A LC-MS/MS method for the specific, sensitive, and simultaneous quantification of 5-aminolevulinic acid and porphobilinogen

  • Zhang J
  • Yasuda M
  • Desnick R
 et al. 
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Abstract

Accurate determinations of 5-aminolevulinic acid (ALA) and porphobilinogen (PBG) in physiologic fluids are required for the diagnosis and therapeutic monitoring of acute porphyrias. Current colorimetric methods are insensitive and over-estimate ALA and PBG due to poor specificity, while LC-MS/MS methods increase sensitivity, but have limited matrices. An LC-MS/MS method was developed to simultaneously determine ALA and PBG concentrations in fluids or tissues which were solid phase extracted, butanol derivatized, and quantitated by selective reaction monitoring using (13)C(5), (15)N-ALA and 2,4-(13)C(2)-PBG internal standards. ALA was separated from interfering compounds on a reverse phase C8-column. For ALA and PBG, the matrix effects (87.3-105%) and process efficiencies (77.6-97.8% and 37.2-41.6%, respectively) were acceptable in plasma and urine matrices. The assay was highly sensitive for ALA and PBG (LLOQ=0.05 muM with 25 muL urine or 100 muL plasma), and required approximately 4 h from extraction to results. ALA and PBG accuracy ranged from 88.2 to 110% (n=10); intra- and inter-assay coefficients of variations were

Author-supplied keywords

  • Adolescent
  • Adult
  • Aminolevulinic Acid/*blood/diagnostic use/*urine
  • Animals
  • Child
  • Child, Preschool
  • Chromatography, Liquid/*methods
  • Disease Models, Animal
  • Female
  • Humans
  • Male
  • Middle Aged
  • Porphobilinogen/*blood/diagnostic use/*urine
  • Porphyria, Acute Intermittent/blood/*diagnosis/uri
  • Sensitivity and Specificity
  • Tandem Mass Spectrometry/*methods
  • Young Adult

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Authors

  • J Zhang

  • M Yasuda

  • R J Desnick

  • M Balwani

  • D Bishop

  • C Yu

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