@article {1342, title = {High-Performance Thin Layer Chromatography Methods for Estimation of Chemical Markers in Stachytarpheta indica Vahl.}, journal = {Pharmacognosy Research}, volume = {17}, year = {2025}, month = {January 2025}, pages = {94-99}, type = {Original Article}, chapter = {94}, abstract = {

Background: Stachytarpheta indica Vahl. (Verbenaceae family), commonly known as Indian Snakeweed, is a valuable medicinal plant. A systematic phytochemical analysis of bioactives of this plant is lacking till date. Objectives: The present study is aimed to develop validated High-Performance Thin-Layer Chromatography (HPTLC) methods for quantification of 6 marker compounds, β-sitosterol, lupeol, ursolic acid, apigenin, caffeic acid and verbascoside in S. indica plant. Materials and Methods: Normal phase chromatography was used to develop mobile phase systems that could aid in the identification and quantification of the phytochemicals in the entire plant. Several enhancements were made to the basic method of thin-layer chromatography to automate the different steps, to increase the resolution achieved and to allow more accurate quantitative measurement. The methods were validated as per the International Council for Harmonization (ICH) guidelines. Results: The linear regression analysis data for the calibration plots showed a good linear relationship (r2=0.9873-0.9979) in the concentration range of 1-5 μg for β-sitosterol, lupeol and ursolic acid; 0.5-5 μg for apigenin, 1-3 μg for caffeic acid; and 3-8 μg per spot for verbascoside with respect to area. The average recoveries for phytoconstituents were 98.32 to 98.97\%, indicating the good reproducibility. Verbascoside 1.3598{\textpm}0.014\% w/w was present at high concentration and ursolic acid 0.0027{\textpm}0.0007\% w/w was present at low concentration in the whole plant powder. Conclusion: The validated TLC-densitometric methods were simple, precise, accurate, specific and can be used for the routine quality control of raw materials of S. indica.

}, keywords = {Apigenin, Caffeic acid, High-performance thin-layer chromatography, Stachytarpheta indica, Ursolic acid, Verbascoside}, doi = {10.5530/pres.20251934}, author = {Vihangi Patel and Krupa Gadhvi and Karuna Modi and Mamta Shah} } @article {171, title = {Comparative Botanical and Phytochemical Studies of Ambiguous Medicinal Plant Species of Wedelia and Eclipta (Fam. Asteraceae) Used in ASU Systems of Medicine with Special Reference to in-silico Screening of Hepatoprotective Potential of Marker Wedelolacto}, journal = {Pharmacognosy Research}, volume = {12}, year = {2020}, month = {August 2020}, pages = {285-298}, type = {Original Article}, chapter = {285}, abstract = {

Context: In traditional medicine, Kesaraja (Ayurveda) or Manjal karisali (Siddha) is effective for jaundice. Aim: Three species of Asteraceae need to be studied for their therapeutic superiority of their intended claim. They are Wedelia chinensis (Osbeck) Merr. Philipp J., Wedelia trilobata (L.) Hitchc. and Eclipta prostrata (L.) L. (Asteraceae). The present study aimed to screen and characterize the potential species for therapeutic purpose. Materials and Methods: The whole plants, W. chinensis (Osbeck) Merr. Philipp J., W. trilobata (L.) Hitchc. and Eclipta prostrata (L.) (Asteraceae) were collected and botanically identified. Preliminary phytochemical analysis and high-performance thin-layer chromatography finger printing with marker wedelolactone were done for the ethanolic extracts of these plants. Botanical and pharmacognostical diagnostic characters of the plants based on macro-morphological, micro-morphological and powder microscopical characterization were worked out. Comparative in-vitro antioxidant potential of ethanolic extracts of these plant species was carried out. Using ADMET SAR software, the pharmacokinetics of wedelolactone were predicted. Using Autodock 4.2 software, the binding energy of wedelolactone on targets of acetaminophen-induced hepatotoxicity namely PPAR-α, AMPK, JNK-1, EGFR, Nrf2, ALT, ALP, GGT, CAR, Frizzled receptor, FXR, ERK1, LXR, mitochondrial glutamate dehydrogenase, p53, mTOR C1, CYP1A2, CYP2E1, 5-lipoxygenase, thrombin, UCP1, GSK1, RXR and PXR was predicted. Results: All the three plant species were pharmacognostically and chemically different. W. chinensis was found to possess more antioxidant potential than the other two plants. The marker compound wedelolactone was not detected in W. trilobata. Wedelolactone passed the Lipinski{\textquoteleft}s rule of five, and the docking analysis of wedelolactone confirmed high binding affinity toward PPAR-α, AMPK, Nrf2, CYP2E1, EGFR, JNK1, UCP-2, thrombin, 5-lipoxygenase, mTORC1, RXR, FXR, LXR, Frizzled receptor, GDH and Erk-1. Conclusion: Based on the above observations, we conclude that the presence of marker compound wedelolactone might have attributed the potency of W. chinensis and E. prostrata in counteracting acetaminophen toxicity when compared with W. trilobata.

}, keywords = {Antioxidant, Ayurveda, Botany, Docking, Eclipta prostrata, High-performance thin-layer chromatography, Macro-morphological, Micro-morphological, Phytochemical, Powder microscopy, Siddha, Wedelia chinensis, Wedelia trilobata, Wedelolactone}, doi = {10.4103/pr.pr_11_20}, author = {C Arunachalam and R Arunadevi and S Murugammal and N Monika and R Susila and S N Sunil Kumar} } @article {82, title = {High-Performance Thin-Layer Chromatography Analysis of Gallic Acid and Other Phytoconstituents of Methanolic Extracts of Myrica nagi Fruit}, journal = {Pharmacognosy Research}, volume = {12}, year = {2020}, month = {May 2020}, pages = {95-101}, type = {Original Article}, chapter = {95}, abstract = {

Background: Myrica nagi Thunb. (family: Myricaceae) is effective against gastric, metabolic, and hepatic disorders. The therapeutic effect of its fruit, which is consumed in North India, has not been confirmed, and detailed chemical profiling of the fruit is thus required. Objectives: The study objective was to develop and optimize a high-performance thin-layer chromatography (HPTLC) method for the characterization of gallic acid, quercetin, myricetin, and caffeic acid in the methanolic extract of M. nagi fruit and the quantification of gallic acid. Materials and Methods: Analyses were performed using HPTLC, and liquid chromatography-mass spectrometry. HPTLC experiments were carried out using an optimized solvent mixture, which enabled the separation and detection (at 254 and 366 nm) of four flavonoid compounds in the dried M. nagi extract. Gallic acid was quantified using calibration curves. Results: The proposed method enabled the detection of gallic acid, quercetin, myricetin, and caffeic acid. Validation took into account the estimation of linearity, limit of detection, limit of quantification, accuracy, and recovery of gallic acid. Gallic acid was quantified at 12.93 μg/mg of dry plant concentrate. Conclusion: This study describes the development of an HPTLC method for the analysis and characterization of phytoconstitutents in the methanolic solution of a dried M. nagi fruit extract. The method was successfully validated for the analysis of gallic acid.

}, keywords = {Flavonoid, Gallic acid, High-performance thin-layer chromatography, Liquid chromatography-mass spectrometry, Myrica nagi}, doi = {10.4103/pr.pr_104_19}, author = {Yash Prashar and Nilesh J Patel} } @article {407, title = {Development of Validated High-performance Thin-layer Chromatography Method for Simultaneous Determination of Quercetin and Kaempferol in Thespesia populnea}, journal = {Pharmacognosy Research}, volume = {9}, year = {2017}, month = {July 2017}, pages = {277-281}, type = {Original Article}, chapter = {277}, abstract = {

Introduction: Thespesia populnea L. (Family: Malvaceae) is a well-known medicinal plant distributed in tropical regions of the world and cultivated in South Gujarat and indicated to be useful in cutaneous affections, psoriasis, ringworm, and eczema. Bark and fruits are indicated in the diseases of skin, urethritis, and gonorrhea. The juice of fruits is employed in treating certain hepatic diseases. The plant is reported to contain flavonoids, quercetin, kaempferol, gossypetin, Kaempferol-3-monoglucoside, β-sitosterol, kaempferol-7-glucoside, and gossypol. T. populnea is a common component of many herbal and Ayurvedic formulation such as Kamilari and Liv-52. Objective: The present study aimed at developing validated and reliable high-performance thin layer chromatography (HPTLC) method for the analysis of quercetin and kaempferol simultaneously in T. populnea. Method: The method employed thin-layer chromatography aluminum sheets precoated with silica gel as the stationary phase and toluene: ethyl acetate: formic acid (6:4:0.3 v/v/v) as the mobile phase, which gave compact bands of quercetin and kaempferol. Result: Linear regression data for the calibration curves of standard quercetin and kaempferol showed a good linear relationship over a concentration range of 100{\textendash}600 ng/spot and 500{\textendash}3000 ng/spot with respect to the area and correlation coefficient (R2) was 0.9955 and 0.9967. The method was evaluated regarding accuracy, precision, selectivity, and robustness. Limits of detection and quantitation were recorded as 32.06 and 85.33 ng/spot and 74.055 and 243.72 ng/spot for quercetin and kaempferol, respectively. Conclusion: We concluded that this method employing HPTLC in the quantitative determination of quercetin and kaempferol is efficient, simple, accurate, and validated.

}, keywords = {High-performance thin-layer chromatography, Kaempferol, Quercetin, Thespesia populnea}, doi = {10.4103/0974-8490.210326}, author = {Hiteksha Panchal and Aeshna Amin and Mamta Shah} }