Polarographic Determination of Serum High Density Lipoprotein-and Low Density Lipoprotein-Cholesterol by Use of Oxygen Analyzer

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Low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and choline containing phospholipids in serum are enzymatically determined by use of a polarographic oxygen analyzer with a long life and high sensitive sensor to determine the low concentration of serum lipids.<BR>The final assay system for cholesterol in the serum, assessed from the oxygen consumption value that we found to be optimum, consists of 1 ml of phosphate buffer (0.1mol/l, pH 7.3), containing 50 μmol of phenol, 0.5% sodium taurocholate as a surfactant, 5U of cholesterol ester hydrolase, 9U of cholesterol oxidase and for phospholipids in serum, consists of 1ml of Trishydrochloride buffer (0.05 mol/l, pH7.6) containing 25μmol of phenol, 0.1 % of Triton X 100 as a surfactant, 5.4 U of phospholipase D and 12 U of cholineoxidase.<BR>Oxygen consumption and the concentration of cholesterol or choline containing phospholipids are lineary related to 2.5 g/l from 0.2 g/l using 50μl of serum within 60 seconds-incubation. Replicated analysis of cholesterol and phospholipids by the present method demonstrated the following inter-run precision; HDL-C, mean (mg/dl)±S. D., 39.0±0.60, c. v., 1.54 %, LDL-C, 116.0±0.70, c. v., 0.60 %, phospholipids, 194.0±6.0, c. v., 3.09.<BR>Bilirubin and ascorbic acid were without effect on the present method unlike the enzymic colorimetric methods.<BR>The correlationship of cholesterol and phospholipids between polarographic method and enzymic colorimetric method was well in normal serum.
Japan Society of Clinical Chemistryの論文

Polarographic Determination of Serum High Density Lipoprotein-and Low Density Lipoprotein-Cholesterol by Use of Oxygen Analyzer

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