微乳液相色谱法同时分离7种水溶性维生素

Simultaneous separation of seven water-soluble vitamins with microemulsion liquid chromatography

采用微乳液相色谱法同时分离7种水溶性维生素(V_(B1)、V_(B2)、V_(B6)、VB_(12)、叶酸、烟酰胺和VC)。考察了微乳流动相体系中表面活性剂、油相、助表面活性剂的种类以及流动相的pH值、柱温等对水溶性维生素分离的影响。优化后微乳体系的组成为:十二烷基硫酸钠(SDS)/聚氧乙烯月桂醇醚(Brij35)/正丁醇/乙酸乙酯/水(质量比为2∶60∶66∶8∶864)。色谱柱为Agilent TC C18(250 mm×4.6 mm,5μm),柱温为30℃,检测波长为254 nm,流速为0.5mL/min。7种水溶性维生素在20 min内达到基线分离。在4~36 mg/L范围内,7种水溶性维生素的质量浓度与峰面积的相关系数均大于0.999 1。不同添加水平下,V_(B1)、V_(B2)、V_(B6)、VC和烟酰胺的平均回收率为93.9%~102.9%。该方法可用于食品和药品中的多种水溶性维生素的分离、鉴别及快速测定。

A novel method was developed for the separation of seven water. soluble vitamins (V-B1, V-B2, V-B6, V-B12, V-C, folic acid and nicotinamide) with microemulsion liquid chromatography. The impacts of following factors on the separation selectivity were investigated: the type of surfactant, the type of oil phase, the type of co-surfactant, the pH of mobile phase and the column temperature. The optimal conditions for the separation of the seven water. soluble vita. mins were as follows: Agilent TC C18 (250 mmx4..6 mm, 5 mu m) was used at 30 degrees C; the micro. emulsion mobile phase was sodium dodecyl sulfate (SDS) / polyoxyethylene lauryl ether (Brij35) / n-butanol / ethyl acetate / water (2 : 60 : 66 : 80 : 864, mass ratio). Baseline separation of the seven water. soluble vitamins was achieved within 20 min with a flow rate of 0..5 mL/min and the detection wavelength of 254 nm. The calibration curve showed a good linearity with the correlation coefficients above 0.999 1 in the linear range from 4 to 36 mg/L. The mean recoveries of VB1, VB2, VB6, VC and nicotinamide were 93.9% - 102.9% at different spiked levels. The optimized and validated method can be used for simultaneous determination of water. soluble vitamins in food and pharmaceuticals.