卫生研究

目的建立反相高效液相色谱法同时测定肉制品中维生素A、3种维生素E异构体及胆固醇的方法,并用此方法测定了几种肉制品中对应营养素的含量。方法样品经皂化和液液萃取,采用Waters Symmetry C₁₈柱(4.6 mm×250 mm, 5μm)进行分离,以甲醇-水(98∶2,V/V)为流动相。二极管阵列检测器在190～350 nm范围内进行3D扫描,选择325、294和210 nm分别作为维生素A、3种维生素E异构体以及胆固醇的特征吸收波长进行检测。以苯并[e]芘作为维生素A、E的内标定量。胆固醇采用外标法定量。结果维生素A、3种生育酚和胆固醇分别在浓度为0.18～9.00、0.76～15.2和0.11～5.50 mg/mL范围内呈良好线性关系,相关系数r>0.995;检出限分别为5、20和330μg/100 g,定量限分别为15、60和990μg/100 g;基质中3种营养素高中低3个浓度水平的测量精密度均小于7.0%、加标回收率为86.3%～105.6%(n=6)。结论该方法灵敏准确,适用于肉制品中维生素A、维生素E以及胆固醇的同时测定。

OBJECTIVE To establish a method for simultaneous analysis of vitamin A, three vitamin E isomers and cholesterol in meat products by reversed-phase high-performance liquid chromatography. This method is used to determine the contents of the corresponding nutrients in several meat products. METHODS The sample was pretreated with saponification and liquid-liquid extraction, then separated on Waters Symmetry C₁₈ column(4.6 mm×250 mm, 5 μm). The mobile phase consisted of methanol(98%) and water(2%). The analytes were detected by photo-diode array(PDA)detector. 325 nm, 294 nm, and 210 nm were selected as the characteristic absorption wavelengths of vitamin A, vitamin E, and cholesterol, respectively by scanning in the range of 190-350 nm in 3 D mode. Benzo[e]pyrene was internal standard for vitamin A and E. Cholesterol was quantified by external standard curve method. RESULTS The concentration of vitamin A, three vitamin E isomers and cholesterol showed good linearity in the range of 0.18-9.00 μg/mL、0.76-5.2 μg/mL and 0.11-5.50 mg/mL, respectively, with the correlation coefficients greater than 0.995. The limits of detections(LODs) were 5、20 and 330 μg/100 g and the limits of quantification(LOQs) for vitamin A, three vitamin E isomers and cholesterol were 15、60 and 990 μg/100 g, respectively. The recoveries at three levels in the matrix were 86.3%-105.6%, and the relative standard deviations(RSDs) were all less than 7.0%(n=6).CONCLUSION The method is sensitive, accurate and suitable for analysis of vitamin A, three vitamin E isomers and cholesterol in meat products.