High Performance Liquid Chromatographic Method for Determination of Ascorbic Acid in Pure Form, Pharmaceutical Formulations, Biological Samples and in Environmental Water Samples

Abstract

A simple, precise, rapid, and accurate reversed–phase high performance liquid
chromatographic method has been developed for the determination of ascorbic acid in pure
from, pharmaceutical formulations, biological samples and in environmental water samples.
Chromatography was carried out on supercar L1 (C18 ) reversed- phase column (25cm×
4.6mm), 5 microns, using a mixture of methanol : water (42: 58v/v) as a mobile phase at a
flow rate of 1.0 ml.min-1.Detection was performed at 254nm at ambient temperature.
The retention time for ascorbic acid was found 2.3 minutes. The calibration curve was
linear (r= 0.9998) over a concentration range from 0.5 to 12.5μg/ml. Limit of detection
(LOD) and limit of quantitation (LOQ) were found 3ng/ml and 9 ng/ml respectively. The
method was validated for its linearity, precision and accuracy.
The proposed method was successfully applied for the determination of ascorbic acid
in pure form, pharmaceutical formulations, and biological samples and in environmental
water samples.