Liquid chromatographic method for determining the percent of olestra in lipid samples

A liquid chromatographic (LC) method has been developed to determine the percent of olestra in lipid samples. To achieve the highest degree of accuracy, this method requires the use of an olestra standard with the same molecular composition as the olestra in the lipid sample to be analyzed. Samples were analyzed by reversed-phase LC using an evaporative light-scattering detector. Chromatography was performed with a 5 microns octadecylsilane-Zorbax column that separates olestra from other lipophilic components. Three types of olestra standards (soybean-oil olestra, unheated cotton-seed-oil olestra, and heated cottonseed-oil olestra), each analyzed in soybean oil, showed linearity when the amount of olestra injected ranged from 20 to 160 micrograms (r = 0.9996). The area under the olestra peak (retention time 3.5 to 4.9 min) was used to quantify the amount of olestra in olestra-lipid samples, by comparing the olestra area for the sample with that of the standard using a curve derived by linear regression. The method was evaluated using 3 types of olestra blended with soybean oil and varying the percent of olestra in the olestra-lipid blend from 5 to 90%. Recovery of olestra from these olestra-lipid blends varied from 99.2 to 106.0%, demonstrating excellent accuracy, with method precision expressed as the coefficient of variation, 0.9%. Each error estimate was derived from 5 parallel determinations. With proper validation (e.g., running an olestra-free blank for each lipid matrix), this method provides a rapid, accurate, and precise technique for measuring the percent of olestra in lipids extracted from olestra-formulated foods and in olestra-lipid blends.