用于尿液中高香草酸和香草扁桃酸的液相色谱-串联质谱法的验证及其在神经母细胞瘤诊断中的应用
Validation of an LC-MS/MS method for urinary homovanillic and vanillylmandelic ACIDS and application to the diagnosis of neuroblastoma.
作者信息
Rossi Lucilla, Matser Yvette A H, Barco Sebastiano, Cafaro Alessia, Pigliasco Federica, Biondi Margherita, Mancin Fabrizio, van der Ham Maria, de Sain-van der Velden Monique G M, Ash Shifra, Popovic Maja Beck, van Kuilenburg André B P, Conte Massimo, Garaventa Alberto, Tytgat Godelieve A M, Cangemi Giuliana
机构信息
Biochemistry, Pharmacology and Newborn Screening Unit, Central Laboratory of Analysis, IRCCS Istituto Giannina Gaslini, Genoa, Italy.
Department of Chemical Sciences - DiSC, University of Padua, Padua, Italy.
出版信息
J Mass Spectrom Adv Clin Lab. 2025 Apr 22;36:73-81. doi: 10.1016/j.jmsacl.2025.04.007. eCollection 2025 Apr.
BACKGROUND
Urinary catecholamine metabolites are well-established biomarkers for neuroblastoma (NB). Homovanillic acid (HVA) and vanillylmandelic acid (VMA) are the most frequently measured metabolites within SIOPEN - Catecholamine Working Group laboratories. Here, we evaluated the performance of a new LC-MS/MS in vitro diagnostic (IVD) kit for HVA and VMA to facilitate inter-laboratory harmonization.
METHODS
HVA and VMA and their deuterated internal standards were analyzed with a commercial method, on a ThermoFisher Quantiva LC-MS/MS. Validation was performed first using internal quality control and external quality assessment (IQC and EQA) samples. Next by clinical validation on 120 samples, previously tested by HPLC-ECD. Finally, 36 samples were exchanged between SIOPEN reference laboratories and analyzed by three methods.
RESULTS
Using QCs and EQA the method was validated in a wide calibration range (4.61-830 µmol/L for HVA and 4.44-800 µmol/L for VMA). Intra-day CVs ( = 5) were 7 and 8 % for HVA and 5 and 6 % for VMA for QC low and QC high, respectively; Inter-day CV% were 7 and 3 % for HVA and 2 and 7 % for VMA at QC low and QC high, respectively. Its application to 120 clinical samples confirmed a high diagnostic accuracy. The inter-laboratory quality control assessment showed interchangeable results ( 0,73 and 0.15 for HVA and VMA, respectively).
CONCLUSION
The LC-MS/MS IVD method could be considered a useful tool for clinical laboratories involved in the measurement of catecholamines, contributing to harmonization efforts.
背景
尿儿茶酚胺代谢产物是神经母细胞瘤（NB）公认的生物标志物。高香草酸（HVA）和香草扁桃酸（VMA）是SIOPEN - 儿茶酚胺工作组实验室中最常检测的代谢产物。在此，我们评估了一种用于HVA和VMA的新型液相色谱 - 串联质谱（LC-MS/MS）体外诊断（IVD）试剂盒的性能，以促进实验室间的一致性。
方法
使用商业方法在ThermoFisher Quantiva LC-MS/MS上分析HVA、VMA及其氘代内标。首先使用内部质量控制和外部质量评估（IQC和EQA）样本进行验证。接下来对120个先前通过高效液相色谱 - 电化学检测（HPLC-ECD）检测的样本进行临床验证。最后，在SIOPEN参考实验室之间交换了36个样本，并通过三种方法进行分析。
结果
使用质量控制（QC）和EQA样本，该方法在较宽的校准范围内得到验证（HVA为4.61 - 830 μmol/L，VMA为4.44 - 800 μmol/L）。低QC和高QC时，日内变异系数（CV，n = 5）对于HVA分别为7%和8%，对于VMA分别为5%和6%；日间CV%对于HVA在低QC和高QC时分别为7%和3%，对于VMA分别为2%和7%。将其应用于120个临床样本证实了高诊断准确性。实验室间质量控制评估显示结果具有互换性（HVA和VMA的组内相关系数分别为0.73和0.15）。
结论
LC-MS/MS IVD方法可被视为参与儿茶酚胺测量的临床实验室的有用工具，有助于协调相关工作。
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