Developed and validated method for the simultaneous estimation of ferulic acid and chlorogenic acid in four Indian ananas comosus merrill cultivars through HPTLC

Souvik Biswas, Arijit Das, Dipan Roy, Tathagata Roy, Debajit Dewan, Rakesh Kumar Paul, Mithun Bhowmick, Mrinmoy Nag*

Bengal College of Pharmaceutical Sciences & Research, Durgapur, West Bengal, India

ABSTRACT

Ananas comosus (L) Merrill fruit is used for the treatment of urinary tract infection, and jaundice. The fruit has anti-obesity activity and used as antihelmithic agent for the treatment of intestinal worms. The Ananas comosus fruit contains many phytoconstituents; among them are Ferulic acid and Chlorogenic acid exhibiting various pharmacological activities. To ensure the content of uniformity of biomarkers, the aim of the present study was to develop a HPTLC method for simultaneous estimation of Ferulic acid and Chlorogenic acid in Ananas comosus fruit. The standard marker compounds and four different fruit extract solutions were applied on the HPTLC plate (20 mm × 10 mm dimension) with the help of Linomat 5 applicator by using WinCATS software. The solvent system toluene, ethyl acetate, methanol, formic acid (4:4:1.2:0.8, v/v) was used as mobile phase. The λmax was set for Ferulic acid and Chlorogenic acid at 280 nm. The calibration range of Ferulic acid was found to be 300-900 ng/ml, with the linear equation Y= 338.485+6.238x with coefficient of determinants (r2) of 0.992 and the calibration range of Chlorogenic acid was found to be 200-800 ng/spot with the linear equation Y= 282.512+ 4.932x with coefficient of determinants (r2) of 0.996. The limit of detection and limit of quantification were found to be for Ferulic acid 14.35, 40.12 ng/spot, and 10.26, 29.51 ng/spot, for Chlorogenic acid respectively. The relative standard deviation of precision and recovery of Ferulic acid and Chlorogenic acid was < 2>

Keywords:

Ananas comosus cultivars, Ferulic acid, Chlorogenic acid, Simultaneous estimation, Method Validation