Application of Gel Permeation Chromatography and Nonaqueous Reverse Phase Chromatography to High Pressure Liquid Chromatographic Determination of Retinyl Palmitate and β-Carotene in Oil and Margarine

Open Access
Publisher Website
Google Scholar
Abstract
A high pressure liquid chromatographic method was developed using high pressure gel permeation chromatography (HP-GPC) and high pressure reverse phase chromatography (RP-HPLC) for quantitation of retinyl palmitate and β-carotene. HP-GPC was used for fractionation of vitamin A active compounds from oil preliminary to quantitation on nonaqueous RP-HPLC. HP-GPC fractionation was completed on oil and margarine dissolved in methylene chloride by 2 elution passes through 2 μStyragel (100Å) columns connected in series with methylene chloride as the mobile phase. RPHPLC separation of retinyl palmitate and β-carotene was achieved on μBondapak C18 (10 μm), using methylene chloride-acetonitrile ( 3 0 + 7 0 ) . Based on 10 repetitive analyses, recoveries of added β-carotene and retinyl palmitate from vegetable oils were 98.6±2.9 and 95.2±2.6%, respectively. The coefficients of variation were 2.9% for β-carotene and 2.7% for retinyl palmitate. The determination of vitamin A activity in 7 margarine brands with label claims of 10% U.S.RDA/serving revealed that all but one of the margarines contained at least 94% of the label claim. Vitamin A activity in the margarines ranged from 90.6 to 110.8% of the label declaration.

This publication has 0 references indexed in Scilit: