Biomedical Chromatography

Dong, Qinghai; An, Yang; Du, Guangguang; Wang, Jia; Liu, Jiayin; Su, Jun; Xie, Hongliu; Liang, Chongyang; Liu, Jihua

doi: 10.1002/bmc.5334pmid: 35045586

Although the chemical components of Panax notoginseng (PN) and Panax ginseng (PG) are similar, their bioactivities are different. In this study, the differential bioactivities of PN and PG were used as the research object. First, the different metabolites in the plasma after oral administration of PN and PG were analyzed using a UPLC‐Q/TOF‐MS‐based metabolomics approach. Afterward, the metabolite‐target‐ pathway network of PN and PG was constructed, and thus the pathways related to different bioactivities were analyzed. As a result, 7 different metabolites were identified in PN group, and 10 different metabolites were identified in the PG group. In the PN group, the metabolite N1 was related to hemostasis, N1 and N3 were related to inhibiting the nerve center, antihypertensive, and abirritation. The metabolites N1, N3, N4, N5, and N6 were related to liver protection. The results showed that the metabolites G1, G2, G3, G5, and G6 in PG group were related to heart protection, and G1, G2, G6, and G9 were related to increased blood pressure. There were 13 signaling pathways related to different biological activities of PN (8 pathways) and PG (5 pathways). These pathways further clarified the mechanism of action that caused the different bioactivities between PN and PG. In summary, metabolomics combined with network pharmacology could be helpful to clarify the material basis of different bioactivities between PN and PG, promoting the research on PN and PG.

Wojciechowska, Marta; Stepnowski, Piotr; Gołębiowski, Marek

doi: 10.1002/bmc.5346pmid: 35066890

This paper presents the effect of insecticides on the composition of the surface compounds of one of the most harmful insects, Tenebrio molitor, by analysis using GC–MS. As a result of the use of insecticides, the composition of the chemical compounds on the surface of insects changes, depending on the insecticides used. The most numerous groups of the marked compounds were fatty acids, alkanes, esters and sterols. The content of the identified compounds in the larvae increased at both 24 and 48 h after the application of insecticides, in comparison with the control samples. The content of identified compounds in the samples taken from the females increased 24, 48 and 72 h after the application of insecticides in comparison with the control samples. By contrast, in samples prepared from males, the content of identified compounds decreased 24 h after the application of insecticides, compared with the control samples. The highest content of chemical compounds was for fatty acids and alkanes after the use of insecticides. The content of fatty acids after the application of the insecticide with deltamethrin was 62.1 ± 3.3–466.9 ± 5.9 μg/g, and after the application of the insecticide with cyfluthrin was 49.9 ± 1.9–458.3 ± 4.2 μg/g. However, the content of alkanes after the use of deltamethrin was 115.6 ± 4.2–4672.0 ± 32.1 μg/g, and after the use of cyfluthrin was 189.4 ± 3.8–3975.0 ± 10.2 μg/g.

Li, Ning; Han, Danhe; Ma, Xuefeng; Qiu, Cheng; Qin, Yuchang; Yao, Ting; Wang, Shanshan; She, Yongxin; Hacımüftüoğlu, Fazil; Abd El‐Aty, A. M.

doi: 10.1002/bmc.5303pmid: 34957590

Biswas, Sayan; Kar, Amit; Chanda, Joydeb; Sharma, Nanaocha; Bharadwaj, Pardeep Kumar; Haldar, Pallab Kanti; Mukherjee, Pulok Kumar

doi: 10.1002/bmc.5328pmid: 34997595

A rapid validated ultra‐fast liquid chromatography–photodiode array detector (UFLC‐PDA) method was developed to identify and quantify ayapanin (AY) and umbelliferone (UM) simultaneously in Ayapana triplinervis Vahl methanolic extract. The method was validated for linearity, limit of detection (LOD; 3:1σ/S), limit of quantification (LOQ; 10:1σ/S), precision, accuracy, specificity and robustness. The response was linear with a good correlation between concentration and mean peak area through a correlation coefficient of 0.9996, y = 7025.7x – 2269.8 and 0.9997, y = y = 16,262x – 946 with LOD of 6.256 ± 0.52 and 3.325 ± 0.36, and LOQ of 18.838 ± 0.18 and 8.870 ± 0.85 for AY (0.67% w/w) and UM (0.18% w/w), respectively. The relative standard deviation (%) of precision and recovery of AY and UM was <2.0%. The proposed method was simple, accurate, specific, precise and reproducible.

Shan, Lian; Jones, Barry R.

doi: 10.1002/bmc.5317pmid: 34981550

Low‐flow chromatography has a rich history of innovation but has yet to reach widespread implementation in bioanalytical applications. Improvements in pump technology, microfluidic connections, and nano‐electrospray sources for MS have laid the groundwork for broader application, and innovation in this space has accelerated in recent years. This article reviews the instrumentation used for nano‐flow LC, the types of columns employed, and strategies for multidimensionality of separations, which are key to the future state of the technique to the high‐throughput needs of modern bioanalysis. An update of the current applications where nano‐LC is widely used, such as proteomics and metabolomics, is discussed. But the trend toward biopharmaceutical development of increasingly complex, targeted, and potent therapeutics for the safe treatment of disease drives the need for ultimate selectivity and sensitivity of our analytical platforms for targeted quantitation in a regulated space. The selectivity needs are best addressed by mass spectrometric detection, especially at high resolutions, and exquisite sensitivity is provided by nano‐electrospray ionization as the technology continues to evolve into an accessible, robust, and easy‐to‐use platform.

Zheng, Xin; Chen, Chen; Gao, Huitao; Sun, Xuefeng; Zhang, Yanbao; Shi, Juhong; Han, Xiaohong

doi: 10.1002/bmc.5306pmid: 34967030

Rivaroxaban is an oral anticoagulant directly inhibiting the activity of Factor Xa, which is widely used for the prophylaxis of thromboembolic disorders. Therapeutic drug monitoring (TDM) is required during therapy for individual dosage adjustment. This study aimed at developing a liquid chromatography/tandem mass spectrometry method that was suitable for rivaroxaban TDM in human plasma and urine and exploring the feasibility of urine drug monitoring in medical care. A 3 min run time of the LC–MS/MS methods was established by employing an Acquity UPLC BEH C18 (2.1 × 50 mm, 1.7 μm) column using gradient elution of 10 mmol/L ammonium acetate containing 0.1% formic acid–0.1% formic acid acetonitrile as a mobile phase at a flow rate of 0.4 ml/min with calibration ranges of 0.5–400 and 10–10,000 ng/ml for human plasma and urine, respectively. Rivaroxaban was detected on a triple quadrupole tandem mass spectrometer with an electrospray ionization source in positive ion mode. The methods showed good linearity within the calibration range. The precision and accuracy, matrix effect, extraction recovery and stability in both human matrices were all validated and meet the international guideline requirements. These validated methods were successfully applied to support the TDM of an aged patient receiving rivaroxaban for therapy.

Giulio, Luca; Padula, Cristina; Pescina, Silvia; Nicoli, Sara; Santi, Patrizia

doi: 10.1002/bmc.5349pmid: 35098556

An innovative and sensitive HPLC–UV method for the extraction and quantification of methotrexate (MTX) in skin layers was developed and validated. Owing to the physico‐chemical characteristics of the drug and the nature of the tissue, it was necessary to use folic acid (FA) as an internal standard for MTX quantification in the dermis. MTX (and FA) analysis was performed on a Phenomenex Jupiter C18 column, using a 50 mm sodium acetate buffer (pH 3.6) and methanol mixture (87:13, v/v) as mobile phase, pumped at 1 ml/min. The absorbance was monitored at 290 nm. The method was selective, linear in the range 0.11–8.49 μg/ml for extraction solvent and 0.05–8.94 μg/ml for pH 7.4 phosphate‐buffered saline, precise and accurate, with lower limits of quantitation of 0.11 μg/ml (extraction solvent) and 0.05 μg/ml (pH 7.4 phosphate‐buffered saline). The method developed is suitable for the quantification of MTX in skin layers at the end of in vitro permeation experiments; the overall mass balance was 96.5 ± 1.4%, in line with the requirements of the Organisation for Economic Co‐operation and Development guideline for the testing of the chemicals (Skin absorption: in vitro method).

Ya, Huiyuan; Li, Huiru; Liu, Xianghui; Chen, Ye; Zhang, Jingxiao; Xie, Yanfu; Wang, Mengshu; Xie, Wanyue; Li, Shipeng

doi: 10.1002/bmc.5331pmid: 35000209

Gleditsiae Spina, the thorn of Gleditsia sinensis Lam., has a long history of being used as a traditional medicine in East Asian countries. However, only a few biologically active substances have been identified from it. In this study, the epidermis, xylem and pith of Gleditsiae Spina, respectively Gs‐E, Gs‐X and Gs‐P, were studied. We used a widely targeted metabolomics method to investigate the chemical composition of Gs‐E, Gs‐X and Gs‐P. A total of 728 putative metabolites were identified from Gleditsiae Spina, including 211 primary metabolites and 517 secondary metabolites. These primary and secondary metabolites could be categorized into more than 10 different classes. Flavonoids, phenolic acids, lipids, amino acids and derivatives, and organic acids constituted the main metabolite groups. Multivariate statistical analysis showed that the Gs‐E, Gs‐X and Gs‐P samples could be clearly separated. Differential accumulated metabolite (DAM) analysis revealed that more than half of the DAMs exhibited the highest relative concentrations in Gs‐E, and most of the DAMs showed the lowest relative concentrations in Gs‐X. Moreover, 11 common differential primary metabolites and 79 common differential secondary metabolites were detected in all comparison groups. These results further our understanding of chemical composition and metabolite accumulation of Gleditsiae Spina.

Garikapati, Kusuma Kumari; Ammu V. V. V., Ravi Kiran; Krishnamurthy, Praveen Thaggikuppe; S. T., Narenderan; B., Babu; Nagappan, Krishnaveni

doi: 10.1002/bmc.5326pmid: 34993979

A bioanalytical method for the quantification of rosiglitazone in rat plasma and tissues (adipose tissue, heart, brain, bone, and kidney) using LC–MS/MS was developed and validated. Chromatographic separation was achieved on a Gemini C18 column (50 × 4.6 mm, 3 μm) using a mobile phase consisting of 10 mM ammonium formate (pH 4.0) and acetonitrile (10:90, v/v) at a flow rate of 0.8 mL/min and injection volume of 10 μL (internal standard: pioglitazone). LC–MS detection was performed with multiple reaction monitoring mode using target ions at m/z → 358.0 and m/z → 357.67 for rosiglitazone and pioglitazone (internal standard), respectively. The calibration curve showed a good correlation coefficient (r2) over the concentration range of 1–10,000 ng/mL. The mean percentage recoveries of rosiglitazone were found to be over the range of 92.54–96.64%, with detection and lower quantification limit of 0.6 and 1.0 ng/mL, respectively. The developed method was validated per U.S. Food and Drug Administration guidelines and successfully utilized to measure rosiglitazone in plasma and tissue samples. Further, the developed method can be utilized for validating specific organ‐targeting delivery systems of rosiglitazone in addition to conventional dosage forms.

doi: 10.1002/bmc.5161pmid: N/A