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Asian-Australas J Anim Sci > Volume 14(10); 2001 > Article
Animal Products
Asian-Australasian Journal of Animal Sciences 2001;14(10): 1465-1469.
DOI: https://doi.org/10.5713/ajas.2001.1465    Published online October 1, 2001.
Determination of Cholesterol in Milk and Dairy Products by High-Performance Liquid Chromatography
H. I. Oh, T. S. Shin, E. J. Chang
Abstract
A sensitive high-performance liquid chromatographic method was developed to determine the content of cholesterol in milk and dairy products. To optimize separation of cholesterol, mobile phases including acetonitrile:2-propanol (8:1, v/v), acetonitrile:methanol (3:1, v/v), and acetonitrile:methanol:2-propanol (7:3:1, v/v/v) were compared. Acetonitrile/methanol/2-propanol was superior to the other mobile phase systems for separating cholesterol. Liquid-liquid extraction (LLE) of cholesterol was simplified using a non-polar solvent, hexane, to remove interfering compounds, and had an excellent recovery (100 1.0%) of cholesterol. A solid phase extraction (SPE) method using Sep-pak C18 was developed and compared with LLE. The SPE method was rapid and highly reproducible. Both extraction methods were useful when used in combination with saponification of esterified cholesterol to facilitate total cholesterol determination. The detection limit of cholesterol was 0.01 g. The newly developed HPLC method was rapid, simple, and accurate, and has advantages over the many methods commonly used.
Keywords: Cholesterol; Milk; Dairy Products; High-Performance Liquid Chromatography


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