摘要
目的观察并比较液相色谱串联质谱技术检测青少年抑郁症患者血清和尿液中色氨酸-犬尿氨酸(TRP-KP)通路代谢物的浓度。方法对该通路中主要代谢物如色氨酸(TRP)、犬尿氨酸(KYN)、犬尿酸(KYNA)和3-羟基犬尿氨酸(3-HK)采用同位素标记法进行定量分析。使用岛津C18色谱柱,0.2%甲酸水溶液和乙腈进行梯度洗脱分离,7 min内采用正离子和多反应监测模式进行检测。考察线性范围、精密度等参数。用该方法检测143例青少年抑郁症患者和110例体检健康者血液和尿液样本并比较。结果以TRP为例,血清和尿液的线性范围分别为0.54~107.84μmol/L和0.74~147.06μmol/L。批内变异系数(CV)≤6.3%,批间CV≤3.22%,实验室总CV≤6.5%。抑郁组和对照组血清和尿液检测结果表明,抑郁组KYNA、KYN和TRP水平低于对照组,而3-HK水平高于对照组,差异具有统计学意义(P<0.01)。除TRP外,其他代谢物水平在尿液中显著高于血清中,差异具有统计学意义(P<0.01)。结论与血清相比,尿液中TRP-KP的代谢物即KYN、KYNA和3-HK浓度更高,由于尿液收集过程是无创的,因此检测尿液中TRP-KP代谢物具有更大的优势。
Objective To observe and compare the concentrations of tryptophan-kynurenine pathway metabolites in serum and urine of adolescents with depression individuals using liquid chromatography-tandem mass spectrometry(LC-MS/MS).Methods The major metabolites in this pathway,such as tryptophan(TRP),kynurenine(KYN),kynurenic acid(KYNA),and 3-hydroxykynurenine(3-HK)were quantitatively analyzed using isotope-labeled internal standards.The separation was achieved using a Shimadzu C18 column with gradient elution of 0.2%aqueous acid and acetonitrile.The detection was performed within 7 minutes using positive ion mode and multiple reaction monitoring(MRM).The parameters,such as linear range and precision were evaluated to assess reliability of the method.Subsequently,this method was applied to detect and compare the results in the samples of serum and urine from 143 adolescents with depression and 110 healthy controls.Results Taking TRP as the example,the linear ranges for serum and urine were 0.54 to 107.84μmol/L and 0.74 to 147.06μmol/L,respectively.The intra-batch coefficient of variation(CV)was≤6.3%,the inter-batch CV was≤3.22%,and the total laboratory CV was≤6.5%.The results showed the KYNA,KYN and TRP levels were lower in the depression group compared to the control group,while 3-HK levels were higher in the depression group with statistical significance(P<0.01).Apart from TRP,the levels of other metabolites were significantly higher in urine than those in serum,with statistical significance(P<0.01).Conclusion Compared to the results of serum,the concentrations of TRP metabolites,including KYN,KYNA and 3-HK were higher than those in urine.The concentrations of TRP-KP metabolites in urine,i.e.,KYN,KYNA and 3-HK were higher than those in serum,and the detection of TRP-KP metabolites in urine may offer a greater advantage because the urine collection process is non-invasive.
作者
马萌萌
滕晓明
陆柳
叶致含
宗明
MA Mengmeng;TENG Xiaoming;LU Liu;YE Zhihan;ZONG Ming(Center of Reproductive Medicine,Shanghai First Maternity and Infant Health Hospital,School of Medicine,Tongji University,Shanghai 201204;Department of Clinical Laboratory,Shanghai East Hospital,School of Medicine,Tongji University,Shanghai 200120,China)
出处
《临床检验杂志》
CAS
2024年第8期561-566,共6页
Chinese Journal of Clinical Laboratory Science
基金
上海市浦东新区卫生系统学科带头人培养计划(PWRd2022-03)。
作者简介
马萌萌,1988年生,女,技师,硕士,主要从事小分子代谢物研究;通信作者:宗明,副主任技师,副教授,E-mail:zongming@tongji.edu.cn。
