Determination of Homovanillic Acid and Vanillylmandelic Acid in Neuroblastoma Screening by Stable Isotope Dilution GC-MS

✍ Scribed by Fauler, G.; Leis, H. J.; Huber, E.; Schellauf, Ch.; Kerbl, R.; Urban, Ch.; Gleispach, H.

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 397 KB
• Volume: 32
• Category: Article
• ISSN: 1076-5174
• DOI: 10.1002/(sici)1096-9888(199705)32:5<507::aid-jms503>3.0.co;2-9

✦ Synopsis

A method for the quantitative determination of homovanillic acid (HVA) and vanillylmandelic acid (VMA), two metabolites of catecholamines, is presented. The assay is based on gas chromatography/electron impact mass spectrometry. The preparation of 13C-labeled VMA from is described. Together with purchased [ 13C 6 ] vanillin deuterated HVA the 13C-labeled VMA is used as an internal standard in stable isotope dilution GC/MS. The method involves elution from soaked Ðlter-papers, determination of creatinine content, extraction of HVA and VMA from eluted and urinary samples and derivatization to the di-and tri(trimethylsilyl) derivatives, respectively. The detection limits were found to be 4.0 pg for HVA and 0.8 pg for VMA. The method was applied to the routine determination of urinary HVA and VMA in a range from 5 to 100 ng HVA and VMA per lg creatinine. The lower limits of pathological concentrations are set at 35 ng lg-1 creatinine for HVA and to 20 ng lg-1 creatinine for VMA, which are in close correlation with the values from other methods, but with the main advantage of reducing the amount of questionable or elevated results from 6.7% (high-performance liquid chromatography (HPLC) alone) to 0.9% (HPLC and GC/MS).