Determination of cholesterol using o-phthalaldehyde
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Updated: Aug 30, 2026

Cholesterol Efflux Assay
Published on: March 6, 2012
1Banting and Best Department of Medical Research, University of Toronto, Toronto, Canada, and Departments of Biochemistry and Pediatrics, University of Arkansas Medical Center, Little Rock, Arkansas 72201.
Abstract:
A simple, rapid method for the determination of cholesterol in plasma and tissue using o-phthalaldehyde is presented. Comparison of this method with the FeCl(3) method gave identical results. However, the o-phthalaldehyde determination is three times more sensitive than the FeCl(3) determination (molar extinction coefficients of 11,610 and 33,440 for FeCl(3) and o-phthalaldehyde, respectively), it takes less time to complete, and the color developed is more stable. The o-phthalaldehyde method can be used to assay free and esterified cholesterol directly after thin-layer chromatographic separation.

