Simultaneous detection of diagnostic biomarkers of alkaptonuria, ornithine carbamoyltransferase deficiency, and neuroblastoma disease by high-performance liquid chromatography/tandem mass spectrometry

Autor: Chien-Chen Lai, Ching Ming Chen, Fuu Jen Tsai, Wei Yi Hsu
Rok vydání: 2013
Předmět:
Zdroj: Clinica Chimica Acta. 420:140-145
ISSN: 0009-8981
DOI: 10.1016/j.cca.2012.10.022
Popis: Background Urinary homovanillic acid (HVA)/vanillylmandelic acid (VMA), orotic acid (OA), and homogentisic acid (HGA) are diagnostic biomarkers of neuroblastoma, ornithine carbamoyl transferase deficiency (OCTD), and alkaptonuria (AKU), respectively. In this study, a high-performance liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed for simultaneous quantification of HVA, VMA, OA, and HGA in urine. Methods After sample preparation, which involved only the dilution procedure, samples were quantified by LC–MS/MS. Full-scan MS/MS mode enabled the urinary markers to be quantified with a high degree of specificity and sensitivity. Rather than using a separate enzymatic method to normalize the concentration of creatinine in urine, we quantified the level of creatinine in urine in one LC–MS run. Results The limits of detection were 10 μg/l for HGA, 25 μg/l for HVA/VMA, and 50 μg/l for OA with a single-to-noise ratio of 3; the limits of quantification were 50 μg/l for HVA and HGA, 100 μg/l for VMA, and 250 μg/l for OA. The linear dynamic range for quantification of the analytes covered 2 to 3 orders of magnitude, depending on the analyte. The relative standard deviation of the developed LC–MS/MS method was less than 4% for the intra-day validation and 10% for the inter-day validation. Conclusions The results show that our LC–MS/MS technique is a highly sensitive and rapid method for screening for biomarkers that are diagnostic of three metabolic diseases.
Databáze: OpenAIRE
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