利用超效能液相層析－－串聯式質譜儀檢測血漿游離態metanephrines用於嗜鉻細胞瘤的診斷

Measurement of Plasma Free Metanephrines by a Ultra-Performance Liquid Chromatography Tandem Mass Spectrometry for Diagnosis of Pheochromocytoma

定量血漿游離態metanephrines被證實為診斷嗜鉻細胞瘤的高敏感度生化檢驗。我們建立以超效能液相層析-串聯式質譜儀(UPLC-MS/MS)定量血漿游離態metanephrines。以96孔盤固相萃取管柱移除血漿蛋白，親水性層析管柱分離metanephrines，以正電電噴灑方式進入UPLC-MS/MS，加上多重反應監控模式偵測可定量normetanephrine及metanephrine，分析範圍分別為0.25~34.21及0.10~32.67 nmol/L。準確度由回收率評估，分別為103%及104%。精密度而言inter-和intra-assay的變異系數皆小於7.2%。本研究建立了一套以UPLC-MS/MS檢測血漿游離態metanephrines的方法可應用於嗜鉻細胞瘤的診斷。

Monitoring plasma free metanephrines has been proven as a highly sensitive biochemical test for the diagnosis of pheochromocytoma. We have developed and validated a ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to determine plasma free metanephrines. Solid phase extraction in a 96-well plate format was used to remove plasma proteins. Metanephrines were separated by hydrophilic interaction chromatography. Detection was performed in multiple-reaction monitoring mode using a tandem mass spectrometry with positive electrospray ionization. The method was validated across the clinically relevant concentration range of 0.25-34.21 and 0.10-32.67 nmol/L for normetanephrine and metanephrine, respectively. Accuracy was evaluated by assessing recovery. The recovery averaged 103 percent for normetanephrine and 104 percent for metanephrine. The intra-assay and inter-assay coefficients of variation were below 7.2 percent for both analytes. In short, we have developed a UPLC-MS/MS method for the detection of plasma free metanephrines to assist diagnosis of pheochromocytoma.