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Simultaneous quantification of withanolides in Withania somnifera by a validated high-performance thin-layer chromatographic method.
J AOAC Int. 2008 Sep-Oct; 91(5):1154-61.JA

Abstract

This paper describes a sensitive, selective, specific, robust, and validated densitometric high-performance thin-layer chromatographic (HPTLC) method for the simultaneous determination of 3 key withanolides, namely, withaferin-A, 12-deoxywithastramonolide, and withanolide-A, in Ashwagandha (Withania somnifera) plant samples. The separation was performed on aluminum-backed silica gel 60F254 HPTLC plates using dichloromethane-methanol-acetone-diethyl ether (15 + 1 + 1 + 1, v/v/v/v) as the mobile phase. The withanolides were quantified by densitometry in the reflection/absorption mode at 230 nm. Precise and accurate quantification could be performed in the linear working concentration range of 66-330 ng/band with good correlation (r2 = 0.997, 0.999, and 0.996, respectively). The method was validated for recovery, precision, accuracy, robustness, limit of detection, limit of quantitation, and specificity according to International Conference on Harmonization guidelines. Specificity of quantification was confirmed using retention factor (Rf) values, UV-Vis spectral correlation, and electrospray ionization mass spectra of marker compounds in sample tracks.

Authors+Show Affiliations

Central Institute of Medicinal and Aromatic Plants, Analytical Chemistry Division, Lucknow, India.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18980134

Citation

Srivastava, Pooja, et al. "Simultaneous Quantification of Withanolides in Withania Somnifera By a Validated High-performance Thin-layer Chromatographic Method." Journal of AOAC International, vol. 91, no. 5, 2008, pp. 1154-61.
Srivastava P, Tiwari N, Yadav AK, et al. Simultaneous quantification of withanolides in Withania somnifera by a validated high-performance thin-layer chromatographic method. J AOAC Int. 2008;91(5):1154-61.
Srivastava, P., Tiwari, N., Yadav, A. K., Kumar, V., Shanker, K., Verma, R. K., Gupta, M. M., Gupta, A. K., & Khanuja, S. P. (2008). Simultaneous quantification of withanolides in Withania somnifera by a validated high-performance thin-layer chromatographic method. Journal of AOAC International, 91(5), 1154-61.
Srivastava P, et al. Simultaneous Quantification of Withanolides in Withania Somnifera By a Validated High-performance Thin-layer Chromatographic Method. J AOAC Int. 2008 Sep-Oct;91(5):1154-61. PubMed PMID: 18980134.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Simultaneous quantification of withanolides in Withania somnifera by a validated high-performance thin-layer chromatographic method. AU - Srivastava,Pooja, AU - Tiwari,Neerja, AU - Yadav,Akhilesh K, AU - Kumar,Vijendra, AU - Shanker,Karuna, AU - Verma,Ram K, AU - Gupta,Madan M, AU - Gupta,Anil K, AU - Khanuja,Suman P S, PY - 2008/11/5/pubmed PY - 2008/12/17/medline PY - 2008/11/5/entrez SP - 1154 EP - 61 JF - Journal of AOAC International JO - J AOAC Int VL - 91 IS - 5 N2 - This paper describes a sensitive, selective, specific, robust, and validated densitometric high-performance thin-layer chromatographic (HPTLC) method for the simultaneous determination of 3 key withanolides, namely, withaferin-A, 12-deoxywithastramonolide, and withanolide-A, in Ashwagandha (Withania somnifera) plant samples. The separation was performed on aluminum-backed silica gel 60F254 HPTLC plates using dichloromethane-methanol-acetone-diethyl ether (15 + 1 + 1 + 1, v/v/v/v) as the mobile phase. The withanolides were quantified by densitometry in the reflection/absorption mode at 230 nm. Precise and accurate quantification could be performed in the linear working concentration range of 66-330 ng/band with good correlation (r2 = 0.997, 0.999, and 0.996, respectively). The method was validated for recovery, precision, accuracy, robustness, limit of detection, limit of quantitation, and specificity according to International Conference on Harmonization guidelines. Specificity of quantification was confirmed using retention factor (Rf) values, UV-Vis spectral correlation, and electrospray ionization mass spectra of marker compounds in sample tracks. SN - 1060-3271 UR - https://www.unboundmedicine.com/medline/citation/18980134/Simultaneous_quantification_of_withanolides_in_Withania_somnifera_by_a_validated_high_performance_thin_layer_chromatographic_method_ L2 - https://antibodies.cancer.gov/detail/CPTC-RRM2-2 DB - PRIME DP - Unbound Medicine ER -