TY - JOUR
AU - Jain,, Vandana
AB - Abstract Bicyclic diterpenoid lactone andrographolide is regarded as a “natural antibiotic” as it is known to exhibit a range of bioactivities including anti-inflammatory, antibacterial, antipyretic, antineoplastic, cardioprotective, hepatoprotective and hypoglycaemic, and is present in Andrographis paniculata. The aim of this article is to review the information on analytical methods for andrographolide in biological samples, pharmaceutical formulations and plant materials. This article includes various techniques such as Spectrophotometry, Chemiluminescence method, Electroanalytical method, Chromatography and various hyphenated techniques. Introduction Andrographis paniculata (Burm.f.) Wall. ex Nees., (English name—King of Bitters) is an herbaceous plant. It belongs to Acanthaceae family. It is native to India and Sri Lanka, and it is also found in abundance in Asian countries such as India, Pakistan, Java, Malaysia and Indonesia (1). It is cultivated extensively in China and Thailand, the East and West Indies, and Mauritius. It is normally grown from seeds and grows in pine, evergreen and deciduous forest areas, and also along roads and in villages (2). Morphology A. paniculata is an annual plant, it is branched, erect and has height about 30–110 cm. It has simple, opposite, lanceolate, glabrous leaves. It has violet to white color flowers; they consist of small, linear 5-particle calyx, tube narrows and corolla about 6-mm long. It also has two stamens and two celled superior ovaries. Capsule seeds are very small and numerous (3, 4). Phytochemistry The leaves of the plant are reported to contain andrographolide, neoandrographolide, andrographiside, homoandrographolide, andrographane, andrographanin, andrographone and andrographosterol. The aerial parts of the plant contain andrographolide, neoandrographolide, andrographiside 14-deoxyandrographolide, 14-deoxy-11,12-didehydroandrographolide, 14-deoxy-11-oxoandrographolide and β-sitosterol. The roots contain 5-hydroxy-7,8,2′3’-tetramethoxyflavone, andrographolide andrographonin, apigenin and 7,4-dioxymethylether (5). Of the above chemicals, andrographolide is the major bioactive constituent responsible for variety of activities. Leaves and stems of plant are used for extracting active phytochemicals; roots are used rarely. The leaves contain about 1.0–2.39% of andrographolide, while the roots, stem, pericarp and seeds contain 0.44,0.20, 0.18 and 0.13% of andrographolide, respectively (6). Andrographolide Bicyclic diterpenoid lactone andrographolide [C20H30O5, (3-[2{decahydro-6hydroxy-5-(hydroxymethyl)-5,8α-dimethyl-2-methylene1-naphthalenyl} ethylidene] dihydro-4-hydroxy-2(3H) furanone)] is concentrated in leaves (1). The structure of andrographolide is shown in Figure 1. Figure 1 Open in new tabDownload slide Structure of andrographolide. Figure 1 Open in new tabDownload slide Structure of andrographolide. Physicochemical properties Molecular weight of andrographolide is 350.4 g/mol (7). Its density is 1.2317 g/cm3. Its melting point is 230–231°C (446–448°F, 503–504 K); it is sparingly soluble in water, soluble in acetone, chloroform, hot ethanol and ether (8). pKa (strongest acidic) is 13.48 and pKa (strongest basic) is −2.8 (9). Andrographolide shows maximum absorption at 223 nm. Bioavailability of andrographolide Andrographolide belongs to Class III drug that is high solubility, low permeability as per Biopharmaceutics Classification System. The drug is metabolized by duodenum and jejunum to a sulfonate 14-deoxy-12-sulfo-andrographolide. It has high-lipophilicity (log P value = 2.632 ± 0.135) and low-aqueous solubility (3.29 ± 0.73 mg/mL), rapid transformation and efflux which together contribute to its poor oral bioavailability property (10). Pharmacological action Traditionally andrographolide is used as an analgesic, anti-inflammatory, antibacterial, antipyretic, antithrombotic, antiviral, antineoplastic, cardioprotective, choleretic, digestive, hepatoprotective, hypoglycaemic, immune enhancer and laxative. It shows immunological benefits in cancer and HIV. It shows preventive effect on common cold and also as antidote against poisons of snakes and insects (5). It is also having ability to treat various ailments such as leprosy, gonorrhoea, scabies, skin eruptions, hypertension, neoplasia, diuresis, dyspepsia and influenza (1). Hepatoprotective activity: A. paniculata is used extensively for the treatment of various liver disorders and jaundice due to the presence of its major constituent andrographolide (11). Andrographolide is effective against liver damage induced by paracetamol or galactosamine. It plays hepatoprotective role also by reducing a lipid peroxidation product malondialdehyde (12). Anti-cancer potential of andrographolide and cytotoxicity against cancer cells: Andrographolide acts both directly and indirectly on the cancer cells (13). Methanolic extract of A. paniculata had shown toxicity against human epidermoid leukaemia and lymphocytic leukaemia cell lines (14). The cytotoxic property of A. paniculata is due to the ability of andrographolide to inhibit proliferation and induce apoptosis in cancer cells (11). Anti-malarial activity: A. paniculata was found to inhibit the multiplication of one of the parasites which transmit malaria Plasmodium berghei (15). Cardiovascular activity: Rise in nitric oxide, cyclic guanosine monophosphate, and activity of superoxide dimutase and declines of lipid peroxide and endothelin, was shown by Andrographis in an atherosclerotic rabbit model. Thus, it suggested that A. paniculata has an antioxidant potential. A. paniculata verified an increase of blood clotting time and is also reported to prevent constriction of blood vessels before angioplasty and after surgery (11). In respiratory system: A. paniculata is used in the treatment of respiratory infections such as common cold, influenza, sore throat, acute and chronic cough, sinusitis, bronchitis and pharyngotonsillitis (16). Anti-inflammatory and anti-angiogenic activity: A. paniculata as well as extract of Andrographis plant are known to have an anti-inflammatory potential (17). Andrographolide treatment inhibits nuclear factor kappa B (NF-kB) binding to deoxyribonucleic acid, reducing the expression of pro-inflammatory proteins such as cyclooxygenase 2 (Cox-2) and nitric-oxide synthase (18). Safety and contraindications: From toxicological studies in animal models and human clinical trials, it was confirmed that andrographolide has very low toxicity. Some people will have allergic reactions to A. paniculata from minor skin rashes to serious anaphylaxis, but it is generally observed at high doses. Antifertility is also one of the side effects of A. paniculata. Also, headache, fatigue, bitter/metallic taste, diarrhea and pruritus are some of the adverse effects of A. paniculata. It is generally suggested to avoid during pregnancy and lactation (6). Analytical methods It is very important to determine drug in various biological samples during drug discovery and drug development as it provides pharmacokinetic information and is necessary to determine dose and safety margins. Quantification of bioactive markers from herbal drugs is a challenging task because of the complexity of phytoconstituents present in it. Quantification of active component is desirable so as to ensure effective pharmacological activity. Development of analytical techniques for the analysis of phytoconstituents in herbal formulations is necessary for their standardization. It is vital for accessing safety, purity and quality in herbal formulations (19). In the present review, we have summarized methods of the separation and quantification of andrographolide by high-performance liquid chromatography (HPLC), rapid resolution liquid chromatography (RRLC), high-performance thin layer chromatography (HPTLC), microemulsion electrokinetic capillary chromatography (MEEKC), micellar electrokinetic capillary chromatography (MEKC), high-speed counter current chromatography (HSCCC), electroanalytical method, ultraviolet spectroscopic (UV), Fourier transform infrared spectroscopy (FTIR), chemiluminescence method and various other hyphenated techniques. Spectroscopic methods UV method Mainly, UV spectroscopy is been used by researchers for the quantification of andrographolide in extract of herbal or polyherbal formulation. It is simple, accurate, robust and cost-effective method. Andrographolide shows solubility in mixture of methanol and water (50:50). Determination of total lactones in A. paniculata was established by using dinitrobenzoic acid and potassium hydroxide solutions as color forming agents, and pink color was formed which was detected at 536 nm. Andrographolide can be estimated based on the condensation of butenolide ring of andrographolide with picric acid, which produced red-orange complex in alkaline solution which could be measured at 494 nm. A typical UV spectrum of andrographolide is shown in Figure 2 and UV method for estimating andrographolide is tabulated in Table I. Figure 2 Open in new tabDownload slide UV spectrum of andrographolide. Figure 2 Open in new tabDownload slide UV spectrum of andrographolide. Table I UV Methods for Estimation of Andrographolide in Plant Materials Sample . λ max . Solvents/reagents . Limit of detection (LOD) and limit of quantification (LOQ) . Ref. . Bulk powder 321 nm Methanol: water (50:50 v/v) - (33) A.paniculata 536 nm 95% ethanol, dinitrobenzoic acid, potassium hydroxide solution. 1.2 and 4.23 μg (34) A. paniculata and AmLycure (Herbal formulation) 494 nm Methanol, picric acid, aqueous sodium hydroxide, Baljet reagent - (35) A. paniculata herb 223 nm Methanol - (36) Sample . λ max . Solvents/reagents . Limit of detection (LOD) and limit of quantification (LOQ) . Ref. . Bulk powder 321 nm Methanol: water (50:50 v/v) - (33) A.paniculata 536 nm 95% ethanol, dinitrobenzoic acid, potassium hydroxide solution. 1.2 and 4.23 μg (34) A. paniculata and AmLycure (Herbal formulation) 494 nm Methanol, picric acid, aqueous sodium hydroxide, Baljet reagent - (35) A. paniculata herb 223 nm Methanol - (36) Open in new tab Table I UV Methods for Estimation of Andrographolide in Plant Materials Sample . λ max . Solvents/reagents . Limit of detection (LOD) and limit of quantification (LOQ) . Ref. . Bulk powder 321 nm Methanol: water (50:50 v/v) - (33) A.paniculata 536 nm 95% ethanol, dinitrobenzoic acid, potassium hydroxide solution. 1.2 and 4.23 μg (34) A. paniculata and AmLycure (Herbal formulation) 494 nm Methanol, picric acid, aqueous sodium hydroxide, Baljet reagent - (35) A. paniculata herb 223 nm Methanol - (36) Sample . λ max . Solvents/reagents . Limit of detection (LOD) and limit of quantification (LOQ) . Ref. . Bulk powder 321 nm Methanol: water (50:50 v/v) - (33) A.paniculata 536 nm 95% ethanol, dinitrobenzoic acid, potassium hydroxide solution. 1.2 and 4.23 μg (34) A. paniculata and AmLycure (Herbal formulation) 494 nm Methanol, picric acid, aqueous sodium hydroxide, Baljet reagent - (35) A. paniculata herb 223 nm Methanol - (36) Open in new tab FT-IR method A novel technique using IR spectroscopy is also been reported for the determination of total lactones in terms of andrographolide. Andrographolide contains a lactone group and of total lactone in terms of andrographolide can be determined using IR spectroscopy. Table II presents IR methods to determine andrographolide. Table II FTIR Methods for the Presence of Andrographolide in Plant Materials Sample . Instrument . Absorption bands . LOD and LOQ . Ref. . Andrographolide powder Perkin Elmer spectrometer with a mercury cadmium telluride A detector and KBr optics Lactone-1727 cm−1 1.5 and 15 μg (37) Andrographolide isolated from fresh plant material CsI on Perkin Elmer 1800 spectrophotometer Lactone-1728 cm−1 1772 cm−1 - (38) Andrographolide Perkin Elmer FTIR spectrometer with a single reflectance horizontal ATR cell Lactone-1722 cm−1 C=C-1674 cm−1 C-O-C-1218 cm−1 Methylene-906 cm−1 1.0 and 3.34 μg/mL (39) Sample . Instrument . Absorption bands . LOD and LOQ . Ref. . Andrographolide powder Perkin Elmer spectrometer with a mercury cadmium telluride A detector and KBr optics Lactone-1727 cm−1 1.5 and 15 μg (37) Andrographolide isolated from fresh plant material CsI on Perkin Elmer 1800 spectrophotometer Lactone-1728 cm−1 1772 cm−1 - (38) Andrographolide Perkin Elmer FTIR spectrometer with a single reflectance horizontal ATR cell Lactone-1722 cm−1 C=C-1674 cm−1 C-O-C-1218 cm−1 Methylene-906 cm−1 1.0 and 3.34 μg/mL (39) Open in new tab Table II FTIR Methods for the Presence of Andrographolide in Plant Materials Sample . Instrument . Absorption bands . LOD and LOQ . Ref. . Andrographolide powder Perkin Elmer spectrometer with a mercury cadmium telluride A detector and KBr optics Lactone-1727 cm−1 1.5 and 15 μg (37) Andrographolide isolated from fresh plant material CsI on Perkin Elmer 1800 spectrophotometer Lactone-1728 cm−1 1772 cm−1 - (38) Andrographolide Perkin Elmer FTIR spectrometer with a single reflectance horizontal ATR cell Lactone-1722 cm−1 C=C-1674 cm−1 C-O-C-1218 cm−1 Methylene-906 cm−1 1.0 and 3.34 μg/mL (39) Sample . Instrument . Absorption bands . LOD and LOQ . Ref. . Andrographolide powder Perkin Elmer spectrometer with a mercury cadmium telluride A detector and KBr optics Lactone-1727 cm−1 1.5 and 15 μg (37) Andrographolide isolated from fresh plant material CsI on Perkin Elmer 1800 spectrophotometer Lactone-1728 cm−1 1772 cm−1 - (38) Andrographolide Perkin Elmer FTIR spectrometer with a single reflectance horizontal ATR cell Lactone-1722 cm−1 C=C-1674 cm−1 C-O-C-1218 cm−1 Methylene-906 cm−1 1.0 and 3.34 μg/mL (39) Open in new tab Chemiluminescence method Flow-injection chemiluminescence method was used for the determination of andrographolide. This method is simple, rapid, low cost and sensitive and has low detection limit (20). IFFM-E mode flow injection chemiluminescence analysis system and UV detector was used. Ammonium cerium (IV) sulfate and sodium sulfite were used in the analysis as they are environment friendly and sample testing just took about 2 min. Chemiluminescence method for the determination of andrographolide is presented in Table III. Table III Chemiluminescence Method Used for Andrographolide Determination in Pharmaceutical Formulations Sample . Instrument . Detector . Solvent . Results . LOD . Ref. . Andrographis tablets IFFM-E mode flow injection chemiluminescence analysis system (Remex Electronic Instrument Co., Ltd. Xi’an, China) LC-20 A HPLC (Shimadzu, Kyoto, Japan) Cary Eclipse Fluorescence Spectrophotometer and Cary 50 Conc UV–visible spectrophotometer (Varian, Palo Alto, CA, USA) UV detector 225 nm Ce/Na2SO3 FI-Cl-2 min 0.0742 (40) Sample . Instrument . Detector . Solvent . Results . LOD . Ref. . Andrographis tablets IFFM-E mode flow injection chemiluminescence analysis system (Remex Electronic Instrument Co., Ltd. Xi’an, China) LC-20 A HPLC (Shimadzu, Kyoto, Japan) Cary Eclipse Fluorescence Spectrophotometer and Cary 50 Conc UV–visible spectrophotometer (Varian, Palo Alto, CA, USA) UV detector 225 nm Ce/Na2SO3 FI-Cl-2 min 0.0742 (40) Open in new tab Table III Chemiluminescence Method Used for Andrographolide Determination in Pharmaceutical Formulations Sample . Instrument . Detector . Solvent . Results . LOD . Ref. . Andrographis tablets IFFM-E mode flow injection chemiluminescence analysis system (Remex Electronic Instrument Co., Ltd. Xi’an, China) LC-20 A HPLC (Shimadzu, Kyoto, Japan) Cary Eclipse Fluorescence Spectrophotometer and Cary 50 Conc UV–visible spectrophotometer (Varian, Palo Alto, CA, USA) UV detector 225 nm Ce/Na2SO3 FI-Cl-2 min 0.0742 (40) Sample . Instrument . Detector . Solvent . Results . LOD . Ref. . Andrographis tablets IFFM-E mode flow injection chemiluminescence analysis system (Remex Electronic Instrument Co., Ltd. Xi’an, China) LC-20 A HPLC (Shimadzu, Kyoto, Japan) Cary Eclipse Fluorescence Spectrophotometer and Cary 50 Conc UV–visible spectrophotometer (Varian, Palo Alto, CA, USA) UV detector 225 nm Ce/Na2SO3 FI-Cl-2 min 0.0742 (40) Open in new tab Electroanalytical method This method has various advantages such as simple, sensitive, inexpensive, low detection limit and environment friendly (21). Electrochemical behavior of andrographolide and its voltammetric determination using differential pulse voltammetry measurements was performed. Boron-doped diamond electrode, Ag/AgCl (saturated Potassium chloride) reference electrode and Platinum counter electrode were employed in the analysis. Table IV presents the method of determination of andrographolide. Table IV Electroanalytical Method for Andrographolide Determination in Biological Samples/Plant Materials Sample . Instrument . Peak . LOD . Ref. . Human urine and oil extract from A. paniculata Electrochemical system AUTOLAB PGSTAT 302 N, Methrom Autolab B.V. (The Netherlands) Boron-doped diamond electrode, pH meter model Orion 1230 Peak was observed at +1.55 V vs. Ag/AgCl electrode in Britton-Robinson buffer solution pH 12 0.75 μM (41) Sample . Instrument . Peak . LOD . Ref. . Human urine and oil extract from A. paniculata Electrochemical system AUTOLAB PGSTAT 302 N, Methrom Autolab B.V. (The Netherlands) Boron-doped diamond electrode, pH meter model Orion 1230 Peak was observed at +1.55 V vs. Ag/AgCl electrode in Britton-Robinson buffer solution pH 12 0.75 μM (41) Open in new tab Table IV Electroanalytical Method for Andrographolide Determination in Biological Samples/Plant Materials Sample . Instrument . Peak . LOD . Ref. . Human urine and oil extract from A. paniculata Electrochemical system AUTOLAB PGSTAT 302 N, Methrom Autolab B.V. (The Netherlands) Boron-doped diamond electrode, pH meter model Orion 1230 Peak was observed at +1.55 V vs. Ag/AgCl electrode in Britton-Robinson buffer solution pH 12 0.75 μM (41) Sample . Instrument . Peak . LOD . Ref. . Human urine and oil extract from A. paniculata Electrochemical system AUTOLAB PGSTAT 302 N, Methrom Autolab B.V. (The Netherlands) Boron-doped diamond electrode, pH meter model Orion 1230 Peak was observed at +1.55 V vs. Ag/AgCl electrode in Britton-Robinson buffer solution pH 12 0.75 μM (41) Open in new tab Chromatographic methods Chromatographic analysis plays an important role for the quantification of markers in any drug or formulation. Chromatographic methods are often more sensitive and precise than other methods such as titrimetry, gravimetry, colorimetry, etc. Chromatography using Green technique Cloud point extraction (CPE) method replaces organic solvents by less toxic substances. Generally, a surfactant is added to the sample at a concentration higher than the critical micelle concentration to yield an organized medium consisting of an anisotropic phase which is often considered as a two-phase system. CPE method is efficient, environment friendly due to the minimal use of organic solvents, rapid and inexpensive. This method was employed for the determination of andrographolide which not only minimizes the use of organic solvents but also combines the extraction and preconcentration in a single step. The non-ionic surfactant Triton X-114 was chosen for extraction. Green method to determine andrographolide is presented in Table V. Table V Chromatographic Green Method for Andrographolide Determination in Biological Sample Sample . CPE . HPLC analysis . Detection . Column . LOD . Ref. . Human plasma Triton X-114 at concentration of 5%(v/v) and 0.45 g of NaCl Agilent 1100 liquid chromatograph M.P.-methanol–acetonitrile–0.5% formic acid aqueous solution (40: 17: 43, v/v/v) 1.0 mL/min 254 nm, UV detector SB C18 column (250 × 4.6 mm i.d., 5 μm) 0.032 μg/mL (42) Sample . CPE . HPLC analysis . Detection . Column . LOD . Ref. . Human plasma Triton X-114 at concentration of 5%(v/v) and 0.45 g of NaCl Agilent 1100 liquid chromatograph M.P.-methanol–acetonitrile–0.5% formic acid aqueous solution (40: 17: 43, v/v/v) 1.0 mL/min 254 nm, UV detector SB C18 column (250 × 4.6 mm i.d., 5 μm) 0.032 μg/mL (42) Open in new tab Table V Chromatographic Green Method for Andrographolide Determination in Biological Sample Sample . CPE . HPLC analysis . Detection . Column . LOD . Ref. . Human plasma Triton X-114 at concentration of 5%(v/v) and 0.45 g of NaCl Agilent 1100 liquid chromatograph M.P.-methanol–acetonitrile–0.5% formic acid aqueous solution (40: 17: 43, v/v/v) 1.0 mL/min 254 nm, UV detector SB C18 column (250 × 4.6 mm i.d., 5 μm) 0.032 μg/mL (42) Sample . CPE . HPLC analysis . Detection . Column . LOD . Ref. . Human plasma Triton X-114 at concentration of 5%(v/v) and 0.45 g of NaCl Agilent 1100 liquid chromatograph M.P.-methanol–acetonitrile–0.5% formic acid aqueous solution (40: 17: 43, v/v/v) 1.0 mL/min 254 nm, UV detector SB C18 column (250 × 4.6 mm i.d., 5 μm) 0.032 μg/mL (42) Open in new tab MEEKC It is a capillary electrokinetic technique wherein microemulsion is used as carrier electrolyte. This technique is used for the separation of neutral as well as charged species (22). Also, separation of lipophilic and hydrophilic substances can be carried out using this technique (23). MDQ capillary electrophoresis system equipped with Photodiode-array (PDA) detector was employed in the analysis by all the researchers. Andrographolide sample has been injected by applying pressure of −0.2 psig for 2 s, while other researcher injected sample by applying 0.5 psig pressure for 3 s. Sodium borate buffer and Sodium dodecyl sulfate (surfactant) were commonly used for the analysis. MEEKC method has better separation efficiency, exceptional resolution, good reproducibility and recovery. MEEKC methods for the determination of andrographolide are tabulated in Table VI. Table VI MEEKC Methods for Andrographolide Determination in Pharmaceutical Formulations/Plant Materials Sample . Instrument . Capillary . Reagents . Detection, voltage and temperature . RT . LOD and LOQ . Ref. . Standards of andrographolide and dehydroandrographolide Chuanxinlian and xiaoyanlidan tablets MDQ CE instrument equipped with PDA detector 60.2 cm × 50 μm i.d. uncoated fused silica capillary having length of 50 cm Heptane 0.81% (w/w), SDS 3.31% (w/w), butan-1-ol 6.61% (w/w) 10-mM sodium tetraborate buffer, pH 9.2 214 nm, 20 kV, 25 and 45°C 15 0.30 and 1.0 μg/mL (43) A. paniculata MDQ CE instrument equipped with PDA detector 31.2 cm × 50 μm i.d. uncoated fused silica capillary 0.5% (w/w) ethyl acetate+0.6% (w/w) SDS+ 97.7–92.9% (w/w) 30-mM sodium borate at pH 9.5 214 nm, −15 kV, 25°C 1.07 - (44) Sample . Instrument . Capillary . Reagents . Detection, voltage and temperature . RT . LOD and LOQ . Ref. . Standards of andrographolide and dehydroandrographolide Chuanxinlian and xiaoyanlidan tablets MDQ CE instrument equipped with PDA detector 60.2 cm × 50 μm i.d. uncoated fused silica capillary having length of 50 cm Heptane 0.81% (w/w), SDS 3.31% (w/w), butan-1-ol 6.61% (w/w) 10-mM sodium tetraborate buffer, pH 9.2 214 nm, 20 kV, 25 and 45°C 15 0.30 and 1.0 μg/mL (43) A. paniculata MDQ CE instrument equipped with PDA detector 31.2 cm × 50 μm i.d. uncoated fused silica capillary 0.5% (w/w) ethyl acetate+0.6% (w/w) SDS+ 97.7–92.9% (w/w) 30-mM sodium borate at pH 9.5 214 nm, −15 kV, 25°C 1.07 - (44) Open in new tab Table VI MEEKC Methods for Andrographolide Determination in Pharmaceutical Formulations/Plant Materials Sample . Instrument . Capillary . Reagents . Detection, voltage and temperature . RT . LOD and LOQ . Ref. . Standards of andrographolide and dehydroandrographolide Chuanxinlian and xiaoyanlidan tablets MDQ CE instrument equipped with PDA detector 60.2 cm × 50 μm i.d. uncoated fused silica capillary having length of 50 cm Heptane 0.81% (w/w), SDS 3.31% (w/w), butan-1-ol 6.61% (w/w) 10-mM sodium tetraborate buffer, pH 9.2 214 nm, 20 kV, 25 and 45°C 15 0.30 and 1.0 μg/mL (43) A. paniculata MDQ CE instrument equipped with PDA detector 31.2 cm × 50 μm i.d. uncoated fused silica capillary 0.5% (w/w) ethyl acetate+0.6% (w/w) SDS+ 97.7–92.9% (w/w) 30-mM sodium borate at pH 9.5 214 nm, −15 kV, 25°C 1.07 - (44) Sample . Instrument . Capillary . Reagents . Detection, voltage and temperature . RT . LOD and LOQ . Ref. . Standards of andrographolide and dehydroandrographolide Chuanxinlian and xiaoyanlidan tablets MDQ CE instrument equipped with PDA detector 60.2 cm × 50 μm i.d. uncoated fused silica capillary having length of 50 cm Heptane 0.81% (w/w), SDS 3.31% (w/w), butan-1-ol 6.61% (w/w) 10-mM sodium tetraborate buffer, pH 9.2 214 nm, 20 kV, 25 and 45°C 15 0.30 and 1.0 μg/mL (43) A. paniculata MDQ CE instrument equipped with PDA detector 31.2 cm × 50 μm i.d. uncoated fused silica capillary 0.5% (w/w) ethyl acetate+0.6% (w/w) SDS+ 97.7–92.9% (w/w) 30-mM sodium borate at pH 9.5 214 nm, −15 kV, 25°C 1.07 - (44) Open in new tab MEKC It is a technique based on partitioning of analytes between the micelles and aqueous phase in the presence of electroosmotic flow. This technique is reliable, efficient, has fast analysis time and requires only minute amount of sample and reagent (24). Table VII presents methods for andrographolide separation using MEKC. Table VII MEKC Methods for Andrographolide Determination in Pharmaceutical Formulations/Plant Materials Sample . Instrument . Capillary . Reagents . Detection, voltage and temperature . RT (min) . LOD . Ref. . A. paniculata, Chuaxinlian and xiaoyan lidan tablets Waters (Milford, MA, USA) Quanta 4000E CE system with fixed wavelength UV detector Fused silica 60 cm × 75 μm i.d. having 53-cm length 15-mM SDS in 30-mM borate buffer at pH 9.5 214 nm, 16 kV, 27°C 8.02 8.64–60.1 mg/l (45) A. paniculata Beckman P/ACE 5000, (Beckman Instruments, Fullerton, CA, USA) Fused silica capillary tube (Supelco, Bellefonte, PA, USA) 20-mM borate buffer, containing 20-mM sodium dodecyl sulphate and 10-mM sodium cholate, adjusted to pH 8.3 214 nm, 25 kV, 35°C 6.44 - (46) Sample . Instrument . Capillary . Reagents . Detection, voltage and temperature . RT (min) . LOD . Ref. . A. paniculata, Chuaxinlian and xiaoyan lidan tablets Waters (Milford, MA, USA) Quanta 4000E CE system with fixed wavelength UV detector Fused silica 60 cm × 75 μm i.d. having 53-cm length 15-mM SDS in 30-mM borate buffer at pH 9.5 214 nm, 16 kV, 27°C 8.02 8.64–60.1 mg/l (45) A. paniculata Beckman P/ACE 5000, (Beckman Instruments, Fullerton, CA, USA) Fused silica capillary tube (Supelco, Bellefonte, PA, USA) 20-mM borate buffer, containing 20-mM sodium dodecyl sulphate and 10-mM sodium cholate, adjusted to pH 8.3 214 nm, 25 kV, 35°C 6.44 - (46) Open in new tab Table VII MEKC Methods for Andrographolide Determination in Pharmaceutical Formulations/Plant Materials Sample . Instrument . Capillary . Reagents . Detection, voltage and temperature . RT (min) . LOD . Ref. . A. paniculata, Chuaxinlian and xiaoyan lidan tablets Waters (Milford, MA, USA) Quanta 4000E CE system with fixed wavelength UV detector Fused silica 60 cm × 75 μm i.d. having 53-cm length 15-mM SDS in 30-mM borate buffer at pH 9.5 214 nm, 16 kV, 27°C 8.02 8.64–60.1 mg/l (45) A. paniculata Beckman P/ACE 5000, (Beckman Instruments, Fullerton, CA, USA) Fused silica capillary tube (Supelco, Bellefonte, PA, USA) 20-mM borate buffer, containing 20-mM sodium dodecyl sulphate and 10-mM sodium cholate, adjusted to pH 8.3 214 nm, 25 kV, 35°C 6.44 - (46) Sample . Instrument . Capillary . Reagents . Detection, voltage and temperature . RT (min) . LOD . Ref. . A. paniculata, Chuaxinlian and xiaoyan lidan tablets Waters (Milford, MA, USA) Quanta 4000E CE system with fixed wavelength UV detector Fused silica 60 cm × 75 μm i.d. having 53-cm length 15-mM SDS in 30-mM borate buffer at pH 9.5 214 nm, 16 kV, 27°C 8.02 8.64–60.1 mg/l (45) A. paniculata Beckman P/ACE 5000, (Beckman Instruments, Fullerton, CA, USA) Fused silica capillary tube (Supelco, Bellefonte, PA, USA) 20-mM borate buffer, containing 20-mM sodium dodecyl sulphate and 10-mM sodium cholate, adjusted to pH 8.3 214 nm, 25 kV, 35°C 6.44 - (46) Open in new tab HSCCC It is a liquid–liquid partitioning chromatographic technique, which does not rely on the use of solid support for separation of solutes. Sample recovery, truncated solvent consumption, efficiency low cost and large sample loading capacity are some of the advantages of CCC (25). Multilayer coil column and UV detector were used by all researchers. Table VIII presents HSCCC method for the separation of andrographolide. Table VIII HSCCC Methods for Andrographolide Determination in Plant Materials Sample . Instrument . Column and Revolution speed . Detector . Solvent system . Peak . Ref. . A. paniculata Multilayer coil counter-current chromatograph manufactured by P.C. (Potomac, MD, USA) 385-mL coil column made of polytetrafluoroethylene tubing (2.6 mm I.D.) Volume = 385 mL 650 rpm 250 nm, UV–vis variable-wavelength Water–methanol-ethyl acetate-n-hexane (2.5:2.5:4:1) 1.5 mL/min 195–285 min (47) A. paniculata TBE-300B HSCCC (Tauto Biotchnique Company, Shanghai, China) Three multilayer coils connected in series having diameter of 1.5 mm, Volume = 300 mL 800 rpm 254 nm, TBD-2000 UV monitor n-hexane/ethyl acetate/methanol/water (2.5:5:2.5:5) 2.0 mL/min 68–78 min (48) A. paniculata TBE-300 A HSCCC equipment (Tauto Biotchnique Company, Shanghai, China) Multilayer column coil of polytetrafluoroethylene (PTFE), Volume = 300 mL 800 rpm 254 nm, 8823A UV detector Petroleum ether-ethyl acetate-methanol-water (3:7:5:5, v/v) 2.0 mL/min - (49) Sample . Instrument . Column and Revolution speed . Detector . Solvent system . Peak . Ref. . A. paniculata Multilayer coil counter-current chromatograph manufactured by P.C. (Potomac, MD, USA) 385-mL coil column made of polytetrafluoroethylene tubing (2.6 mm I.D.) Volume = 385 mL 650 rpm 250 nm, UV–vis variable-wavelength Water–methanol-ethyl acetate-n-hexane (2.5:2.5:4:1) 1.5 mL/min 195–285 min (47) A. paniculata TBE-300B HSCCC (Tauto Biotchnique Company, Shanghai, China) Three multilayer coils connected in series having diameter of 1.5 mm, Volume = 300 mL 800 rpm 254 nm, TBD-2000 UV monitor n-hexane/ethyl acetate/methanol/water (2.5:5:2.5:5) 2.0 mL/min 68–78 min (48) A. paniculata TBE-300 A HSCCC equipment (Tauto Biotchnique Company, Shanghai, China) Multilayer column coil of polytetrafluoroethylene (PTFE), Volume = 300 mL 800 rpm 254 nm, 8823A UV detector Petroleum ether-ethyl acetate-methanol-water (3:7:5:5, v/v) 2.0 mL/min - (49) Open in new tab Table VIII HSCCC Methods for Andrographolide Determination in Plant Materials Sample . Instrument . Column and Revolution speed . Detector . Solvent system . Peak . Ref. . A. paniculata Multilayer coil counter-current chromatograph manufactured by P.C. (Potomac, MD, USA) 385-mL coil column made of polytetrafluoroethylene tubing (2.6 mm I.D.) Volume = 385 mL 650 rpm 250 nm, UV–vis variable-wavelength Water–methanol-ethyl acetate-n-hexane (2.5:2.5:4:1) 1.5 mL/min 195–285 min (47) A. paniculata TBE-300B HSCCC (Tauto Biotchnique Company, Shanghai, China) Three multilayer coils connected in series having diameter of 1.5 mm, Volume = 300 mL 800 rpm 254 nm, TBD-2000 UV monitor n-hexane/ethyl acetate/methanol/water (2.5:5:2.5:5) 2.0 mL/min 68–78 min (48) A. paniculata TBE-300 A HSCCC equipment (Tauto Biotchnique Company, Shanghai, China) Multilayer column coil of polytetrafluoroethylene (PTFE), Volume = 300 mL 800 rpm 254 nm, 8823A UV detector Petroleum ether-ethyl acetate-methanol-water (3:7:5:5, v/v) 2.0 mL/min - (49) Sample . Instrument . Column and Revolution speed . Detector . Solvent system . Peak . Ref. . A. paniculata Multilayer coil counter-current chromatograph manufactured by P.C. (Potomac, MD, USA) 385-mL coil column made of polytetrafluoroethylene tubing (2.6 mm I.D.) Volume = 385 mL 650 rpm 250 nm, UV–vis variable-wavelength Water–methanol-ethyl acetate-n-hexane (2.5:2.5:4:1) 1.5 mL/min 195–285 min (47) A. paniculata TBE-300B HSCCC (Tauto Biotchnique Company, Shanghai, China) Three multilayer coils connected in series having diameter of 1.5 mm, Volume = 300 mL 800 rpm 254 nm, TBD-2000 UV monitor n-hexane/ethyl acetate/methanol/water (2.5:5:2.5:5) 2.0 mL/min 68–78 min (48) A. paniculata TBE-300 A HSCCC equipment (Tauto Biotchnique Company, Shanghai, China) Multilayer column coil of polytetrafluoroethylene (PTFE), Volume = 300 mL 800 rpm 254 nm, 8823A UV detector Petroleum ether-ethyl acetate-methanol-water (3:7:5:5, v/v) 2.0 mL/min - (49) Open in new tab HPTLC HPTLC is based on thin layer chromatography, but with enhancements to increase the resolution of compounds (26). It is rapid, simple, robust method requires low cost, sample preparation is simple and better resolution can be obtained. In HPTLC of andrographolide, mostly aluminium plates pre-coated with silica gel were used as stationary phase. Chloroform, toluene and methanol were used as mobile phase by most of the researchers, while ethyl acetate, formic acid and acetone were also used in some of HPTLC determination. Detection was carried out at between 230 and 254 nm. Table IX presents the methods for analysis and estimation of andrographolide using HPTLC. Table IX HPTLC Methods Used for Andrographolide Determination in Pharmaceutical Formulations/Plant Materials Sample . λ Max . Stationary phase . Solvent system . Rf. value . LOD and LOQ . Ref. . Andrographolide 231 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (7:1 v/v) 0.41 30 and 100 ng (50) Kalmegh Navayas Loha 235 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: ethyl acetate: formic acid (5:4.5:0.5 v/v) 0.38 - (51) A. paniculata 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (7:1) 0.35 - (52) A. paniculata 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (27: 3 v/v) 0.44 40 and 100 ng/spot (53) Polyherbal formulations Amylcure (Brand-A), Livomyn (Brand-B), Livcompound (Brand-C) 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (9:1, v/v) 0.64 3.05 and 18.28 ng/spot (54) Estimation of andrographolide and wedelolactone in herbal formulations 254 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: acetone: formic acid (9:6:1) 0.52 26.16 and 79.28 ng/spot (55) Herbal extract and polyherbal formulations 229 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: toluene: methanol (66:26:8, v/v/v) 0.49 3.5 and 11.7 ng (56) Polyherbal formulation (Livogat capsule) 254 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: acetone: formic acid [9: 7: 1 v/v/v] 0.51 62.91 and 209.7 ng/spot (57) Determination of Andrographolide in Bulk Drug and A. paniculata Formulations 230 nm TLC Aluminium plates precoated with silica gel 60 F254 Dichloromethane-toluene-ethanol (6:3:1, v/v/v) 0.36 34–109 and 112–363 ng/spot (58) Toluene-ethyl acetate-formic acid (5:3.5:1.5, v/v/v) 0.43 111–125 ng/spot and 370–420 ng/spot Sample . λ Max . Stationary phase . Solvent system . Rf. value . LOD and LOQ . Ref. . Andrographolide 231 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (7:1 v/v) 0.41 30 and 100 ng (50) Kalmegh Navayas Loha 235 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: ethyl acetate: formic acid (5:4.5:0.5 v/v) 0.38 - (51) A. paniculata 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (7:1) 0.35 - (52) A. paniculata 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (27: 3 v/v) 0.44 40 and 100 ng/spot (53) Polyherbal formulations Amylcure (Brand-A), Livomyn (Brand-B), Livcompound (Brand-C) 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (9:1, v/v) 0.64 3.05 and 18.28 ng/spot (54) Estimation of andrographolide and wedelolactone in herbal formulations 254 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: acetone: formic acid (9:6:1) 0.52 26.16 and 79.28 ng/spot (55) Herbal extract and polyherbal formulations 229 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: toluene: methanol (66:26:8, v/v/v) 0.49 3.5 and 11.7 ng (56) Polyherbal formulation (Livogat capsule) 254 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: acetone: formic acid [9: 7: 1 v/v/v] 0.51 62.91 and 209.7 ng/spot (57) Determination of Andrographolide in Bulk Drug and A. paniculata Formulations 230 nm TLC Aluminium plates precoated with silica gel 60 F254 Dichloromethane-toluene-ethanol (6:3:1, v/v/v) 0.36 34–109 and 112–363 ng/spot (58) Toluene-ethyl acetate-formic acid (5:3.5:1.5, v/v/v) 0.43 111–125 ng/spot and 370–420 ng/spot Open in new tab Table IX HPTLC Methods Used for Andrographolide Determination in Pharmaceutical Formulations/Plant Materials Sample . λ Max . Stationary phase . Solvent system . Rf. value . LOD and LOQ . Ref. . Andrographolide 231 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (7:1 v/v) 0.41 30 and 100 ng (50) Kalmegh Navayas Loha 235 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: ethyl acetate: formic acid (5:4.5:0.5 v/v) 0.38 - (51) A. paniculata 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (7:1) 0.35 - (52) A. paniculata 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (27: 3 v/v) 0.44 40 and 100 ng/spot (53) Polyherbal formulations Amylcure (Brand-A), Livomyn (Brand-B), Livcompound (Brand-C) 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (9:1, v/v) 0.64 3.05 and 18.28 ng/spot (54) Estimation of andrographolide and wedelolactone in herbal formulations 254 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: acetone: formic acid (9:6:1) 0.52 26.16 and 79.28 ng/spot (55) Herbal extract and polyherbal formulations 229 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: toluene: methanol (66:26:8, v/v/v) 0.49 3.5 and 11.7 ng (56) Polyherbal formulation (Livogat capsule) 254 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: acetone: formic acid [9: 7: 1 v/v/v] 0.51 62.91 and 209.7 ng/spot (57) Determination of Andrographolide in Bulk Drug and A. paniculata Formulations 230 nm TLC Aluminium plates precoated with silica gel 60 F254 Dichloromethane-toluene-ethanol (6:3:1, v/v/v) 0.36 34–109 and 112–363 ng/spot (58) Toluene-ethyl acetate-formic acid (5:3.5:1.5, v/v/v) 0.43 111–125 ng/spot and 370–420 ng/spot Sample . λ Max . Stationary phase . Solvent system . Rf. value . LOD and LOQ . Ref. . Andrographolide 231 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (7:1 v/v) 0.41 30 and 100 ng (50) Kalmegh Navayas Loha 235 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: ethyl acetate: formic acid (5:4.5:0.5 v/v) 0.38 - (51) A. paniculata 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (7:1) 0.35 - (52) A. paniculata 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (27: 3 v/v) 0.44 40 and 100 ng/spot (53) Polyherbal formulations Amylcure (Brand-A), Livomyn (Brand-B), Livcompound (Brand-C) 232 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: methanol (9:1, v/v) 0.64 3.05 and 18.28 ng/spot (54) Estimation of andrographolide and wedelolactone in herbal formulations 254 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: acetone: formic acid (9:6:1) 0.52 26.16 and 79.28 ng/spot (55) Herbal extract and polyherbal formulations 229 nm TLC Aluminium plates precoated with silica gel 60 F254 Chloroform: toluene: methanol (66:26:8, v/v/v) 0.49 3.5 and 11.7 ng (56) Polyherbal formulation (Livogat capsule) 254 nm TLC Aluminium plates precoated with silica gel 60 F254 Toluene: acetone: formic acid [9: 7: 1 v/v/v] 0.51 62.91 and 209.7 ng/spot (57) Determination of Andrographolide in Bulk Drug and A. paniculata Formulations 230 nm TLC Aluminium plates precoated with silica gel 60 F254 Dichloromethane-toluene-ethanol (6:3:1, v/v/v) 0.36 34–109 and 112–363 ng/spot (58) Toluene-ethyl acetate-formic acid (5:3.5:1.5, v/v/v) 0.43 111–125 ng/spot and 370–420 ng/spot Open in new tab HPLC It is an advanced technique of liquid chromatography, which is used for the separation, identification and quantification of each constituent of mixture (27). High speed, high resolution, sensitivity, accuracy and reproducibility are some of the advantages of HPLC. The HPLC method generally uses C18 column as stationary phase. Indian Pharmacopoeia recommends liquid chromatographic method for the determination of andrographolide by employing following method: Mobile phase: (i) a potassium dihydrogen orthophosphate buffer and (ii) acetonitrile, column: Octadecyl-silica(ODS) 5 μm, 25 cm × 4.6 mm, flow rate:1.5 mL/min, spectrophotometer set at 223 nm and injection volume: 20 μL (28). Indian Herbal Pharmacopoeia also recommends HPLC analysis by two methods: Method I:—mobile phase: chloroform: methanol (9:1), flow rate: 0.7 mL/min, column: 5-μm spherical silica (3.0 mm × 15 cm), detector: UV at 254 nm and the retention time (RT) was found to be 3 min. Method II:—mobile phase: methanol: water (65:35), flow rate: 1.0 mL/min, column C18 ODS, symmetry, 3 μ, 25 cm × 4 mm, detector: 223 nm and RT was 4.5 min (29). In many of the cases, combination of chloroform, methanol, acetonitrile and water was used widely, while isopropyl alcohol, formic acid and orthophosphoric acid were also used as mobile phase. Detection of andrographolide was performed between 205 and 254 nm with most of the analysis at 223 nm. Flow rate used in the analysis was in the range of 0.7–2 mL/min; mostly 1 mL/min was kept. UV–Vis detectors were used commonly in the detection. The following Tables X and XI present methods for the estimation and quantification of andrographolide in various samples using HPLC. Most of the chromatographic studies have been performed using methanol as extracting solvent and diluent. Table X HPLC Methods for Andrographolide Determination in Pharmaceutical Formulations/Plant Materials Sample . λ Max and detector . Column . HPLC mobile phase and flow rate . RT (min) . LOD and LOQ . Ref. . A. paniculata herb, extracts and dosage forms. 223-nm, Photodiode array detector Phenomenex Luna C18, 5 μm, (250 × 4.6 mm) Isocratic—acetonitrile: 0.1% (v/v) phosphoric acid in water (40: 60, v/v) 1 mL/min 5.8 0.001 and 0.004 μg/20 μL (59) Estimation of andrographolide (dried A. paniculata plant) and piperine 223 nm, UV–Vis detector C18 column (250 × 4 mm), 5 μL (waters Phenomenex) Isocratic—methanol: water (65:35) 2.45 - (60) Powdered plant of A. paniculata 210 nm, Photodiode array detector RP-18 column (250 × 4 mm, 5 mm Merck, Mumbai, India) Gradient—acetonitrile (15%, solvent A) methanol–water, 60:40 (85%, solvent B) 0.6 mL/min 5.41 5 and 10 μg/mL (61) Chuanxinlian tablets and A. paniculata 205 nm, UV–Vis detector C 18 column (5 μm, 150 mm × 4.6 mm i.d.) Isocratic-methanol–water (55:45, v/v) + silver ions (0.005 mol(L-1)) 5.5 0.05 and 0.25 μg/mL (62) A. paniculata 223 nm, UV–Vis detector Sunfire C-18 column (4.6 × 250 mm, 0.005 mm) Isocratic-methanol: water (65:35 v/v) 1 mL/min 5.6 - (63) Toxiroak Premix, A polyherbal mycotoxin inhibitor 226 nm, Photodiode array detector Phenomenex luna C18 column (250 mm × 4.6 mm, 5 μm) Isocratic-acetonitrile: ortho-phosphoric acid (0.1%),40:60 v/v 1.0 mL/min 5.19 0.06 and 0.2 μg/mL (63) Mixed herbal extract of Sambiloto (A. paniculata) leaves and meniran herbs 230-nm, Photodiode array detector Cosmosil (150 × 4.6 mm, 5 μm) Isocratic-acetonitrile:0.1% phosphoric acid (40:60) 1 mL/min 3 0.102 and 0.339 mg/mL (64) A. paniculata and andrographolide capsule 223 nm, UV-Vis detector C-18 column Isocratic-water: methanol (35:65) 0.7 mL/min 7.55 - (65) A. paniculata 229 nm, UV-Vis detector Sunfire C18 column Isocratic-methanol: water (6:4, v/v) 4.34 - (66) A. paniculata 223 nm, G1314B variable wavelength detector Zorbax Eclipse XDB-C18 column (4.6 mm × 150 mm, 5 μm) Isocratic-isopropyl alcohol: formic acid: water (70:10:20 v/v) 1.0 mL/min 3.67 0.06 and 0.17 μg/mL (67) Analysis of scopoletine, andrographolide, quercetin and luteolin in Antihypertension Jamu formulation 250 nm, UV–Vis detector LiChoCART column Gradient-formic acid (1%), methanol 2 mL/min 15.95 0.102 and 0.339 mg/mL (68) Analysis of andrographolide, phyllanthin and hypophyllanthin from herbal hepatoprotective formulation 230 nm, Photodiode Array detector Symmetry C8 column (250 mm × 4.6 mm, 5 μm) Gradient—0.1% orthophosphoric acid (solvent A) and (1:1) acetonitrile: methanol (solvent B) 1 mL/min 10.07 20 and 60 ng (69) A. paniculata 223 nm, Photodiode Array detector X Bridge TM C18 column (4.6 mm × 250 mm, 5.0 μm) Gradient-water(A) and mixture of methanol and reagent alcohol(B)-(1:1) 10 - (70) Andrographolide and its interaction with Curcumin and Artesunate 254 nm C18 column (250 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (1:1) 1 mL/min 3.75 - (71) Analysis of andrographolide in A. paniculata at two different stages of life cycle of plant 223 nm, UV–Vis detector C18 column (250 nm × 4.6 mm) Isocratic-methanol: water (65: 35) 1.5 mL/min 2.87 - (72) Quantification of andrographolide in Self-Nano Emulsifying Drug Delivery System (Snedds) 229 nm, UV–Vis detector Xterra MS C 18 column (150 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (70:30) 0.8 mL/min 2.72 1.95 and 3.13 μg/mL (73) A. paniculata 230 nm, UV–Vis detector C18 column (250 mm × 4.6 mm) Isocratic-methanol 1 mL/min 2.82 - (74) A. paniculata 254 nm, UV–Vis detector Spherical silica (3.9 mm × 15 cm,5 μm) column Isocratic -chloroform: methanol (90: 10) 0.7 mL/min 2.90 - (75) A. paniculata 220 nm, Photodiode array detector Merck RP18e Chromolith performance RP column (100 × 4.6 mm, i.d.) Gradient-acetonitrile: water (50:50) for hexane extract and acetonitrile: water (25:75) for chloroform, methanol or water extract 1.2 mL/min 3.85 - (76) Andrographis herb 225 nm, Photodiode array detector Waters XBridge™ Shield RP 18 column (250 mm × 4.6 mm i.d., 3.5 μm) Water (A) Acetonitrile (B) 0–25 min, 20–55% B. 1 mL/min 12.4 0.06 and 0.21 μg/mL (77) A. paniculata 230 nm, UV–Vis detector Thermo C-18 column (4.6 × 250 mm, 5 μ particle size) Isocratic-methanol (1 mL/min) 7.97 - (78) Andrographolide 224 nm, UV–Vis detector Sunfire C-18 (150 × 4.6 mm, 5 μ) column Isocratic-methanol: water (67:33) (1 mL/min) 2.58 1.0 and 3.34 μg/mL (39) Plant materials and commercial products of A. paniculata 225 nm, Photodiode array detector Supelco Discovery RP-18 column (250 × 4.6 mm, 5 μ particle size Gradient-water and ACN with ACN varying from 20 to 50% for 40 min 14.78 100 and 200 ng/mL (79) Commercial andrographis products Evaporative light scattering detector (ELSD) Reverse-phase column (Supelco Discovery C18) Gradient-water (deionized) (A) and acetonitrile (B) 18.59 LOD-50 ng (80) A. paniculata planted in various seasons and regions in Thailand 220 nm, UV–Vis detector μ-bondapack C18 column (30 cm × 0.39 mm i.d) Isocratic-42:58 methanol: water (2 mL/min) 1.45 - (81) Sample . λ Max and detector . Column . HPLC mobile phase and flow rate . RT (min) . LOD and LOQ . Ref. . A. paniculata herb, extracts and dosage forms. 223-nm, Photodiode array detector Phenomenex Luna C18, 5 μm, (250 × 4.6 mm) Isocratic—acetonitrile: 0.1% (v/v) phosphoric acid in water (40: 60, v/v) 1 mL/min 5.8 0.001 and 0.004 μg/20 μL (59) Estimation of andrographolide (dried A. paniculata plant) and piperine 223 nm, UV–Vis detector C18 column (250 × 4 mm), 5 μL (waters Phenomenex) Isocratic—methanol: water (65:35) 2.45 - (60) Powdered plant of A. paniculata 210 nm, Photodiode array detector RP-18 column (250 × 4 mm, 5 mm Merck, Mumbai, India) Gradient—acetonitrile (15%, solvent A) methanol–water, 60:40 (85%, solvent B) 0.6 mL/min 5.41 5 and 10 μg/mL (61) Chuanxinlian tablets and A. paniculata 205 nm, UV–Vis detector C 18 column (5 μm, 150 mm × 4.6 mm i.d.) Isocratic-methanol–water (55:45, v/v) + silver ions (0.005 mol(L-1)) 5.5 0.05 and 0.25 μg/mL (62) A. paniculata 223 nm, UV–Vis detector Sunfire C-18 column (4.6 × 250 mm, 0.005 mm) Isocratic-methanol: water (65:35 v/v) 1 mL/min 5.6 - (63) Toxiroak Premix, A polyherbal mycotoxin inhibitor 226 nm, Photodiode array detector Phenomenex luna C18 column (250 mm × 4.6 mm, 5 μm) Isocratic-acetonitrile: ortho-phosphoric acid (0.1%),40:60 v/v 1.0 mL/min 5.19 0.06 and 0.2 μg/mL (63) Mixed herbal extract of Sambiloto (A. paniculata) leaves and meniran herbs 230-nm, Photodiode array detector Cosmosil (150 × 4.6 mm, 5 μm) Isocratic-acetonitrile:0.1% phosphoric acid (40:60) 1 mL/min 3 0.102 and 0.339 mg/mL (64) A. paniculata and andrographolide capsule 223 nm, UV-Vis detector C-18 column Isocratic-water: methanol (35:65) 0.7 mL/min 7.55 - (65) A. paniculata 229 nm, UV-Vis detector Sunfire C18 column Isocratic-methanol: water (6:4, v/v) 4.34 - (66) A. paniculata 223 nm, G1314B variable wavelength detector Zorbax Eclipse XDB-C18 column (4.6 mm × 150 mm, 5 μm) Isocratic-isopropyl alcohol: formic acid: water (70:10:20 v/v) 1.0 mL/min 3.67 0.06 and 0.17 μg/mL (67) Analysis of scopoletine, andrographolide, quercetin and luteolin in Antihypertension Jamu formulation 250 nm, UV–Vis detector LiChoCART column Gradient-formic acid (1%), methanol 2 mL/min 15.95 0.102 and 0.339 mg/mL (68) Analysis of andrographolide, phyllanthin and hypophyllanthin from herbal hepatoprotective formulation 230 nm, Photodiode Array detector Symmetry C8 column (250 mm × 4.6 mm, 5 μm) Gradient—0.1% orthophosphoric acid (solvent A) and (1:1) acetonitrile: methanol (solvent B) 1 mL/min 10.07 20 and 60 ng (69) A. paniculata 223 nm, Photodiode Array detector X Bridge TM C18 column (4.6 mm × 250 mm, 5.0 μm) Gradient-water(A) and mixture of methanol and reagent alcohol(B)-(1:1) 10 - (70) Andrographolide and its interaction with Curcumin and Artesunate 254 nm C18 column (250 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (1:1) 1 mL/min 3.75 - (71) Analysis of andrographolide in A. paniculata at two different stages of life cycle of plant 223 nm, UV–Vis detector C18 column (250 nm × 4.6 mm) Isocratic-methanol: water (65: 35) 1.5 mL/min 2.87 - (72) Quantification of andrographolide in Self-Nano Emulsifying Drug Delivery System (Snedds) 229 nm, UV–Vis detector Xterra MS C 18 column (150 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (70:30) 0.8 mL/min 2.72 1.95 and 3.13 μg/mL (73) A. paniculata 230 nm, UV–Vis detector C18 column (250 mm × 4.6 mm) Isocratic-methanol 1 mL/min 2.82 - (74) A. paniculata 254 nm, UV–Vis detector Spherical silica (3.9 mm × 15 cm,5 μm) column Isocratic -chloroform: methanol (90: 10) 0.7 mL/min 2.90 - (75) A. paniculata 220 nm, Photodiode array detector Merck RP18e Chromolith performance RP column (100 × 4.6 mm, i.d.) Gradient-acetonitrile: water (50:50) for hexane extract and acetonitrile: water (25:75) for chloroform, methanol or water extract 1.2 mL/min 3.85 - (76) Andrographis herb 225 nm, Photodiode array detector Waters XBridge™ Shield RP 18 column (250 mm × 4.6 mm i.d., 3.5 μm) Water (A) Acetonitrile (B) 0–25 min, 20–55% B. 1 mL/min 12.4 0.06 and 0.21 μg/mL (77) A. paniculata 230 nm, UV–Vis detector Thermo C-18 column (4.6 × 250 mm, 5 μ particle size) Isocratic-methanol (1 mL/min) 7.97 - (78) Andrographolide 224 nm, UV–Vis detector Sunfire C-18 (150 × 4.6 mm, 5 μ) column Isocratic-methanol: water (67:33) (1 mL/min) 2.58 1.0 and 3.34 μg/mL (39) Plant materials and commercial products of A. paniculata 225 nm, Photodiode array detector Supelco Discovery RP-18 column (250 × 4.6 mm, 5 μ particle size Gradient-water and ACN with ACN varying from 20 to 50% for 40 min 14.78 100 and 200 ng/mL (79) Commercial andrographis products Evaporative light scattering detector (ELSD) Reverse-phase column (Supelco Discovery C18) Gradient-water (deionized) (A) and acetonitrile (B) 18.59 LOD-50 ng (80) A. paniculata planted in various seasons and regions in Thailand 220 nm, UV–Vis detector μ-bondapack C18 column (30 cm × 0.39 mm i.d) Isocratic-42:58 methanol: water (2 mL/min) 1.45 - (81) Open in new tab Table X HPLC Methods for Andrographolide Determination in Pharmaceutical Formulations/Plant Materials Sample . λ Max and detector . Column . HPLC mobile phase and flow rate . RT (min) . LOD and LOQ . Ref. . A. paniculata herb, extracts and dosage forms. 223-nm, Photodiode array detector Phenomenex Luna C18, 5 μm, (250 × 4.6 mm) Isocratic—acetonitrile: 0.1% (v/v) phosphoric acid in water (40: 60, v/v) 1 mL/min 5.8 0.001 and 0.004 μg/20 μL (59) Estimation of andrographolide (dried A. paniculata plant) and piperine 223 nm, UV–Vis detector C18 column (250 × 4 mm), 5 μL (waters Phenomenex) Isocratic—methanol: water (65:35) 2.45 - (60) Powdered plant of A. paniculata 210 nm, Photodiode array detector RP-18 column (250 × 4 mm, 5 mm Merck, Mumbai, India) Gradient—acetonitrile (15%, solvent A) methanol–water, 60:40 (85%, solvent B) 0.6 mL/min 5.41 5 and 10 μg/mL (61) Chuanxinlian tablets and A. paniculata 205 nm, UV–Vis detector C 18 column (5 μm, 150 mm × 4.6 mm i.d.) Isocratic-methanol–water (55:45, v/v) + silver ions (0.005 mol(L-1)) 5.5 0.05 and 0.25 μg/mL (62) A. paniculata 223 nm, UV–Vis detector Sunfire C-18 column (4.6 × 250 mm, 0.005 mm) Isocratic-methanol: water (65:35 v/v) 1 mL/min 5.6 - (63) Toxiroak Premix, A polyherbal mycotoxin inhibitor 226 nm, Photodiode array detector Phenomenex luna C18 column (250 mm × 4.6 mm, 5 μm) Isocratic-acetonitrile: ortho-phosphoric acid (0.1%),40:60 v/v 1.0 mL/min 5.19 0.06 and 0.2 μg/mL (63) Mixed herbal extract of Sambiloto (A. paniculata) leaves and meniran herbs 230-nm, Photodiode array detector Cosmosil (150 × 4.6 mm, 5 μm) Isocratic-acetonitrile:0.1% phosphoric acid (40:60) 1 mL/min 3 0.102 and 0.339 mg/mL (64) A. paniculata and andrographolide capsule 223 nm, UV-Vis detector C-18 column Isocratic-water: methanol (35:65) 0.7 mL/min 7.55 - (65) A. paniculata 229 nm, UV-Vis detector Sunfire C18 column Isocratic-methanol: water (6:4, v/v) 4.34 - (66) A. paniculata 223 nm, G1314B variable wavelength detector Zorbax Eclipse XDB-C18 column (4.6 mm × 150 mm, 5 μm) Isocratic-isopropyl alcohol: formic acid: water (70:10:20 v/v) 1.0 mL/min 3.67 0.06 and 0.17 μg/mL (67) Analysis of scopoletine, andrographolide, quercetin and luteolin in Antihypertension Jamu formulation 250 nm, UV–Vis detector LiChoCART column Gradient-formic acid (1%), methanol 2 mL/min 15.95 0.102 and 0.339 mg/mL (68) Analysis of andrographolide, phyllanthin and hypophyllanthin from herbal hepatoprotective formulation 230 nm, Photodiode Array detector Symmetry C8 column (250 mm × 4.6 mm, 5 μm) Gradient—0.1% orthophosphoric acid (solvent A) and (1:1) acetonitrile: methanol (solvent B) 1 mL/min 10.07 20 and 60 ng (69) A. paniculata 223 nm, Photodiode Array detector X Bridge TM C18 column (4.6 mm × 250 mm, 5.0 μm) Gradient-water(A) and mixture of methanol and reagent alcohol(B)-(1:1) 10 - (70) Andrographolide and its interaction with Curcumin and Artesunate 254 nm C18 column (250 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (1:1) 1 mL/min 3.75 - (71) Analysis of andrographolide in A. paniculata at two different stages of life cycle of plant 223 nm, UV–Vis detector C18 column (250 nm × 4.6 mm) Isocratic-methanol: water (65: 35) 1.5 mL/min 2.87 - (72) Quantification of andrographolide in Self-Nano Emulsifying Drug Delivery System (Snedds) 229 nm, UV–Vis detector Xterra MS C 18 column (150 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (70:30) 0.8 mL/min 2.72 1.95 and 3.13 μg/mL (73) A. paniculata 230 nm, UV–Vis detector C18 column (250 mm × 4.6 mm) Isocratic-methanol 1 mL/min 2.82 - (74) A. paniculata 254 nm, UV–Vis detector Spherical silica (3.9 mm × 15 cm,5 μm) column Isocratic -chloroform: methanol (90: 10) 0.7 mL/min 2.90 - (75) A. paniculata 220 nm, Photodiode array detector Merck RP18e Chromolith performance RP column (100 × 4.6 mm, i.d.) Gradient-acetonitrile: water (50:50) for hexane extract and acetonitrile: water (25:75) for chloroform, methanol or water extract 1.2 mL/min 3.85 - (76) Andrographis herb 225 nm, Photodiode array detector Waters XBridge™ Shield RP 18 column (250 mm × 4.6 mm i.d., 3.5 μm) Water (A) Acetonitrile (B) 0–25 min, 20–55% B. 1 mL/min 12.4 0.06 and 0.21 μg/mL (77) A. paniculata 230 nm, UV–Vis detector Thermo C-18 column (4.6 × 250 mm, 5 μ particle size) Isocratic-methanol (1 mL/min) 7.97 - (78) Andrographolide 224 nm, UV–Vis detector Sunfire C-18 (150 × 4.6 mm, 5 μ) column Isocratic-methanol: water (67:33) (1 mL/min) 2.58 1.0 and 3.34 μg/mL (39) Plant materials and commercial products of A. paniculata 225 nm, Photodiode array detector Supelco Discovery RP-18 column (250 × 4.6 mm, 5 μ particle size Gradient-water and ACN with ACN varying from 20 to 50% for 40 min 14.78 100 and 200 ng/mL (79) Commercial andrographis products Evaporative light scattering detector (ELSD) Reverse-phase column (Supelco Discovery C18) Gradient-water (deionized) (A) and acetonitrile (B) 18.59 LOD-50 ng (80) A. paniculata planted in various seasons and regions in Thailand 220 nm, UV–Vis detector μ-bondapack C18 column (30 cm × 0.39 mm i.d) Isocratic-42:58 methanol: water (2 mL/min) 1.45 - (81) Sample . λ Max and detector . Column . HPLC mobile phase and flow rate . RT (min) . LOD and LOQ . Ref. . A. paniculata herb, extracts and dosage forms. 223-nm, Photodiode array detector Phenomenex Luna C18, 5 μm, (250 × 4.6 mm) Isocratic—acetonitrile: 0.1% (v/v) phosphoric acid in water (40: 60, v/v) 1 mL/min 5.8 0.001 and 0.004 μg/20 μL (59) Estimation of andrographolide (dried A. paniculata plant) and piperine 223 nm, UV–Vis detector C18 column (250 × 4 mm), 5 μL (waters Phenomenex) Isocratic—methanol: water (65:35) 2.45 - (60) Powdered plant of A. paniculata 210 nm, Photodiode array detector RP-18 column (250 × 4 mm, 5 mm Merck, Mumbai, India) Gradient—acetonitrile (15%, solvent A) methanol–water, 60:40 (85%, solvent B) 0.6 mL/min 5.41 5 and 10 μg/mL (61) Chuanxinlian tablets and A. paniculata 205 nm, UV–Vis detector C 18 column (5 μm, 150 mm × 4.6 mm i.d.) Isocratic-methanol–water (55:45, v/v) + silver ions (0.005 mol(L-1)) 5.5 0.05 and 0.25 μg/mL (62) A. paniculata 223 nm, UV–Vis detector Sunfire C-18 column (4.6 × 250 mm, 0.005 mm) Isocratic-methanol: water (65:35 v/v) 1 mL/min 5.6 - (63) Toxiroak Premix, A polyherbal mycotoxin inhibitor 226 nm, Photodiode array detector Phenomenex luna C18 column (250 mm × 4.6 mm, 5 μm) Isocratic-acetonitrile: ortho-phosphoric acid (0.1%),40:60 v/v 1.0 mL/min 5.19 0.06 and 0.2 μg/mL (63) Mixed herbal extract of Sambiloto (A. paniculata) leaves and meniran herbs 230-nm, Photodiode array detector Cosmosil (150 × 4.6 mm, 5 μm) Isocratic-acetonitrile:0.1% phosphoric acid (40:60) 1 mL/min 3 0.102 and 0.339 mg/mL (64) A. paniculata and andrographolide capsule 223 nm, UV-Vis detector C-18 column Isocratic-water: methanol (35:65) 0.7 mL/min 7.55 - (65) A. paniculata 229 nm, UV-Vis detector Sunfire C18 column Isocratic-methanol: water (6:4, v/v) 4.34 - (66) A. paniculata 223 nm, G1314B variable wavelength detector Zorbax Eclipse XDB-C18 column (4.6 mm × 150 mm, 5 μm) Isocratic-isopropyl alcohol: formic acid: water (70:10:20 v/v) 1.0 mL/min 3.67 0.06 and 0.17 μg/mL (67) Analysis of scopoletine, andrographolide, quercetin and luteolin in Antihypertension Jamu formulation 250 nm, UV–Vis detector LiChoCART column Gradient-formic acid (1%), methanol 2 mL/min 15.95 0.102 and 0.339 mg/mL (68) Analysis of andrographolide, phyllanthin and hypophyllanthin from herbal hepatoprotective formulation 230 nm, Photodiode Array detector Symmetry C8 column (250 mm × 4.6 mm, 5 μm) Gradient—0.1% orthophosphoric acid (solvent A) and (1:1) acetonitrile: methanol (solvent B) 1 mL/min 10.07 20 and 60 ng (69) A. paniculata 223 nm, Photodiode Array detector X Bridge TM C18 column (4.6 mm × 250 mm, 5.0 μm) Gradient-water(A) and mixture of methanol and reagent alcohol(B)-(1:1) 10 - (70) Andrographolide and its interaction with Curcumin and Artesunate 254 nm C18 column (250 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (1:1) 1 mL/min 3.75 - (71) Analysis of andrographolide in A. paniculata at two different stages of life cycle of plant 223 nm, UV–Vis detector C18 column (250 nm × 4.6 mm) Isocratic-methanol: water (65: 35) 1.5 mL/min 2.87 - (72) Quantification of andrographolide in Self-Nano Emulsifying Drug Delivery System (Snedds) 229 nm, UV–Vis detector Xterra MS C 18 column (150 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (70:30) 0.8 mL/min 2.72 1.95 and 3.13 μg/mL (73) A. paniculata 230 nm, UV–Vis detector C18 column (250 mm × 4.6 mm) Isocratic-methanol 1 mL/min 2.82 - (74) A. paniculata 254 nm, UV–Vis detector Spherical silica (3.9 mm × 15 cm,5 μm) column Isocratic -chloroform: methanol (90: 10) 0.7 mL/min 2.90 - (75) A. paniculata 220 nm, Photodiode array detector Merck RP18e Chromolith performance RP column (100 × 4.6 mm, i.d.) Gradient-acetonitrile: water (50:50) for hexane extract and acetonitrile: water (25:75) for chloroform, methanol or water extract 1.2 mL/min 3.85 - (76) Andrographis herb 225 nm, Photodiode array detector Waters XBridge™ Shield RP 18 column (250 mm × 4.6 mm i.d., 3.5 μm) Water (A) Acetonitrile (B) 0–25 min, 20–55% B. 1 mL/min 12.4 0.06 and 0.21 μg/mL (77) A. paniculata 230 nm, UV–Vis detector Thermo C-18 column (4.6 × 250 mm, 5 μ particle size) Isocratic-methanol (1 mL/min) 7.97 - (78) Andrographolide 224 nm, UV–Vis detector Sunfire C-18 (150 × 4.6 mm, 5 μ) column Isocratic-methanol: water (67:33) (1 mL/min) 2.58 1.0 and 3.34 μg/mL (39) Plant materials and commercial products of A. paniculata 225 nm, Photodiode array detector Supelco Discovery RP-18 column (250 × 4.6 mm, 5 μ particle size Gradient-water and ACN with ACN varying from 20 to 50% for 40 min 14.78 100 and 200 ng/mL (79) Commercial andrographis products Evaporative light scattering detector (ELSD) Reverse-phase column (Supelco Discovery C18) Gradient-water (deionized) (A) and acetonitrile (B) 18.59 LOD-50 ng (80) A. paniculata planted in various seasons and regions in Thailand 220 nm, UV–Vis detector μ-bondapack C18 column (30 cm × 0.39 mm i.d) Isocratic-42:58 methanol: water (2 mL/min) 1.45 - (81) Open in new tab Table XI HPLC Methods Used for Determining the Presence of Andrographolide in Biological Samples Sample . λ Max and detector . Column . HPLC mobile phase . RT (min) . LOD and LOQ . Ref. . Rabbit serum (rabbit was administered andrographolide isolated from dried A. paniculata) 223 nm, UV–Vis detector Neuclosil C18 octadecyl silane (ODS), 5 μm, (250 × 4.6 mm Hypersil) Isocratic-methanol: water (65:35) 1.0 mL/min 3.69 - (82) Rabbit plasma (Oral administration of A. paniculata extract to rabbit) 225 nm, UV–Vis detector C18 column (250 mm × 4.6 mm i.d.,5 μm) Isocratic-methanol: acetonitrile: water (50:10:40, v/v) 4.3 0.019 μg/ mL (83) A23187-INDUCED New Zealand rabbit’s (treated with andrographolide) urine and faeces 227 nm, UV–Vis detector C 18 column (250 mm in, 5-μm and 120 Å pore size) Isocratic-methanol–water (55:45) 1 mL/min 0.5 mL/min 5.5 1.87 and 5.45 μg/mL (84) Plasma of male white rabbits of New Zealand strain 229 nm, UV–Vis detector Xterra MS C 18 column (150 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (60:40) Flow rate-0.8 mL/min 3.6 - (85) Serum of Nicobari fowl feeded with Kalmegh extract 223 nm, UV–Vis detector C-18 column (250 × 4 mm, 5 μm) Isocratic-methanol: water (65:35) 1 mL/min 4.37 - (86) Liposomes and tablets containing andrographolide were administered to rat and its whole blood was analyzed 225-nm, Diode Array Detector Chromasil ODS column (250 × 4.6 mm, 5 μm) Isocratic-methanol: water (52: 48 v/v) 0.8 mL/min 10.63 0.015 and 0.053 μg/mL (87) A. paniculata, Andrographis extract and tablet RRLC-LC2010 HPLC system coupled with a Waters Premier MS equipped with electrospray ionization and Time of flight Mass Spectrometer ACE Excel 3 Super C18 column (100 mm × 2.1 mm i.d., 3.0 μm) Acetonitrile and water mobile phase additive-0.1% formic acid 0.20 mL/min 5 0.06 and 0.2 μg/mL (88) Sample . λ Max and detector . Column . HPLC mobile phase . RT (min) . LOD and LOQ . Ref. . Rabbit serum (rabbit was administered andrographolide isolated from dried A. paniculata) 223 nm, UV–Vis detector Neuclosil C18 octadecyl silane (ODS), 5 μm, (250 × 4.6 mm Hypersil) Isocratic-methanol: water (65:35) 1.0 mL/min 3.69 - (82) Rabbit plasma (Oral administration of A. paniculata extract to rabbit) 225 nm, UV–Vis detector C18 column (250 mm × 4.6 mm i.d.,5 μm) Isocratic-methanol: acetonitrile: water (50:10:40, v/v) 4.3 0.019 μg/ mL (83) A23187-INDUCED New Zealand rabbit’s (treated with andrographolide) urine and faeces 227 nm, UV–Vis detector C 18 column (250 mm in, 5-μm and 120 Å pore size) Isocratic-methanol–water (55:45) 1 mL/min 0.5 mL/min 5.5 1.87 and 5.45 μg/mL (84) Plasma of male white rabbits of New Zealand strain 229 nm, UV–Vis detector Xterra MS C 18 column (150 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (60:40) Flow rate-0.8 mL/min 3.6 - (85) Serum of Nicobari fowl feeded with Kalmegh extract 223 nm, UV–Vis detector C-18 column (250 × 4 mm, 5 μm) Isocratic-methanol: water (65:35) 1 mL/min 4.37 - (86) Liposomes and tablets containing andrographolide were administered to rat and its whole blood was analyzed 225-nm, Diode Array Detector Chromasil ODS column (250 × 4.6 mm, 5 μm) Isocratic-methanol: water (52: 48 v/v) 0.8 mL/min 10.63 0.015 and 0.053 μg/mL (87) A. paniculata, Andrographis extract and tablet RRLC-LC2010 HPLC system coupled with a Waters Premier MS equipped with electrospray ionization and Time of flight Mass Spectrometer ACE Excel 3 Super C18 column (100 mm × 2.1 mm i.d., 3.0 μm) Acetonitrile and water mobile phase additive-0.1% formic acid 0.20 mL/min 5 0.06 and 0.2 μg/mL (88) Open in new tab Table XI HPLC Methods Used for Determining the Presence of Andrographolide in Biological Samples Sample . λ Max and detector . Column . HPLC mobile phase . RT (min) . LOD and LOQ . Ref. . Rabbit serum (rabbit was administered andrographolide isolated from dried A. paniculata) 223 nm, UV–Vis detector Neuclosil C18 octadecyl silane (ODS), 5 μm, (250 × 4.6 mm Hypersil) Isocratic-methanol: water (65:35) 1.0 mL/min 3.69 - (82) Rabbit plasma (Oral administration of A. paniculata extract to rabbit) 225 nm, UV–Vis detector C18 column (250 mm × 4.6 mm i.d.,5 μm) Isocratic-methanol: acetonitrile: water (50:10:40, v/v) 4.3 0.019 μg/ mL (83) A23187-INDUCED New Zealand rabbit’s (treated with andrographolide) urine and faeces 227 nm, UV–Vis detector C 18 column (250 mm in, 5-μm and 120 Å pore size) Isocratic-methanol–water (55:45) 1 mL/min 0.5 mL/min 5.5 1.87 and 5.45 μg/mL (84) Plasma of male white rabbits of New Zealand strain 229 nm, UV–Vis detector Xterra MS C 18 column (150 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (60:40) Flow rate-0.8 mL/min 3.6 - (85) Serum of Nicobari fowl feeded with Kalmegh extract 223 nm, UV–Vis detector C-18 column (250 × 4 mm, 5 μm) Isocratic-methanol: water (65:35) 1 mL/min 4.37 - (86) Liposomes and tablets containing andrographolide were administered to rat and its whole blood was analyzed 225-nm, Diode Array Detector Chromasil ODS column (250 × 4.6 mm, 5 μm) Isocratic-methanol: water (52: 48 v/v) 0.8 mL/min 10.63 0.015 and 0.053 μg/mL (87) A. paniculata, Andrographis extract and tablet RRLC-LC2010 HPLC system coupled with a Waters Premier MS equipped with electrospray ionization and Time of flight Mass Spectrometer ACE Excel 3 Super C18 column (100 mm × 2.1 mm i.d., 3.0 μm) Acetonitrile and water mobile phase additive-0.1% formic acid 0.20 mL/min 5 0.06 and 0.2 μg/mL (88) Sample . λ Max and detector . Column . HPLC mobile phase . RT (min) . LOD and LOQ . Ref. . Rabbit serum (rabbit was administered andrographolide isolated from dried A. paniculata) 223 nm, UV–Vis detector Neuclosil C18 octadecyl silane (ODS), 5 μm, (250 × 4.6 mm Hypersil) Isocratic-methanol: water (65:35) 1.0 mL/min 3.69 - (82) Rabbit plasma (Oral administration of A. paniculata extract to rabbit) 225 nm, UV–Vis detector C18 column (250 mm × 4.6 mm i.d.,5 μm) Isocratic-methanol: acetonitrile: water (50:10:40, v/v) 4.3 0.019 μg/ mL (83) A23187-INDUCED New Zealand rabbit’s (treated with andrographolide) urine and faeces 227 nm, UV–Vis detector C 18 column (250 mm in, 5-μm and 120 Å pore size) Isocratic-methanol–water (55:45) 1 mL/min 0.5 mL/min 5.5 1.87 and 5.45 μg/mL (84) Plasma of male white rabbits of New Zealand strain 229 nm, UV–Vis detector Xterra MS C 18 column (150 mm × 4.6 mm, 5 μm) Isocratic-methanol: water (60:40) Flow rate-0.8 mL/min 3.6 - (85) Serum of Nicobari fowl feeded with Kalmegh extract 223 nm, UV–Vis detector C-18 column (250 × 4 mm, 5 μm) Isocratic-methanol: water (65:35) 1 mL/min 4.37 - (86) Liposomes and tablets containing andrographolide were administered to rat and its whole blood was analyzed 225-nm, Diode Array Detector Chromasil ODS column (250 × 4.6 mm, 5 μm) Isocratic-methanol: water (52: 48 v/v) 0.8 mL/min 10.63 0.015 and 0.053 μg/mL (87) A. paniculata, Andrographis extract and tablet RRLC-LC2010 HPLC system coupled with a Waters Premier MS equipped with electrospray ionization and Time of flight Mass Spectrometer ACE Excel 3 Super C18 column (100 mm × 2.1 mm i.d., 3.0 μm) Acetonitrile and water mobile phase additive-0.1% formic acid 0.20 mL/min 5 0.06 and 0.2 μg/mL (88) Open in new tab R‌RLC This method was developed for the qualitative and quantitative analysis of chemical constituents in A. paniculata including diterpenoid lactones. RRLC is a variant of HPLC using columns having particle size less than 2 μm (typically, 1.8 μm). These short columns (50 mm × 4.6 mm) are operated at high flow rate and they show better performance as compared with conventional columns having 5-μm porous particles (250 mm × 4.6 mm). The separation was better and analysis time was also reduced by using short columns (30). Ethanol exhibits relatively higher extraction efficiency for andrographolide; hence, it was applied as in this experiment. Hyphenated techniques Hyphenated techniques have various advantages such as fast and accurate analysis, higher degree of automation, better reproducibility, reduction of contamination, etc. (31). Quantification of andrographolide in biological samples using Liquid Chromatography–tandem mass spectrometry (LC–MS/MS) is becoming the method of choice as it offers limiting time-consuming, water-free derivatization, intense analyte clean up and concentration steps. Ultra performance liquid chromatography—tandem mass spectrometer (UPLC–MS/MS) allows the detection of analytes of interest at very low concentration due to its sensitivity, selectivity, shorter run time and low consumption of solvents (32). Various hyphenated techniques used for andrographolide determination are presented in Tables XII and XIII. An overview of analytical methods for andrographolide estimation is presented in Figure 3. Table XII Hyphenated Techniques Used for Determining the Presence of Andrographolide in Pharmaceutical Formulations/Plant Materials Sample . Instrument . Column . Mobile phase/solvents/detection wavelength . RT (min) . LOD and LOQ . Ref. . Fresh leaves and stems of A. paniculata UPLC™ (Waters, Milford, MA) Mass spectrometer (API 4000 QTRAPT™ MS/MS system from AB Sciex, Concord, ON, Canada) Acquity BEH C18 (2.1 mm × 50 mm, 1.7 μm) Gradient:0.1% formic acid in water(A) 0.1% formic acid in acetonitrile(B) 0.3 mL/min 2.10 0.18 and 0.75 ng/mL (89) A. paniculata tablets Hewlett Packard LC-1100 gradient liquid chromatography instrument Hypersil ODS column (250 × 40 mm, 5 μm) Gradient-solvent A (0.025% trifluoroacetic acid in acetonitrile) and solvent B (0.025% trifluoroacetic acid in water) 1 mL/min 27 - (90) A. paniculata Dynamic microwave-assisted extraction coupled on-line with HPLC TM010 microwave resonance cavity DAD detector Pinnacle 11 C18 (250 mm × 4.6 mm i.d.,5 μm) column (Restok, Bellefonte, PA, USA) Methanol 1 mL/min 225 nm 3.75 0.06 and 0.2 μg/mL (91) Sample . Instrument . Column . Mobile phase/solvents/detection wavelength . RT (min) . LOD and LOQ . Ref. . Fresh leaves and stems of A. paniculata UPLC™ (Waters, Milford, MA) Mass spectrometer (API 4000 QTRAPT™ MS/MS system from AB Sciex, Concord, ON, Canada) Acquity BEH C18 (2.1 mm × 50 mm, 1.7 μm) Gradient:0.1% formic acid in water(A) 0.1% formic acid in acetonitrile(B) 0.3 mL/min 2.10 0.18 and 0.75 ng/mL (89) A. paniculata tablets Hewlett Packard LC-1100 gradient liquid chromatography instrument Hypersil ODS column (250 × 40 mm, 5 μm) Gradient-solvent A (0.025% trifluoroacetic acid in acetonitrile) and solvent B (0.025% trifluoroacetic acid in water) 1 mL/min 27 - (90) A. paniculata Dynamic microwave-assisted extraction coupled on-line with HPLC TM010 microwave resonance cavity DAD detector Pinnacle 11 C18 (250 mm × 4.6 mm i.d.,5 μm) column (Restok, Bellefonte, PA, USA) Methanol 1 mL/min 225 nm 3.75 0.06 and 0.2 μg/mL (91) Open in new tab Table XII Hyphenated Techniques Used for Determining the Presence of Andrographolide in Pharmaceutical Formulations/Plant Materials Sample . Instrument . Column . Mobile phase/solvents/detection wavelength . RT (min) . LOD and LOQ . Ref. . Fresh leaves and stems of A. paniculata UPLC™ (Waters, Milford, MA) Mass spectrometer (API 4000 QTRAPT™ MS/MS system from AB Sciex, Concord, ON, Canada) Acquity BEH C18 (2.1 mm × 50 mm, 1.7 μm) Gradient:0.1% formic acid in water(A) 0.1% formic acid in acetonitrile(B) 0.3 mL/min 2.10 0.18 and 0.75 ng/mL (89) A. paniculata tablets Hewlett Packard LC-1100 gradient liquid chromatography instrument Hypersil ODS column (250 × 40 mm, 5 μm) Gradient-solvent A (0.025% trifluoroacetic acid in acetonitrile) and solvent B (0.025% trifluoroacetic acid in water) 1 mL/min 27 - (90) A. paniculata Dynamic microwave-assisted extraction coupled on-line with HPLC TM010 microwave resonance cavity DAD detector Pinnacle 11 C18 (250 mm × 4.6 mm i.d.,5 μm) column (Restok, Bellefonte, PA, USA) Methanol 1 mL/min 225 nm 3.75 0.06 and 0.2 μg/mL (91) Sample . Instrument . Column . Mobile phase/solvents/detection wavelength . RT (min) . LOD and LOQ . Ref. . Fresh leaves and stems of A. paniculata UPLC™ (Waters, Milford, MA) Mass spectrometer (API 4000 QTRAPT™ MS/MS system from AB Sciex, Concord, ON, Canada) Acquity BEH C18 (2.1 mm × 50 mm, 1.7 μm) Gradient:0.1% formic acid in water(A) 0.1% formic acid in acetonitrile(B) 0.3 mL/min 2.10 0.18 and 0.75 ng/mL (89) A. paniculata tablets Hewlett Packard LC-1100 gradient liquid chromatography instrument Hypersil ODS column (250 × 40 mm, 5 μm) Gradient-solvent A (0.025% trifluoroacetic acid in acetonitrile) and solvent B (0.025% trifluoroacetic acid in water) 1 mL/min 27 - (90) A. paniculata Dynamic microwave-assisted extraction coupled on-line with HPLC TM010 microwave resonance cavity DAD detector Pinnacle 11 C18 (250 mm × 4.6 mm i.d.,5 μm) column (Restok, Bellefonte, PA, USA) Methanol 1 mL/min 225 nm 3.75 0.06 and 0.2 μg/mL (91) Open in new tab Table XIII Hyphenated Techniques Used for Determining the Presence of Andrographolide in Biological Samples Sample . Instrument . Column . Mobile phase/ solvents/detection wavelength . RT/Results . LOD and LOQ . Ref. . Urine and faeces sample of rat administered with A. paniculata extract HPLC system (Waters Corp., USA) Agilent-1100 HPLC/6520 Q-TOF-MS system (Agilent Corp., USA) Waters Symmetry Shield TM RP C18 column (3.9 × 150 mm, 5 μm) and Nova-Pak C18 Guard-Pak TM column Gradient—0.2% formic acid (A) acetonitrile (B) 0.5 mL/min 275 nm 18.3 min Precursor ion-699.1579 Fragment ion-687.2453 - (92) Pharmacokinetic analysis and tissue distribution of andrographolide in rat LC system consisting of LC-20 ADvp (Shimadzu, Kyoto, Japan) Mass spectrometer composed of turbo ion spray with atmospheric pressure ionization source (API-4000, AB Sciex Instruments, Foster City, CA) C18 column (2 × 30 mm, 5 μm) from phenomenex (Torrance, CA) Gradient—2-mM ammonium acetate buffer (A) and a mixture of acetonitrile and solvent A (80:20, v/v) (B) 0.8 mL/min 1.55 min [MH]− ion at m/z = 349.20 LOQ- 3.91 ng/mL (93) Determination of four major active diterpenoids from A.paniculata in human Plasma LC–MS/MS (6490 Triple Quadrupole, Agilent Technologies) Kinetex column (4.6 × 150 mm, 2.6 μm, Phenomenex) Gradient-water (solvent A) acetonitrile(B) 0.5 mL/min 5.6 min m/z-331.1 LOQ-2.50 ng/mL (94) Determination of andrographolide in human plasma Waters (Milford, MA, USA) Alliance 2695 liquid chromatographic system and Micro-mass ZQ 2000 ESI mass spectrometer Johnson Spherigel C18 column (200 mm × 4.6 mm, 5 μm) Isocratic-methanol: aqueous solution (70:30, v/v) 4.7 min m/z-331 LOQ- 9.9 ng/mL (95) Human Plasma HPLC-ESI-MS/MS equipment. Finigan TSQ Discovery Max Tandem mass spectrometer (Thermo Electron Corporation, San Jose, CA, USA) with ESI ion source Hanbon C18 (4.6 mm × 150 mm, 5 μm, Jiangsu Hanbon Science and Technology Co.Ltd., China) Isocratic-methanol: water (70:30, v/v) 0.7 mL/min 3.27 min m/z-331.1 1.0 ng/mL (96) Rat plasma Shimadzu LC-10AT (Shimadzu Corporation, Kyoto, Japan). SPD-10A UV/Vis detector. Neuclosil C18(250 × 4.6 mm i.d., 5 μm, Phenomenex) Isocratic-methanol: water (65:35, v/v) 210 nm 1 mL/min 4.35 min 15 ng (97) Rat serum Surveyor HPLC system (Thermo-Finnigan, San Jose, CA, USA) Zorbax SB-18(150 × 3.0 mm, 3.5 μm, Agilent) Isocratic-methanol–water (60:40) 0.3 mL/min 6.94 min - (98) Finnigan LCQ DEC XPplus ion-trap mass spectrometer 5 kV Auxilliary nitrogen gas 300°C m/z-331 - UPLC Waters Acquitytm UPLC system (Waters Corp., Milford, MA, USA) Acquity UPLC BEH C18 column (50 × 2.1 mm i.d., 1.7 μm) Gradient-water-acetonitrile 0.3 mL/min 2.56 min - Rat plasma Triple quadrupole mass spectrometer (API 4000, AB Sciex, Foster City, CA, USA) Agilent 1200 Series liquid chromatography system Waters Xbridge Shield RP C18 column (50 mm × 2.1 mm i.d., 5 μm) (Waters Corp., Mildford, MA, USA) Gradient-A (0.1% acetic acid in water) B (50% acetonitrile in methanol) 0.4 mL/min 2.6 min m/z- 331 - (99) Sample . Instrument . Column . Mobile phase/ solvents/detection wavelength . RT/Results . LOD and LOQ . Ref. . Urine and faeces sample of rat administered with A. paniculata extract HPLC system (Waters Corp., USA) Agilent-1100 HPLC/6520 Q-TOF-MS system (Agilent Corp., USA) Waters Symmetry Shield TM RP C18 column (3.9 × 150 mm, 5 μm) and Nova-Pak C18 Guard-Pak TM column Gradient—0.2% formic acid (A) acetonitrile (B) 0.5 mL/min 275 nm 18.3 min Precursor ion-699.1579 Fragment ion-687.2453 - (92) Pharmacokinetic analysis and tissue distribution of andrographolide in rat LC system consisting of LC-20 ADvp (Shimadzu, Kyoto, Japan) Mass spectrometer composed of turbo ion spray with atmospheric pressure ionization source (API-4000, AB Sciex Instruments, Foster City, CA) C18 column (2 × 30 mm, 5 μm) from phenomenex (Torrance, CA) Gradient—2-mM ammonium acetate buffer (A) and a mixture of acetonitrile and solvent A (80:20, v/v) (B) 0.8 mL/min 1.55 min [MH]− ion at m/z = 349.20 LOQ- 3.91 ng/mL (93) Determination of four major active diterpenoids from A.paniculata in human Plasma LC–MS/MS (6490 Triple Quadrupole, Agilent Technologies) Kinetex column (4.6 × 150 mm, 2.6 μm, Phenomenex) Gradient-water (solvent A) acetonitrile(B) 0.5 mL/min 5.6 min m/z-331.1 LOQ-2.50 ng/mL (94) Determination of andrographolide in human plasma Waters (Milford, MA, USA) Alliance 2695 liquid chromatographic system and Micro-mass ZQ 2000 ESI mass spectrometer Johnson Spherigel C18 column (200 mm × 4.6 mm, 5 μm) Isocratic-methanol: aqueous solution (70:30, v/v) 4.7 min m/z-331 LOQ- 9.9 ng/mL (95) Human Plasma HPLC-ESI-MS/MS equipment. Finigan TSQ Discovery Max Tandem mass spectrometer (Thermo Electron Corporation, San Jose, CA, USA) with ESI ion source Hanbon C18 (4.6 mm × 150 mm, 5 μm, Jiangsu Hanbon Science and Technology Co.Ltd., China) Isocratic-methanol: water (70:30, v/v) 0.7 mL/min 3.27 min m/z-331.1 1.0 ng/mL (96) Rat plasma Shimadzu LC-10AT (Shimadzu Corporation, Kyoto, Japan). SPD-10A UV/Vis detector. Neuclosil C18(250 × 4.6 mm i.d., 5 μm, Phenomenex) Isocratic-methanol: water (65:35, v/v) 210 nm 1 mL/min 4.35 min 15 ng (97) Rat serum Surveyor HPLC system (Thermo-Finnigan, San Jose, CA, USA) Zorbax SB-18(150 × 3.0 mm, 3.5 μm, Agilent) Isocratic-methanol–water (60:40) 0.3 mL/min 6.94 min - (98) Finnigan LCQ DEC XPplus ion-trap mass spectrometer 5 kV Auxilliary nitrogen gas 300°C m/z-331 - UPLC Waters Acquitytm UPLC system (Waters Corp., Milford, MA, USA) Acquity UPLC BEH C18 column (50 × 2.1 mm i.d., 1.7 μm) Gradient-water-acetonitrile 0.3 mL/min 2.56 min - Rat plasma Triple quadrupole mass spectrometer (API 4000, AB Sciex, Foster City, CA, USA) Agilent 1200 Series liquid chromatography system Waters Xbridge Shield RP C18 column (50 mm × 2.1 mm i.d., 5 μm) (Waters Corp., Mildford, MA, USA) Gradient-A (0.1% acetic acid in water) B (50% acetonitrile in methanol) 0.4 mL/min 2.6 min m/z- 331 - (99) Open in new tab Table XIII Hyphenated Techniques Used for Determining the Presence of Andrographolide in Biological Samples Sample . Instrument . Column . Mobile phase/ solvents/detection wavelength . RT/Results . LOD and LOQ . Ref. . Urine and faeces sample of rat administered with A. paniculata extract HPLC system (Waters Corp., USA) Agilent-1100 HPLC/6520 Q-TOF-MS system (Agilent Corp., USA) Waters Symmetry Shield TM RP C18 column (3.9 × 150 mm, 5 μm) and Nova-Pak C18 Guard-Pak TM column Gradient—0.2% formic acid (A) acetonitrile (B) 0.5 mL/min 275 nm 18.3 min Precursor ion-699.1579 Fragment ion-687.2453 - (92) Pharmacokinetic analysis and tissue distribution of andrographolide in rat LC system consisting of LC-20 ADvp (Shimadzu, Kyoto, Japan) Mass spectrometer composed of turbo ion spray with atmospheric pressure ionization source (API-4000, AB Sciex Instruments, Foster City, CA) C18 column (2 × 30 mm, 5 μm) from phenomenex (Torrance, CA) Gradient—2-mM ammonium acetate buffer (A) and a mixture of acetonitrile and solvent A (80:20, v/v) (B) 0.8 mL/min 1.55 min [MH]− ion at m/z = 349.20 LOQ- 3.91 ng/mL (93) Determination of four major active diterpenoids from A.paniculata in human Plasma LC–MS/MS (6490 Triple Quadrupole, Agilent Technologies) Kinetex column (4.6 × 150 mm, 2.6 μm, Phenomenex) Gradient-water (solvent A) acetonitrile(B) 0.5 mL/min 5.6 min m/z-331.1 LOQ-2.50 ng/mL (94) Determination of andrographolide in human plasma Waters (Milford, MA, USA) Alliance 2695 liquid chromatographic system and Micro-mass ZQ 2000 ESI mass spectrometer Johnson Spherigel C18 column (200 mm × 4.6 mm, 5 μm) Isocratic-methanol: aqueous solution (70:30, v/v) 4.7 min m/z-331 LOQ- 9.9 ng/mL (95) Human Plasma HPLC-ESI-MS/MS equipment. Finigan TSQ Discovery Max Tandem mass spectrometer (Thermo Electron Corporation, San Jose, CA, USA) with ESI ion source Hanbon C18 (4.6 mm × 150 mm, 5 μm, Jiangsu Hanbon Science and Technology Co.Ltd., China) Isocratic-methanol: water (70:30, v/v) 0.7 mL/min 3.27 min m/z-331.1 1.0 ng/mL (96) Rat plasma Shimadzu LC-10AT (Shimadzu Corporation, Kyoto, Japan). SPD-10A UV/Vis detector. Neuclosil C18(250 × 4.6 mm i.d., 5 μm, Phenomenex) Isocratic-methanol: water (65:35, v/v) 210 nm 1 mL/min 4.35 min 15 ng (97) Rat serum Surveyor HPLC system (Thermo-Finnigan, San Jose, CA, USA) Zorbax SB-18(150 × 3.0 mm, 3.5 μm, Agilent) Isocratic-methanol–water (60:40) 0.3 mL/min 6.94 min - (98) Finnigan LCQ DEC XPplus ion-trap mass spectrometer 5 kV Auxilliary nitrogen gas 300°C m/z-331 - UPLC Waters Acquitytm UPLC system (Waters Corp., Milford, MA, USA) Acquity UPLC BEH C18 column (50 × 2.1 mm i.d., 1.7 μm) Gradient-water-acetonitrile 0.3 mL/min 2.56 min - Rat plasma Triple quadrupole mass spectrometer (API 4000, AB Sciex, Foster City, CA, USA) Agilent 1200 Series liquid chromatography system Waters Xbridge Shield RP C18 column (50 mm × 2.1 mm i.d., 5 μm) (Waters Corp., Mildford, MA, USA) Gradient-A (0.1% acetic acid in water) B (50% acetonitrile in methanol) 0.4 mL/min 2.6 min m/z- 331 - (99) Sample . Instrument . Column . Mobile phase/ solvents/detection wavelength . RT/Results . LOD and LOQ . Ref. . Urine and faeces sample of rat administered with A. paniculata extract HPLC system (Waters Corp., USA) Agilent-1100 HPLC/6520 Q-TOF-MS system (Agilent Corp., USA) Waters Symmetry Shield TM RP C18 column (3.9 × 150 mm, 5 μm) and Nova-Pak C18 Guard-Pak TM column Gradient—0.2% formic acid (A) acetonitrile (B) 0.5 mL/min 275 nm 18.3 min Precursor ion-699.1579 Fragment ion-687.2453 - (92) Pharmacokinetic analysis and tissue distribution of andrographolide in rat LC system consisting of LC-20 ADvp (Shimadzu, Kyoto, Japan) Mass spectrometer composed of turbo ion spray with atmospheric pressure ionization source (API-4000, AB Sciex Instruments, Foster City, CA) C18 column (2 × 30 mm, 5 μm) from phenomenex (Torrance, CA) Gradient—2-mM ammonium acetate buffer (A) and a mixture of acetonitrile and solvent A (80:20, v/v) (B) 0.8 mL/min 1.55 min [MH]− ion at m/z = 349.20 LOQ- 3.91 ng/mL (93) Determination of four major active diterpenoids from A.paniculata in human Plasma LC–MS/MS (6490 Triple Quadrupole, Agilent Technologies) Kinetex column (4.6 × 150 mm, 2.6 μm, Phenomenex) Gradient-water (solvent A) acetonitrile(B) 0.5 mL/min 5.6 min m/z-331.1 LOQ-2.50 ng/mL (94) Determination of andrographolide in human plasma Waters (Milford, MA, USA) Alliance 2695 liquid chromatographic system and Micro-mass ZQ 2000 ESI mass spectrometer Johnson Spherigel C18 column (200 mm × 4.6 mm, 5 μm) Isocratic-methanol: aqueous solution (70:30, v/v) 4.7 min m/z-331 LOQ- 9.9 ng/mL (95) Human Plasma HPLC-ESI-MS/MS equipment. Finigan TSQ Discovery Max Tandem mass spectrometer (Thermo Electron Corporation, San Jose, CA, USA) with ESI ion source Hanbon C18 (4.6 mm × 150 mm, 5 μm, Jiangsu Hanbon Science and Technology Co.Ltd., China) Isocratic-methanol: water (70:30, v/v) 0.7 mL/min 3.27 min m/z-331.1 1.0 ng/mL (96) Rat plasma Shimadzu LC-10AT (Shimadzu Corporation, Kyoto, Japan). SPD-10A UV/Vis detector. Neuclosil C18(250 × 4.6 mm i.d., 5 μm, Phenomenex) Isocratic-methanol: water (65:35, v/v) 210 nm 1 mL/min 4.35 min 15 ng (97) Rat serum Surveyor HPLC system (Thermo-Finnigan, San Jose, CA, USA) Zorbax SB-18(150 × 3.0 mm, 3.5 μm, Agilent) Isocratic-methanol–water (60:40) 0.3 mL/min 6.94 min - (98) Finnigan LCQ DEC XPplus ion-trap mass spectrometer 5 kV Auxilliary nitrogen gas 300°C m/z-331 - UPLC Waters Acquitytm UPLC system (Waters Corp., Milford, MA, USA) Acquity UPLC BEH C18 column (50 × 2.1 mm i.d., 1.7 μm) Gradient-water-acetonitrile 0.3 mL/min 2.56 min - Rat plasma Triple quadrupole mass spectrometer (API 4000, AB Sciex, Foster City, CA, USA) Agilent 1200 Series liquid chromatography system Waters Xbridge Shield RP C18 column (50 mm × 2.1 mm i.d., 5 μm) (Waters Corp., Mildford, MA, USA) Gradient-A (0.1% acetic acid in water) B (50% acetonitrile in methanol) 0.4 mL/min 2.6 min m/z- 331 - (99) Open in new tab Figure 3 Open in new tabDownload slide Overview of analytical methods for estimation of andrographolide. Figure 3 Open in new tabDownload slide Overview of analytical methods for estimation of andrographolide. Discussion The abovementioned methods have application in the determination of drug in various pharmaceutical formulations such as tablets, capsules, liposomes, extracts and self-nano emulsifying drug delivery systems. Human plasma, urine, rat plasma, tissue, urine and faeces samples, rabbit plasma and fowl serum have been analyzed for the andrographolide by the abovementioned methods. These methods can be used for the routine analysis of andrographolide. These methods have been used for the separation and quantification of andrographolide in various biological samples and pharmaceutical preparations. All the methods are reported to be accurate, sensitive, reproducible, robust and reliable. Conclusion This review has summarized the analytical methods for determining the presence of bicyclic diterpenoid lactone andrographolide. It possesses many biological activities and is of great importance; hence, different analytical methods are needed for its estimation, quantification and separation in various samples. A broad range of analytical methods are available for the analysis of andrographolide in various pharmaceutical formulations and biological samples. The analysis of the compiled data revealed that the HPLC was extensively used for the determination of andrographolide. The method based on CPE coupled with high-performance liquid chromatography separation and ultraviolet (UV) detection (Green method) was found to be efficient, environmentally friendly, rapid and inexpensive. Many hyphenated techniques have also been applied for bioanalytical studies. This review would help analysts knowing the combination of solvents and instruments employed in the analysis of andrographolide. Conflict of Interest The authors declare that this article content has no conflicts of interest. References [1] Joseph , S.M. ; Scientific aspects of the therapeutic use of A.paniculata (kalmegh): A review ; Int J Pharm Sci Rev Res , ( 2014 ); 27 ( 1 ): 10 – 13 . Google Scholar OpenURL Placeholder Text WorldCat [2] Mishra , S.K. , Sangwan , N.S., Sangwan , R.S.; Phcog rev.: Plant review A.paniculata (Kalmegh): A review ; Pharmacogn Rev , ( 2007 ); 1 ( 2 ): 283 – 296 . Google Scholar OpenURL Placeholder Text WorldCat [3] Dhiman , A. , Goyal , J., Sharma , K., Nanda , A., Dhiman , S.; A review on medicinal prospectives of A.paniculata nees ; J Pharm Sci Innovation , ( 2012 ); 1 ( 1 ): 1 – 4 . Google Scholar OpenURL Placeholder Text WorldCat [4] Sharma , S. , Sharma , Y.P., Bhardwaj , C.; HPLC quantification of andrographolide in different parts of A.paniculata (Burm.F.) wall. Ex Nees ; J Pharmacogn Phytochem , ( 2018 ); 7 ( 3 ): 168 – 171 . Google Scholar OpenURL Placeholder Text WorldCat [5] Joselin , J. , Jeeva , S.; Andrographis paniculata: A review of its traditional uses, phytochemistry and pharmacology ; Med Aromat Plants , ( 2014 ); 3 ( 4 ): 1 – 15 . Google Scholar OpenURL Placeholder Text WorldCat [6] Benoy , G.K. , Animesh , D.K., Mandal , A., Dubey , K.P., Halder , S.; An overview on A.paniculata (Burm.F.) Nees ; Int J Res Ayurveda Pharm , ( 2012 ); 3 ( 6 ): 752 – 760 . Google Scholar Crossref Search ADS WorldCat [7] National Center for Biotechnology Information . PubChem Compound Database, CID=5318517 . https://pubchem.ncbi.nlm.nih.gov/compound/5318517 ( accessed February 2, 2019 ). [8] Thanasekaran , J. , Hsieh , C.-Y., Lee , J.-J., Sheu , J.R.; Experimental and clinical pharmacology of A.paniculata and its major bioactive phytoconstituent andrographolide ; Evid Based Complement Alternat Med , ( 2013 ); 846740 : 1 – 16 . Google Scholar OpenURL Placeholder Text WorldCat [9] Andrographolide - DrugBank . https://www.drugbank.ca/drugs/DB05767 ( accessed December 11, 2019 ) [10] Pandey , G. , Rao , C.H.; Andrographolide: Its pharmacology, natural bioavailability and current approaches to increase its content in Andrographis paniculata ; Int J Complement Alt Med , ( 2018 ); 11 ( 6 ): 355 – 360 . Google Scholar OpenURL Placeholder Text WorldCat [11] Biswa , D.B. , Sharma , P.K., Kumar , N., Dudhe , R., Bansal , V.; Pharmacological activity of A.paniculata: A brief review ; Pharmacol Ther , ( 2011 ); 2 : 1 – 10 . Google Scholar OpenURL Placeholder Text WorldCat [12] Handa , S.S. , Sharma , A.; Hepatoprotective activity of andrographolide against galoctosomine and paracetamole intoxication in rats ; Indian J Med Res , ( 1990 ); 92 : 284 – 292 . Google Scholar PubMed OpenURL Placeholder Text WorldCat [13] Vojdani , A. , Erde , J.; Regulatory T cells, a potent immunoregulatory target for CAM researchers: Modulating tumor immunity, autoimmunity and alloreactive immunity (III) ; Evid Based Complement Alternat Med , ( 2006 ); 3 ( 3 ): 309 – 316 . Google Scholar Crossref Search ADS PubMed WorldCat [14] Siripong P ; Konkathip B; Preechanukool K; Picha P; Tunsuwan K; Taylor WC.; Cytotoxic diterpenoid constituents from A.paniculata Nees. leaves; J Sci Soc Thailand, ( 1992); 18 : 187 – 194 . [15] Misra , P. , Pal , N.L., Guru , P.Y., Katiyar , J.C., Srivastava , V., Tandon , J.S.; Antimalarial activity of A.paniculata (Kalmegh) against Plasmodium berghei NK65 in mastomys natalensis ; Int J Pharm , ( 1992 ); 30 : 263 – 274 . Google Scholar OpenURL Placeholder Text WorldCat [16] ThamLikitkul , V. , Dechatiwongse , T., Theerapong , S., Chantrakul , C., Boonroj , P., Punkrut , W. et al. ; Efficacy of A.paniculata, Nees for pharyngotonsillitis in adults ; J Med Assoc Thai , ( 1991 ); 74 ( 10 ): 437 – 442 . Google Scholar PubMed OpenURL Placeholder Text WorldCat [17] Chiou , W.F. , Chen , C.F., Lin , J.J.; Mechanisms of suppression of inducible nitric oxide synthase (iNOS) expression in RAW 264.7 cells by andrographolide ; Br J Pharmacol , ( 2000 ); 129 : 1553 – 1560 . Google Scholar Crossref Search ADS PubMed WorldCat [18] Hidalgo , M.A. , Romero , A., Figueroa , J., Cortes , P., Concha , I.I., Hancke , J.L. et al. ; Andrographolide interferes with binding of nuclear factor-kb to DNA in HL-60-derived neutrophilic cells ; Br J Pharmacol , ( 2005 ); 144 : 680 – 686 . Google Scholar Crossref Search ADS PubMed WorldCat [19] Shaikh , S. , Jain , V.; Development and validation of novel RP-HPLC method for the simultaneous estimation of ellagic acid and quercetin in an ayurvedic formulation ; Int J Appl Pharm , ( 2018 ); 10 ( 4 ): 111 – 116 . Google Scholar Crossref Search ADS WorldCat [20] Asghar , M. , Yaqoob , M., Nabi , A.; Chemiluminescent determination of cyromazine in milk samples using copper (III) chelate-triton X-100 by flow injection analysis ; Chem Res Chin Univ , ( 2017 ); 33 ( 3 ): 354 – 359 . Google Scholar Crossref Search ADS WorldCat [21] Stankovic , D. , Mehmeti , E., Svorc , L., Kalcher , K.; New electrochemical method for the determination of β-carboline alkaloids, harmalol and harmine, in human urine samples and in Banisteriopsis caapi ; Microchem J , ( 2015 ); 118 : 95 – 100 . Google Scholar Crossref Search ADS WorldCat [22] Buchberger , W.W. ; Microemulsion electrokinetic chromatography ; Methods Mol Biol , ( 2008 ); 1483 : 91 – 109 . Google Scholar Crossref Search ADS WorldCat [23] Altria , K.D. ; Highly efficient and selective separations of a wide range of analytes obtained by an optimised microemulsion electrokinetic chromatography method ; Chromatographia , ( 1999 ); 49 ( 7–8 ): 457 – 464 . Google Scholar Crossref Search ADS WorldCat [24] Riekkola , M.-L. ; Micellar electrokinetic capillary chromatography ; 1st ed. Academic Press, Finland, ( 2000 ); pp. 1280 – 1286 . [25] Song , K. , Lee , K.J., Kim , Y.S.; Development of an efficient fractionation method for the preparative separation of sesquiterpenoids from Tussilago farfara by counter-current chromatography ; J Chromatogr A , ( 2017 ); 1489 : 107 – 114 . Google Scholar Crossref Search ADS PubMed WorldCat [26] Attimarad , M. , Ahmed , K.K.M., Aldhubaib , B.E., Harsha , S.; High-performance thin layer chromatography: A powerful analytical technique in pharmaceutical drug discovery ; Pharm Methods , ( 2011 ); 2 ( 2 ): 71 – 75 . Google Scholar Crossref Search ADS PubMed WorldCat [27] Thammana , M. ; A review on high performance liquid chromatography (HPLC) ; Res Rev: J Pharm Anal , ( 2016 ); 5 ( 2 ): 22 – 28 . Google Scholar OpenURL Placeholder Text WorldCat [28] Indian Pharmacopoeia Volume III . Published by The Indian Pharmacopoeia Commission , Ghaziabad , ( 2018 ), p. 3813 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC [29] Indian Herbal Pharmacopoeia Revised New Edition , Vol 2002 . Published by Indian Drug Manufacturers Association , Ebenezer Printing House, Mumbai , ( 2002 ), pp. 57 – 66 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC [30] Yoshida , T. , Majors , R.E.; High-speed analyses using rapid resolution liquid chromatography on 1.8 μm porous particles ; J Sep Sci , ( 2006 ); 29 ( 16 ): 2421 – 2432 . Google Scholar Crossref Search ADS PubMed WorldCat [31] Nagajyothi , S. , Swetha , Y., Neeharika , J., Suresh , P.V., Ramarao , N.; Hyphenated techniques-a comprehensive review ; Int J Adv Res Dev , ( 2017 ); 2 ( 4 ): 63 – 71 . Google Scholar OpenURL Placeholder Text WorldCat [32] Zhang , J. , Tang , C., Oberly , P.J., Minnigh , M.B., Achilles , S.L., Poloyac , S.M.; A sensitive and robust UPLC–MS/MS method for quantitation of estrogens and progestogens in human serum ; Contraception , ( 2019 ); 99 : 244 – 250 . Google Scholar Crossref Search ADS PubMed WorldCat [33] Pancham , Y. , Patil , N., B , G., Mannur , V.; Development and validation of analytical method for determination of andrographolide in bulk powder ; Int J Pharma Res Health Sci , ( 2019 ); 7 ( 1 ): 2899 – 2903 . Google Scholar Crossref Search ADS WorldCat [34] Aromdee , C. , Wichitchote , P., Jantakun , N.; Spectrophotometric determination of total lactones in A.paniculata Nees; Songklanakarin ; J Sci Technol , ( 2005 ); 27 ( 6 ): 1227 – 1231 . Google Scholar OpenURL Placeholder Text WorldCat [35] Shah , K. , Trivedi , P., Shivprakash, Pundarikakshudu , K.; Spectrophotometric determination of andrographolides in Andrographis paniculata Nees and its formulation ; Indian J Pharm Sci , ( 2007 ); 69 ( 3 ): 457 – 458 . Google Scholar Crossref Search ADS WorldCat [36] Ahamad , J. , Ameen , M.S.M., Anwer , E.T., Uthirapathy , S., Porwal , O.; Qualitative and quantitative standardization of Andrographis paniculata by TLC technique and UV method ; AMDHS , ( 2019 ); 2 ( 3 ): 29 – 32 . Google Scholar Crossref Search ADS WorldCat [37] Shivali , G. , Praful , L., Vijay , V.; A validated fourier transform infrared spectroscopy method for quantification of total lactones in Inula racemose and A.paniculata ; Phytochem Anal , ( 2012 ); 23 : 171 – 176 . Google Scholar Crossref Search ADS PubMed WorldCat [38] Singh , P.K. , Hasan , T., Prasad , O., Sinha , L., Raj , K., Mishra , N.; FT-IR spectra and vibrational spectroscopy of Andrographolide ; Spectroscopy , ( 2006 ); 20 : 275 – 283 . Google Scholar Crossref Search ADS WorldCat [39] Indrati , O. , Martien , R., Rohman , A., Nugroho , A.K.; Employment of ATR-FTIR and HPLC-UV method for detection and quantification of Andrographolide ; Int J App Pharm , ( 2018 ); 10 ( 6 ): 135 – 138 . Google Scholar Crossref Search ADS WorldCat [40] Jiang , Z. , Hao , Z., Wu , Q., Li , Y., Liu , H., Yan , L.; A novel flow injection chemiluminescence method for determination of andrographolide in Andrographis tablets ; Drug Test Anal , ( 2012 ); 5 ( 5 ): 340 – 345 . Google Scholar Crossref Search ADS PubMed WorldCat [41] Stankovic , D.M. , Samphao , A., Kuzmanovic , D., Kalcher , K.; Novel electroanalyical method for the determination of andrographolide from A.paniculata extract and urine sample ; Microchem J , ( 2015 ); 122 : 16 – 19 . Google Scholar Crossref Search ADS WorldCat [42] Zhao , Q. , Ding , J., Jin , H., Ding , L., Ren , N.; A green method using a micellar system for determination of andrographolide and dehydroandrographolide in human plasma ; J Chromatogr Sci , ( 2013 ); 51 ( 4 ): 341 – 348 . Google Scholar Crossref Search ADS PubMed WorldCat [43] Yangfang , Z. , Xingping , L., Zongde , Z., Liren , C., Yongmin , L.; Simultaneous determination of andrographolide and dehydroandrographolide in Andrographis paniculate and Chinese medicinal preparations by microemulsion electrokinetic chromatography ; J Pharm Biomed Anal , ( 2006 ); 40 ( 1 ): 157 – 161 . Google Scholar Crossref Search ADS PubMed WorldCat [44] Zhao , Y. ; Ming , Y.; Zhang , H.; Luo , X.; Chen , L.; Yongmin, rapid determination of diterpenoids in A.paniculata by microemulsion electrokinetic capillary chromatography with short-end injection ; Chromatographia, ( 2005); 62 ( 11/12 ): 611 - 615 . Google Scholar Crossref Search ADS WorldCat [45] Zhao , J. , Yang , G., Liu , H., Wang , D., Song , X., Chen , Y.; Determination of andrographolide, deoxyandrographolide and neoandrographolide in the chinese herb A.paniculata by micellar electrokinetic capillary chromatography ; PhytochemAnal , ( 2002 ); 13 ( 4 ): 222 – 227 . Google Scholar OpenURL Placeholder Text WorldCat [46] Cheung , H.Y. , Cheung , C.S., Kong , C.K.; Determination of bioactive diterpenoids from A.paniculata by micellar electrokinetic chromatography ; J Chromatogr A , ( 2001 ); 930 ( 1-2 ): 171 – 176 . Google Scholar Crossref Search ADS PubMed WorldCat [47] Du , Q. , Jerz , G., Winterhalter , P.; Separation of andrographolide and neoandrographolide from the leaves of A.paniculata using high-speed counter current chromatography ; J Chromatogr A , ( 2003 ); 984 ( 1 ): 147 – 151 . Google Scholar Crossref Search ADS PubMed WorldCat [48] Zhang , Y.Q. , Wang , S.-S., Han , C., Xu , J.-F., Luo , J.-G., Kong , L.-Y.; Online hyphenation of extraction, Sephadex LH-20 column chromatography and high-speed counter current chromatography: A highly efficient strategy for the preparative separation of andrographolide from A.paniculata in a single step ; J Sep Sci , ( 2017 ); 40 ( 24 ): 4865 – 4871 . Google Scholar Crossref Search ADS PubMed WorldCat [49] Sun , X. , Yan , H., Zhang , Y., Wang , X., Qin , D., Yu , J.; Preparative separation of diterpene lactones and flavones from A.paniculata using off-lines two-dimensional high-speed counter-current chromatography ; Molecules , ( 2019 ); 24 ( 3 ): 620 . Google Scholar Crossref Search ADS WorldCat [50] Mamatha , A. ; Quantitative HPTLC analysis of andrographolide in A.paniculata obtained from different geographical sources (India) ; Int J Pharm Pharm Sci , ( 2011 ); 3 ( 2 ): 42 – 44 . Google Scholar OpenURL Placeholder Text WorldCat [51] Pawar , R.K. , Sharma , S., Singh , K.C., Sharma , K.R.; Development and validation of HPTLC method for the determination of andrographolide in kalmegh navayas Loha an ayurvedic formulation ; Int J Pharm Pharm Sci , ( 2011 ); 3 ( 2 ): 85 – 89 . Google Scholar OpenURL Placeholder Text WorldCat [52] Raina , A.P. , Kumar , A., Pareek , S.K.; HPTLC analysis of hepatoprotective diterpenoid andrographolide from A.paniculata Nees (Kalmegh) ; Indian J Pharm Sci , ( 2007 ); 69 ( 3 ): 473 – 475 . Google Scholar Crossref Search ADS WorldCat [53] Misra , H. , Soni , M., Mehta , D., Mehta , B.K., Jain , D.C.; An improved HPTLC – UV method for rapid estimation of andrographolide in A.paniculata (Burm. F) Nees ; InPharm Communique (Suppl) , ( 2009 ); 2 ( 2 ): 51 – 53 . Google Scholar OpenURL Placeholder Text WorldCat [54] Zade , S.R. , Priya , B., Bagmar , U.R., Ganjiwale , R.O.; Development and validation of HPTLC method for estimation of hepatoprotective diterpenoid andrographolide in polyherbal formulations ; Int J Pharm Pharm Sci , ( 2013 ); 5 ( 3 ): 976 – 980 . Google Scholar OpenURL Placeholder Text WorldCat [55] Patel , M.B. , Kadakia , V.M., Mishra , S.H.; Simultaneous estimation of andrographolide and wedelolactone in herbal formulations ; Indian J Pharm Sci , ( 2008 ); 70 ( 5 ): 689 – 693 . Google Scholar Crossref Search ADS PubMed WorldCat [56] Jain , P.K. , Ravichandran , V., Jain , P.K., Agrawal , R.K.; High-performance thin layer chromatography method for estimation of andrographolide in herbal extract and polyherbal formulations ; J Saudi Chem Soc , ( 2010 ); 14 ( 4 ): 383 – 389 . Google Scholar Crossref Search ADS WorldCat [57] Shingala , D. , Gaudani , R., Nayak , B., Ghodasara , T.; Development and validation of HPTLC method for estimation of gallic acid and andrographolide in polyherbal formulation ; Int J Pharm Sci , ( 2012 ); 3 ( 4 ) Suppl-1 : 2673 – 2680 . Google Scholar OpenURL Placeholder Text WorldCat [58] Phattanawasin , P. , Burana-Osot , J., Sotanaphun , U., Kumsum , A.; Stability-indicating TLC-image analysis method for determination of andrographolide in bulk drug and A.paniculata formulations ; Acta Chromatogr , ( 2016 ); 28 ( 4 ): 525 – 540 . Google Scholar Crossref Search ADS WorldCat [59] Vijaykumar , K. , Murthy , P.B.S., Kannababu , S., Syamasundar , B., Subbaraju , G.V.; Estimation of andrographolide in A.paniculata herb, extracts and dosage forms ; Int J Appl Sci Eng , ( 2007 ); 5 ( 1 ): 27 – 39 . Google Scholar OpenURL Placeholder Text WorldCat [60] Karpakavalli , M. , Sini , K.R., Arthi , I.; Microwave assisted extraction and estimation of piperine, andrographolide using HPLC techniques ; Pharm Globale , ( 2012 ); 5 ( 03 ): 1 – 5 . Google Scholar OpenURL Placeholder Text WorldCat [61] Kumar , S. , Dhanani , T., Shah , S.; Extraction of three bioactive diterpenoids from A.paniculata: Effect of the extraction techniques on extract composition and quantification of three andrographolides using high-performance liquid chromatography ; J Chromatogr Sci , ( 2014 ); 52 ( 9 ): 1043 – 1050 . Google Scholar Crossref Search ADS PubMed WorldCat [62] Xu , T. , Pan , J., Zhao , L.; Simultaneous determination of four andrographolides in Andrographis paniculate Nees by silver ion reversed phase high-performance liquid chromatography ; J Chromatogr Sci , ( 2008 ); 46 ( 8 ): 747 – 750 . Google Scholar Crossref Search ADS PubMed WorldCat [63] Kotagiri , R. , Kanaaujia , A., Singh , P., Thakur , D.; Validated RP-HPLC method for the quantification of andrographolide in toxiroak premix, a polyherbal mycotoxin inhibitor ; Int J Pharm Sci Res , ( 2013 ); 4 ( 7 ): 2623 – 2628 . Google Scholar OpenURL Placeholder Text WorldCat [64] Da’i , M. , Wikantyasning , E.R., Suhendi , A., Hairunisa , I.; Validated HPLC method for determination of andrographolide in mixed herbal extract ; Int J Pharm Sci Rev Res , ( 2015 ); 35 ( 2 ): 140 – 143 . Google Scholar OpenURL Placeholder Text WorldCat [65] Sajeeb , B.K. , Kumar , U., Halder , S., Bachar , S.C.; Identification and quantification of andrographolide from A.paniculata (Burm. F.) wall. Ex Nees by RP-HPLC method and standardization of its market preparations ; Dhaka Univ J Pharm Sci , ( 2015 ); 14 ( 1 ): 71 – 78 . Google Scholar Crossref Search ADS WorldCat [66] Syukri , Y. , Nugroho , A.E., Martien , R., Lukitaningsih , E.; Validation for the quantification of andrographolide isolated from A.paniculata Nees plant using HPLC ; Jurnal Sains Farmasi dan Klinis , ( 2015 ); 2 ( 1 ): 8 – 14 . Google Scholar Crossref Search ADS WorldCat [67] Jadhao , D.P. , Thorat , B.N.; Quantitative estimation of andrographolide by reverse phase high liquid chromatography method from A.paniculata Nees ; Int J Phytopharm , ( 2012 ); 2 ( 5 ): 149 – 153 . Google Scholar Crossref Search ADS WorldCat [68] Fidrianny , I. , Rukoyah , U., Ruslan , W.K.; Method development for simultaneous analysis of marker scopoletine, andrographolide, quercetin and luteolin in antihypertension Jamu formulation using RP-HPLC ; Int J Pharm Pharm Sci , ( 2015 ); 7 ( 3 ): 332 – 336 . Google Scholar OpenURL Placeholder Text WorldCat [69] Bhope , S.G. , Kuber , V.V., Nagore , D.H., Gaikwad , P.S., Patil , M.J.; Development and validation of RP-HPLC method for simultaneous analysis of andrographolide, phyllanthin, and hypophyllanthin from herbal hepatoprotective formulation ; Acta Chromatogr , ( 2013 ); 25 ( 1 ): 159 – 169 . Google Scholar Crossref Search ADS WorldCat [70] Tanguturi , H. , Nallamotu , H.R., Ganta , S.R., Sastry , K.P.; Quantification of andrographolide in Kalmegh (A.paniculata Nees.) influenced by time of planting and harvesting ; Int J Pure App Biosci , ( 2017 ); 5 ( 4 ): 59 – 63 . Google Scholar Crossref Search ADS WorldCat [71] Mishra , K. , Dash , A.P., Dey , N.; Andrographolide: A novel antimalarial diterpene lactone compound from A.paniculata and its interaction with curcumin and artesunate ; J Trop Med , ( 2011 ); 2011 , 579518: 1 – 6 . Google Scholar Crossref Search ADS WorldCat [72] Sharma , M. , Sharma , A., Tyagi , S.; Quantitative HPLC analysis of andrographolide in A.paniculata at two different stages of life cycle of plant ; Acta Chim Pharm, Indica , ( 2012 ); 2 ( 1 ): 1 – 7 . Google Scholar OpenURL Placeholder Text WorldCat [73] Syukri , Y. , Afetma , D.W., Sirin , M., Fajri , R., Ningrum , A.D.K., Setiawan , S.D. et al. ; Validation of a simple HPLC-UV method for the quantification of andrographolide in self-nano emulsifying drug delivery system (Snedds) for dissolution study ; IJDDT , ( 2017 ); 7 ( 4 ): 239 – 242 . Google Scholar OpenURL Placeholder Text WorldCat [74] Majee , C. , Gupta , B.K., Mazumder , R., Chakraborthy , G.S.; HPLC method development and characterization of bio-active molecule isolated from Andrographis paniculate ; Int J PharmTech Res , ( 2011 ); 3 ( 3 ): 1586 – 1592 . Google Scholar OpenURL Placeholder Text WorldCat [75] Sharma , A. , Lal , K., Handa , S.S.; Standardization of the Indian crude drug Kalmegh by high pressure liquid chromatographic determination of andrographolide ; Phytochem Anal , ( 1992 ); 3 ( 3 ): 129 – 131 . Google Scholar Crossref Search ADS WorldCat [76] Srivastava , A. , Misra , H., Verma , R.K., Gupta , M.M.; Chemical fingerprinting of A.paniculata using HPLC, HPTLC and densitometry ; Phytochem Anal , ( 2004 ); 15 ( 5 ): 280 – 285 . Google Scholar Crossref Search ADS PubMed WorldCat [77] Zhao , Y. , Kao , C.-P., Wu , K.-C., Liao , C.-R., Ho , Y.-L., Chang , Y.-S.; Chemical compositions, chromatographic fingerprints and antioxidant activities of Andrographis Herba ; Molecules , ( 2014 ); 19 : 18332 – 18350 . Google Scholar Crossref Search ADS PubMed WorldCat [78] Kushram , A. , Masih , S.K., Mir , S.A.; Variation of andrographolide content in A.paniculata from different sites of Balaghat region of Jabalpur (M.P.) ; Int J Curr Res Rev , ( 2017 ); 9 ( 22 ): 1 – 3 . Google Scholar OpenURL Placeholder Text WorldCat [79] Li , W. , Fitzloff , J.F.; HPLC-PDA determination of bioactive diterpenoids from plant materials and commercial products of Andrographis paniculate ; J Liq Chromatogr Relat Technol , ( 2004 ); 27 ( 15 ): 2407 – 2420 . Google Scholar Crossref Search ADS WorldCat [80] Li , W. , Fitzloff , J.F.; Determination of andrographolide in commercial Andrographis (Andrographis paniculate) products using HPLC with evaporative light scattering detection ; J Liq Chromatogr Relat Technol , ( 2002 ); 25 ( 9 ): 1335 – 1343 . Google Scholar Crossref Search ADS WorldCat [81] Patarapanich , C. , Laungcholatan , S., Mahaverawat , N., Chaichantipayuth , C., Pummangura , S.; HPLC determination of active diterpene lactones from Andrographis paniculate Nees planted in various seasons and regions in Thailand ; Thai J Pharm Sci , ( 2007 ); 31 : 91 – 99 . Google Scholar OpenURL Placeholder Text WorldCat [82] Kumaran , K.S. , Thirugnaasambantham , P., Viswanathan , S., Murthy , M.S.R.; An HPLC method for the estimation of andrographolide in rabbit serum ; Indian J Pharm , ( 2003 ); 35 : 109 – 112 . Google Scholar OpenURL Placeholder Text WorldCat [83] Chen , L. , Yu , A., Zhuang , X., Zhang , K., Wang , X., Ding , L. et al. ; Determination of andrographolide and dehydroandrographolide in rabbit plasma by on-line soild phase extraction of high-performance liquid chromatography ; Talanta , ( 2007 ); 74 ( 1 ): 146 – 152 . Google Scholar Crossref Search ADS PubMed WorldCat [84] Levita , J. , Juwita , T., Ramadhani , S., Saptarini , N.M., Mutakin , M.; Chromatogram profiles of andrographolide in A23187-induced New Zealand rabbit’s urine and faeces ; J Appl Pharm Sci , ( 2017 ); 7 ( 1 ): 156 – 159 . Google Scholar Crossref Search ADS WorldCat [85] Syukri , Y. , Widarno , I.S., Adewiyah , A., Wibowo , A., Martien , R., Lukitaningsih , E. et al. ; Development and validation of a simple HPLC-UV method for the quantification of andrographolide in rabbit plasma ; IJDDT , ( 2017 ); 7 ( 1 ): 22 – 26 . Google Scholar OpenURL Placeholder Text WorldCat [86] Sujatha , T. , Sunder , J., Lal , S.; Development of HPLC based method to study the bioactive component of Andrographolide paniculata in serum of Nicobari fowl ; Indian J Poult Sci , ( 2015 ); 50 ( 3 ): 33 – 335 . Google Scholar OpenURL Placeholder Text WorldCat [87] Suo , X.-B. , Zhang , H., Wang , Y.-Q.; HPLC determination of andrographolide in rat whole blood: Study on the pharmacokinetics of andrographolide incorporated in liposomes and tablets ; Biomed Chromatogr , ( 2007 ); 21 ( 7 ): 730 – 734 . Google Scholar Crossref Search ADS PubMed WorldCat [88] Song , Y.-X. , Liu , S.-P., Jin , Z., Qin , J.-F., Jian , Z.-Y.; Qualitative and quantitative analysis of Andrographis paniculate by rapid resolution liquid chromatography/time-of-flight mass spectrometry ; Molecules , ( 2013 ); 18 : 12192 – 12207 . Google Scholar Crossref Search ADS PubMed WorldCat [89] Chandra , P. , Kannujia , R., Pandey , R., Shukla , S., Bahadur , L., Pal , M. et al. ; Rapid quantitative analysis of multi-components in Andrographis paniculate using UPLC-QqQLIT- MS/MS: Application to soil sodicity and organic farming ; Ind Crop Prod , ( 2016 ); 83 : 423 – 430 . Google Scholar Crossref Search ADS WorldCat [90] Wangboonskul , J. , Daodee , S., Jarukamjorn , K., Sripanidkulchai , B.-O.; Pharmacokinetic study of Andrographis paniculate tablets in healthy Thai male volunteers ; Thai Pharm Health Sci J , ( 2006 ); 1 ( 3 ): 209 – 218 . Google Scholar OpenURL Placeholder Text WorldCat [91] Chen , L. , Jin , H., Ding , L., Zhang , H., Wang , X., Wang , Z. et al. ; On-line coupling of dynamic microwave-assisted extraction with high-performance liquid chromatography for determination of andrographolide and dehydroandrographolide in Andrographis paniculate Nees ; J Chromatogr A , ( 2007 ); 1140 ( 1-2 ): 71 – 77 . Google Scholar Crossref Search ADS PubMed WorldCat [92] Li , W. , Sun , X., Xu , Y., Wang , X., Bai , J., Ji , Y.; Multi-analysis strategy for metabolism of Andrographis paniculate in rat using liquid chromatography/ quadrupole time-of-flight mass spectrometry ; Biomed Chromatogr , ( 2014 ); 29 ( 7 ): 1016 – 1026 . Google Scholar Crossref Search ADS PubMed WorldCat [93] Bera , R. , Ahmed , S.K.M., Sarkar , L., Sen , S., Karmakar , S.; Pharmacokinetic analysis and tissue distribution of andrographolide in rat by a validated LC-MS/MS method ; Pharm Biol , ( 2014 ); 52 ( 3 ): 321 – 329 . Google Scholar Crossref Search ADS PubMed WorldCat [94] Pholphana , N. , Panomvana , D., Rangkadilok , N., Suriyo , T., Ungtrakul , T., Pongpun , W. et al. ; A simple and sensitive LC-MS/MS method for determination of four major active diterpenoids from Andrographis paniculate in human plasma and its application to a pilot study ; Planta Med , ( 2015 ); 82 ( 01/02 ): 113 – 120 . Google Scholar Crossref Search ADS PubMed WorldCat [95] Gu , Y. , Ma , J., Lu , Y., Chen , B., Yao , S.; Determination of andrographolide in human plasma by high performance liquid chromatography/mass spectrometry ; J Chromatogr B , ( 2007 ); 854 ( 1-2 ): 328 – 331 . Google Scholar Crossref Search ADS WorldCat [96] Xu , L. , Xiao , D., Lou , S., Zou , J., Zhu , Y., Fan , H. et al. ; A simple and sensitive HPLC-ESI-MS/MS method for determination of andrographolide in human plasma ; J Chromatogr B , ( 2009 ); 877 ( 5-6 ): 502 – 506 . Google Scholar Crossref Search ADS WorldCat [97] Naidu , S.R. , Ameer , O.Z., Salman , I.M., Venkatesh , G., Sadikun , A., Asmawi , M.Z.; Pharmacokinetic study of A.paniculata chloroform extract in rats ; Pharmacologyonline , ( 2009 ); 1 : 309 – 319 . Google Scholar OpenURL Placeholder Text WorldCat [98] Yang , T. , Xu , C., Wang , Z.T., Wang , C.H.; Comparative pharmacokinetic studies of andrographolide and its metabolite of 14-deoxy-12-hydroxy-andrographolide in rat by ultra-performance liquid chromatography-mass spectrometry ; Biomed Chromatogr , ( 2013 ); 27 ( 7 ): 931 – 937 . Google Scholar Crossref Search ADS PubMed WorldCat [99] Wang , J. , Yang , W., Wang , G., Tang , P., Sai , Y.; Determination of six components of A.paniculata extract and one major metabolite of andrographolide in rat plasma by liquid chromatography- tandem mass spectrometry ; J Chromatogr B , ( 2014 ); 951–952 : 78 – 88 . Google Scholar Crossref Search ADS WorldCat © The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)
TI - Andrographolide: A Review of Analytical Methods
JF - Journal of Chromatographic Science
DO - 10.1093/chromsci/bmaa091
DA - 2021-01-14
UR - https://www.deepdyve.com/lp/oxford-university-press/andrographolide-a-review-of-analytical-methods-yLq0uqEDhg
SP - 191
EP - 203
VL - 59
IS - 2
DP - DeepDyve
ER -