HPTLC method development and validation for the quantification of gallic acid in poly herbal formulation

John PP, Banurekha J, Kumar M, Venkateswarlu BS

Nirmala college of Health Science, Nirmala Group of Institutions, Nutmeg Gardens, P.O. Kunnapilly, Meloor, Chalakudy, Thrissur, India.

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John PP, Banurekha J, Kumar M, Venkateswarlu BS, 2023. HPTLC method development and validation for the quantification of gallic acid in polyherbal formulation. Journal of medical pharmaceutical and allied sciences, V 12 - I 2, Pages - 5730 – 5735. Doi: 10.55522/jmpas.V12I2.4963.

ABSTRACT

HPTLC method is developed to estimate gallic acid in the Poly herbal formulation. Developed methods were statistically evaluated according to ICH guidelines. Ethyl Acetate: Toluene: Methanol: Formic acid (3:2:1:5 v/v) was selected as a solvent system for HPTLC. The formulation was prepared by mixing all the ingredients in equal ratios and extracting them with Alcohol, Petroleum ether, Ethyl acetate, and chloroform. Pure Gallic acid is given a strong spot at Rf 0.65 in this solvent system and absorption maxima comes out to be 254 nm. Ethanolic extract of Salacia reticulata (EESR) and Ethanolic Extract of local formulation (EELF) showed spot of Gallic acid at Rf 0.65 along with nine other spots in this solvent system. The standard curve was plotted between different concentrations of Gallic acid and peak area. The method was found to be linear in the concentration range of 200-2000ng with a correlation coefficient of 0.985. In precision studies, % RSD (Relative standard deviation) values of intraday were shown from 0.98-1.2 and the inter-day study was between 1.04-1.53. The percent recovery was between 98.40-99.27%. The limit of Detection (LOD) and Limit of Quantification (LOQ) were found to be 50.1556 ng/ml and 167.1853ng/spot for Gallic acid. Purity of each spot which is scanned at wavelength range 200-800nm with the value of r (s, m) within the range 0.980-0.999 and r (m, e) within the range 0.984-0.999. EESR was found to pose a maximum amount of Gallic acid of 3.3346 (μg/gm) while EELF was found to contain a minimum amount of Gallic acid 2.8117 (μg/gm). The method used was found to be liner, accurate and precise.

Keywords:

HPTLC, Validation, Gallic acid, Marker, Local formulation, Salacia reticulata.

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