Determination of "free" glycerol in human serum reference materials by isotope-dilution gas chromatography-mass spectrometry

Serum free glycerol analyses are an important part of the preparation and evaluation of human serum reference materials used for the quality assurance of triglyceride assays. However, enzymatic kits for free glycerol analysis obtained from different vendors have, on occasion, provided different results for a given sample. In an effort to establish the "target" glycerol content of selected reference materials, we have established a method for the analysis of serum free glycerol by using isotope-dilution gas chromatography-mass spectrometry, incorporating [1,3-13C2]glycerol as the internal standard. The use of a simplified serum extraction and clean-up procedure resulted in (uncorrected) recoveries of glycerol averaging about 90% before derivatization, and in estimated concentrations for spiked serum pools that corresponded closely to the expected values. A comparison of enzymatic and gas chromatographic-mass spectrometric results for several reference serum pools suggest that the latter method is of value in evaluating and validating routine enzymatic methods for free glycerol analysis.