title: Direct and simultaneous quantitation of 5-aminolaevulinic acid and porphobilinogen in human serum or plasma by hydrophilic interaction liquid chromatography-atmospheric pressure chemical ionization/tandem mass spectrometry.
abstract: Serum/plasma concentrations of 5-aminolaevulinic acid (ALA) and porphobilinogen (PBG) are elevated in patients with acute hepatic porphyrias, especially during acute attacks. Current assays require lengthy sample pre-treatment and derivatization steps. We report here a rapid, sensitive and specific hydrophilic interaction liquid chromatography-tandem mass spectrometry method for the direct and simultaneous quantitation of ALA and PBG in serum or plasma following simple protein precipitation with acetonitrile and centrifugation prior to injection. ALA and PBG were detected using selected reaction monitoring mode, following positive atmospheric pressure chemical ionization. Calibration was linear from 0.05 to 50 µmol/L for ALA and PBG. For both analytes, imprecision (relative standard deviation) was <13% and accuracy (percentage nominal concentrations) was between 92 and 107%. The method was successfully applied to the measurement of ALA and PBG in serum or plasma samples for the screening, biochemical diagnosis and treatment monitoring of patients with acute hepatic porphyrias.
authors.affiliation: Clinical Biochemistry, King's College Hospital, Denmark Hill, London, UK.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Benton
authors.forename: Lewis
authors.initials: L
authors.lastname: Couchman
authors.forename: Joanne T
authors.initials: JT
authors.lastname: Marsden
authors.forename: David C
authors.initials: DC
authors.lastname: Rees
authors.forename: Caje
authors.initials: C
authors.lastname: Moniz
authors.forename: Chang Kee
authors.initials: CK
authors.lastname: Lim
mesh_terms: D000622:Aminolevulinic Acid / Q000097:blood* / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D016018:Least-Squares Analysis; D011162:Porphobilinogen / Q000097:blood* / Q000737:chemistry; D017118:Porphyria, Acute Intermittent / Q000097:blood; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D011162:Porphobilinogen; D000622:Aminolevulinic Acid