靶向超高效液相色谱-串联质谱联用方法同时测定结直肠癌患者血浆中32种氨基酸含量

引用本文: 杨阳, 高守红, 张凤, 仲人前, 王志鹏, 陈万生. 靶向超高效液相色谱-串联质谱联用方法同时测定结直肠癌患者血浆中32种氨基酸含量[J]. 分析化学, 2022, 50(7): 1083-1092. doi: 10.19756/j.issn.0253-3820.211255 shu

Citation: YANG Yang, GAO Shou-Hong, ZHANG Feng, ZHONG Ren-Qian, WANG Zhi-Peng, CHEN Wan-Sheng. Simultaneous Determination of 32 Kinds of Amino Acids in Plasma from Colorectal Cancer Patients Based on Targeted Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry[J]. Chinese Journal of Analytical Chemistry, 2022, 50(7): 1083-1092. doi: 10.19756/j.issn.0253-3820.211255 shu

靶向超高效液相色谱-串联质谱联用方法同时测定结直肠癌患者血浆中32种氨基酸含量

杨阳 ^1,2, ,
高守红 ^2, ,
张凤 ^2, ,
仲人前 ^3, ,
王志鹏 ^{2,
,} ,
陈万生 ^{2,
,}

1.
徐州医科大学附属淮海医院/中国人民解放军陆军第七十一集团军医院药剂科, 徐州 221004;
2.
中国人民解放军海军军医大学附属长征医院医疗保障中心药剂科, 上海 200003;
3.
中国人民解放军海军军医大学附属长征医院临床实验诊断科, 上海 200003

通讯作者: 王志鹏,E-mail:wangzhipeng@smmu.edu.cn; 陈万生,E-mail:chenwansheng@smmu.edu.cn

基金项目:
科技部国际科技合作专项项目(No.2015DFA31810)、上海市科学技术委员会科研基金项目(No.17411972400)、中国人民解放军陆军科技英才培养工程项目和徐州市医学青年后备人才工程项目资助。

摘要: 建立了靶向超高效液相色谱-串联质谱联用方法(UHPLC-MS/MS)用于同时测定结直肠癌患者血浆中32种氨基酸含量。采用Agilent Zorbax SB-C₁₈色谱柱(150 mm×3.0 mm,5 μm)、二元梯度洗脱系统(流动相A为0.02%七氟丁酸-0.2%甲酸溶液,流动相B为甲醇)进行色谱分离,运行时间为10 min。采用蛋白沉淀法进行样品前处理,选择多反应监测模式,以电喷雾电离正离子模式获取数据。32种氨基酸和3种内标氨基酸的提取回收率为50.4%~182.4%,基质效应为41.0%~136.4%;32种氨基酸在其相应线性范围内的线性相关系数均大于0.99,日内准确度和日间准确度分别为-14.3%~13.7%(RSD≤8.8%)和-14.4%~13.4%(RSD≤12.2%),不同条件下的稳定性偏差均在±15%之内。应用本方法同时测定了6名结直肠癌患者血浆中32种氨基酸含量。

关键词:

English

Simultaneous Determination of 32 Kinds of Amino Acids in Plasma from Colorectal Cancer Patients Based on Targeted Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

1.
Department of Pharmacy, the Affiliated Huaihai Hospital of Xuzhou Medical University, the 71st Group Army Hospital of CPLA Army, Xuzhou 221004, China;
2.
Department of Pharmacy, Medical Service Center, the Affiliated Changzheng Hospital, Naval Medical University of CPLA, Shanghai 200003, China;
3.
Department of Clinical Laboratory Diagnostics, the Affiliated Changzheng Hospital, Naval Medical University of CPLA, Shanghai 200003, China

Corresponding author: WANG Zhi-Peng, ; CHEN Wan-Sheng,

Received Date: 27 March 2021
Revised Date: 31 March 2022
Fund Project:
Supported by the International Science and Technology Cooperation and Communication Special Fund of China (No.2015DFA31810), the Scientific Research Fund of Shanghai Science and Technology Committee (No.17411972400), the Science and Technology Talent Development Project of CPLA Army, and the Xuzhou Medical Young Talent Project.

Abstract: A targeted ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for simultaneous determination of 32 kinds of amino acids in plasma from colorectal cancer (CRC) patients. The chromatographic separation was performed on an Agilent Zorbax SB-C₁₈ column (150 mm×3.0 mm, 5 μm) with a binary gradient elution system (Mobile phase A:0.02% heptafluorobutyric acid and 0.2% formic acid in water; mobile phase B:methanol). The run time was 10 min. The multiple reaction monitoring mode was chosen with an electrospray ionization source operating in the positive ionization mode for data acquisition. The sample was pretreated based on protein precipitation. The average recoveries and matrix effects for 32 kinds of amino acids and 3 kinds of internal standards were 50.4%-182.4% and 41.0%-136.4%, respectively. The linear correlation coefficients were above 0.99 for 32 kinds amino acids. The intra- and inter-day accuracy for 32 kinds of amino acids ranged from -14.3% to 13.7% (RSD ≤ 8.8%) and from -14.4% to 13.4% (RSD ≤ 12.2%), respectively, and the deviations of stability under different conditions were within ±15%. This method was successfully utilized to quantify 32 kinds of amino acids in plasma samples from 6 CRC patients simultaneously.

Key words:

Amino acid
/ Colorectal cancer
/ Plasma
/ Ultra-high performance liquid chromatography-tandem mass spectrometry

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

靶向超高效液相色谱-串联质谱联用方法同时测定结直肠癌患者血浆中32种氨基酸含量

通讯作者: 王志鹏,E-mail:wangzhipeng@smmu.edu.cn; 陈万生,E-mail:chenwansheng@smmu.edu.cn

English

Simultaneous Determination of 32 Kinds of Amino Acids in Plasma from Colorectal Cancer Patients Based on Targeted Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

Corresponding author: WANG Zhi-Peng, ; CHEN Wan-Sheng,

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

靶向超高效液相色谱-串联质谱联用方法同时测定结直肠癌患者血浆中32种氨基酸含量

通讯作者: 王志鹏,E-mail:wangzhipeng@smmu.edu.cn; 陈万生,E-mail:chenwansheng@smmu.edu.cn

English

Simultaneous Determination of 32 Kinds of Amino Acids in Plasma from Colorectal Cancer Patients Based on Targeted Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

Corresponding author: WANG Zhi-Peng, ; CHEN Wan-Sheng,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

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