Research Article: Major bioactive phenolics in Bergenia species from the Indian Himalayan region: Method development, validation and quantitative estimation using UHPLC-QqQLIT-MS/MS

Date Published: July 27, 2017

Publisher: Public Library of Science

Author(s): Renu Pandey, Brijesh Kumar, Baleshwar Meena, Mukesh Srivastava, Tripti Mishra, Vandana Tiwari, Mahesh Pal, Narayanan K. Nair, Dalip K. Upreti, Tikam Singh Rana, Sanjay B. Jadhao.


Bergenia species are important medicinal plants used in indigenous systems of medicine for their antilithiatic and diuretic properties. An ultra-high performance liquid chromatography coupled to hybrid linear ion trap triple quadrupole mass spectrometry (UHPLC-QqQLIT-MS/MS) method has been developed and validated for the estimation of quantitative variation of eight major bioactive phenolics in the rhizomes (150 samples) of four species of this herb, Bergenia (B. ciliata, B. ligulata, B. purpurascens and B. stracheyi). Chromatographic separation was obtained on a Waters ACQUITY UPLCTM BEH (ethylene bridged hybrid) C18 column with a mobile phase consisting of 0.1% (v/v) formic acid aqueous solution and acetonitrile under a gradient elution manner. A hybrid linear ion trap triple quadrupole mass spectrometer was operated in negative electrospray ionization mode with multiple reactions monitoring for detection and quantification of the eight compounds. The validated method demonstrated good linearity (r2 ≥ 0.9991), precision (RSD ≤ 1.87%) and accuracy (95.16–102.11%, RSD ≤ 1.83%) for all reference analytes. The quantitative results revealed that B. ligulata contains the highest amount of the major active marker-bergenin. The results also suggest that sensitive UHPLC-QqQLIT-MS/MS method, a sensitive, accurate and convenient one, could be helpful in identification of potential accession(s), rapid quality control and establishing authenticity of Bergenia species as raw material for pharmaceutical industries.

Partial Text

The genus Bergenia Moench (Saxifragaceae) consists of about 10 species of perennial rhizomatous herbs distributed chiefly in the temperate and subtropical regions of Central and East Asia [1]. The four species of Bergenia viz., B. ciliata (Haw.) Sternb., B. ligulata (Wall.) Engl., B. stracheyi (Hook.f. & Thoms.) Engl., and B. purpurascens (Hook.f. & Thoms.) Engl. are predominantly found in the Himalayan regions of India [2].

The newly standardized method we developed and applied in this study has not only facilitated rapid and accurate determination of target analytes but also fulfilled all the criteria of a validated method as per ICH guidelines in terms of sensitivity, precision and accuracy. Results indicated that B. ligulata collected from Jammu and Kashmir contained highest total content of eight phenolics and the maximum amount of major active marker (Bergenin) which might be medicinally potent regarding cytotoxic and antioxidant activity, whereas B. stracheyi collected from Kullu (HP) contained the highest quantity of arbutin. However, direct activity relation of phenolics to the geographic location in a large scale sample is a subject of future study. The present study could be helpful in identifying potential accession(s) as a raw material of Bergenia species for pharmaceutical industries for preparation of herbal formulations and sustainable utilization of natural resources. The established method also provided a sensitive, accurate and convenient approach for rapid quality control and the establishment of authenticity of Bergenia species.




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