Abstract
Summary: We assessed the reliability of a method designed for common electron-impact GC-MS systems to determine in a single run most organic acids and glycine conjugates of clinical interest in amniotic fluid. Suitable sensitivity was achieved by dividing the selected-ion chromatogram into 12 time segments during which the monitoring dwelt on specific ions. Twelve metabolites were simultaneously quantified in amniotic fluid, with performances ranging from very good to clinically acceptable. The total coefficient of variation was 2.5–14.1% and the detection limit was well below the lower value of the physiological range. For five other metabolites, the precision was lower and/or the detection limit was near the physiological range. The method was clinically assessed by the prenatal detection of three cases of tyrosinaemia type I and one case of propionic acidaemia. Analytical and clinical evaluation of the method showed that GC-MS with electron-impact fragmentation can be an informative analytical approach for low-level organic acids in physiological fluids. Apart from the case of glycine conjugates, the method shows a fair reliability for amniotic fluid analysis, which might warrant its use for prenatal diagnosis of organic acidurias. However, this method cannot replace procedures using isotopic internal standards, nor GC-MS based on chemical ionization fragmentation, which remain confirmatory analytical methods of choice.
Similar content being viewed by others
REFERENCES
Baric I, Wagner L, Feyh P, Liesert M, Buckel W, Hoffmann GF (1999) Sensitivity and speci-ficity of free and total glutaric acid and 3-hydroxyglutaric acid measurements by stable-isotope dilution assays for the diagnosis of glutaric aciduria type I. J InheritMetab Dis 22: 867–881.
Chalmers RA, Lawson AM (1982) Organic Acids in Man. London: Chapman and Hall.
Coude M, Chadefaux B, Rabier D, Kamoun P (1990) Early amniocentesis and amniotic fluid organic acid levels in the prenatal diagnosis of organic acidemias. Clin Chim Acta 187: 329–332.
Duez P, Kumps A, Mardens Y (1996) GC-MS pro?ling of urinary organic acids evaluated as a quantitative method. Clin Chem 42: 1609–1615.
Gibson KM, Aramaki S, Sweetman L, et al (1990) Stable isotope dilution analysis of 4-hydroxybutyric acid: an accurate method for quantification in physiological fluids and the prenatal diagnosis of 4-hydroxybutyric aciduria. Biomed Environ Mass Spectrom 19: 89–93.
Gibson KM, ten Brink HJ, Schor DS, et al (1993) Stable-isotope dilution analysis of D-and L-2-hydroxyglutaric acid: application to the detection and prenatal diagnosis of D-and L-2-hydroxyglutaric acidemias. Pediatr Res 34: 277–280.
Hagenfeldt L, Hagenfeldt K (1972) Gas chromatographic-mass spectrometric analysis of organic acids in amniotic fluid. Clin Chim Acta 42: 219–220.
Hine DG, Hack AM, Goodman SI, Tanaka K (1986) Stable isotope dilution analysis of isovalerylglycine in amniotic fluid and urine and its application for the prenatal diagnosis of isovaleric acidemia. Pediatr Res 20: 222–226.
Hoffmann G, Aramaki S, Blum-Hoffmann E, Nyhan WL, Sweetman L (1989) Quantitative analysis for organic acids in biological samples: batch isolation followed by gas chromatographic-mass spectrometric analysis. Clin Chem 35: 587–595.
Holm J, Ponders L, Sweetman L (1989) Prenatal diagnosis of propionic and methylmalonic acidaemia by stable isotope dilution analysis of amniotic fluid. J Inherit Metab Dis 12: 271–273.
Inoue Y, Kuhara T (2002) Rapid and sensitive method for prenatal diagnosis of propionic acidemia using stable isotope dilution gas chromatography-mass spectrometry and urease pre-treatment. J Chromatogr B Anal Technol Biomed Life Sci 776: 71–77.
Jakobs C (1994) Antenatal diagnosis of inherited metabolic disorders using gas chromatography-mass spectrometry. In Farriaux J-P, Dhondt J-L, eds. New Horizon in Neonatal Screening. Amsterdam: Excerpta Medica, 357–364.
Jakobs C, Sweetman L, Nyhan WL (1984a) Chemical analysis of succinylacetone and 4-hydroxyphenyllactate in amniotic fluid using selective ion monitoring. Prenat Diagn 4: 187–194.
Jakobs C, Sweetman L, Nyhan WL (1984b) Hydroxy acid metabolites of branched-chain amino acids in amniotic fluid. Clin Chim Acta 140: 157–166.
Jakobs C, Sweetman L, Nyhan WL, Gruenke L, Craig JC, Wadman SK (1984c) Stable isotope dilution analysis of orotic acid and uracil in amniotic fluid. Clin Chim Acta 143: 123–133.
Jakobs C, Sweetman L, Nyhan WL, Packman S (1984d) Stable isotope dilution analysis of 3-hydroxyisovaleric acid in amniotic fluid: contribution to the prenatal diagnosis of inherited disorders of leucine catabolism. J Inherit Metab Dis 7: 15–20.
Jakobs C, Sweetman L, Wadman SK, Duran M, Saudubray JM, NyhanWL (1984e) Prenatal diagnosis of glutaric aciduria type II by direct chemical analysis of dicarboxylic acids in amniotic fluid. Eur J Pediatr 141: 153–157.
Jakobs C, Dorland L, Wikkerink B, Kok RM, de Jong AP, Wadman SK (1988) Stable isotope dilution analysis of succinylacetone using electron capture negative ion mass fragmentography: an accurate approach to the pre-and neonatal diagnosis of hereditary tyrosinemia type I. Clin Chim Acta 171: 223–231.
Jakobs C, Ten Brink HJ, Stellaard F (1990) Prenatal diagnosis of inherited metabolic disorders by quantitation of characteristic metabolites in amniotic fluid: facts and future. Prenat Diagn 10: 265–271.
Jakobs C, ten Brink HJ, Langelaar SA, et al (1991) Stable isotope dilution analysis of N-acetylaspartic acid in CSF, blood, urine and amniotic fluid: accurate postnatal diagnosis and the potential for prenatal diagnosis of Canavan disease. J Inherit Metab Dis 14: 653–660.
Kelley RI, Stamas JN (1993) Quanti?cation of N-acetyl-aspartic acid in urine by isotope dilution gas chromatography-mass spectrometry. J Inherit Metab Dis 16: 918–919.
Kleijer WJ, van der Kraan M, Huijmans JG, van den Heuvel CM, Jakobs C (1995) Prenatal diagnosis of isovaleric acidaemia by enzyme and metabolite assay in the ?rst and second trimesters. Prenat Diagn 15: 527–533.
Kretschmer RE, Bachmann C (1988) Methylcitric acid determination in amniotic fluid by electron-impact mass fragmentography. J Clin Chem Clin Biochem 26: 345–348.
Kumps A, Duez P, Genin J, Mardens Y (1999) Gas chromatography-mass spectrometry analysis of organic acids: altered quantitative response for aqueous calibrators and dilute urine specimens. Clin Chem 45: 1297–1300.
Matalon R, Michals-Matalon K (1999) Prenatal diagnosis of Canavan disease. Prenat Diagn 19: 669–706.
Naylor G, Sweetman L, Nyhan WL, et al (1980) Isotope dilution analysis of methylcitric acid in amniotic fluid for the prenatal diagnosis of propionic and methylmalonic acidemia. Clin Chim Acta 107: 175–183.
NCCLS (1992) NCCLS Evaluation Protocols SC1-B. Villanova, PA: National Committee for Clinical Laboratory Standards.
Ng KJ, Andresen BD, Bianchine JR, et al (1982) Capillary gas chromatographic-mass spectrometric pro?les of trimethylsilyl derivatives of organic acids from amniotic fluids of different gestational age. J Chromatogr 228: 43–50.
Nicholls TM, Hahnel R, Wilkinson SP (1978) Organic acids in amniotic fluid. Clin Chim Acta 84: 11–17.
Niwa T (1986) Metabolic pro?ling with gas chromatography-mass spectrometry and its application to clinical medicine. J Chromatogr 379: 313–345.
Rinaldo P, O'Shea JJ, Coates PM, Hale DE, Stanley CA, Tanaka K (1988) Medium-chain acyl-CoA dehydrogenase de?ciency. Diagnosis by stable-isotope dilution measurement of urinary n-hexanoylglycine and 3-phenylpropionylglycine. NEngl J Med 319: 1308–1313.
Schor DS, Verhoeven NM, Struys EA, ten Brink HJ, Jakobs C (2002) Quanti?cation of 3-hydroxyglutaric acid in urine, plasma, cerebrospinal fluid and amniotic fluid by stable-isotope dilution negative chemical ionization gas chromatography-mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 780: 199–204.
Shigematsu Y, Hata I, Nakai A, et al (1996) Prenatal diagnosis of organic acidemias based on amniotic fluid levels of acylcarnitines. Pediatr Res 39: 680–684.
Sweetman L (1984) Prenatal diagnosis of the organic acidurias. J InheritMetab Dis 7: 18–22.
Sweetman L (1991) Organic acid analysis. In Hommes FA, ed. Techniques in Diagnostic Human Biochemical Genetics. A Laboratory Manual. New York: Wiley-Liss, 143–176.
Trefz FK, Schmidt H, Tauscher B, et al (1981) Improved prenatal diagnosis of methylmalonic acidemia: mass fragmentography of methylmalonic acid in amniotic fluid and maternal urine. Eur J Pediatr 137: 261–266.
Williams VP, Ching DK, Cederbaum SD (1979) Adsorption of organic acids from amniotic fluid and urine onto silica gel before analysis by gas chromatography and combined gas chromatography/mass spectrometry. Clin Chem 25: 1814–1820.
Yamaguchi S, Shimizu N, Orii T, et al (1991) Prenatal diagnosis and neonatal monitoring of a fetus with glutaric aciduria type II due to electron transfer ?avoprotein (beta-subunit) de?ciency. Pediatr Res 30: 439–443.
Zinn AB, Hine DG, Mahoney MJ, Tanaka K (1982) The stable isotope dilution method for measurement of methylmalonic acid: a highly accurate approach to the prenatal diagnosis of methylmalonic acidemia. Pediatr Res 16: 740–745.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kumps, A., Vamos, E., Mardens, Y. et al. Assessment of an electron-impact GC-MS method for organic acids and glycine conjugates in amniotic fluid. J Inherit Metab Dis 27, 567–579 (2004). https://doi.org/10.1023/B:BOLI.0000042981.52186.a9
Issue Date:
DOI: https://doi.org/10.1023/B:BOLI.0000042981.52186.a9