液相色谱-串联质谱法测定咀嚼片中B12含量
目的 建立液相色谱-串联质谱法(LC-MS/MS)测定多种维生素咀嚼片(儿童型)中维生素B12的分析方法, 为维生素B12及其制剂的含量测定提供参考。方法 样品中的维生素B12用10 mmol/L乙酸铵超声提取, 三氯甲烷沉淀蛋白质, 以10 mmol/L乙酸铵-甲醇作为流动相经SUPELCO Discovery ?HS C18柱(4.6 mm×150 mm, 5 μm)梯度洗脱分离, 流速0.6 mL/min, 采用LC-MS/MS在正离子模式下检测, 外标法定量。结果 LC-MS/MS法在30～150 ng/mL范围内, 维生素B12的浓度和峰面积的线性良好, 相关系数r=0.993, 精密度为2.4%, 在不同的添加水平下, 方法的回收率85.9%～95.7%, 检出限为5 ng/g。结论 液相色谱-串联质谱法简单快速、灵敏度高、准确性强, 适用于保健食品中维生素B12的分析。
Objective To establish a method for determination of vitamin B12 in chewable multivitamins (childhood type) by liquid chromatography-tandem mass spectrometry(LC-MS/MS), and provide a reference for determination of vitamin B12 and its preparation. Methods Samples were ultrasonically extracted by 10 mmol/L ammonium acetate, and protein were precipitated by chloroform; then the samples were separated by SUPELCO Discovery ?HS C18 column(4.6 mm×150 mm, 5 μm)on a mobile phase of 10 mmol/L ammonium acetate-methanol as gradient elution solution, and the flow rate at 0.6 mL/min. LC-MS/MS was applied for the detection in the (ESI+)-MRM mode under the external standard method. Results The linear relationship between vitamin B12 concentration and peak area were good within 30～150 ng/mL range (r=0.993), and the precision was 2.4%; the recovery of different adding levels was 85.9%～95.7%, and the limit of quantification was 5 ng/g. Conclusion Liquid chromatography-tandem mass spectrometry method is simple, effective and sensitive, and it is applicable to the analysis of vitamin B12 in health foods.
标题：液相色谱-串联质谱法测定咀嚼片中B12含量
英文标题：Determination of vitamin B12 in chewable tablets by liquid chromatography-tandem mass spectrometry
作者：
潘拾朝 汤臣倍健股份有限公司
黄康惠 汤臣倍健股份有限公司
张喜金 汤臣倍健股份有限公司
叶少文 汤臣倍健股份有限公司
中文关键词:液相色谱-串联质谱法,维生素B12,保健食品,
英文关键词:liquid chromatography-tandem mass spectrometry,vitamin B12,health foods,
发表日期：2015-06-09
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