柱前衍生化结合UHPLC-MS/MS技术测定抑郁症患者血浆中4种色氨酸代谢物的价值

Determination of four tryptophan metabolites in plasma of patients with depression by pre-column derivatization combined with UHPLC-MS/MS

目的建立抑郁症患者血浆中色氨酸、5-羟色胺、5-羟基吲哚乙酸及犬尿氨酸的UHLC-MS/MS分析方法,探讨重症抑郁患者血浆中色氨酸及其代谢物的变化情况。方法将23例重症抑郁患者设为研究组,12例健康体检者设为对照组。采集两组血浆,以咖啡酸为内标,苯甲酰氯作为衍生化试剂,将离心后血清注入LC-MS/MS检测,比较两组血浆中色氨酸及其代谢物浓度。结果色氨酸、5-羟色胺、5-羟基吲哚乙酸及犬尿氨酸的线性关系良好,方法精密度及准确度符合规定,提取回收率较为稳定,符合生物样品分析要求。研究组5-羟色胺水平显著低于对照组(P<0.05),5-羟基吲哚乙酸和犬尿氨酸水平显著高于对照组(P<0.05或0.01)。结论UHLC-MS/MS检测方法可以简便、快速、有效评价重症抑郁患者的疾病严重程度。

Objective To establish a UHLC-MS/MS method for determination of tryptophan, 5-hydroxytryptamine, 5-hydroxyindole acetic acid and kynurenine in human plasma and investigate the changes of tryptophan and its metabolites in plasma of patients with severe depression. Methods 23 cases of patients with severe depression were included in the study group, and 12 healthy subjects were selected as the control group. Collect the plasma of the two groups, using caffeic acid as internal standard and benzoyl chloride as derivatization reagent. After centrifugation, serum was injected into LC-MS/MS for detection. The concentrations of tryptophan and its metabolites in plasma were compared between the two groups. Results The linear relationship among TRP, 5-HT, 5-HIAA and KYN was good. The precision and accuracy of the method were in compliance with the regulations. The extraction recovery rate was stable and meet the requirements of biological sample analysis. The 5-HT level of the study group was significantly lower than that of the control group (P<0.05), and the levels of 5-HIAA and KYN were significantly higher than those of the control group (F<0. 05 or 0. 01). Conclusion The UHLC-MS/MS method can be used to evaluate the severity of disease in patients with severe depression easily, quickly and effectively.