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Clinical Chemistry 25: 273-278, 1979;
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Clinical Chemistry, Vol 25, 273-278, Copyright © 1979 by American Association for Clinical Chemistry

Manual and continuous-flow colorimetry of triacylglycerols by a fully enzymic method

RE Megraw, DE Dunn and HG Biggs

We describe a fully enzymic method for manual and continuous-flow colorimetric assay of triacylglycerols (triglycerides) in serum. Triglycerides are enzymically hydrolyzed in 10 min by lipase and microbial esterase. The resulting free glycerol is measured enzymically by glycerol kinase and glycerol-3-phosphate dehydrogenase. The NADH so formed is oxidized by coupling with a tetrazolium salt/diaphorase system. The test follows Beer's law to 8 g/L, and the final color is stable for at least 1 h for serum, 15 min for aqueous triolein standards. The manual assay requires only 25 microliter of serum and few manipulations. A specific triolein standard was developed for calibrating the manual method. For the continuous-flow method, calibration is made with four concentrations of glycerol standard. The procedure is sensitive, has good precision and accuracy, and gives results that compare well with chemical and enzymic commercial kit methods.


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Copyright © 1979 by the American Association for Clinical Chemistry.