Enzymatic determinations of cholesterol in high-density-lipoprotein fractions prepared by a precipitation technique.

Bernard W Steele, D. F. Koehler, M. M. Azar, T. P. Blaszkowski, K. Kuba, M. E. Dempsey

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Abstract

An enzymatic method for cholesterol in serum [Clin. Chem. 20, 470 (1974)] was initially found to be unsatisfactory for measuring cholesterol in high-density-lipoprotein fractions prepared by precipitation with Mn2+. A fine precipitate formed in the cuvette and cholesterol values were falsely increased. We describe a simple, convenient method for circumventing these problems. An ethylenediaminetetraacetate solution is used to reconstitute the enzymatic reagent. Cholesterol values by this procedure correlated with those obtained by the Lipid Research Clinic's procedure for the same lipoprotein fraction preparations (regression slope, .998; Y-intercept, 8.9 mg/liter; correlation coefficient, .984; standard error of the estimate, 16.8 mg/liter). Precision of the assay, including the precipitation step, was calculated. The SDwithin day was 9.7 mg/liter and SDoverall was 23.7 mg/liter. Results for total cholesterol with the modified reagent were linearly related to concentrations exceeding 4 g/liter, thereby permitting determination of high-density-lipoproteins and total cholesterol in a single run.

Original languageEnglish
Pages (from-to)98-101
Number of pages4
JournalClinical Chemistry
Volume22
Issue number1
StatePublished - Jan 1 1976
Externally publishedYes

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HDL Lipoproteins
HDL Cholesterol
Cholesterol
Lipoproteins
Lipids
Precipitates
Assays
Serum
Research

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Steele, B. W., Koehler, D. F., Azar, M. M., Blaszkowski, T. P., Kuba, K., & Dempsey, M. E. (1976). Enzymatic determinations of cholesterol in high-density-lipoprotein fractions prepared by a precipitation technique. Clinical Chemistry, 22(1), 98-101.

Enzymatic determinations of cholesterol in high-density-lipoprotein fractions prepared by a precipitation technique. / Steele, Bernard W; Koehler, D. F.; Azar, M. M.; Blaszkowski, T. P.; Kuba, K.; Dempsey, M. E.

In: Clinical Chemistry, Vol. 22, No. 1, 01.01.1976, p. 98-101.

Research output: Contribution to journalArticle

Steele, BW, Koehler, DF, Azar, MM, Blaszkowski, TP, Kuba, K & Dempsey, ME 1976, 'Enzymatic determinations of cholesterol in high-density-lipoprotein fractions prepared by a precipitation technique.', Clinical Chemistry, vol. 22, no. 1, pp. 98-101.
Steele BW, Koehler DF, Azar MM, Blaszkowski TP, Kuba K, Dempsey ME. Enzymatic determinations of cholesterol in high-density-lipoprotein fractions prepared by a precipitation technique. Clinical Chemistry. 1976 Jan 1;22(1):98-101.
Steele, Bernard W ; Koehler, D. F. ; Azar, M. M. ; Blaszkowski, T. P. ; Kuba, K. ; Dempsey, M. E. / Enzymatic determinations of cholesterol in high-density-lipoprotein fractions prepared by a precipitation technique. In: Clinical Chemistry. 1976 ; Vol. 22, No. 1. pp. 98-101.
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N2 - An enzymatic method for cholesterol in serum [Clin. Chem. 20, 470 (1974)] was initially found to be unsatisfactory for measuring cholesterol in high-density-lipoprotein fractions prepared by precipitation with Mn2+. A fine precipitate formed in the cuvette and cholesterol values were falsely increased. We describe a simple, convenient method for circumventing these problems. An ethylenediaminetetraacetate solution is used to reconstitute the enzymatic reagent. Cholesterol values by this procedure correlated with those obtained by the Lipid Research Clinic's procedure for the same lipoprotein fraction preparations (regression slope, .998; Y-intercept, 8.9 mg/liter; correlation coefficient, .984; standard error of the estimate, 16.8 mg/liter). Precision of the assay, including the precipitation step, was calculated. The SDwithin day was 9.7 mg/liter and SDoverall was 23.7 mg/liter. Results for total cholesterol with the modified reagent were linearly related to concentrations exceeding 4 g/liter, thereby permitting determination of high-density-lipoproteins and total cholesterol in a single run.

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