AUTOMATED FLUOROMETRIC-DETERMINATION OF CELLULAR CHOLESTEROL

We developed a completely automated fluorimetric method for the determ ination of cellular cholesterol, consisting of enzymatic hydrolysis of cholesteryl ester to free cholesterol and enzymatic oxidation of free cholesterol in the presence of an indicator substrate to produce a fl uorescent product. For control preparations of monocytes, the mean det ection limit was 2.57 mu mol/5 x 10(5) cells and the mean within-batch coefficients of variation were 9.30, 6.00 and 3.73% at mean cholester ol concentrations of 1.94, 9.05 and 12.49 mu mol/5 x 10(5) cells, resp ectively. The results correlated well with those obtained by gas-liqui d chromatography.