超高效液相色谱-串联质谱法检测2型糖尿病伴胰岛素抵抗患者血浆儿茶酚胺及其代谢物含量

The relationship between catecholamine and its metabolites determined by UPLC-MS/MS and type 2 diabetes mellitus with insulin resistance

目的应用超高效液相色谱-串联质谱法(UPLC-MS/MS)检测儿茶酚胺(CAs)及其代谢物在2型糖尿病(T2DM)伴胰岛素抵抗(IR)患者血浆中的表达水平,研究CAs及其代谢物与T2DM患者IR的相关性。方法选取2019年10月至2020年10月北部战区总医院就诊的年龄>18岁的T2DM伴IR患者(IR组)105例,同期健康对照者(CO组)95例,应用HPLC-MS/MS法检测血浆中CAs及其代谢物浓度:多巴胺(DA)、肾上腺素(E)、去甲肾上腺素(NE)、变肾上腺素(MN)、去甲变肾上腺素(NMN)、高香草酸(HVA)、香草扁桃酸(VMA)。结果应用UPLC-MS/MS检测CAs及其代谢物浓度范围在1.1~326.49 nmol/L之间线性关系良好,R值均大于0.99。各指标定量下限在0.11~0.15 nmol/L,批内和批间精密度变异系数(CV)均在15%以内,平均回收率88%~105%,符合临床检测方法学准确定量标准。IR组E、NE、MN、NMN、HVA、VMA等浓度较CO组有明显升高,差异有统计学意义(P<0.05)。IR组DA浓度与CO组差异无统计学意义(P>0.05)。结论UPLC-MS/MS法检测血浆中CAs及其代谢物具有简单、准确、特异性高等优点。T2DM伴IR患者血浆中CAs及其代谢产物明显高于正常人群。

Objective To detect catecholamines(CAs)and its metabolites in type 2 diabetes mellitus(T2DM)patients with insulin resistance(IR)by ultra-performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS).To study the correlation between CAs and its metabolites and IR in patients with T2DM.Methods One hundred and five T2DM patients with IR(IR group)and 95 healthy controls(CO group)were selected from October 2019 to October 2020 in General Hospital of Northern Theater Command.The concentrations of dopamine(DA),epinephrine(E),norepinephrine(NE),metanephrine(MN),norepinephrine(NMN),homovanillic acid(HVA)and vanillic mandelic acid(VMA)were determined by HPLC-MS/MS.Results UPLC-MS/MS was used to detect CAs and their metabolites with good linear relationship in the range from 1.1-326.49 nmol/L(R>0.99).The lower limit of quantification of each index is 0.11-0.15 nmol/L,the coefficient of variation(CV)of intra and inter day were all less than 15%and the average recovery rate was 88%-105%,which well meets the accurate quantitative standard of clinical testing methodology.The concentrations of E,NE,MN,NMN,HVA and VMA in IR group were significantly higher than that in CO group(P<0.05).The concentration of DA in IR group was higher than that in CO group,but the difference was not statistically significant(P>0.05).Conclusion UPLC-MS/MS is simple,accurate and specific for the detection of CAs and its metabolites.The levels of CAs and its metabolites in T2DM patients with IR are significantly higher than those in control people.