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Simultaneous quantitative determination of multiple bioactive markers in Ocimum sanctum obtained from different locations and its marketed herbal formulations using UPLC‐ESI‐MS/MS combined with principal component analysis
Author(s) -
Pandey Renu,
Chandra Preeti,
Srivastava Mukesh,
Mishra D. K.,
Kumar Brijesh
Publication year - 2015
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.2551
Subject(s) - chemistry , chromatography , formic acid , ocimum , coptisine , high performance liquid chromatography , traditional medicine , palmatine , medicine
Ocimum sanctum L., with phenolic acids, flavonoids, propenyl phenols and terpenoids as active pharmacological constituents, is a popular medicinal herb and is present as an ingredient in many herbal formulations. Therefore, development of a reliable analytical method for simultaneous determination of the pharmacologically active constituents of O. sanctum is of high importance. Objective To develop and validate a new, rapid, sensitive and selective UPLC–ESI/MS/MS method for simultaneous determination of 23 bioactive markers including phenolic acids, flavonoids, propenyl phenol and terpenoid in the leaf extract and marketed herbal formulations of O. sanctum . Methods An UPLC–ESI/MS/MS method using negative electrospray ionisation (ESI) in multiple‐reaction‐monitoring (MRM) mode was used for simultaneous determination. Chromatographic separation was achieved on an Acquity UPLC BEH C 18 ‐column using a gradient elution with 0.1% formic acid in water and 0.1% formic acid in acetonitrile. Principal component analysis (PCA) was applied to correlate and discriminate eight geographical collections of O. sanctum based on quantitative data of the analytes. Results The developed method was validated as per International Conference on Harmonization guidelines and found to be accurate, with overall recovery in the range 95.09–104.84% (RSD ≤ 1.85%), precise (RSD ≤ 1.98%) and linear (r 2 ≥ 0.9971) over the concentration range of 0.5–1000 ng/mL. Ursolic acid was found to be the most abundant marker in all the samples investigated, except for the marketed tablet. Conclusion The method established is simple, rapid and sensitive, hence it can be reliably utilised for the quality control of O. sanctum and derived herbal formulations. Copyright © 2015 John Wiley & Sons, Ltd.