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

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

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72h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—40μg／mL的范围内表现出良好的线性关系（R=0．9999），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。

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, Pcg, and HCC827 were cultured for 24, 48, and 72 h. A HILIC-MS/MS method based on the surrogate analyte of Wrosine 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.