上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

基于替代分析物的亲水作用色谱串联质谱法定量检测细胞培养液中酪氨酸含量

李文彬， 董江锴*，孟爽，雷绘敏，王聪慧，沈瑛，唐亚斌，朱亮   

  1. 上海交通大学 基础医学院药理学教研室， 上海 200025
  • 出版日期:2016-07-28 发布日期:2016-08-31
  • 作者简介:李文彬（1992—）， 女， 硕士生; 电子信箱: liwenbin9282@126.com。*为并列第一作者。
  • 基金资助:

    国家自然科学基金（81173084, 81573018）；上海市科委基金（15YF1406700）

Surrogate analyte-based quantitative measurement of the tyrosine level in cell culture media by HILIC-MS/MS

LI Wen-bin, DONG Jiang-kai*, MENG Shuang, LEI Hui-min, WANG Cong-hui, SHEN Ying, TANG Ya-bin, ZHU Liang   

  1. Department of Pharmacology, Basic Medicine Faculty of Shanghai Jiao Tong University, Shanghai 200025, China
  • Online:2016-07-28 Published:2016-08-31
  • Supported by:

    National Natural Science Foundation of China, 81173084, 81573018; Foundation of Shanghai Municipal Science and Technology Oommission, 15YF1406700

PDF (PC)

1718

摘要：

目的 通过准确测定细胞培养液中酪氨酸含量，无损地动态监测肺癌细胞系相较于非肿瘤细胞系的异常酪氨酸代谢。方法 收集16HBE、PC9和HCC827各细胞系培养24、48和72 h后的细胞培养液样本，通过建立基于酪氨酸替代分析物的HILICMS/MS方法, 定量检测不同时间点各细胞培养液中的酪氨酸含量。结果 该方法在1~40 μg/mL的范围内表现出良好的线性关系（R=0.999 9），相对回收率为93.02%~102.53%，重复性好（3.91%~8.69%）。结论 该方法适用于细胞培养液中酪氨酸的测定，可用于观察肺癌细胞系的特异性酪氨酸代谢。

关键词: 酪氨酸, 替代分析物, 亲水作用色谱串联质谱法, 细胞培养液

Abstract:

Objective To non-invasively and dynamically monitor abnormal tyrosine metabolism in lung cancer cell lines by accurately measuring the tyrosine level in cell culture media. Methods The cell culture media samples were collected after cell lines 16HBE, PC9, and HCC827 were cultured for 24, 48, and 72 h. A HILIC-MS/MS method based on the surrogate analyte of tyrosine was developed to quantitatively measure the tyrosine level in cell culture media at different time points. Results This method showed a good linear correlation within the concentration range of 1-40 μg/mL (R=0.999 9) with relative recovery rates of 93.02%-102.53% and good repeatability (3.91%-8.69%). Conclusion This method is suitable for measuring the tyrosine level in cell culture media and can be used to observe specific tyrosine metabolism in lung cancer cell lines.

Key words: tyrosine, surrogate analyte, HILIC-MS/MS, cell culture media