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Clinical Chemistry 46: 63-72, 2000;
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Right arrow Lipids, Lipoproteins, and Cardiovascular Risk Factors
(Clinical Chemistry. 2000;46:63-72.)
© 2000 American Association for Clinical Chemistry, Inc.

Articles

Assessment of Between-Instrument Variations in a HPLC Method for Serum Lipoproteins and Its Traceability to Reference Methods for Total Cholesterol and HDL-Cholesterol

Shinichi Usui1, Masakazu Nakamura2, Kazuhiro Jitsukata3, Masayuki Nara4, Seijin Hosaki1 and Mitsuyo Okazaki4,a

1 School of Allied Health Science, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8519, Japan.

2 Department of Epidemiology and Mass Examination for CVD, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka 537-8511, Japan.

3 Department of Clinical Chemistry, SRL Inc., 51 Komiya-cho, Hachiohji-shi, Tokyo 192-8535, Japan.

4 Laboratory of Chemistry, College of Liberal Arts and Sciences, Tokyo Medical and Dental University, 2-8-30, Kohnodai, Ichikawa-shi, Chiba 272-0827, Japan.
a Author for correspondence. Fax 81-47-300-7100; e-mail okazaki.cul{at}cul.tmd.ac.jp

Background: The main purpose of this study was to evaluate the between-instrument variation of the HPLC method for the measurement of total cholesterol (TC), HDL-cholesterol (HDL-C), LDL-cholesterol (LDL-C), VLDL-cholesterol (VLDL-C), chylomicron cholesterol (CM-C), LDL size, and HDL size. Furthermore, the accuracy of the HPLC was assessed for the determination of TC and HDL-C, compared with CDC reference methods.

Methods: We used four HPLC instruments with different column-load numbers from 250 to 5000. For accuracy assessment of TC and HDL-C, we used the reference methods recommended by the CDC.

Results: The values measured by the four instruments were highly correlated with each other (mean r = 0.965), and the absolute mean differences were 4–43 mg/L for TC, 4–30 mg/L for HDL-C, 0–48 mg/L for LDL-C, 7–66 mg/L for VLDL-C, 0–7 mg/L for CM-C, 0.1–0.3 nm for LDL size, and 0–0.1 nm for HDL size. For TC, the HPLC instruments showed high correlation and good agreement with the reference method: r = 0.997; total error <6.6%; absolute mean bias <1.2%. For HDL-C, the results from the HPLC method were significantly higher (10.8% absolute mean bias) than those of the CDC reference method, in spite of good correlation between the two methods (r = 0.998).

Conclusions: The between-instrument variation in serum lipoprotein analysis by HPLC was confirmed to be very small. This method met the US National Cholesterol Education Program’s performance criteria for TC but not for HDL-C.




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