碱性离子液体水解-双水相胶束体系提取-超高效液相色谱法测定水产品中胆固醇的含量  

Determination of Cholesterol in Aquatic Products by UHPLC after Hydrolysis with Alkaline Ionic Liquid and Extraction with Aqueous Two-Phase Micellar System

作　　者：宋玉凤 朱俊向[3] 于丁一 张小军[2] 马剑锋 SONG Yufeng;ZHU Junxiang;YU Dingyi;ZHANG Xiaojun;MA Jianfeng(College of Food and Pharmacy,Zhejiang Ocean University,Zhoushan 316021,China;Laboratory of Aquatic Product Processing and Quality Safety,Marine Fisheries Research Institute of Zhejiang,Zhoushan 316021,China;College of Food Science and Engineering,Qingdao Agricultural University,Qingdao 266109,China;Chongqing International Certificate Testing Technology Co.,Ltd.,Chongqing 400000,China)

机构地区：[1]浙江海洋大学食品与药学学院,舟山316021 [2]浙江省海洋水产研究所水产品加工与质量安全研究室,舟山316021 [3]青岛农业大学食品科学与工程学院,青岛266109 [4]重庆艾世特检测技术服务有限公司,重庆400000

出　　处：《理化检验（化学分册）》2022年第2期152-156,共5页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

基　　金：浙江省分析测试科技计划项目(2018C37023)。

摘　　要：基于碱性离子液体水解以及双水相胶束体系(ATPMS)提取,以超高效液相色谱法(UHPLC)测定水产品中胆固醇的含量,并以分配系数为指标对ATPMS组成进行了优化。称取0.10 g肉糜状水产品,加入0.36 mol·L^(-1)氢氧化1-丁基-3-甲基咪唑盐([C_(4)mim]OH)碱性离子液体1 mL,于90℃加热30 min,冷却后以5000 r·min;转速离心5 min。取全部上清液,加入3 mL由0.5%(质量分数,下同)1-丁基-3-甲基咪唑乙酸盐[C_(4)mim]Ac离子液体和5.0%(质量分数,下同)Triton X-114组成的ATPMS[介质为柠檬酸-磷酸盐缓冲液(pH 7.0)],加热至浊点,保持3 min,取出下层相,用甲醇定容至5 mL,涡旋30 s,超声5 min。分取1 mL过0.22μm滤膜,滤液供UHPLC分析。结果显示:以由0.5%1-丁基-3-甲基咪唑咪唑盐([C_(4)mim]Im)或[C_(4)mim]Ac和5.0%Triton X-114组成的ATPMS提取胆固醇时,所得分配系数较高;胆固醇的质量浓度在5~200 mg·L^(-1)内与其对应的峰面积呈线性关系,检出限(3S/N)为7.5μg·g^(-1);由[C_(4)mim]Ac组成的ATPMS所得的回收率(84.1%~89.9%)略大于由[C_(4)mim]Im组成的ATPMS的(80.0%~83.9%),且和参考GB 5009.128-2016设置的对照组(催化剂为氢氧化钾,提取剂为甲醇)的接近,3组试验测定值的相对标准偏差(n=5)基本一致,均小于3.0%。Method for determination of cholesterol in aquatic products was proposed by UHPLC based on alkaline ionic liquid hydrolysis and aqueous two-phase micellar system(ATPMS)extraction,and the composition of ATPMS was optimised in terms of partition coefficient.Minced meat packed aquatic product of 0.10 g was taken and mixed with 0.36 mol·L^(-1) hydroxide 1-butyl-3-methylimidazole salt([C_(4)mim]OH)alkaline ionic liquid(1 mL),and the mixture was heated at 90℃ for 30 min.After cooling and centrifuging at 5000 r·min;for 5 min,the entire supernatant was taken,3 mL of ATPMS[citric acid-phosphate buffer(pH 7.0)as media]consisting of 0.5%(mass fraction,the same below)1-butyl-3-methyl imidazole acetate([C_(4)mim]Ac)and 5.0%(mass fraction,the same below)Triton X-114 was added,and the mixture was heated to the cloud point,keeping for 3 min.The lower phase was taken,diluted to 5 mL with methanol,vortexed for 30 s and sonicated for 5 min.An aliquot(1 mL)of the above solution was taken and passed through 0.22μm filter membrane and the filtrate was used for UHPLC analysis.It was shown by results that the ATPMS consisting of 0.5%[C_(4)mim]Im or[C_(4)mim]Ac and 5.0%Triton X-114 gave a high partition coefficient for extraction of cholesterol.Linear relationship between the mass concentration of cholesterol and respective peak area was kept in the range of 5-200 mg·L^(-1),with detection limit(3 S/N)of 7.5μg·g^(-1);the results(84.1%-89.9%)of recovery obtained from the ATPMS containing[C_(4)mim]Ac were slightly higher than those(80.0%-83.9%)obtained from the ATPMS containing[C_(4)mim]Im,and were similar to those obtained from the control group with reference to GB 5009.128-2016(KOH as catalyst and methanol as extractant).RSDs(n=5)of the determined values of the three groups were basically the same and less than 3.0%.

关 键 词：碱性离子液体 双水相胶束体系 超高效液相色谱法 胆固醇 水产品 

分 类 号：O657.7[理学—分析化学]