Improved Thin-Layer Chromatographic Method for the Separation of Cholesterol, Egg Phosphatidylcholine, and Their Degradation Products

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Degradation products of egg phosphatidylcholine (EPC) and cholesterol were analyzed with different normal- and reversed-phase thin-layer chromatography (TLC) systems. The best separation, in terms of the highest number of degradation products from both analytes, was obtained with a reversed-phase system, using butanol–methanol–water–96–98% (v/v) acetic acid (40 + 40 + 20 + 4, v/v/v/v) as the mobile phase after overnight saturation at 25°C. A special development technique was used. After a first development, the plate was dried and a second development was performed in the same direction. This method enabled us to separate lysophosphatidylcholine, several free fatty acids and hydroperoxides, and several undefined degradation products of EPC and cholesterol. All products were visualized after the plate was dipped in a 1% (v/v) solution of 4-methoxybenzaldehyde in 98% sulfuric acid–96–98% (v/v) acetic acid–ethanol– water (2 + 10 + 60 + 30), presenting a blue color or a white spot against a colored background. After activation at 110°C, a stable color for both analytes was reached after 12 min. Precision of <5% was obtained at 2 levels of analysis. Good linearity was obtained in the range of 5–30 μg for EPC (r = 0.991) and 5–40 μg for cholesterol (r = 0.991). These results show that TLC can be an inexpensive and easy alternative for the analysis of EPC and cholesterol.

Document Type: Research Article

Affiliations: 1: VUB Pharmaceutical Technology and Physical Pharmacy, Laarbeeklaan 103, 1090 Brussels, Belgium. 2: VUB Anesthesiology, Laarbeeklaan 101, 1090 Brussels, Belgium. 3: VUB Pharmaceutical Technology and Physical Pharmacy, Laarbeeklaan 101, 1090 Brussels, Belgium.

Publication date: November 1, 2002

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  • The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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