高效液相色谱法同时测定植物油中角鲨烯、生育酚和甾醇烯  

Simultaneous Determination of Squalene, Tocopherols and Steradienes in Vegetable Oils by HPLC

作　　者：徐向华 张欣[2] 于瑞祥[3] 方晓明[4] 丁卓平[2] 

机构地区：[1]中国检验认证集团上海有限公司检测中心,上海201201 [2]上海海洋大学食品学院,上海201306 [3]上海市计量测试技术研究院,上海201203 [4]上海出入境检验检疫局,上海200135

出　　处：《食品科学》2015年第16期141-147,共7页Food Science

基　　金：国家质检总局科研项目(2012IK184);上海市科委工程中心建设项目(11DZ2280300);上海市技术性贸易措施应对专项(12TBT010)

摘　　要：建立高效液相色谱法同时测定植物油中角鲨烯、4种生育酚和4种甾醇烯类化合物的方法。样品经正己烷提取、硅胶柱净化,首次采用C30柱（250 mm×4.6 mm i.d.,5μm）同时分离9种组分,流动相为乙腈-叔丁基甲醚（70∶30,V/V）溶液和水,梯度洗脱,流速1.0 m L/min。紫外检测器和荧光检测器串联使用,角鲨烯和甾醇烯的紫外检测波长分别为210 nm和235 nm,生育酚的荧光检测激发波长为290 nm,发射波长为340 nm。结果表明：9种化合物在其线性范围（生育酚为0.05~100 mg/L、角鲨烯为5~500 mg/L、甾醇烯为0.02~1.0 mg/L）内的相关系数均大于0.999,在3种添加水平（生育酚为1、100、500 mg/kg,角鲨烯为10、100、500 mg/kg,甾醇烯为0.075、0.15、0.50 mg/kg）时其平均回收率为85.5%~103.8%,相对标准偏差小于10%。方法的检出限分别为：生育酚0.03 mg/kg、角鲨烯3 mg/kg、甾醇烯0.012 mg/kg;定量限分别为：生育酚0.1 mg/kg、角鲨烯10 mg/kg、甾醇烯0.04 mg/kg。本方法灵敏、准确、可靠,已用于植物油中9种脂溶性化合物的同时检测。A method for the simultaneous determination of squalene, four tocopherols and four steradienes in vegetable oils by high-performance liquid chromatography（HPLC） coupled with UV detector and fluorescence detector was developed. The sample was dissolved in n-hexane and cleaned up on a silica gel column. Nine analytes were first separated on a C30 column（250 mm × 4.6 mm i.d., 5 μm） by gradient elution, at 25 ℃, with a mobile phase consisting of acetonitrile/methyl-tert-butyl ether（70：30, V/V） and water at a flow rate of 1.0 m L/min. The wavelength of UV detection was set at 210 nm for squalene and 235 nm for steradienes, respectively. The excitation and emission wavelengths of fluorescence detection were set at 290 nm and 340 nm for tocopherols. The method showed linear response for concentrations in the range of 0.05–100.0 mg/L for tocopherols, 5–500 mg/L for squalene, and 0.02–1.0 mg/L for steradienes, and the correlation coefficients（γ） were greater than 0.999. The average recovery rates of nine analytes in vegetable oils ranged from 85.5% to 103.8% at three spiked levels of 1, 100 and 500 mg/kg for tocopherols, 10, 100 and 500 mg/kg for squalene, and 0.075, 0.15 and 0.50 mg/kg for steradienes, and the relative standard deviations（RSDs） were less than 10%. The limits of detection（LOD） were 0.03 mg/kg for tocopherols, 3 mg/kg for squalene, and 0.012 mg/kg for steradienes. The limits of quantification（LOQ） were 0.1 mg/kg for tocopherols, 10 mg/kg for squalene, and 0.04 mg/kg for steradienes. The proposed method is sensitive, accurate and reliable, and has been successfully applied to real sample analysis.

关 键 词：角鲨烯 生育酚 甾醇烯 高效液相色谱 植物油 

分 类 号：O657.7[理学—分析化学]