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We established a high-precision selective determination method for free N-acetylneuraminic acid in human serum, involving capillary gas chromatography-mass spectrometry with selected ion monitoring and specific separation on a Sep Pak silica cartridge. In contrast to the value of 800 ng/ml of N-acetylneuraminic acid previously reported by Haverkamp et al. (J. Haverkamp, R. Schauer, and M. Wember (1976) Hoppe-Seyler's Z. Physiol. Chem. 357, 1699), who used the thiobarbituric acid method, the present method gave a value of 194 +/- 96 ng/ml for 22 serum samples from normal Japanese male volunteers aged between 20 and 30 years. The mass fragmentogram of serum showed a good signal/noise ratio, and the measurement was very specific, accurate, and reproducible.
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