J. Straczek et al., QUANTIFICATION OF METHYLMALONIC ACID IN SERUM MEASURED BY CAPILLARY GAS-CHROMATOGRAPHY MASS-SPECTROMETRY AS TERT-BUTYLDIMETHYLSILYL DERIVATIVES, Journal of chromatography. Biomedical applications, 620(1), 1993, pp. 1-7
Quantification of methylmalonic acid in serum is considered one of the
most sensitive parameters for diagnosis of cobalamin deficiency. Meth
ylmalonic acid was measured as tert.-butyldimethylsilyl derivatives fo
rmed in the presence of dimethylformamide. Under these conditions the
derivative was quantified using gas chromatography-mass spectrometry w
ith selected-ion monitoring at 403 and 406 for methylmalonic acid and
methyl-d3-malonic acid, respectively. The characteristics of the metho
d were: linearity from 0.04 to 1.7 mumol/l with linear regression equa
tion y = -0.0199 + 0.727 x (r = 0.999); recovery 95.5 +/- 6.8%; detect
ion threshold 17 fmol injected; within-day variation coefficient rangi
ng from 3. 10% to 5.6% according to sample concentration; and day-to-d
ay coefficient of variation of 6.8% and 5.3% for methylmalonic acid co
ncentrations of 0.103 mumol/l and 0.360 mumol/l. respectively. The nor
mal range after log transformation was estimated at 0.03-0.26 mumol/l,
0.06-0.33 mumol/l and 0.02-0.40 mumol/l in children of 4-14 years (n
= 39), in healthy subjects at 20-40 years (n = 70) and in healthy elde
rly persons older than 60 years (n = 14), respectively. The normal ran
ge in children was significantly lower than that in adults and, in con
trast, normal values in the elderly were significantly higher.