were adopted by Strano Rossi et al. (143); moreover, HS-SPDE was performed twice (117, 141). A PDMS fiber was mainly used for these highly lipophilic substances, although with different film thicknesses. Cannabinoids in oral fluid were mainly detected by direct immersion of the fiber without derivatization, except for Gentili et al. (145), who used HS-SPME after in-matrix silyl derivatization. On the other hand, hair samples that required previous decontamination and hydrolysis were examined by DI-SPME (115), HS-SPME (95, 97, 101, 104, 105, 118, 119, 144) and HS-SPDE (117, 141), with (95, 97, 101, 105, 117, 141, 144) and without derivatization (104, 115, 118, 119). Fully automated analysis of the three cannabinoids by HS-SPME or HS-SPDE directly on hair hydrolysate followed by on-coating derivatization with N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) enabled the detection limits of 0.05–0.14 ng/mg (105, 117). A lower detection limit (0.003 ng/mg) has been achieved by Pragst et al. (97) using in-sample silyl derivatization of hair. Fully automated analysis of the three cannabinoids THC, CBD and CBN on hair without derivatization (115, 118, 119) showed detection limits of 0.1, 0.07 and 0.031 ng/mg, respectively. Finally, 14 of the 18 studies performed validation of the method (95, 97, 98, 101, 102, 115, 117–119, 140, 141, 143–145), and deuterated IS has been predominantly used. Opioids A total of 17 of the 89 papers considered methadone and heroin metabolites in the SPME analysis, mainly for the detection of methadone and its primary metabolite (66, 126–130, 132), often with simultaneous identification of other substances (96, 102, 104, 139–142, 144, 145). Only three studies were devoted to the analysis of morphine (MOR), codeine (COD) and monoacetylmorphine (MAM); the first (131) aimed to detect them in hair with in-matrix trimethylsilyl derivatization by using a PDMS fiber in the HS-SPME mode with an LOD as low as 0.05 ng/mg for MOR and MAM. The second (145) and third (96) studies examined many drugs of abuse, including MOR, COD, MAM and methadone in oral fluid and hair, respectively, by using a PDMS fiber and HS-SPME GC–MS after in-matrix derivatization. Regarding the matrix investigated, it can be noted that of the 17 papers, five analyzed oral fluid, nine examined hair, and three explored sweat, breath or plasma. In the studies reviewed, HS-SPME has been performed for the detection of opiates in hair (66, 96, 104, 129, 131, 142, 144), in oral fluid (145) and in sweat (102). DI-SPME was also used for the detection of opiates in oral fluid (128, 130, 139, 140), hair (126) and plasma (127). HS-SPDE has been performed for the simultaneous detection of methadone with other illicit drugs in hair (141). A different experiment was carried out by Beck et al. (132): they sampled the exhaled breath of three patients in a methadone maintenance program, directly trapping breath by SPME. An 85-μm PA fiber with 10 minutes of sampling time was used, and subsequent liquid chromatography tandem–mass spectrometry (LC–MS-MS) analysis was performed. This experiment, based on the trapping of volatile analytes from air, showed promising results, but the trapped amount was very low; in fact, the amount of methadone trapped on the fiber was < 1 pg/min. All the studies reviewed, except Beck et al. (132), performed validation; the samples have always been examined with GC–MS, except Strano-Rossi et al. (143), who used GC with nitrogen phosphorous detector (GC-NPD), Lachenmeier et al. (141), who employed GC–MS-MS, and Beck et al. (132), who applied LC–MS-MS. Finally, Raikos et al. (138) determined fentanyl by HS-SPME and GC-NPD in the urine of patients with coronary bypass, obtaining an LOD as low as 1.4 ng/mL. Cocaine and metabolites After the intake of COC, some metabolites can arise, which can be still active, such as CE, or without any pharmacological activity (benzoylecgonine [BE]); the detection of BE requires derivatization to achieve a suitable chromatographic separation; hence, their detection requires strategies to obtain satisfactory results. In the papers reviewed (15/89), some authors chose to detect COC itself, while others preferred to simultaneously identify COC together with its metabolites. Hall et al. (120) analyzed BE in urine after in-matrix derivatization with hexyl chloroformate and extraction by SPME; GC–MS determination allowed them to achieve an LOD of 30 ng/mL. BE was also detected by de Toledo et al. (74) in the hair of a victim together with COC and CE after in-matrix derivatization with butyl chloroformate, DI-SPME and GC–MS; Aleksa et al. (96) detected these substances with other illicit drugs in the hair samples collected from the hospital of sick children using in-matrix derivatization with N,O-bis (trimethylsilyl) trifluoroacetamide (BSTFA) and then HS-SPME, followed by GC–MS. COC and CE have been detected together in sweat (121), hair (122) and plasma (123) by DI-SPME. When a simultaneous analysis of multiple illicit drugs was performed, COC has been studied in hair (141, 142), while COC and CE together were examined in sweat (102), oral fluid (139, 140, 145) and hair (144); HS-SPME (102, 142, 144, 145) as well as DI-SPME (121–125, 139, 140) and HS-SPDE (141) were employed. Fucci et al. (124) and Gambelunghe et al. (125) examined not only COC and CE but also other COC congeners, such as tropacocaine and norcocaine, in hair samples of people addicted to COC. Validation was performed by all the studies about COC and metabolites reviewed. γ-Hydroxybutyrate SPME proved to be a suitable technique for detecting γ-hydroxybutyrate (GHB) in plasma (135) and urine (90, 135). Frison et al. (135) detected GHB after its conversion to the lactone form, γ-butyrolactone, employing HS-SPME with a CW/TPR fiber; they applied the method to plasma and urine samples of a patient under treatment with GHB. The analysis by GC/positive chemical ionization MS allowed us to reach an LOD value as low as 0.05 μg/mL and 0.1 μg/mL in plasma and urine, respectively. Blair et al. (90) applied DI-SPME by the use of a PDMS fiber directly to GHB after in-matrix derivatization with hexyl chloroformate on urine samples of four people suspected of abusing GHB and an accidental overdose. The results obtained were compared with a conventional method, with good agreement. Both authors used deuterated ISs and validated the methodology. Various xenobiotics Several different xenobiotics have been identified by SPME, including volatile compounds, pesticides and drugs, sometimes in attempted suicide or homicide and sometimes when accidental poisoning was suggested. The blood matrix was investigated to identify dichlorobenzene (DCB) (87) and strychnine (92) in intoxicated persons and toluene in glue-sniffers (89). Liu et al. (87) described that the three isomers of DCB were still present after 36 h from hospital admission. Barroso et al. (92) reported the quantitative determination of strychnine in all three cases of intoxication. Lee et al. (133) detected tricyclic antidepressants by HS-SPME from a blood matrix with an LOD of 8 ng/mL for imipramine. Namera et al. (86, 88) performed HS-SPME on serum samples to detect two herbicides (propanil and nereistoxin) in people hospitalized due to attempted suicide. Propanil was detected until 60 h after admission (86), while nereistoxin was detected until 24-h post-admission at a concentration of 90 ng/mL (88). Polychlorinated biphenyl (PCB) congeners and 1,1,1-trichloro-2,2-bis(4-chloro phenyl) ethane (DDT) have been studied on the hair matrix of children living in urban and rural regions using HS-SPME and 1,2,3,4-tetrachloronaphtalene as the IS (99). The authors documented that 94.4% of the samples were positive for at least one of the examined DDTs, especially for rural areas. Frison et al. (91) referred to a sexual assault (DFSA) committed after the deep sedation of the victim with barbiturates. Scalp and pubic hair have been used; a decrease in pentobarbital and thiopental concentration from proximal to distal hair was also observed, with positive results in pubic hair. Sporkert et al. (104) performed a simultaneous analysis of many drugs and drugs of abuse in the scalp hair of dead and living people; in nine cases, amitriptyline, doxepin, nicotine and trimipramine were reported, although the authors did not specify whether the population was living or dead. Zeng et al. (94) performed DI-SPME (an 85-μm PA fiber was employed) coupled to GC/flame thermoionic detector on urine specimens of three intoxicated patients to detect tetramine with an LOD of 0.02 ng/mL. The peak time concentration profiling has been described at 90 h. Benzodiazepines have been examined by DI-SPME in urine (134) and plasma (136). Guan et al. (134) performed the analysis after the acid hydrolysis of flunitrazepam and oxazolam with the synthesis of the respective benzophenones and then analyzed them by GC with electron capture detector (LODs of 2 and 6 ng/mL, respectively). Frison et al. (136) analyzed midazolam after deproteinization of plasma using a PA fiber and GC–MS and obtained an LOD of 1 ng/mL. Strano-Rossi et al. (143) developed an SPME method for detecting 59 stimulants and narcotics from the urine of two people to prevent and protect health in sporting environments. Post-mortem toxicology The literature reviewed disclosed several studies about post-mortem analytical toxicology. In the studies reviewed herein, several post-mortem matrices have been investigated, including gastric content and body tissues (59, 63, 64, 73, 75, 77–80, 82, 83), blood for illicit as well as therapeutic drugs (57, 60, 61, 65, 69, 70, 81), herbicides, malathion (58), methyl parathion (75) and VOCs (63, 64, 68, 72, 73, 76–80, 82, 83, 85). SPME has been performed on urine collected post-mortem to detect VOCs (59, 64, 72) and illicit (71) and therapeutic drugs (69). Hair of dead people has been used as method validation for lidocaine (62) and several illicit drugs (66, 74). The applicability of the method was demonstrated in some actual hair specimens obtained from both dead and living people who abused stimulants (73) to measure MA distribution. Finally, cadaveric gases were used to investigate VOC markers for PMI evaluation (84). In some of the studies reviewed, the validation was not carried out (59, 68, 73, 77, 78, 83, 84), and no quantitative data were reported (68, 69, 80, 83–85). Driving under the influence of alcohol or illicit drugs In 1997, SPME was used to confirm the presence of volatile compounds in two road accident victims (59). In the first case, ethanol and methylene chloride were detected in urine, while the second case involved the analysis of volatile petroleum products in gastric content (59). Schulz et al. (23-25) focused on the HS-SPME GC–MS technique to detect beverage flavoring compounds, including eugenol, anethole and menthone, in serum as markers of alcohol intake on driver populations—not post-offense drinking claims, but simply standard traffic control cases of drivers. Anzillotti et al. (98) employed DI-SPME to test for cannabinoids in oral fluid as an alternative matrix to blood. The authors confirmed that oral fluid is a reliable matrix for assessing ongoing psycho-physical alteration because of the easy and non-invasive collection procedures. Cannabinoids were also confirmed in hair samples of people examined for driving liability by SPME and GC–MS analysis (105). Gentili et al. (102) validated a procedure for the simultaneous analysis of 11 illicit drugs (COC, AP, methadone, cannabis and ketamine) using HS-SPME and GC–MS (PDMS fiber) on sweat as a biological specimen. Analytical performances The SPME technique is preferably employed in the headspace mode, particularly to detect VOCs and ethanol biomarkers; the latter compounds in fact are always investigated by HS-SPME, except for González-Illán et al. (35), who performed DI-SPME. In the remaining 92 of the 133 papers regarding the detection of drugs and drugs of abuse in various scenarios, 59 of the 92 articles preferred the HS mode, and 24 of the 92 articles used SPME with direct immersion. Only three articles used SPDE (112, 117, 141), examining hair samples, and one author applied the total vaporization of the sample coupled with SPME (TV-SPME), which is an alternative approach for the detection of nicotine in hair (100). The remaining studies did not specify the modality. Validation of the method is an important step; it is unavoidable for the detection and particularly for the quantitation of compounds in biological specimens. According to the Analytical Procedure Guidelines (148), most of the papers validated the method elaborated to harmonize the scientific approaches of procedure development. The present review highlighted that methods developed for the determination of alcohol biomarkers were always validated with few exceptions: Pragst et al. (13) who firstly developed the method, Dumitrascu et al. (40) whose aim was the evaluation of cosmetic treatments, Zelner et al. (28) and Natekar et al. (36). Methods developed to qualitatively detect VOCs for odor profiling were never validated; only Kusano et al. (48) considered the LOD and LOQ for selected, commonly found compounds. Regarding fatalities, validation was sometimes not performed (59, 68, 73, 77, 83, 84), most likely because of the case reports (73, 77, 83) and VOC detection (59, 68, 84); in particular, Perrault et al. (84) aimed to study PMI. The other groups mainly validated their methods. Quantitative detection is often reported in the searched literature; qualitative results are generally available for volatile compounds, in few cases for alcohol markers (13, 16, 28) and in 30–40% of papers summarized in Tables 3–5. GC–MS was more frequently adopted, probably for the well-established coupling of SPME with this separation technique, which however often requires derivatization prior to separation to improve chromatographic performance. Various kinds of derivatization methods were searched and applied due to physicochemical characteristics of some substances examined. Three different strategies can be considered: the reaction may occur in the sample, on the fiber or in the injection port. The present review disclosed that a total of 24 of the 133 studies performed derivatization. In the case of “in-matrix derivatization.” the reagent is added directly to the sample, and the derivative is formed before SPME; hence, the performance of SPME does not change. Overall, 15 of the 24 papers preferred this approach. As the reagent is stable for hydrolysis, since SPME is performed from aqueous samples, alkyl chloroformates are often used (71, 74, 109, 113, 120, 135). In-sample derivatization can also be achieved with heptafluorobutyric anhydride (HFBA) (29, 36, 37), pentafluorobenzyl bromide (70), BSTFA (95–97, 131) or MSTFA (145). The “on-fiber derivatization” is usually performed with low-volatility reagents deposited on the fiber prior to HS-SPME or subsequent to extraction; after SPME, the fiber is exposed to the vapor phase above the derivatization reagent in a separate vessel. Our review found seven of the 24 papers performing this type of derivatization using MSTFA (105, 117, 144), BSTFA (101, 141) or N-methylbis (trifluoroacetamide) for AP (111, 112) as the reagent. Complications of the on-fiber version can be loss of substance or derivative in the derivatization vial and carryover. The last kind of derivatization can be performed in the injection port of the gas chromatograph; we found only two papers (57, 65) using HFBA for AP. Only a few papers adopted a statistical approach, which were limited to VOC analysis and markers of alcohol abuse. Statistical evaluation for VOCs in scent was mainly performed using Spearman (47, 50, 51) and PCA analysis (47, 49–51, 56); Chilcote et al. (56) also performed HCA and Hudson et al. (45) achieved the statistical evaluation using three-dimensional covariance mapping. Receiver operating characteristic (20, 30, 31), sensitivity and specificity (26, 28, 36) were evaluated for alcohol markers. A statistical study of EtG was performed by Agius et al. (37). Conclusions The present systematic review disclosed that SPME technology applications cover several routine and research forensic toxicology areas concerning both dead and living people. SPME was preferably employed in the headspace mode, particularly to detect VOCs for the “primary odor profile” and ethanol biomarkers. This strategy can be considered a promising complementary, not alternative, tool to the use of trained dogs as forensic evidence in courts of justice. Some studies in fact characterized VOCs to better understand primary odor to track single individual and to recognize scents from different parts of the deceased body. The chance to detect minor ethanol metabolites by SPME is another well-explored forensic field highlighted by the studies herein reviewed. The results of most of the studies have been adopted by SoHT consensus (149) that suggested specific cutoffs to demonstrate abstinence over a pre-defined time period and to assess chronic excessive alcohol consumption. Most of the papers investigated FAEEs mainly as the sum of four ethyl esters, although SoHT (149, 150) recommended the use of ethyl palmitate alone for interpretation of data. Ethyl palmitate has the advantage of better analytical performance, particularly in the low concentration range, and no disadvantages concerning discrimination power, sensitivity and specificity. Despite of the more complex biological material respect to living people, good outcomes had been obtained by SPME applications to cadaveric samples; these results encourage the implementation of this technique for forensic purposes in post-mortem context. In conclusion, applications of SPME in many fields of forensic sciences highlight the versatility of this fast and solventless alternative to conventional extraction techniques. GENERAL ABBREVIATIONS AND REAGENTS AAD Autoerotic accidental death BAC Blood alcohol concentration BSTFA N,O-bis (trimethylsilyl)trifluoroacetamide CI Chemical ionization DFSA Drug-facilitated sexual assault DI-SPME Direct immersion solid phase microextraction F Female GC Gas chromatography GC–MS Gas chromatography–mass spectrometry GC–MS-MS Gas chromatography–tandem mass spectrometry GC-NPD Gas chromatography with nitrogen phosphorous detector GCxGC 2nd dimentional gas chromatography GTFCh Society of Toxicological and Forensic Chemistry h Hour HCA Hierarchical cluster analysis HFBA Heptafluorobutyric anhydride HS-GC Headspace gas chromatography HS-SPME Headspace solid phase microextraction ICU Intensive care unit IS Internal standard LC Liquid chromatography LC–MS Liquid chromatography mass spectrometry LC–MS-MS Liquid chromatography–tandem mass spectrometry LLE Liquid–liquid extraction LOD Limit of detection LOQ Limit of quantitation M Male MS Mass spectrometry MSTFA N-Methyl-N-trimethylsilyltrifluoroacetamide n.r. Not reported NPD Nitrogen phosphorous detector OF Oral fluid PCA Principal component analysis PMI Postmortem interval ROC Receiver operating characteristic SPDE Solid phase dynamic extraction SPME Solid-phase microextraction STU Scent transfer unit TOFMS Time-of-flight mass spectrometry TV-SPME Total vaporization solid-phase microextraction VOC volatile organic compound WDT Workplace drug testing SUBSTANCES INVESTIGATED ACH Acetone cyanohydrin ALT Alanine aminotransferase AP Amphetamine AST Aspartate transaminase BDB 3,4-Methylendioxyphenyl-2-butanamine BDZ Benzodiazepines BE Benzoylecgonine C2Cl4 Tetrachloroethylene CBD Cannabidiol CBN Cannabinol CDT Carbohydrate deficient transferrin CE Cocaethylene CN Cianyde COC Cocaine COD Codeine DCB Dichlorobenzene DDT 1,1,1-Trichloro-2,2-bis(4-chloro phenyl) ethane DHN S,S′-dimethyl dihydronereistoxin. EDDP 2-Ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine EMDP 2-Ethyl-5-methyl-3,3-diphenylpyrroline FCH Formaldehyde cyanohydrin GGT γ-Glutamyltransferase GHB γ-Hydroxy butyrate KET Ketamine LPG Liquefied petroleum gas MA Methamphetamine 6-MAM 6-Monoacetylmorphine MBDB N-Methyl-1-(3,4-methylendioxyphenyl)-2-butanamine MDA 3,4-Methylendioxyamphetamine MDEA 3,4-Methylendioxyethylamphetamine MDMA 3,4-Methylendioxymethamphetamine MDPA 3,4-Methylendioxypropylamphetamine METH Methadone MMTEA N-methyl-N-(2-methyl-thio-1-methylthiomethyl)ethylamine MOR Morphine MTEA 2-Methyl-thio-1-methylthiomethylethylamine Nor-COC Norcocaine PCB Polychlorinated biphenyls Peth Phosphatidylethanol SQ Squalene TCE Trichloroethylene THC D9-tetrahydrocannabinol THCA Tetrahydrocannabinolic acid THPI Tetrahydrophtalimide TNT 2,4,6-Trinitrotoluene VHC Volatile hydrocarbon compounds VOC Volatile organic compound VSA Volatile substance abuse KIND OF FIBERS AC activated carbon CAR Carboxen CW Carbowax DVB Divinylbenzene PA Polyacrylate PDMS Polydimethyldisiloxane TPR Templated resin References 1. 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TI - A Systematic Review of Solid-Phase Microextraction Applications in the Forensic Context
JO - Journal of Analytical Toxicology
DO - 10.1093/jat/bkz077
DA - 2020-04-30
UR - https://www.deepdyve.com/lp/oxford-university-press/a-systematic-review-of-solid-phase-microextraction-applications-in-the-GopyjrrdfC
SP - 1
VL - Advance Article
IS - 
DP - DeepDyve
ER -