蒙古族传统奶酪中8种脂溶性维生素和β-胡萝卜素高效液相色谱检测方法的建立

目的] 建立同时测定蒙古族传统奶酪中VA、VD₂、VD₃、α-生育酚、β-生育酚、γ-生育酚、δ-生育酚、VK₁共8种脂溶性维生素及天然色素β-胡萝卜素含量的高效液相色谱方法（high performance liquid chromatography,HPLC）。方法] 选取蒙古族传统奶酪为试样,经无水乙醇提取,酶解后用正己烷萃取,旋蒸浓缩,复溶过滤膜。经Agilent poroshell 120 EC-C18（150 mm×4.6 mm,4 μm）色谱柱分离,以甲醇和水（体积比为95∶5）为流动相,采用FLD和VWD检测器检测,外标法定量。结果] VA、α-生育酚、β-生育酚、γ-生育酚、δ-生育酚、VK₁、β-胡萝卜素在0.1~1.0 μg/mL浓度范围内线性关系良好,VD₂、VD₃在0.05~1.00 μg/mL浓度范围内线性关系良好,相关系数R²均大于0.998。结论] 建立的HPLC方法具有简便、快捷、灵敏度高、回收率高、分析时间短等优点,可为蒙古族传统乳制品中多种脂溶性维生素及天然色素的快速检测提供技术支撑。

Establishment of an HPLC Method for Simultaneous Determination of 8 Fat-soluble Vitamins and β-carotene in Mongolian Traditional Cheese

Objective] The present study was conducted to establish a high performance liquid chromatography （HPLC） method for simultaneous determination of 8 fat-soluble vitamins （vitamin A, vitamin D₂, vitamin D₃, α-tocopherol, β-tocopherol, γ-tocopherol, δ-tocopherol and vitamin K₁） and the natural pigment β-carotene in Mongolian traditional cheese. Method] The cheese samples were extracted with absolute ethanol and then enzymatically hydrolyzed. After extraction with n-hexane and concentration by rotary evaporator, the products were redissolved and filtered. Subsequently, they were separated by Agilent poroshell 120 EC-C18 chromatographic column （150 mm×4.6 mm, 4 μm） using the mobile phases of methanol and water （V/V=95∶5）. FLD and VWD were used as detectors, and the external reference method was used for quantification. Result] The standard curves of vitamin A, α-tocopherol, β-tocopherol, γ-tocopherol, δ-tocopherol, vitamin K₁ and β-carotene all exhibited ideal linear relation in the concentration range of 0.1-1.0 μg/mL （R²>0.998）, and those of vitamin D₂ and vitamin D₃ had good linear relation in the concentration range of 0.05-1.00 μg/mL （R²>0.998）. Conclusion] The established HPLC method has the advantages of simplicity, rapidity, high sensitivity, high recovery rate and short detection time, which provides technical support for the rapid detection of multiple fat-soluble vitamins and natural pigments in Mongolian traditional dairy products.