超高效液相色谱-串联质谱法测定蛋白胨中6种脂溶性维生素

Determination of 6 kinds of fat-soluble vitamins in peptone using ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  脂溶性维生素  蛋白胨  超高效液相色谱-串联质谱法

英文关键词:fat-soluble vitamins  peptone  ultra performance liquid chromatography-tandem mass spectrometry

中文摘要:

      目的 建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)同时测定蛋白胨中6种脂溶性维生素的分析方法。方法 样品经甲醇-正己烷(8:2, V/V)混合溶液提取, Syncronis C18色谱柱(100 mm×2.1 mm, 1.7 μm)分离, 以0.1%(V/V)甲酸甲醇溶液(含10 mmol/L甲酸铵)为流动相进行洗脱, 流速为0.2 mL/min。采用电喷雾电离源、正离子扫描、多反应监测模式(multiple reaction monitoring, MRM)进行检测。结果 6种脂溶性维生素在各自质量浓度范围内相关系数(r2)均大于0.999, 线性关系良好, 检出限为0.026~1.109 mg/kg, 定量限为0.085~3.696 mg/kg, 相对标准偏差均<3%(n=6), 且在不同添加水平下的回收率为81.5%~114.8%。结论 该方法操作简单, 重现性好, 准确度高, 适用于蛋白胨中6种脂溶性维生素的测定。

英文摘要:

      Objective To establish a method for the simultaneous determination of 6 kinds of fat-soluble vitamins in peptone using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods The samples were extracted by methanol hexane (8:2, V:V) solution, separated by Syncronis C18 column (100 mm×2.1 mm, 1.7 μm), eluted with 0.1% (V:V) formic acid methanol solution (containing 10 mmol/L ammonium formate) as mobile phase, and the flow rate was 0.2 mL/min. Positive electrospray ionization (ESI+) and multiple reaction monitoring mode were used for detection. Results The correlation coefficients (r2) of the 6 fat-soluble vitamins in their linear ranges were greater than 0.999. The limits of detection were in the range of 0.026?1.109 mg/kg, and the limits of quantitation were 0.085?3.696 mg/kg. The relative standard deviations of the method were less than 3% (n=6), and the average recoveries were 81.5%?114.8% at different spiked levels. Conclusion This method is simple, sensitive and accurate, which is suitable for the determination of 6 fat-soluble vitamins in peptone.