Automated method for the direct determination of total serum cholesterol

Fluorometric analysis

“…Fluorometric analyses of cholesterol in blood made use of the fluorescence developed in chloroform solution by treatment with acetyl choride-zinc chloride reagent (654) or acetic anhydride followed by sulfuric acid (740); an automated procedure has also been described (335). Sterol acetate spots on thin layer chromatograms fluoresce brightly after spraying with 0.2% ethanolic dibromofluorescein (138).…”

Variables in the technicon‐N‐24 method for the determination of serum cholesterol

“…I N RECENT YEARS several procedures have been proposed for the automated determination of serum cholesterol using the AutoAnalyzer. These methods have been based on the reaction of cholesterol with p-toluenesulfonic acid (1,2) or ferric chloride (3-6'). The AntoAnalyzer-N-24 method (4) which is based on the ferric chloride reaction, is one of the more widely used procedures.…”

Notes on the Quality of Performance of Serum Cholesterol Assays

“…The colour reactions given by cholesterol with Lewis acids, first studied by Salkowski, and then by Liebermann and by Burchard, form the basis of all commonly used manual and automated assays (Table I). Simple direct colorimetric AutoAnalyzer methods, using Liebermann-Burchard reagents, have been described (Kenny and Jamieson, 1964;Levine et al, 1967;Annan and Isherwood, 1969), but although good concordance with extraction methods can be obtained with normal sera, it is difficult to account for the effects of haernolysis or lipaemia (Moline and Barron, 1969) or of hyperbilirubinaemia (Zak et al 1970). Not surprisingly, the response of cholesterol esters markedly depends on the temperature and period of incubation with the acid reagent (Siegel and Bowdoin, 1971).…”