高效液相色谱-质谱法同时测定婴幼儿配方奶粉中叶酸、VB_(12)和生物素

Simultaneous Determination of Folic Acid,Vitamin B_(12) and Biotin in Infant Formula by HPLC-MS-MS

建立高效液相色谱．质谱法对婴幼儿配方奶粉中叶酸、VB12和生物素同时定性和定量分析方法。采用等电点沉淀（pH1．90／4．70）法提取样品，SB．Aq（2．1mm×100mm，1．8μm）色谱柱分离，0．1％（V／n甲酸溶液和乙腈作流动相，等度（16．5：83．5，V/V）洗脱，质谱MRM检测。结果表明：叶酸、VB12和生物素在5min内得到很好的分离。线性范围分别为3．5-250ng／mL、0．8～5．0ng／mL、0．7-50ng／mL，加样回收率平均值分别为97．8％、102．3％、96．9％，RSD分别为1．33％、0．9％、1．37％。该方法具有较好的精密度和准确性、灵敏度高、操作简单快速，可以应用于婴幼儿配方奶粉中叶酸、VB12和生物素的含量同时测定。

A method using high performance liquid chromatography tandem mass spectrometry （HPLC-MS-MS） for simultaneous determination of folic acid, vitamin B12 and biotin in infant milk power was presented. Samples were sequentially extracted and then subjected to isoelectric precipitation （pH 1.90/4.70）. The analysis was carried out on an SB-Aq column （2.1 mm × 100 mm, 1.8 μm） with isocratic elution using a mobile phase made up of 0.1% （V/V） aqueous formic acid and acetonitrile （16.5：83.5, V/V）. The analytes were detected in the MRM mode. Folic acid, vitamin B12 and biotin in samples could be separately well within 5 min. The calibration curves were linear over the concentration range of 3.5 - 250 ng/mL for folic acid, 0.8 - 5 ng/mL for vitamin Bt2, and 0.7 - 50 ng/mL for biotin. The average recovery rates were 97.8%, 102.3% and 96.9%, with RSD of 1.33%, 0.9% and 1.37% for folic acid, vitamin B12 and biotin, respectively. The developed method is precise, accurate, sensitive, fast and simple, which can thereforebe applied for the analysis of folic acid, vitamin BI2 and biotin in infant formula.