J. Lipid Res.
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Originally published In Press as doi:10.1194/jlr.D600009-JLR200 on June 20, 2006

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Journal of Lipid Research, Vol. 47, 2089-2096, September 2006
Copyright © 2006 by American Society for Biochemistry and Molecular Biology

Methods

Measurement of serum total glycerides and free glycerol by high-performance liquid chromatography1

Hongxia Li*, Jun Dong*, Wenxiang Chen2,*,{dagger}, Shu Wang*, Hanbang Guo*, Yong Man*, Peisheng Mo{dagger} and Jianzhai Li*

* Beijing Hospital Institute of Geriatrics, Beijing, 100730 China
{dagger} National Center for Clinical Laboratories, Beijing, 100730 China

1 An abstract of part of this study was presented at the 18th International Congress of Clinical Chemistry and Laboratory Medicine, Kyoto, Japan, 2002.

Published, JLR Papers in Press, June 20, 2006.

2 To whom correspondence should be addressed. e-mail: chenwenxiang{at}263.net

Serum levels of total glycerides and free glycerol are important indices of lipid metabolism and cardiovascular disease risk. Convenient enzymatic methods of measurement have been available, but they are susceptible to interference. Situations exist in both research and clinical laboratories in which more specific and precise methods are needed. We developed HPLC methods for the measurement of serum total glycerides and free glycerol. For total glycerides, serum was mixed with an internal standard (1,2,4-butanetriol) and treated with alcoholic sodium hydroxide to hydrolyze glycerides to glycerol. After deproteinization with tungstic acid, the glycerol was benzoylated with an optimized Schotten-Baumann reaction and analyzed by HPLC. For free glycerol, serum was equilibrated with the internal standard and deproteinized with tungstic acid to remove the glycerides. The glycerol was benzoylated and analyzed as for total glycerol. Various factors were investigated, and no significant sources of interference were detected. The total coefficients of variation ranged from 0.7% to 2.0% for total glycerides and from 1.7% to 3.2% for free glycerol. The analytical recoveries ranged from 98.5% to 101.6%. In conclusion, simple and reliable HPLC methods for serum total glycerides and free glycerol have been developed. The methods may also be used for the analyses of glycerol or glycerides in other biological samples.

Supplementary key words triglycerides • Schotten-Baumann reaction • benzoylation • butanetriol


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Copyright © 2006 by the American Society for Biochemistry and Molecular Biology.