液相色谱－串联质谱检测血浆变肾上腺素和去甲变肾上腺素方法的建立

Development of a LC-MS/MS method for the simultaneous determination of metanephrine and norepinephrine in human plasma

目的：建立一种稳定的血浆变肾上腺素（ MN）和去甲变肾上腺素（ NMN）的液相色谱－串联质谱检测方法（ LC－MS／MS）。方法方法学建立。自2012年1月至12月于复旦大学附属中山医院体检中心招募126名健康人，同时在中山医院就诊的泌尿外科和内分泌科患者中入组疾病组21例（嗜铬细胞瘤17例，异位嗜铬细胞瘤4例），高血压组108例，疾病对照组84例。以同位素氘代作为内标，采用BEH HILIC色谱柱进行分离。流动相为甲酸铵缓冲液（100 mmol／L，pH3）和纯乙腈，梯度洗脱；柱温：35℃；使用Waters Xevo^TM TQ MS ACQUITY UPLC System建立方法，并对该方法的线性，回收率，精密度，最低检测下限进行性能评价。比较各组同健康人组MN和NMN水平的差异。结果该方法通过了线性，回收率，精密度和最低检测下限的性能评价。嗜铬细胞瘤组同健康人组的MN和NMN水平有显著统计学差异。结论建立的LC－MS／MS方法检测MN和NMN，适合于临床应用。（中华检验医学杂志，2015，38：605－608）

Objective The aim of our study was to develop a robust LC-MS/MS method for determination of MN and NMN in blood plasma.Methods A liquid chromatography -tandem mass spectrometric （ LC-MS/MS） method was used, with signal linearity, lower limits of quantitation, precision and accuracy being evaluated.The study recruited 126 healthy volunteers, and MN and NMN in blood plasma were determined.At the same time samples from 21 patients （ 17 pheochromocytoma, 4 ectopic pheochromocytoma） , a hypertension group of 108 persons, and a control group of 84 persons were analyzed. A paired T test was used to compare the MN and NMN levels between the different groups.Results The performance characteristics for the method in terms of linearity, lower limits of quantitation, precision and accuracy were verified.Significant differences were found between the concentration levels of MN and NMN in the diseased and healthy groups.Conclusion A robust and reliable LC-MS/MS method for the determination of MN and NMN in blood plasma has been developed and was shown to be suitable for clinical application.