van Zundert Sofie K M, Griffioen Pieter H, van Rossem Lenie, Willemsen Sten P, de Rijke Yolanda B, van Schaik Ron H N, Steegers-Theunissen Régine P M, Mirzaian Mina
Department of Clinical Chemistry, Erasmus MC, University Medical Center, Rotterdam, the Netherlands.
Department of Obstetrics and Gynecology, Erasmus MC, University Medical Center, Rotterdam, the Netherlands.
Clin Chem Lab Med. 2022 Dec 5;61(3):442-451. doi: 10.1515/cclm-2022-0790. Print 2023 Feb 23.
In this study we describe the development and validation of a liquid chromatography mass spectrometry method (LC-MS/MS) to quantify five tryptophan (TRP) metabolites within the kynurenine- and serotonin pathway and apply the method to serum samples of women in the first trimester of pregnancy. A secondary aim was to investigate the correlation between body mass index (BMI) and the five analytes.
A LC-MS/MS was developed for the analysis of TRP, kynurenine (KYN), 5-hydroxytryptophan (5-HTP), hydroxytryptamine (5-HT), and 5-hydroxyindole acetic acid (5-HIAA). Serum samples (n=374) were analyzed of pregnant women (median gestational age: 8 ± 2 weeks) participating in a subcohort of the Rotterdam Periconceptional Cohort (Predict study).
The LC-MS/MS method provided satisfactory separation of the five analytes (7 min run). For all analytes R was >0.995. Within- and between-run accuracies were 72-97% and 79-104%, and the precisions were all <15% except for the between-run precisions of the low QC-samples of 5-HTP and 5-HT (both 16%). Analyte concentrations were determined in serum samples of pregnant women (median (IQR)); TRP (µmol/L): 57.5 (13.4), KYN (µmol/L): 1.4 (0.4), 5-HTP (nmol/L): 4.1 (1.2), 5-HT (nmol/L): 615 (323.1), and 5-HIAA (nmol/L): 39.9 (17.0). BMI was negatively correlated with TRP, 5-HTP, and 5-HIAA (TRP: r=-0.18, p<0.001; 5-HTP: r=-0.13, p=0.02; natural log of 5-HIAA: r=-0.11, p=0.04), and positively with KYN (r=0.11, p=0.04).
The LC-MS/MS method is able to accurately quantify kynurenine- and serotonin pathway metabolites in pregnant women, providing an opportunity to investigate the role of the TRP metabolism in the (patho)physiology of pregnancy.
在本研究中，我们描述了一种液相色谱-质谱联用方法（LC-MS/MS）的开发与验证，该方法用于定量犬尿氨酸和血清素途径中的五种色氨酸（TRP）代谢物，并将该方法应用于妊娠早期妇女的血清样本。次要目的是研究体重指数（BMI）与这五种分析物之间的相关性。
开发了一种LC-MS/MS方法用于分析TRP、犬尿氨酸（KYN）、5-羟色氨酸（5-HTP）、羟色胺（5-HT）和5-羟吲哚乙酸（5-HIAA）。对参与鹿特丹围孕期队列（预测研究）一个亚队列的孕妇（中位孕周：8±2周）的血清样本（n = 374）进行了分析。
LC-MS/MS方法对这五种分析物实现了令人满意的分离（运行时间7分钟）。所有分析物的R均>0.995。批内和批间准确度分别为72 - 97%和79 - 104%，除5-HTP和5-HT低质量控制样本的批间精密度为16%外，其余精密度均<15%。测定了孕妇血清样本中分析物的浓度（中位数（四分位间距））；TRP（μmol/L）：57.5（13.4），KYN（μmol/L）：1.4（0.4），5-HTP（nmol/L）：4.1（1.2），5-HT（nmol/L）：615（323.1），5-HIAA（nmol/L）：39.9（17.0）。BMI与TRP、5-HTP和5-HIAA呈负相关（TRP：r = -0.18，p<0.001；5-HTP：r = -0.13，p = 0.02；5-HIAA的自然对数：r = -0.11，p = 0.04），与KYN呈正相关（r = 0.11，p = 0.04）。
LC-MS/MS方法能够准确地定量孕妇体内犬尿氨酸和血清素途径的代谢物，为研究TRP代谢在妊娠（病理）生理学中的作用提供了机会。
