血清中葡萄糖的测量比对-ID-LC—MS／MS法

利用同位素稀释-液相色谱-质谱法-（ID-LC-MS／MS）准确测定血清中葡萄糖含量。以[13C6]葡萄糖为内标，用重量法准确地与血清混合，离心沉淀蛋白后在碱性条件下与1．苯基-3-甲基-5-吡唑酮反应，以ZORBAXRX-SIL色谱柱分离，以乙腈-水（体积此25：75）为流动相，使用电喷雾三重四极杆串联质谱多重反应监测模式（MRM）测定，同位素稀释的括号法进行定量。采用美国NIST的血清标准物质SRM965b进行了确证，并用该方法参加JCTLM关于血清中葡萄糖含量的国际比对，测量值与均值的相对偏差分别为1．1％（A样）．1．8％（B样），结果在等效范围内。

International Comparison on the Determination of Serum Glucose by ID-LC-MS/MS

The determination of glucose in human serum by liquid chromatography–isotope dilution–mass spectrometry was described.glucose as internal standard was added to serum samples and equilibrated with endogenous glucose.Serum proteins were removed by a precipitation with anhydrous ethanol.Serum glucose and the internal standard were then reacted with l-phenyl-3-methyl-5-pyrazolone,separated by ZORBAX RX–SIL column using isocratic mobile phase of acetonitrile–water(volum ratio was 25∶75).A tandem mass spectrometer equipped with an electrospray ionization source operated in multiple reaction monitoring(MRM) mode,and the results of determination were calculated from peak area ratios by the bracketing method based on the theory of isotope dilution analysis.The method was validated by analyzing NIST serum reference material SRM 965b.International comparision of JCTLM for glucose content in serum was carried out by the method.The relative deviation between measured value and the mean were 1.1%(A sample),1.8%(B sample).