蔬菜中胡萝卜素测定方法的改良  

Optimization for determination methods of α-,β- carotene in vegetables

作　　者：曹梦锦 张雪松[1] 王晓婧[1] 沈湘[1] 王竹[1] 

机构地区：[1]中国疾病预防控制中心营养与健康所,北京100050

出　　处：《卫生研究》2016年第3期477-482,共6页Journal of Hygiene Research

基　　金：科技部十二五科技支撑项目(No.2012BAD33B01);卫生标委会:国家食品安全标准食品中β-胡萝卜素的测定

摘　　要：目的为有效分离α-、β-胡萝卜素,对胡萝卜素测定方法进行改良。方法样品均质后,采用乙醇60℃浸提,53℃皂化,石油醚萃取,水洗,干燥,浓缩。采用YMC Carotenoid C_(30)色谱柱(3μm,4.6 mm×150 mm),流动相A(甲醇-乙腈-水,73.5∶24.5∶2)、流动相B(甲基叔丁基醚),进行梯度洗脱。检测波长450 nm,高效液相色谱法测定。结果分离α-、β-胡萝卜素及其顺反异构体。多因素方差分析显示,皂化前采用乙醇加热浸提对α-、β-胡萝卜素的提取效果最佳。α-及全反式-β-胡萝卜素分别在0.495~9.90μg/m L、0.552~10.4μg/m L范围内呈良好的线性关系,r^2均大于0.999;加标回收率为81.6%~93.3%,精密度<10%(n=6);当称样量为5 g时,检出限为0.5μg/100 g,定量限为1.5μg/100 g。结论该法有效提高了胡萝卜素的检测技术,灵敏度和准确度较高,可用于蔬菜中α-及β-胡萝卜素含量的测定。Objective To optimize the determination method of carotene in order to separate the alpha and beta carotene. Methods The homogenized samples were extracted with ethanol before saponification,and then extracted with petroleum ether followed by wash、dry and concentration. At last, the separation was performed on the high performance liquid chromatographic with YMC Carotenoid C（30） column（ 3 μm,4. 6 mm ×150 mm）. The mobile phase was consisted of A（ methanol ∶ acetonitrile ∶ water = 73. 5 ∶24. 5∶ 2） and B（ methyl tert-butyl ether）,with a linear gradient elution. The detection wavelength was 450 nm. Results The C（30）-HPLC column was used throughout the experiment exhibiting a significant benefit for the separation of carotenes and isomers.Multifactor analysis of variance showed that heating by the anhydrous ethyl alcohol before the saponification was more suitable for extracting the carotenoids from vegetables. Under the optimal condition,the linear range of α-carotene was 0. 495- 9. 90 μg /mL,and alltrans-β-carotene was 0. 552- 10. 4 μg / m L,with the linear correlation coefficient were above 0. 999. The relative standard deviation for the alpha and beta carotene were less than 10%（ n = 6）. The average spike recoveries for alpha and beta carotene in vegetableswere 81. 6%- 93. 3%. The detection and quantification limits for alpha and beta carotene were 0. 5 μg /100 g and 1. 5 μg /100 g,respectively,when taking 5g samples. Conclusion The method is accurate,reliable,and fit for analyzing α-carotene and β- carotene in fresh vegetables.

关 键 词：α-胡萝卜素 Β-胡萝卜素 C30色谱柱 高效液相色谱法 

分 类 号：R151.3[医药卫生—营养与食品卫生学]