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October 1, 1976Clinical Chemistry

Cholesterol oxidase-based determination, by continuous-flow analysis, of total and free cholesterol in serum.

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Authors

RLR F LieJSJoan M SchmitzNational Institutes of HealthKPK J PierreSRI International

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Implication

Laboratory evaluation demonstrates accurate continuous-flow enzymatic quantification of total and free serum cholesterol, suggesting reliable automation for clinical lipid profiling.

Key Points

  • To develop and evaluate an automated, continuous-flow enzymatic assay using cholesterol oxidase to measure total and free cholesterol in serum.
  • Total cholesterol was measured using enzymatic hydrolysis of cholesterol esters, oxidation via cholesterol oxidase, and colorimetric detection of liberated peroxide using 4-aminoantipyrine, phenol, and peroxidase on an AutoAnalyzer II system.
  • Free cholesterol was quantified on the same manifold by omitting cholesterol esterase from the reagent mixture.
  • Calibrations utilized pre-assayed cholesterol-in-serum materials to bypass solvent incompatibility, with results compared against standard Liebermann-Burchard reaction methods.
  • Total cholesterol measured enzymatically correlated strongly with the isopropanol extraction and Liebermann-Burchard method (total cholesterol, g/L: y_enz = 0.991x_lb + 0.05; r = 0.996).
  • Free cholesterol measurements aligned closely with traditional digitonide precipitation and Liebermann-Burchard colorimetric detection (free cholesterol, %: y_enz = 0.982x_dig - 0.7; r = 0.956).

Cite This Study

Lie et al. (1976) studied this question.

synapsesocial.com/papers/6a70d952a2d7cf2e39c2b97fhttps://doi.org/10.1093/clinchem/22.10.1627
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