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Cell-free Biochemical Fluorometric Enzymatic Assay for High-throughput Measurement of Lipid Peroxidation in High Density Lipoprotein
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Published on: October 12, 2017

Direct and simple fluorescence detection method for oxidized lipoproteins.

Takeshi Ikeda1, Makoto Seo, Ikuo Inoue

  • 1Department of Chemistry, Graduate School of Science, Rikkyo University, 3-34-1, Nishi-Ikebukuro, Toshima-ku, Tokyo, 171-8501, Japan.

Analytical Chemistry
|January 2, 2010
PubMed
Summary

A new fluorescence spectrometry method directly quantifies oxidized lipoproteins, offering a cost-effective clinical tool. This technique differentiates oxidized LDL and HDL levels in patients with dyslipidemia, aiding coronary heart disease risk assessment.

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Area of Science:

  • Biochemistry
  • Clinical Chemistry
  • Analytical Chemistry

Background:

  • Low-density lipoprotein (LDL) and high-density lipoprotein (HDL) quantification are key for metabolic syndrome assessment.
  • Small dense LDL and oxidized LDL/HDL particles are increasingly recognized as significant coronary heart disease risk factors.
  • Current methods for characterizing oxidized lipoproteins are often expensive for routine clinical use.

Purpose of the Study:

  • To develop a straightforward and cost-effective fluorescence spectrometry method for direct quantitation of oxidized lipoproteins.
  • To enable clinical measurement of oxidized LDL and HDL particles in patients with dyslipidemia.

Main Methods:

  • Direct quantitation of oxidized lipoproteins using fluorescence spectrometry.
  • Excitation in UV (365 +/- 10 nm) or visible (470 +/- 10 nm) ranges.
  • Emission detection at 450 +/- 30 nm or 535 +/- 15 nm.
  • Analysis of 1 microL of 1 mg/mL lipoprotein samples without expensive antibodies.

Main Results:

  • Biological samples from dyslipidemia patients showed higher fluorescence for oxidized LDL and light HDL compared to normal subjects.
  • Samples from dyslipidemia patients exhibited lower fluorescence for oxidized heavy HDL compared to normal subjects.
  • The method demonstrated potential for clinical application in dyslipidemia patients.

Conclusions:

  • A simple, direct fluorescence spectrometry method for oxidized lipoprotein quantitation has been established.
  • This technique offers a cost-effective alternative for clinical assessment of oxidized lipoproteins.
  • The findings highlight differential oxidized LDL and HDL patterns in dyslipidemia, relevant to coronary heart disease risk.