title: Determination of Withanolides inWithania somniferaby Liquid Chromatography: Single-Laboratory Validation, First Action 2015.17.
abstract: An LC method was developed and validated in 2007 for analyzing Withania somnifera raw material (root) and dried extracts for withanolide content, including withanoside IV, withanoside V, withaferin A, 12-deoxywithastromonolide, withanolide A, and withanolide B. The method involved the extraction of the analytes with methanol, their subsequent filtration, and then analysis on a C18 column with an acetonitrile gradient and UV detection. Single-laboratory validation yielded linearity generally in the range of 20 to 200 μg/mL for each analyte, with a repeatability precision of RSD < 3% in most cases, and recovery in the range of 90 to 105%. These results compare well with the performance criteria recently detailed in AOAC Standard Method Performance Requirement 2015.007. The method was shown to be rugged with respect to different analysts, equipment, and days of analysis, and the sample solution was shown to be stable for 24 h at room temperature after extraction. The method was reviewed by the AOAC Expert Review Panel on Dietary Supplements (Set 2 Ingredients) and approved for First Action Official MethodSM status.
authors.affiliation: Natural Remedies Private Ltd, 5B, Veerasandra Industrial Area, Hosur Rd, Bangalore 560 100, India.
authors.forename: Rojison
authors.initials: R
authors.lastname: Koshy
authors.affiliation: Natural Remedies Private Ltd, 5B, Veerasandra Industrial Area, Hosur Rd, Bangalore 560 100, India.
authors.forename: Mayachari Shivanand
authors.initials: MS
authors.lastname: Anand
authors.affiliation: Natural Remedies Private Ltd, 5B, Veerasandra Industrial Area, Hosur Rd, Bangalore 560 100, India.
authors.forename: Balasubramanian
authors.initials: B
authors.lastname: Murali
authors.affiliation: Natural Remedies Private Ltd, 5B, Veerasandra Industrial Area, Hosur Rd, Bangalore 560 100, India.
authors.forename: Sharon L
authors.initials: SL
authors.lastname: Brunelle
mesh_terms: D002853:Chromatography, Liquid; D007753:Laboratories; D010936:Plant Extracts / Q000737:chemistry; D032341:Withania / Q000737:chemistry*; D054358:Withanolides / Q000032:analysis*
chemical_list: D010936:Plant Extracts; D054358:Withanolides