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.
BACKGROUND
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 C18 -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.


CONCLUSIONS
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.
Sophisticated Analytical Instrument Facility, CSIR-Central Drug Research Institute, Lucknow -, 226031, Uttar Pradesh, India.|Academy of Scientific and Innovative Research (AcSIR), New Delhi -, 110025, India.
Renu
R
Pandey
Sophisticated Analytical Instrument Facility, CSIR-Central Drug Research Institute, Lucknow -, 226031, Uttar Pradesh, India.|Academy of Scientific and Innovative Research (AcSIR), New Delhi -, 110025, India.
Preeti
P
Chandra
Academy of Scientific and Innovative Research (AcSIR), New Delhi -, 110025, India.|Biometry and Statistics Division, CSIR-Central Drug Research Institute, Lucknow -, 226031, Uttar Pradesh, India.
Mukesh
M
Srivastava
Academy of Scientific and Innovative Research (AcSIR), New Delhi -, 110025, India.|Botany Division, CSIR-Central Drug Research Institute, Lucknow -, 226031, Uttar Pradesh, India.
D K
DK
Mishra
Sophisticated Analytical Instrument Facility, CSIR-Central Drug Research Institute, Lucknow -, 226031, Uttar Pradesh, India.|Academy of Scientific and Innovative Research (AcSIR), New Delhi -, 110025, India.
Brijesh
B
Kumar
D007194:India; D008515:Medicine, Ayurvedic; D031339:Ocimum / Q000737:chemistry* / Q000145:classification; D010936:Plant Extracts / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry; D010946:Plants, Medicinal / Q000737:chemistry*; D025341:Principal Component Analysis; D011786:Quality Control; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D010936:Plant Extracts
Ocimum sanctum; UPLC-ESI/MS/MS; multiple reaction monitoring (MRM)