Estimation of quercetin by highperformance thin-layer chromatography and in vitro anti-inflammatory activity of Leptadenia reticulata (Wight & Arn.) from different parts of India

Abstract

Background: Leptadenia reticulata (Wight & Arn.) has made important contributions to both traditional and
modern medicine. Aims and Objectives: The study aims to develop and validate an high-performance thinlayer
chromatography (HPTLC) method for the determination of quercetin content in L. reticulata collected from
various parts of the country and to assess its anti-inflammatory activity using the bovine serum albumin (BSA)
inhibition method. Materials and Methods: Five samples were obtained from different geographical locations
in India, and no significant variation was found in the morpho-anatomical study. HPTLC method developed
using a toluene: chloroform: ethyl acetate: formic acid (2.5: 2: 4.5: 1) mobile phase at a wavelength of 300 nm.
Second, anti-inflammatory activity of L. reticulata extract was assessed using the BSA inhibition approach.
Results: For the standard and sample, Rf value of quercetin (0.49) was observed. Concentration range of quercetin
(60–160 ng/spot) was determined to be linear for the linearity approach, with a regression coefficient for quercetin
of 0.999 and an optimal recovery of 101.70%. The limits of detection and limits of quantification were found to
be 0.78 and 2.36 ng, respectively. Study revealed quercetin content differed among L. reticulata collected from
different geographical regions. The highest and lowest quercetin contents in L. reticulata were detected in LR-103
and LR-099, respectively. From the results obtained, sample LR-103 showed effective anti-inflammatory activity
compared to LR-099 at certain concentrations. Conclusion: The developed method is suitable for the detection
and quantification of the lowest content of quercetin in L. reticulata. The obtained data indicate that geographic
origin and climatic conditions may be major factors in determining the phytocompounds and biological activities
of L. reticulata plant species.