Abstract

METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF ACEPHATE BY USING REVERSED PHASE HPLC

Popat R R*, Adhao V S, Barde L N, Mahajan N M, Thenge R R and Ajmire P V

ABSTRACT

A reversed-phase high performance liquid chromatography (RP-HPLC) method was developed and validated for the estimation of Acephate in bulk form. The separation was achieved on C18 Hypersil BDS column (250mm × 4.6 mm i.d., 5.0 μm) using water and methanol in the ratio 80:20 v/v as mobile phase and at a flow rate of 1.2mL/min. Detection was carried out using a PDA detector at215 nm. The total chromatographic analysis time per sample was about 8.0min with Acephate eluting at retention timeof about 4.8min. The method was validated for accuracy, precision, specificity, linearity and sensitivity. Validation studies demonstrated that this HPLC method is simple,specific, rapid, reliable and reproducible. The standard curve was linear over the concentration range of 1-50μg/mL withR2 close to one (0.9994). The limit of detection (LOD) and limit of Quantitation (LOQ) obtained for Acephate were 0.015 μg/ml and 0.04μg/mL, respectively. The developed and validated method was successfully applied for the quantitative analysis of Acephate in the bulk form. The high recovery and low relartive standard deviation confirm the suitability of the proposed method for the determination of Acephate inbulk form.

Keywords: Organophosphorus, Pesticides, Quantitative.