Clinical Chemistry
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Clinical Chemistry 45: 696-699, 1999;
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(Clinical Chemistry. 1999;45:696-699.)
© 1999 American Association for Clinical Chemistry, Inc.

Technical Briefs

High-Throughput Measurement of Oxidizability of Low-Density Lipoproteins Suitable for Use in Clinical Trials,1

Thomas G. Colea and Nilima Parikh

1 This work was presented previously in abstract form: Cole TG, Parikh N. High-throughput analysis of oxidizability of LDL suitable for use in clinical studies [Abstract]. Clin Chem 1998;44:A84.
a author for correspondence: fax 314-362-4782, e-mail Thom@imgate.wustl.edu

The susceptibility of lipoproteins to oxidation is thought to be a critical step in a complex process culminating in arterial lipid deposition; however, the role of the oxidizability of lipoproteins in atherosclerosis has not been demonstrated in any large-scale clinical trials (1)(2)(3)(4). Although several measurements of the oxidizability of LDL have been developed, the most commonly used method is the measurement of the rate of formation of conjugated hydroperoxides of polyunsaturated fatty acids in response to exposure to copper ions, which leads to the formation of conjugated dienes (CDs) (5). Three major parameters are used to describe the oxidizability of LDL by the CD assay: (a) the lag time (LT) during which lipoprotein-associated antioxidants are consumed, thereby sparing the LDL lipids from oxidation; (b) the maximum rate of CD formation (Max V); and (c) the total amount of CD formed ({Delta}CD).

In practice, the CD assay requires the sequential measurement of absorbance at 234 nm in a spectrophotometer over a prolonged period of time, often >3 h. Because the automated cell carrier of most spectrophotometers holds a maximum of only six cuvettes, the analysis of large numbers of specimens, such as for large-scale clinical trials, is difficult and time-consuming. In addition, the analysis of replicate specimens or the inclusion of quality-control materials generally is precluded by the small number of specimens that can be analyzed simultaneously. We have developed methods that allow the isolation of LDL from a large number of specimens and the analysis of the oxidizability of this isolated LDL. Several factors were necessary for these goals to be attained: (a) the use of an ultracentrifuge rotor with capacity for a large number of specimens (Type 25; Beckman Instruments); (b) the recent availability of a 96-well microtiter . . . [Full Text of this Article]


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The following articles in journals at HighWire Press have cited this article:


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[Abstract] [Full Text] [PDF]


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