Summary
Succinic semialdehyde (SSA) accumulates in the inborn error of meta- bolism succinic semialdehyde dehydrogenase deficiency owing to impaired enzymatic conversion to succinic acid. We developed a stable-isotope dilution liquid chromato- graphy–tandem mass spectrometry method for the determination of SSA in urine and cerebrospinal fluid samples. Stable-isotope-labelled [13C4]SSA, serving as internal standard, was prepared by reaction of ninhydrin with L-[13C5]glutamic acid. SSA in body fluids was converted to its dinitrophenylhydrazine (DNPH) derivative, without sample purification prior to the derivatization procedure. The DNPH derivative of SSA was injected onto a C18 analytical column and chromatography was performed by isocratic elution. Detection was accomplished by tandem mass spectrometry operating in the negative multiple-reaction monitoring mode. The limit of detection was 10 nmol/L and the calibration curves over the range 0–500 pmol of SSA showed good linearity (r2 > 0.99). The intra-day coefficient of variation (n = 10) for urine was 2.7% and inter-day coefficient of variation (n = 5) for urine was 8.5%. The average recoveries performed on two levels by enriching urine and cerebrospinal fluid samples ranged between 85 and 115%, with coefficients of variation < 8%. The method enabled the first determination of normal values for SSA in urine and pathological values of SSA in urine and cerebrospinal fluid samples derived from patients with succinic semialdehyde dehydrogenase deficiency.
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References
Andreoli R, Manini P, Corradi M, et al (2003) Determination of patterns of biologically relevant aldehydes in exhaled breath condensate of healthy subjects by liquid chromatography/atmospheric chemical ionisation tandem mass spectrometry. Rapid Commun Mass Spectrom 17: 637–645.
Di Padova C, Alderman J, Lieber CS (1986) Improved methods for the measurements of acetaldehyde concentrations in plasma and red blood cells. Alcohol Clin Exp Res 10: 86–89.
Gibson KM, Jakobs C (2001) Disorders of β- and γ-amino acids in free and peptide-linked forms. In: Scriver CR, Beaudet AL, Sly WS, Valle D, eds; Childs B, Kinzler KW, Vogelstein B, assoc. eds. The Metabolic and Molecular Bases of Inherited Metabolic Disease, 8th edn. New York: McGraw-Hill, 2079–2105.
Jakobs C, Bojasch M, Monch E, et al (1981) Urinary excretion of gamma-hydroxybutyric acid in a patient with neurological abnormalities. The probability of a new inborn error of metabolism. Clin Chim Acta 111: 169–178.
Kaufman EE, Nelson T, Fales HM, et al (1988) Isolation and characterization of a hydroxyacid–oxoacid transhydrogenase from rat kidney mitochondria. J Biol Chem 263: 16872–16879.
Loscher W (2002) Basic pharmacology of valproate: a review after 35 years of clinical use for the treatment of epilepsy. CNS Drugs 16: 669–694.
Lujan R, Shigemoto R, Lopez-Bendito G (2005) Glutamate and GABA receptor signaling in the developing brain. Neuroscience 130: 567–580.
Luo XP, Yazdanpanah M, Bhooi N, et al (1995) Determination of aldehydes and other lipid peroxidation products in biological samples by gas chromatography–mass spectrometry. Anal Biochem 228: 294–298.
Maitre M (1997) The gamma-hydroxybutyrate signaling system in brain: organization and functional implications. Prog Neurobiol 51: 337–361.
Nagy K, Pollreisz F, Takats Z, et al (2004) Atmospheric pressure chemical ionization mass spectrometry in biological matrices. Rapid Commun Mass Spectrom 18: 2473–2478.
Nicholson KL, Balster RL (2001) GHB: a new and novel drug of abuse. Drug Alcohol Depend 63: 1–22.
Pearl PL, Gibson KM (2004) Clinical aspects of the disorders of GABA metabolism in children. Curr Opin Neurol 17: 107–113.
Tomita M, Okuyama T, Kawai S (1990) Determination of malonaldehyde in oxidised biological materials by high-performance liquid chromatography. J Chromatogr 515: 391–397.
Wong CG, Chan KF, Gibson KM, et al (2004) Gamma-hydroxybutyric acid: neurobiology and toxicology of a recreational drug. Toxicol Rev 23: 3–20.
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Struys, E.A., Jansen, E.E.W., Gibson, K.M. et al. Determination of the GABA analogue succinic semialdehyde in urine and cerebrospinal fluid by dinitrophenylhydrazine derivatization and liquid chromatography–tandem mass spectrometry: Application to SSADH deficiency. J Inherit Metab Dis 28, 913–920 (2005). https://doi.org/10.1007/s10545-005-0111-0
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DOI: https://doi.org/10.1007/s10545-005-0111-0