采用 UPLC-MS/MS 分析血清中天冬氨酸和香草扁桃酸用于神经母细胞瘤的诊断检测。

Analysis of vanillylmandelic acid and homovanillic acid by UPLC-MS/MS in serum for diagnostic testing for neuroblastoma.

机构信息

Seattle Children's Hospital, Department of Laboratories, 4800 Sand Point Way NE, Seattle, WA 98105, United States.

出版信息

Clin Chim Acta. 2013 Sep 23;424:253-7. doi: 10.1016/j.cca.2013.06.024. Epub 2013 Jul 3.

DOI:10.1016/j.cca.2013.06.024

PMID:23830883

Abstract

BACKGROUND

Vanillylmandelic acid (VMA) and homovanillic acid (HVA) are typically measured in urine for the diagnosis and monitoring of neuroblastoma, a tumor in children <5 y. A protocol for evaluation of serum VMA and HVA has been utilized at our institution for approximately 25 y, originally validated using high performance liquid chromatography (HPLC) with an electrochemical detector. We recently validated a serum VMA/HVA method by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).

METHODS

After solvent extraction and clean up with Ultrafree centrifugal filters, samples were analyzed by UPLC-MS/MS in multiple reaction monitoring mode.

RESULTS

The assay was linear between 2 and 1000 ng/ml for VMA and HVA. Within run and run to run CVs were <5% for VMA at all levels, <10% for HVA at high levels, and <20% at low levels. Correlation with the HPLC method was acceptable with a constant bias. The reference interval for VMA by UPLC-MS/MS was determined to be ≤20 ng/ml, and HVA≤30 ng/ml. Original patient data comparing urine to serum showed diagnostic agreement >80% for both VMA and HVA.

CONCLUSION

Correlation of VMA and HVA was acceptable after adjustment of reference intervals. Collection of a single serum sample instead of 24-h urine collection saves time and improves accuracy of measurement due to difficulty of collecting a 24-h urine sample in infants and young children. UPLC-MS/MS also offers improved analyte specificity, improved signal to noise, and rapid analysis time.

摘要

背景

香草扁桃酸（VMA）和高香草酸（HVA）通常在尿液中测量，用于诊断和监测儿童<5 岁的神经母细胞瘤，这是一种肿瘤。我们机构大约 25 年来一直在使用评估血清 VMA 和 HVA 的方案，最初使用高效液相色谱法（HPLC）和电化学检测器进行验证。我们最近通过超高效液相色谱-串联质谱法（UPLC-MS/MS）验证了血清 VMA/HVA 方法。

方法

在使用 Ultrafree 离心过滤器进行溶剂提取和净化后，通过 UPLC-MS/MS 在多重反应监测模式下分析样品。

结果

VMA 和 HVA 的测定在 2 至 1000ng/ml 之间呈线性关系。在所有水平下，VMA 的日内和日间变异系数（CV）均<5%，高水平下的 HVA<10%，低水平下的 HVA<20%。与 HPLC 方法的相关性可接受，具有恒定偏差。UPLC-MS/MS 确定的 VMA 参考区间为≤20ng/ml，HVA≤30ng/ml。比较尿液和血清的原始患者数据表明，VMA 和 HVA 的诊断一致性均>80%。