@article {1320, title = {High-Performance Thin Layer Chromatography (HPTLC) Analysis for the Quantification of Constituents in Ficus religiosa}, journal = {Pharmacognosy Research}, volume = {16}, year = {2024}, month = {October 2024}, pages = {929-934}, type = {Original Article}, chapter = {929}, abstract = {

Background: Ficus religiosa, revered for its medicinal properties in traditional medicine, has drawn scientific interest due to its diverse chemical composition. This study aimed to develop a densitometric High-Performance Thin Layer Chromatography (HPTLC) technique to estimate phenolic compounds present in the hydroalcoholic extract of dried root bark of Ficus religiosa. Materials and Methods: This study utilized HPTLC to identify the constituents within Ficus religiosa root bark. The isolated constituent was evaluated by various analytical methods like phytochemical tests, UV, HPTLC etc. Subsequently, the HPTLC method underwent rigorous validation, confirming its reliability and accuracy in separating and quantifying the constituents within the root bark of Ficus religiosa. Results: Isolation of phytoconstituents was done by column chromatographic technique using toluene: ethyl acetate: methanol (3:3:2). The isolated component was confirmed as a flavonoid moiety by various chemical tests and by analytical evaluation. Also, HPTLC analysis of the isolate revealed the compound was Quercetin a flavonoid moiety or its derivative. Conclusion: These findings underscore the presence of main phytoconstituents in Ficus religiosa root bark which is responsible for pharmacological effects and offers insights into its application in both traditional and contemporary medicinal practices.

}, keywords = {Ficus religiosa, Flavonoids, HPTLC, Quantification, Validation}, doi = {10.5530/pres.16.4.106}, author = {Jyothisree Gangadharan and Umadevi Sankararajan} } @article {231, title = {Analytical Method Development and Validation for the Simultaneous Estimation of Abacavir and Lamivudine by Reversed-phase High-performance Liquid Chromatography in Bulk and Tablet Dosage Forms}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {February 2018}, pages = {92-97}, type = {Original Article}, chapter = {92}, abstract = {

Objective: A simple rapid, accurate, precise, and reproducible validated reverse phase high performance liquid chromatography (HPLC) method was developed for the determination of Abacavir (ABAC) and Lamivudine (LAMI) in bulk and tablet dosage forms. Methods: The quantification was carried out using Symmetry Premsil C18 (250 mm {\texttimes} 4.6 mm, 5 μm) column run in isocratic way using mobile phase comprising methanol: water (0.05\% orthophosphoric acid with pH 3) 83:17 v/v and a detection wavelength of 245 nm and injection volume of 20 μl, with a flow rate of 1 ml/min. Results: In the developed method, the retention times of ABAC and LAMI were found to be 3.5 min and 7.4 min, respectively. The method was validated in terms of linearity, precision, accuracy, limits of detection, limits of quantitation, and robustness in accordance with the International Conference on Harmonization guidelines. Conclusion: The assay of the proposed method was found to be 99\% {\textendash} 101\%. The recovery studies were also carried out and mean \% recovery was found to be 99\% {\textendash} 101\%. The \% relative standard deviation from reproducibility was found to be \<2\%. The proposed method was statistically evaluated and can be applied for routine quality control analysis of ABAC and LAMI in bulk and in tablet dosage form.

}, keywords = {Abacavir, Dosage forms, Lamivudine, Method development, Reverse phase high performance liquid chromatography, Validation}, doi = {10.4103/pr.pr_96_17}, author = {Sufiyan Ahmad Raees Ahmad and Lalit Patil and Mohammed Rageeb Mohammed Usman and Mohammad Imran and Rashid Akhtar} } @article {431, title = {Validation of High-Performance Liquid Chromatography Method for Quantification of Costunolide Content in Laurus nobilis}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {October 2018}, pages = {422-425}, type = {Original Article}, chapter = {422}, abstract = {

Objective: To establish a method to determine the content of costunolide in Laurus nobilis leaves by high-performance liquid chromatography (HPLC). Materials and Methods: The separation was performed on a reversed-phase C18 column (100 {\r A}, 150 mm {\texttimes} 4.6 mm, 5 μm pore size) using a mobile phase composed of water:acetonitrile (40:60) at a flow rate of 1.0 ml/min. The detection was carried out on a ultraviolet detector at 210 nm. The developed method was validated according to the requirements for International Conference on Harmonisation guidelines. Results: The proposed method for costunolide was validated for linearity with excellent correlation coefficient (r2 \> 0.999). The relative standard deviation (RSD) is less than 1\% in precision (i.e repeatability and intermediate) of the method. The recovery rate for costunolide was within 100.54\%{\textendash}102.62\%. The limit of detection and limit of quantitation were 2.29 and 6.64 parts per million, respectively. Conclusion: The developed HPLC method is simple, rapid, precisely, accurately, and widely accepted and it is recommended for efficient assays in routine work.

}, keywords = {Costunolide, Laurus nobilis, Linearity, Precision, Recovery, Validation}, doi = {10.4103/pr.pr_31_18}, author = {Puttaswamy Naveen and Harakanahalli Basavegowda Lingaraju1 and Siddappa Chandrappa1 and Koretekera Shekharappa Priya, and Kodimule Shyam Prasad} } @article {422, title = {High Performance Thin layer Chromatography: Densitometry Method for Determination of Rubraxanthone in the Stem Bark Extract of Garcinia cowa Roxb}, journal = {Pharmacognosy Research}, volume = {9}, year = {2017}, month = {July 2017}, pages = {230-233}, type = {Original Article}, chapter = {230}, abstract = {

Context: Garcinia cowa is a medicinal plant widely grown in Southeast Asia and tropical countries. Various parts of this plant have been used in traditional folk medicine. The bark, latex, and root have been used as an antipyretic agent, while fruit and leaves have been used as an expectorant, for indigestion and improvement of blood circulation. Aims: This study aims to determine the concentration of rubraxanthone found in ethyl acetate extract of the stem bark of G. cowa by the high-performance thin-layer chromatography (HPTLC). Materials and Methods: HPTLC method was performed on precoated silica gel G 60 F254 plates using an HPTLC system with a developed mobile-phase system of chloroform: ethyl acetate: methanol: formic acid (86:6:3:5). A volume of 5 μL of standard and sample solutions was applied to the chromatographic plates. The plates were developed in saturated mode of twin trough chamber at room temperature. The method was validated based on linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and specificity. The spots were observed at ultraviolet 243 nm. Results: The linearity of rubraxanthone was obtained between 52.5 and 157.5 ppm/spot. The LOD and LOQ were found to be 4.03 and 13.42 ppm/spot, respectively. Conclusion: The proposed method showed good linearity, precision, accuracy, and high sensitivity. Therefore, it may be applied for the quantification of rubraxanthone in ethyl acetate extract of the stem bark of G. cowa.

}, keywords = {Chromatography, Densitometry, Garcinia cowa Roxb., High-performance thin-layer chromatography, Rubraxanthone, Validation}, doi = {10.4103/pr.pr_144_16}, author = {Dachriyanus Hamidi and Hilyatul Aulia and Meri Susanti} } @article {367, title = {Rapid Development and Validation of Improved Reversed-Phase High-performance Liquid Chromatography Method for the Quantification of Mangiferin, a Polyphenol Xanthone Glycoside in Mangifera indica}, journal = {Pharmacognosy Research}, volume = {9}, year = {2017}, month = {April 2017}, pages = {215-219}, type = {Short Communication}, chapter = {215-}, abstract = {

Mangiferin, a polyphenolic xanthone glycoside from Mangifera indica, is used as traditional medicine for the treatment of numerous diseases. The present study was aimed to develop and validate a reversed-phase high-performance liquid chromatography (RP-HPLC) method for the quantification of mangiferin from the bark extract of M. indica. RP-HPLC analysis was performed by isocratic elution with a low-pressure gradient using 0.1\% formic acid: acetonitrile (87:13) as a mobile phase with a flow rate of 1.5 ml/min. The separation was done at 26{\textdegree}C using a Kinetex XB-C18 column as stationary phase and the detection wavelength at 256 nm. The proposed method was validated for linearity, precision, accuracy, limit of detection, limit of quantification, and robustness by the International Conference on Harmonisation guidelines. In linearity, the excellent correlation coefficient more than 0.999 indicated good fitting of the curve and also good linearity. The intra- and inter-day precision showed \<1\% of relative standard deviation of peak area indicated high reliability and reproducibility of the method. The recovery values at three different levels (50\%, 100\%, and 150\%) of spiked samples were found to be 100.47, 100.89, and 100.99, respectively, and low standard deviation value \<1\% shows high accuracy of the method. In robustness, the results remain unaffected by small variation in the analytical parameters, which shows the robustness of the method. Liquid chromatography{\textendash}mass spectrometry analysis confirmed the presence of mangiferin with M/Z value of 421. The assay developed by HPLC method is a simple, rapid, and reliable for the determination of mangiferin from M. indica.

}, keywords = {Liquid chromatography{\textendash}mass spectrometry, Mangifera indica, Mangiferin, Reversed-phase high-performance liquid chromatography, Validation}, doi = {10.4103/0974-8490.204652}, author = {P Naveen and HB Lingaraju and K Shyam Prasad} } @article {394, title = {Spectrophotometric Quantification of Flavonoids in Herbal Material, Crude Extract, and Fractions from Leaves of Eugenia uniflora Linn}, journal = {Pharmacognosy Research}, volume = {9}, year = {2017}, month = {July 2017}, pages = {253-260}, type = {Original Article}, chapter = {253}, abstract = {

Background: The traditional use of Eugenia uniflora L. ({\textquotedblleft}Pitanga{\textquotedblright}) is reported due to several properties, which have often been related to its flavonoid content. Objective: The aim was to evaluate analytical procedures for quantification of total flavonoids content (TFCs) by ultraviolet-visible (UV-Vis) spectrophotometry in the herbal material (HM), crude extract (CE), and fractions from leaves of E. uniflora. Materials and Methods: The method for quantification of flavonoids after complexation with aluminum chloride (AlCl3) was evaluated: amount of sample (0.25{\textendash}1.5 g); solvent (40\%{\textendash}80\% ethanol); reaction time and AlCl3concentration (2.5\%{\textendash}7.5\%). The procedures by direct dilution (DD) and after acid hydrolysis (AH) were used and validated for HM and CE and applied to the aqueous fraction (AqF), hexane fraction, and ethyl acetate fractions (EAF). Results: The ideal conditions of analysis were ethanol 80\% as solvent; 0.5 g of sample; λmax of 408 (DD) and 425 nm (AH); 25 min after addition of AlCl3 5\%. The procedures validated for standards and samples showed linearity (R2 \> 0.99) with limit of detection and limit of quantification between 0.01 and 0.17 mg/mL (rutin and quercetin); and 0.03 and 0.09 mg/mL (quercetin), for DD and AH, respectively. The procedures were accurate (detect, practice, and repair \<5\% and recovery \>90\%), and stable under robustness conditions (luminosity, storage, reagents, and equipment). The TFCs in AqF and EAF were 0.65 g\% and 17.72 g\%, calculated as rutin. Conclusions: UV-Vis methods for quantification of TFC in HM, CE, and fractions from leaves of E. uniflora were suitably validated. Regarding the analysis of fractions, the EAF achieved enrichment of about nine times in the content of flavonoids.

}, keywords = {Eugenia uniflora L, Fractions, Total flavonoids content, Ultraviolet-visible, Validation}, doi = {10.4103/pr.pr_143_16}, author = {Rhayanne T. M. Ramos and Isabelle C. F. Bezerra and Magda R. A. Ferreira and Luiz Alberto Lira Soares} } @article {510, title = {High-performance Thin-layer Chromatography Method Development, Validation, and Simultaneous Quantification of Four Compounds Identified in Standardized Extracts of Orthosiphon stamineus}, journal = {Pharmacognosy Research}, volume = {8}, year = {2016}, month = {August 2016}, pages = {238-243}, type = {Original Article}, chapter = {238}, abstract = {

Context: Orthosiphon stamineus is a medicinal herb widely grown in Southeast Asia and tropical countries. It has been used traditionally as a diuretic, abdominal pain, kidney and bladder inflammation, gout, and hypertension. Aims: This study aims to develop and validate the high-performance thin layer chromatography (HPTLC) method for quantification of rosmarinic acid (RA), 3{\textquoteright}-hydroxy-5,6,7,4{\textquoteright}-tetramethoxyflavone (TMF), sinensitin (SIN) and eupatorin (EUP) found in ethanol, 50\% ethanol and water extract of O. stamineus leaves. Materials and Methods: HPTLC method was conducted using an HPTLC system with a developed mobile phase system of toluene: ethyl acetate: formic acid (3:7:0.1) performed on precoated silica gel 60 F254 TLC plates. The method was validated based on linearity, accuracy, precision, limit of detection, limit of quantification (LOQ), and specificity, respectively. The detection of spots was observed at ultraviolet 254 nm and 366 nm. Results: The linearity of RA, TMF, SIN, and EUP were obtained between 10 and 100 ng/spot with high correlation coefficient value (R2) of more than 0.986. The limit of detection was found to be 122.47 {\textpm} 3.95 (RA), 43.38 {\textpm} 0.79 (SIN), 17.26 {\textpm} 1.16 (TMF), and 46.80 {\textpm} 1.33 ng/spot (EUP), respectively. Whereas the LOQ was found to be 376.44 {\textpm} 6.70 (RA), 131.45 {\textpm} 2.39 (SIN), 52.30 {\textpm} 2.01 (TMF), and 141.82 {\textpm} 1.58 ng/spot (EUP), respectively. Conclusion: The proposed method showed good linearity, precision, accuracy, and high sensitivity. Hence, it may be applied in a routine quantification of RA, SIN, TMF, and EUP found in ethanol, 50\% of ethanol and water extract of O. stamineus leaves.

}, keywords = {Eupatorin, High-performance thin-layer chromatography, Orthosiphon stamineus, Rosmarinic acid, Sinensitin, Validation}, doi = {10.4103/0974-8490.188872}, author = {Suzana Hashim and Hooi Kheng Beh and Mohamad Shahrul Ridzuan Hamil and Zhari Ismail and Amin Malik Shah Abdul Majid} }