Gupta and sushil kumar central institute of medicinal and aromatic plants, po cimap, lucknow 226015, india a rapid and simple reversephase high performance liquid chromatographic method has been developed for. The detector wavelength was set at the absorbance maximum of. Centella extract showed a peak with the same wavelength and retention time as that of the asiaticoside standard, while the retention time of boldine was 2. An overview of applications of precolumn derivatization. Separation and determination of asiaticoside, asiaticoside b and madecassoside in centella asiatica total triterpenoid saponins by hplc. High performance liquid chromatographic analysis of. High performance liquid chromatography is basically a highly improved form of column. Hence, to determine the concentration of each compound present in the distinctive parts of centella asiatica, high performance liquid chromatography hplc method is carried out. Thin layer chromatography tlc and high performance liquid chromatography hplc analysis showed the presence of asiaticoside in the in vitro grown leaves, callus and cell suspension cultured cells. Hplc analysis an agilent 1100 hplc system was used for routine analysis, but this isocratic method is capable of being executed on virtually any functional hplc with a uv detector. A simple, accurate, precise, faster reverse phase hplc method has been developed and. At the end of each chapter there is a list of references andor further reading which will help the reader to develop their expertise in the technique. Effects of solvents and extraction methods wan joo kim 1, jae duck kim 1, bambang veriansyah 1, jaehoon kim 1 seung geun oh 2 and raymond r tjandrawinata 3.
A highperformance liquid chromatography analysis showed that the elicitation with 100 m concentration of methyl jasmonate enhanced asiaticoside content by 69fold in callus culture, 39fold in shoot cultures, and ata by 1. Estimation of asiaticoside by using rphplc and fame analysis of. For the analysis of asiaticoside, reverse phase high performance liquid chromatography rphplc was used. The concentration of asiaticacoside per gram extracts were range from 10. Hplc standard operating procedure last revised march, 3, 2016 this document is intended to be a guide for operating the garner labs hplc systems and is only intended to provide instructions for users who are already generally familiar with hplc operation. Hplc, for comparison with a reference extract of centella and a standard solution of asiaticoside.
The chromatographic conditions are included in the caption to figure 3. On behalf of the organizing committee, it is my pleasure to invite you to the 2018 first asia international symposium for highperformance thinlayer chromatography hptlc asia 2018 which will take place in november 2830, 2018, bangkok, thailand. Aflatoxin analysis in food by hplc and uhplc jasco. Biomedical analysis research unit, faculty of pharmaceutical sciences. Urban article pdf available in chiang mai journal of science 353 september 2008 with 2,938 reads. Asiaticoside is hydrolytic cleavage of the sugar moiety to become asiatic acid which is responsible for the therapeutic effects 1617. Elicitation effect on the production of asiaticoside and. Separation and determination of asiaticoside, asiaticosideb and. The present work is aimed to perform phytochemical analysis and estimation of antioxidant activity by dpph assay. Validated high performance liquid chromatography for simultaneous. Establishment of callus and cell suspension cultures of. Validation of hplc method for the determination of retinol in.
Methods for induction of callus and cell suspension cultures have been developed for the medicinally important herb centella asiatica l. Identification of centella asiatica extract in a cosmetic cream. Asiaticoside analyticalstandard 16830152 c48h78o19. Traditional chinese medicine hplc timehonored remedies. Determination of madecassoside and asiaticoside contents of c. Application note food testing and agriculture abstract agilent captiva premium syringe filters were tested thoroughly for sample preparation in the analysis of food additives in beverages. The simultaneous quantification of madecassoside, asiaticoside, madecassic acid and asiatic acid in. Asiaticoside, centella asiatica, hplc, isomer, madecassoside.
Hplc analysis of asiaticoside standard showed a single peak at 204 nm with a retention time of 0. High performance liquid chromatographic analysis of asiaticoside in. A normal phase tlc analysis was performed using silicagel as a stationary phase and a mobile phase containing water. Hplc analysis of each compound after 4, 6, 8 and 10h n3. Most methods for the analysis of the total triterpenes use gradient elution reversedphase highperformance liquid chromatography rp hplc with lowwavelength uv detection 203205 nm because the triterpenes do not strongly absorb above 205 nm. Separation and determination of asiaticoside, asiaticosideb. The concentration of asiaticoside found in centella extract. The present study reports the development and application of an hplc coupled pulsed amperometric detection method to determine the madecassoside and asiaticoside contents of centella asiatica leaf and of commercial c. Quantitative determination of triterpenes in centella extract and. Pdf hplc separation and isolation of asiaticoside from centella. An extensive survey of literature reveals that analysis of madecassic, asiatic acid, medecassoside and asiaticoside has been done by hplc where the retention time for all these constituents ranges between 1524 min3. The formulation was diluted in water, sonicated, and adjusted to 100 mgml before analysis. The remaining madecassoside and asiaticoside were compared to an initial time. Analysis of food additives in beverages using syringe filter filtration and hplc author limian zhao agilent technologies, inc.
Packaging 10 mg in glass bottle 50 mg in poly bottle biochemphysiol actions madecassoside is a triterpenoid compound found in centella asiatica gotu kola, a medicinal herb used in traditional chinese medicine tcm, ayurvedic medicine, and traditional african medicine. Pdf hplc separation and isolation of asiaticoside from. High performance liquid chromatography madecassoside a b s t r a c t the objective of the work was to validate the high performance liquid chromatography for simultaneous determination of stability of madecassoside and asiaticoside in centella asiatica l. A high performance liquid chromatographic method was developed for. High performance liquid chromatographic analysis of asiaticoside in centella asiatica l. Hplc separation and isolation of asiaticoside from. Pdf a simple, sensitive high performance liquid chromatographic method was developed for the determination of asiaticoside in centella. Effects of some culture conditions on production of asiaticoside from centella centella asiatica l. Urban, herba emahmpc2911772009 page 644 the substances of therapeutic interest are the saponin containing triterpene acids and their sugar.
Thus, elicitation with methyl jasmonate is an effective. Validated high performance liquid chromatography for simultaneous determination of stability of madecassoside and asiaticoside in film forming polymeric. Asiaticoside, one of the principle terpenoids in c. Highperformance liquid chromatographic determination of asiatic. Pdf estimation of asiaticoside by using rphplc and fame. Journal of chromatography b an improved hplcuv method for the.
Helmi yousif alfarra and muhammd nor omar, hplc separation and isolation of asiaticoside from centella asiatica and its biotransformation by a. For the analysis of asiaticoside, reverse phase high performance liquid chromatography rp hplc was used. Most methods for the analysis of the total triterpenes use gradient elution reversedphase highperformance liquid chromatography rphplc with lowwavelength uv detection 203205 nm because the triterpenes do not strongly absorb above 205 nm. An rp18 column and a linear gradient were used in the hplc method. The presence of madecassoside was also determined by tlc.
High performance liquid chromatography hplc analysis triterpene standard asiaticoside stock was made using 5 mg sigma, malaysia dissolved in 10 ml hplc grade methanol and then diluted further to obtain concentrations of 50, 100 and 150 ugml, respectively. The most appropriate soxhlet extraction time was determined using these data. The study was carried out to develop a rapid and accurate hplc method to isolate the major bioactive phytochemical asiaticoside in centella asiatica and to transform this metabolite using microorganism. Welcome to the 1st asia international symposium for highperformance thinlayer chromatography. Analysis of food additives in beverages using syringe filter. In addition to providing an introduction to hplc for pharmaceutical analysis it is intended that this book will be a useful resource. The separation is completed within 12 minutes by conventional hplc and within the 3. The hplc gradient method suing a pda detector at 205 nm is able to. Estimation of asiaticoside by using rphplc and fame analysis of medicinally important plant centella asiatica. Hplc separation and isolation of asiaticoside from centella.
Urban cells cultured in 5l bioreactor was investigated. Asiaticoside madecassic acid asiatic acid method validation quanti. Pdf high performance liquid chromatographic analysis of. Hplc analysis carried out with perkinelmer, flexar, pda plus detector. This study reports presence of asiaticoside in samples of centella cream both by tlc and hplc. High performance liquid chromatography analysis the analysis is preceded with standard calibration curves that are constructed from the chromatograms for the. This method breaks down complex mixtures into individual compounds, which in turn are identified and quantified by suitable detectors. Asiaticoside content was determined by hplc analysis. An improved hplcuv method for the simultaneous quanti.
High performance liquid chromatography is a powerful tool in analysis. Estimation of asiaticoside by using rp hplc and fame analysis of medicinally important plant centella asiatica article pdf available september 2017 with 267 reads how we measure reads. Reversephase high performance liquid chromatography of asiaticoside in centella asiatica r. Hplc analysis the % of asiaticoside and madecassoside in the methanolic extract of leaf callus was found to be 1. Quantification of asiatic acid, asiaticoside and madecassoside in the samples of c. Asiaticoside was separated using an optimized hplc method using phenomenex, luna, 5 m c18 150 mm x 4. Hplc method validation for simultaneous estimation of.
Madecassoside 1 and asiaticoside 2 analysis was performed by reversedphase hplc agilent 1260 infinity, agilent, california, usa. Abstract a high performance liquid chromatographic method was developed for the separation and determination of asiaticoside and madecassoside isomers. Aa was determined by a novel precolumn derivatization rp hplc method with uvvis detection for the quantitative determination of total concentration in beagle dog plasma. We report an efficient reversedphasehplc rp hplc procedure to resolve. Reversephase high performance liquid chromatography of. Highperformance liquid chromatography hplc is used increasingly in the analysis of food samples to separate and detect additives and contaminants. Quantitative estimation of asiatic acid, asiaticoside.
522 56 648 594 359 1290 1342 890 162 1483 777 1071 701 1322 912 25 986 1331 1352 718 695 918 1432 636 681 712 369 1092 1369 570 81 1432 505 218 1142 531 1329 1296 657 1091 683 706