Dglycoside bonds of cellulose followed by a twophase acid hydrolysis to supplement the diosgenin extraction. This study aimed to determine diosgenin content of costus speciosus water extract cs. Hplc is a useful and commonly used quantification method. Many hplc methods have been reported for determination of diosgenin in various species of genus dioscorea. Green production of diosgenin from discorea nipponica. Both methods were precise and specific and there was no statistical. Saponins concentration the change in the saponins diosgenindiglucoside, diosgenin glucosiderhamnoside, diosgeninglucoside in pdzw in the ssf process was determined by hplc mass spectrometry. Chromatographic method for determination of the amino acid. Diosgenin and tigogenin could be separated also by hplc method of analysis by using sperisorb ssw and ultrasphere ods with nhexaneethanol and watedmethanol solvents systems 19. Analysis of amino acids by hplcanalysis of amino acids by. Costaceae, commonly called as spiral ginger or crepe. Quantification of diosgenin concentration in the fsa extract. Yams, trigonella foenumgraecum and costus speciosus are some of its source.
The separation of filtered plant extracts as well as a mixture of authentic standard saponin samples of gitogenin, diosgenin. Diosgenin was identified by gas chromatographymass spectrometry gcms. Dioscorea is a genus of flowering plants, and some dioscorea species are known and used as a source for the steroidal sapogenin diosgenin. One of the chemical constituents that is responsible for some of the biological activity is diosgenin. Elephantopus scaber linn is an important medicinal plant. Gas chromatography gc and high performance liquid chromatography hplc methods were developed for the simultaneous analysis of phytosterols and diosgenin in two representative taiwanese yams. The diosgenin quantification in leaves and rhizome samples of costus speciosus, costus pictus and costus igneus was performed using hplc analysis.
To screen potential resource from dioscorea species and related medicinal plants for diosgenin extraction, a rapid method to compare the contents of diosgenin in various plants is crucial. Regression curve equation and correlation coefficient. Highperformance liquid chromatographic hplc techniques have been reported for determination of various materials, including glycyrrhizin9,2 cholesterol autoxidation productsz, potato glycoalkaloids2z and monoterpenes23. A satisfactory result with sharp well defined and resolved peak with minimum tailing was achieved using a mobile phase consisting acetonitrile. Hplc method has also been extensively used for quantification of secondary metabolites and natural products li et al. Shimandzu lc20 at prominence solvent delivery module, a manual. The concentrations of diosgenin in different samples were. As shown in figure 1, hptlc of the hplc purified compound d confirmed the single compound purity at r f value 0. Gas chromatography gc and high performance liquid chromatography hplc methods were developed for the simultaneous analysis of phytosterols and diosgenin in two representative taiwanese yams, dioscorea pseudojaponica yamamoto and dioscorea alata l. Spectrophotometric method diosgenin can be reacted with 70% perchloric acid at a room temp. Parameter diosgenin linearity range gml 210 slope 15. High pressure liquid chromatography 20 chem 4 introduction chromatography can be described as a mass transfer process involving adsorption using a nonpolar stationary phase and a mobile polar phase titrating through the column.
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. Pdf diosgenin quantification by hplc in a dioscorea. Hplc analysis of diosgenin in three species of costus. Determination of diosgenin in asparagus officinalis. Determination of diosgenin in dioscoreo zingiberensis cell by rp hplc. Comparative analysis of diosgenin in dioscorea species and related medicinal plants by uplcdadms bmc biochemistry, aug 2014 tao yi, lanlan fan, hongli chen, guoyuan zhu, hauman suen, yina tang, lin zhu, chu chu, zhongzhen zhao, hubiao chen. Wright by couplingcellulose enzymolysis and twophase acid hydrolysis in tandem with hplc uv. Among them, spectrophotometry and high performance liquid chromatography are the common approaches for diosgenin analysis. With uvvis spectrophotometric analysis, the peaks of samples and standard with same. Highperformance liquid chromatographic method for the. An introduction to high performance liquid chromatography.
Then the calibration curve was constructed within the concentration ranging from 1. In the present study, reversed phase hplc method was developed for the estimation of ldopa from mucuna prurines, its extract and in aphrodisiac formulation. The separation of diosgenin by hplc has also been reported 21. Results the retention factor r f of diosgenin and sarsasapogenin on tlc plate were 0.
Research article open access comparative analysis of. Chromatographic conditions hplc separation was performed with a jasco system. After calculation from the regression equation of the standard curve, the contents of diosgenin and sarsasapogenin in the a. Hormonal treatment after four weeks of subculture, the in vitro cultures were moved into. An ultraperformance liquid chromatography uplc coupled. The active component of the column, the sorbent or the stationary phase, is typically a granular. Pdf validation of a method for diosgenin extraction from. All the samples showed characteristic peaks of diosgenin at the same retention time as that of standard diosgenin. The yellow color remains constant for atleast 3 minutes. Pdf hplc analysis of diosgenin in three species of costus.
Simultaneous determination of diosgenin and guggulsterone. Analysis of amino acids by hplcanalysis of amino acids by hplc rita steed agilent technologies, inc. Ethylene treatment improves diosgenin accumulation in in vitro cultures of dioscorea zingiberensis via upregulation of cas and hmgr. Antioxidant activity and rp hplc analysis of diosgenin from the callus oftribulus terrestris corresponding author. A simple, accurate and sensitive hplc method was developed for estimation. Pdf diosgenin quantification, characterisation and.
The assay of diosgenin was performed by hplc method. General description diosgenin is a triterpene and a glycone derivative of dioscin. Determination of diosgenin in dioscoreo zingiberensis cell by rphplc. Research article development and validation of hptlc. Thus, the present work describes development and validation of a new, simple and sensitive reversed phase hplc method. Water extract of cs was obtained with infundation method. Recent highlights on pharmacology and analytical methodology mafalda jesus,1 ana p. Quantification of anti fertility compounddiosgenin. It was successfully applied for the analysis of the drug in marketed formulation and could be effectively used for the routine analysis of formulation containing the drug without any alteration in the chromatography conditions. Diosgenin yield estimation and purity analysis by hplc method, along with accuracy and precision analysis, is presented.
Additionally, hplc segments are made with smaller sorbent particles 2. Determination of diosgenin content in medicinal plants with. Hptlc and hplc for the determination of diosgenin in fenugreek seeds. Hplc analysis showed that the retention time for both isolated compound d and the standard diosgenin was 7. To establish a method for determining the content of diosgenin in asparagus officinalis byproduct by rphplc. Quantitative analysis of diosgenin and trigonelline compounds by high performance liquid chromatography hplc according of li et al. Diosgenin quantification by hplc in a dioscorea polygonoides. Hplc, hptlc, and tlc have been reported for analysis of diosgenin or quercetin in fenugreek individually. The diosgenin content was determined by rphplc, and rphplc conditions were.
High performance liquid chromatography hplc method. Agilent 1100 workstation was used for hplc analysis. The diosgenin concentrations in a dioscorea polygonoides tuber collection from the eje cafetero ec, colombia were determined by hplc and their percentages ranged from 0. Rapid quantitative analysis of diosgenin in the tubers of dioscorea zingiberensis c. However, literature survey shows no single quantitate method available for simultaneous estimation of diosgenin and quercetin from fenugreek or any other plant. The high performance thin layer chromatography was carried out for the estimation of diosgenin content in elephantopus. Diosgenin, a steroidal sapogenin, occurs abundantly in plants such as dioscorea alata, smilax china, and trigonella foenum graecum.
Qualitative and quantitative analysis the tested amino acids were identified by comparing their retention times with that of standards used. Pdf quantitative analysis for diosgenin content in. Qualitative and quantitative analysis of unknown mixtures determining what is there, and how much. The linearity of diosgenin was determine by applying standard solution of different concentrations ranging from 0. Ebscohost serves thousands of libraries with premium essays, articles and other content including a validated rphplc method for the estimation of diosgenin from polyherbal formulation containing tribulus terrestris linn. This gives hplc high determining or resolving power the capacity to recognize components while isolating mixtures. Diosgenin, hplc, steroidal sapogenin, trigonella foenum graecum, validation.
In this study, a sensitive, accurate and reproducible high performance liquid chromatographic method was developed for determination of diosgenin from fenugreek trigonella foenumgraecum leaves in 22 ecotypes of iranian fenugreek. This bioactive phytochemical not only is used as an important starting material for the preparation of several steroidal drugs in the pharmaceutical industry, but has revealed also high potential and interest in the treatment of various types of disorders such. Diosgenin yield estimation and purity analysis by hplc method, along. Summery of validation parameter for the proposed hplc method. An introduction to high performance liquid chromatography high performance liquid chromatography, or hplc, is the most common analytical separation tool and is used in many aspects of drug manufacture and research. For this, the cellulase enzyme promoted the release of dioscin from plant cells through the breaking of. Chromatographic evaluation of steroidal saponin diosgenin from leaves of the medicinal plant by hptlc. The diosgenin concentration was determined by highpressure liquid chromatography hplc according to the method described by huang et al. Lrhamnopyranosyl12,14betadglucopyranoside, polyphyllin iii cas number 19057604.
Regression analysis of calibration curve for diosgenin for the proposed hplc method. Research article open access comparative analysis of diosgenin in dioscorea species and related medicinal plants by uplcdadms tao yi1, lanlan fan2, hongli chen3, guoyuan zhu4, hauman suen1, yina tang1, lin zhu1, chu chu5, zhongzhen zhao1 and hubiao chen1 abstract. Medicinal plants form an essential part of indigenous pharmaceutical system. Ethylene treatment improves diosgenin accumulation in in. Antioxidant activity and rphplc analysis of diosgenin from the callus oftribulus terrestris corresponding author. For hplc analysis, diosgenin was conducted at detectors uv vis, 210 nm on a reversedphase agilent tcc18 column ods 25 cm 4. Simultaneous analysis of diosgenin and sarsasapogenin in. The diosgenin standard was purchased from sigma aldrich scientific chemicals. In addition to providing an introduction to hplc for pharmaceutical analysis it is intended that this book will be a useful resource. Normally diosgenin and sarsasapogenin are analysed separately by thin. Hplc chromatograms of diosgenin standard, gokshur powder and rasayana churna are shown in figure 4, figure 5. Five different fenugreek varieties were subjected to anewly developed diosgenin extraction method, and an hplc chromatogram showed a single peak corresponding to diosgenin. Simultaneous determination of diosgenin and guggulsteronez in gokshuradi guggulu tablet by highperformance thinlayer chromatography the objective of this work was to develop and validate a high performance thin layer chromatography method for simultaneous determination of guggulosteronez and diosgenin in combined dosage form.
Diosgenin in the callus of costus speciosus zingiberaceae was also analyzed by performing tlc analysis10. Comparative analysis of diosgenin in dioscorea species and. Development of validated rphplc method for the estimation. Hplc analysis was performed on c 18 column using a mixture of water. Chromatogram from the hplc analysis showed the peak area for diosgenin. The whole plant is a good source of various bioactive phytochemicals. Reversed phase high performance liquid chromatography rphplc with uv detector was done for the analysis of four saponins of achyranthes aspera and cissus quadrangularis. Hplc chromatograph of extract of dioscorea bulbifera tuber. Then, rp hplc uv allowed the analysis of diosgenin of the prepared sample. Quantitative determination of diosgenin from dioscorea. Production of diosgenin from dioscorea zingiberensis with. Wright by couplingcellulose enzymolysis and twophase acid hydrolysis in tandem with hplcuv. Extraction of diosgenin, a bioactive compound from natural.
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