Abstract
Neuroblastoma and other neural crest tumors can be characterized by the increased production and excretion of catecholamines and their metabolites. Homovanillic acid (HVA) and vanillylmandelic acid (VMA) are important catecholamine metabolites that can be measured to provide relatively rapid laboratory diagnosis and clinical follow-up of neuroblastoma. We present a procedure to quantify HVA and VMA in urine samples which have been diluted to a creatinine concentration of 2 mg/dL. Diluted samples are spiked with deuterated internal standards, acidified, and extracted with an organic solvent. A bis(trimethylsilyl)trifluoroacetamide (BSTFA) with 1% trimethylchlorosilane (TMCS) and pyridine mixture is added to the dried extract to create trimethylsilyl derivatives of HVA and VMA. The derivatized compounds are measured using gas chromatography–mass spectrometry (GC/MS).
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References
Swift CC, Eklund MJ, Kraveka JM, Alazraki AL (2018) Updates in diagnosis, management, and treatment of neuroblastoma. Radiographics 38(2):566–580
Williams CM, Greer M (1963) Homovanillic acid and vanilmandelic acid in diagnosis of neuroblastoma. JAMA 183:836–840
Czarnecka M, Tilan J, Kitlinska J (2012) Sympathetic neurotransmitters in neuroblastoma – between physiology and pathology. In: Neuroblastoma – present and future. Intech, pp 137–152
Strenger V, Kerbl R, Dornbusch HJ, Ladenstein R, Ambros PF, Ambros IM, Urban C (2007) Diagnostic and prognostic impact of urinary catecholamines in neuroblastoma patients. Pediatr Blood Cancer 48(5):504–509
Allenbrand R, Garg U (2010) Quantitation of homovanillic acid (HVA) and vanillylmandelic acid (VMA) in urine using gas chromatography-mass spectrometry (GC/MS). Methods Mol Biol 603:261–269
Fauler G, Leis HJ, Huber E, Schellauf C, Kerbl R, Urban C, Gleispach H (1997) Determination of homovanillic acid and vanillylmandelic acid in neuroblastoma screening by stable isotope dilution GC-MS. J Mass Spectrom 32(5):507–514
Hwang N, Chong E, Oh H, Cho HW, Lee JW, Sung KW, Lee SY (2021) Application of an LC-MS/MS method for the simultaneous quantification of homovanillic acid and vanillylmandelic acid for the diagnosis and follow-up of neuroblastoma in 357 patients. Molecules 26(11):27–33
Seviour JA, McGill AC, Dale G, Craft AW (1988) Method of measurement of urinary homovanillic acid and vanillylmandelic acid by gas chromatography-mass spectrometry suitable for neuroblastoma screening. J Chromatogr 432:273–277
Soldin SJ, Hill JG (1981) Liquid-chromatographic analysis for urinary 4-hydroxy-3-methoxymandelic acid and 4-hydroxy-3-methoxyphenylacetic acid, and its use in investigation of neural crest tumors. Clin Chem 27(3):502–503
Soldin SJ, Lam G, Pollard A, Allen LC, Logan AG (1980) High performance liquid chromatographic analysis of urinary catecholamines employing amperometric detection: references values and use in laboratory diagnosis of neural crest tumors. Clin Biochem 13(6):285–291
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Beals, M., Ramoo, B., Clinton Frazee, C., Garg, U. (2022). Quantitation of Neuroblastoma Markers Homovanillic Acid (HVA) and Vanillylmandelic Acid (VMA) in Urine by Gas Chromatography–Mass Spectrometry (GC/MS). In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Biomolecular Analysis. Methods in Molecular Biology, vol 2546. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2565-1_17
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DOI: https://doi.org/10.1007/978-1-0716-2565-1_17
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Publisher Name: Humana, New York, NY
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