一种 3 分钟 UPLC-MS/MS 法同时测定血浆儿茶酚胺及其代谢物：方法验证和诊断效率。
A 3-min UPLC-MS/MS method for the simultaneous determination of plasma catecholamines and their metabolites: Method verification and diagnostic efficiency.
机构信息
Department of Laboratory Medicine, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu 610072, Sichuan, China.
School of Medicine, University of Electronic Science and Technology of China, Chengdu 610054, Sichuan, China.
出版信息
Clin Biochem. 2021 Jan;87:67-73. doi: 10.1016/j.clinbiochem.2020.10.009. Epub 2020 Oct 21.
OBJECTIVE
To verify a rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the quantification of catecholamines and their metabolites, and to validate its efficiency for the diagnosis of phaeochromocytomas and paragangliomas (PPGLs).
METHODS
Plasma samples were pretreated with solid-phase extraction, followed by a 3-min UPLC-MS/MS analysis to quantify epinephrine (E), norepinephrine (NE), dopamine (DA), metanephrine (MN), normetanephrine (NMN) and 3-methoxytyramine (3-MT), simultaneously. The UPLC-MS/MS method was comprehensively verified and its diagnostic efficiency on PPGLs was tested using 7 PPGLs and 408 non-PPGLs patient plasma samples.
RESULTS
Using the developed method, the limit of detections (LODs) of the 6 analytes ranged from 0.0002 nmol/L (MN) to 0.0250 nmol/L (NE), while the lower limit of measuring intervals (LLMIs) ranged from 0.05 nmol/L (E, MN and NMN) to 0.10 nmol/L (NE and DA). The reportable ranges were 0.05-30.00 nmol/L for E, MN and NMN, 0.10-30.00 nmol/L for NE and DA, 1.00-300.00 pg/mL for 3-MT. No significant matrix effect was detected after correcting using internal standard. Besides, intra-day and inter-day precision were also within acceptance criteria with coefficient of variations (CVs) ≤ 15% and recoveries ranged from 95% to 115% for all the 6 analytes. The carryover effect was lower than 10%. Its diagnostic efficiency for PPGLs was significantly increased, the areas under the receiver operating characteristic (ROC) curves were increased from 68.7% to 89.1% (using E, NE and DA) to 75.2%-99.9% (using MN, NMN and 3-MT).
CONCLUSION
This study verified a rapid UPLC-MS/MS method for the determination of catecholamines and their metabolites in human plasma. It showed high diagnostic efficiency and will serve as an important tool to avoid the risk for missing patients with PPGLs.
目的
验证一种用于定量检测儿茶酚胺及其代谢物的超高效液相色谱-串联质谱（UPLC-MS/MS）方法的快速灵敏性，并验证其在嗜铬细胞瘤和副神经节瘤（PPGLs）诊断中的有效性。
方法
采用固相萃取法预处理血浆样品，然后进行 3 分钟 UPLC-MS/MS 分析，同时定量检测肾上腺素（E）、去甲肾上腺素（NE）、多巴胺（DA）、甲氧基肾上腺素（MN）、去甲甲氧基肾上腺素（NMN）和 3-甲氧基酪胺（3-MT）。综合验证 UPLC-MS/MS 方法，并采用 7 例 PPGLs 和 408 例非 PPGLs 患者的血浆样本检测其对 PPGLs 的诊断效率。
结果
采用所建立的方法，6 种分析物的检测限（LOD）范围为 0.0002nmol/L（MN）至 0.0250nmol/L（NE），而测量下限（LLMI）范围为 0.05nmol/L（E、MN 和 NMN）至 0.10nmol/L（NE 和 DA）。报告范围为 0.05-30.00nmol/L 用于 E、MN 和 NMN，0.10-30.00nmol/L 用于 NE 和 DA，1.00-300.00pg/mL 用于 3-MT。用内标校正后，未检测到明显的基质效应。此外，所有 6 种分析物的日内和日间精密度均符合要求，变异系数（CV）≤15%，回收率在 95%-115%之间。交叉污染效应低于 10%。其对 PPGLs 的诊断效率显著提高，ROC 曲线下面积从 68.7%增加到 89.1%（使用 E、NE 和 DA）至 75.2%-99.9%（使用 MN、NMN 和 3-MT）。
结论
本研究验证了一种用于检测人血浆中儿茶酚胺及其代谢物的快速 UPLC-MS/MS 方法。该方法具有较高的诊断效率，将成为避免漏诊 PPGLs 患者风险的重要工具。
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