同位素稀释液相色谱串联质谱法测定血清总甘油

目的 建立同位素稀释液相色谱串联质谱测定血清总甘油的方法.方法 以13C3]-甘油作内标,用氢氧化钾异丙醇溶液水解血清甘油酯为游离甘油,将游离甘油转化为苯甲酸酯,用同位素稀释液相色谱串联质谱(LC/MS/MS)分离检测,用标准曲线法定量.结果 甘油/内标峰面积比与甘油浓度(0.565～4.517 mmol/L)线性相关系数大于0.999 9;测定不同浓度血清总甘油批内变异系数(CV)平均为0.52%(范围0.21%～2.62%),总CV平均为1.15%(范围0.62%～2.00%);分析国际和国家标准物质,测定值与认证值的偏倚小于1%(-0.20%～1.06%).结论 建立同位素稀释液相色谱串联质谱测定血清总甘油方法,方法特异、精密、准确,可望用作血清总甘油测定参考方法.

Determination of serum total glycerol by isotope dilution liquid chromatography tandem mass spectrometry

Objective To develop a method for measuring serum total glycerol by isotope dilution (ID)liquid chromatography tandem mass spectrometry(LC/MS/MS).Methods Serum samples were mixed with 13C3]-glycerol(internal standard)and treated with alcoholic potassium hydroxide to hydrolyze serum glycerides to glycerol.The natural glycerol and the internal standard were benzoylated with the Schotten-Baumann reaction and the benzoates were analyzed by LC/MS/MS.Results The correlation coefficients between the peak area ratios and glycerol concentrations were 0.999 9 and higher.The withinrun coefficients of variation(CV)for serum total glycerol analysis averaged 0.94%(ranged 0.21%～2.62%)and the total CV 1.15%(0.62%～2.00%).Results on certified reference materials(SRM 909b Level Ⅰ and LevelⅡ;GBW 09146 and GBW 09147)showed an averaged bias of 0.20%(-0.20%～1.06%).Conclusions An ID-LC/MS/MS method for serum total glycerol has been developed.The method iS highly specific and precise and may be used as a candidate reference method for serum total glycerol measurements.