Determination of 37 Amino Acids in Serum by Liquid Chromatography-tandem Mass Spectrometry

ZHAO Yun-xia; LI Zi-ying; Chen Xin; ZHANG Yu-xin; XIAO Wei-min; LIU Ai-ping; LI Mei-fang; JIN Yi-bao; DU Qing-ping; WANG Qi; SU Jia-ting; YANG Guo-wu; FANG Wen

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Determination of 37 Amino Acids in Serum by Liquid Chromatography-tandem Mass Spectrometry

更新时间：2025-06-04

- Determination of 37 Amino Acids in Serum by Liquid Chromatography-tandem Mass Spectrometry
- Journal of Instrumental Analysis (2025)
- 作者机构：
  
  1.贵州医科大学医学检验学院临床生物化学教研室，贵州贵阳550004
  2.深圳市计量质量检测研究院，广东深圳518131
  3.深圳深检集团医学检验实验室，广东深圳518131
  4.广东工业大学生态环境与资源学院，广东广州510006
  6.深圳市宝安区公共卫生服务中心，广东深圳518108
  6.深圳市药品检验研究院，广东深圳518057
- 作者简介：
- 基金信息：
- DOI：
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ZHAO Yun-xia, LI Zi-ying, Chen Xin, ZHANG Yu-xin, XIAO Wei-min, LIU Ai-ping, LI Mei-fang, JIN Yi-bao, DU Qing-ping, WANG Qi, SU Jia-ting, YANG Guo-wu, FANG Wen. Determination of 37 Amino Acids in Serum by Liquid Chromatography-tandem Mass Spectrometry[J/OL]. Journal of instrumental analysis, 2025.
DOI：

ZHAO Yun-xia, LI Zi-ying, Chen Xin, ZHANG Yu-xin, XIAO Wei-min, LIU Ai-ping, LI Mei-fang, JIN Yi-bao, DU Qing-ping, WANG Qi, SU Jia-ting, YANG Guo-wu, FANG Wen. Determination of 37 Amino Acids in Serum by Liquid Chromatography-tandem Mass Spectrometry[J/OL]. Journal of instrumental analysis, 2025. DOI：

摘要

本研究建立了一种基于液相色谱-串联质谱法（LC-MS/MS）定量血清中37种氨基酸的方法。样本经过10%磺基水杨酸沉淀蛋白处理后，使用Phenomenex Kinetex F5（250 mm × 4.6 mm，5 μm）色谱柱分离，以0.02%甲酸水和乙腈作为流动相梯度洗脱，在电喷雾离子源正离子（ESI）和多反应监测模式（MRM）下检测。对检测方法进行方法学验证，结果表明，37种氨基酸在检出限至1 000 ng/mL范围内线性良好，相关系数均大于0.999，各氨基酸的保留时间在4.60~9.16 min之间，检出限在0.2~10 ng/mL之间，定量限在0.5~20 ng/mL，不同加标浓度的平均回收率在83.21~117.57%之间，相对标准偏差在0.85~9.39%之间。应用该方法检测49例血清样本发现，鹅肌肽与胱硫醚未检出，同型半胱氨酸检出率为10.20%，其余34种氨基酸的检出率均大于90%。该方法具有高的准确性、灵敏度和特异性，可用于血清样本中的氨基酸含量测定，为与氨基酸代谢紊乱相关的疾病提供更全面的检测数据。

Abstract

This study developed a method for the determination of 37 amino acids in serum using liquid chromatography-tandem mass spectrometry. Serum sample was treated with 10% sulfosalicylic acid to achieve protein precipitation. The sepernatant was chromatographically separated using a Phenomenex Kinetex F5 (250 mm × 4.6 mm

5 μm) column with a gradient elution consisting of 0.02% formic acid in water and acetonitrile. Detected under electrospray ion source positive ion (ESI) and multiple reaction monitoring mode (MRM). The results showed that the developed method exhibited good sensitivity and linearity for the selected 37 amino acids within the concentration range from the limit of detection to 1

000 ng/mL

with correlation coefficients exceeding 0.999. The retention time for individual amino acid was ranged from 4.60-9.16 min

while the LOD and limit of quantification was in the ranges of 0.2-10 ng/mL and 0.5-20 ng/mL

respectively. The recovery rate across different spike levels was 83.21-117.57%

and the relative standard deviation varied between 0.85-9.39%. The method was applied to quantify 37 amino acids in 49 serum samples. The results indicated that

with the exception of Anserine and Cystathionine

which were not detected

and Homocysteine

which had a detection rate of 10.20%

the detection rate of the other 34 amino acids were greater than 90%. This method demonstrates good detection performance and can be used for the determination of amino acid in serum samples

providing more comprehensive data for diseases related to amino acid metabolism disorders.

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