title: Simultaneous Quantification of Syringic Acid and Kaempferol in Extracts of Bergenia Species Using Validated High-Performance Thin-Layer Chromatographic-Densitometric Method.
abstract: A rapid, sensitive, selective and robust quantitative densitometric high-performance thin-layer chromatographic method was developed and validated for separation and quantification of syringic acid (SYA) and kaempferol (KML) in the hydrolyzed extracts of Bergenia ciliata and Bergenia stracheyi. The separation was performed on silica gel 60F254 high-performance thin-layer chromatography plates using toluene : ethyl acetate : formic acid (5 : 4: 1, v/v/v) as the mobile phase. The quantification of SYA and KML was carried out using a densitometric reflection/absorption mode at 290 nm. A dense spot of SYA and KML appeared on the developed plate at a retention factor value of 0.61 ± 0.02 and 0.70 ± 0.01. A precise and accurate quantification was performed using linear regression analysis by plotting the peak area vs concentration 100-600 ng/band (correlation coefficient: r = 0.997, regression coefficient: R(2) = 0.996) for SYA and 100-600 ng/band (correlation coefficient: r = 0.995, regression coefficient: R(2) = 0.991) for KML. The developed method was validated in terms of accuracy, recovery and inter- and intraday study as per International Conference on Harmonisation guidelines. The limit of detection and limit of quantification of SYA and KML were determined, respectively, as 91.63, 142.26 and 277.67, 431.09 ng. The statistical data analysis showed that the method is reproducible and selective for the estimation of SYA and KML in extracts of B. ciliata and B. stracheyi.
authors.affiliation: Pharmacognosy and Ethnopharmacology Division, CSIR-National Botanical Research Institute, Lucknow 226001, India Department of Chemistry, Integral University, Lucknow 226001, India.
authors.forename: Nishi
authors.initials: N
authors.lastname: Srivastava
authors.affiliation: Pharmacognosy and Ethnopharmacology Division, CSIR-National Botanical Research Institute, Lucknow 226001, India.
authors.forename: Amit
authors.initials: A
authors.lastname: Srivastava
authors.affiliation: Pharmacognosy and Ethnopharmacology Division, CSIR-National Botanical Research Institute, Lucknow 226001, India.
authors.forename: Sharad
authors.initials: S
authors.lastname: Srivastava
authors.affiliation: Pharmacognosy and Ethnopharmacology Division, CSIR-National Botanical Research Institute, Lucknow 226001, India pharmacognosy1@rediffmail.com nishisri13jan@gmail.com.
authors.forename: Ajay Kumar Singh
authors.initials: AK
authors.lastname: Rawat
authors.affiliation: Department of Chemistry, Integral University, Lucknow 226001, India.
authors.forename: Abdul Rahman
authors.initials: AR
authors.lastname: Khan
mesh_terms: D000085:Acetates; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D005561:Formates; D005707:Gallic Acid / Q000031:analogs & derivatives* / Q000302:isolation & purification; D006868:Hydrolysis; D044949:Kaempferols / Q000302:isolation & purification*; D057230:Limit of Detection; D010936:Plant Extracts / Q000737:chemistry*; D012044:Regression Analysis; D015203:Reproducibility of Results; D027343:Rhizome / Q000737:chemistry; D031995:Saxifragaceae / Q000737:chemistry*; D012997:Solvents; D014050:Toluene
chemical_list: D000085:Acetates; D005561:Formates; D044949:Kaempferols; D010936:Plant Extracts; D012997:Solvents; C030544:formic acid; D014050:Toluene; D005707:Gallic Acid; C006552:kaempferol; C007650:ethyl acetate; C001945:syringic acid