suberylglycine and 3-phenylpropionylglycine

In order to determine which are useful early diagnostic markers for medium-chain acyl-CoA dehydrogenase (MCAD) deficiency, we have analysed urine from an asymptomatic neonate. Profiling of urinary organic acids followed by peak confirmation by electron impact mass spectrometry revealed a high suberate/adipate ratio (greater than 1.0) and the presence of n-hexanoylglycine (HG). Acylcarnitine analysis by fast atom bombardment mass spectrometry (FAB-MS) was inconclusive, but FAB-MS/MS (tandem mass spectrometry) revealed diagnostic amounts of octanoylcarnitine and hexanoylcarnitine. Quantitative analysis of acylglycines by stable isotope dilution and chemical ionization mass spectrometry revealed a 30-fold increase in HG and increased suberylglycine, but no increase in 3-phenylpropionylglycine.

Topics: Acyl-CoA Dehydrogenase; Acyl-CoA Dehydrogenases; Carnitine; Child, Preschool; Creatinine; Dicarboxylic Acids; Gas Chromatography-Mass Spectrometry; Genetic Markers; Glycine; Humans; Infant, Newborn; Male; Trimethylsilyl Compounds

We describe a gas chromatographic/mass spectrometric method for the accurate determination of n-hexanoylglycine, 3-phenylpropionylglycine and suberylglycine in urine for the diagnosis of hereditary medium-chain acyl-CoA dehydrogenase (MCAD) deficiency. These acylglycines had previously been detected in urine from patients with MCAD deficiency, but their diagnostic values were unknown because of a lack of appropriate analytical methods. n-Hexanoyl(1,2-13C)glycine, 3-phenylpropionyl(2-13C,15N)glycine and suberyl(2-13C,15N)glycine were synthesized and used as internal standards. Ammonia chemical ionization was utilized to generate intense [M + H]+ ions for selected-ion monitoring quantification. The whole procedure is fast and can be performed by a low-resolution gas chromatographic/mass spectrometric system, giving accurate results over a range of three orders of magnitude (0.0167-16.7 micrograms/ml). The results from the analyses of 54 urine samples from 21 MCAD-deficient patients and various control samples using this method established that n-hexanoyglycine and 3-phenylpropionylglycine were highly diagnostic for this disease, while suberylglycine was found less specific.

Topics: Chemical Phenomena; Chemistry; Child; Chromatography, Ion Exchange; Dicarboxylic Acids; Gas Chromatography-Mass Spectrometry; Glycine; Humans

Medium-chain acyl-CoA dehydrogenase (MCAD) deficiency, one of the most common inherited metabolic disorders, is often mistaken for the sudden infant death syndrome or Reye's syndrome. Diagnosing it has been difficult because of a lack of fast and reliable diagnostic methods. We developed a stable-isotope dilution method to measure urinary n-hexanoylglycine, 3-phenylpropionylglycine, and suberylglycine, and we retrospectively tested its accuracy in diagnosing MCAD deficiency. We measured the concentrations of these three acylglycines in 54 urine samples from 21 patients with confirmed MCAD deficiency during the acute and asymptomatic phases of the illness and compared the results with the concentrations in 98 samples from healthy controls and patient controls with various diseases. The levels of urinary hexanoylglycine and phenylpropionylglycine were significantly increased in all samples from the patients with MCAD deficiency, clearly distinguishing them from both groups of controls. Although urinary suberylglycine was increased in the patients, the range of values in the normal controls who were receiving formula containing medium-chain triglycerides was very wide, overlapping somewhat with the values in the patients with asymptomatic MCAD deficiency. These results indicate that the measurement of urinary hexanoylglycine and phenylpropionylglycine by our method is highly specific for the diagnosis of MCAD deficiency. The method is fast and can be applied to random urine specimens, without any pretreatment of patients.

Topics: Acyl-CoA Dehydrogenase; Acyl-CoA Dehydrogenases; Child; Child, Preschool; Diagnosis, Differential; Dicarboxylic Acids; Glycine; Humans; Infant; Infant, Newborn; Methods; Radioisotope Dilution Technique; Retrospective Studies; Reye Syndrome; Sudden Infant Death