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Clinical Chemistry 21: 1575-1581, 1975;
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Clinical Chemistry, Vol 21, 1575-1581, Copyright © 1975 by the American Association for Clinical Chemistry

Manual and Semi-Automated Procedures for Measurement of Triglycerides in Serum

Joseph L. Giegel 1, Anna Bell Ham 1, and William Clema 1

1 Clinical Chemistry Research and Development Laboratory, DADE Division American Hospital Supply Corp., Box 520672, Miami, Fla. 33152

We describe manual and semi-automated procedures for serum triglyceride determinations, in which lipids are partitioned between a water/isopropanol phase and a nonane phase. More than 99% of the triglyceride is extracted into the nonane phase, as determined by recovery of 3H-labeled triolein. Studies with 14C-labeled lecithin demonstrate that less than 1.3% is extracted into the nonane phase at concentrations up to 2.5 g/liter. A novel feature of the method is that glycerol can be liberated from triglyceride by sodium hydroxide at room temperature in less than 5 min. Glycerol is oxidized by periodate in 1-2 min at 25 °C; the formaldehyde produced is reacted with 2,4-pentanedione to yield 3,5-diacetyl-1,3-dihydrolutidine. The manual procedure requires less than 20 min; the semi-automated method requires 7 min from sampling to readout. The procedure may be run at 30-40 samples/h, with stable baseline and less than 2.0% carryover. Both methods are linear to 0.50 g (5.6 mol) of triolein per liter. Analytical recoveries at several concentrations ranged from 97-101% (mean, 100%)


Key Words: solvent partition method • normal (reference) values • continuous-flow analysis • stability of triglycerides in serum • hyperlipemia

Submitted on March 31, 1975
Accepted on July 2, 1975




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Copyright © 1975 by the American Association for Clinical Chemistry.