摘要
目的 基于脂肪酶对脂肪的酶解作用,构建了一种室温皂化的前处理方式,将植物油中结合态的甾醇酯高效游离出来,利用气相色谱-串联质谱法(gas chromatography-tandem mass spectrometry,GC-MS/MS)建立植物油中的菜籽甾醇、β-谷甾醇、芸薹甾醇、豆甾醇、环阿屯醇5种甾醇的检测方法。方法 植物油经脂肪酶酶解、碳酸钾-乙醇溶液皂化后将甾醇游离出来,利用正己烷萃取,采用HP-5MS毛细管色谱柱(30 m×0.25 mm,0.25μm)分离,GC-MS/MS多反应监测(multiple reaction monitoring,MRM)模式进行检测外标法定量。结果 在弱极性色谱柱HP-5MS上5种甾醇色谱行为良好;缓冲溶液pH为9.0、脂肪酶用量为0.05 g、酶解20 min时酶解效果最好,室温条件进行皂化,正己烷作为萃取溶剂,β-谷甾醇及其他4种甾醇的线性范围分别为0.05~50.00 mg/L和0.05~20.00 mg/L,相关系数均大于0.999,方法的加标回收率在84.7%~101.6%之间,相对标准偏差为1.4%~4.1%,5种甾醇的检出限为3.0 mg/kg,定量限为10.0 mg/kg。结论 本研究开发的方法简便快速,便于植物油中甾醇的快速测定。
Objective To develop a pretreatment method of saponification at room temperature based on the enzymatic hydrolysis of lipase on fat to effectively separate the bound sterol esters from vegetable oil, and develop the determination methods of 5 kinds of phytosterols(namely brassicasterol, campesterol, stigmasterol, β-sitosterol and cycloartenol) in vegetable oils by gas chromatography-tandem mass spectrometry(GC-MS/MS). Methods The oil samples were hydrolyzed by lipase, saponified using potassium carbonate-ethanol system, the sterols were extracted by n-hexane, separated by HP-5MS capillary column(30 m×0.25 mm, 0.25 μm), detected by GC-MS/MS with multiple reaction monitoring(MRM) mode, and quantified by external standard method. Results The chromatographic behavior of 5 kinds of phytosterols was good on HP-5MS. When pH of buffer solution was 9.0,lipase dosage was 0.05 g and enzymatic hydrolysis was 20 min, the enzymatic hydrolysis effect was the best,saponification was carried out at room temperature, n-hexane was used as extraction solvent, the linear range of β-sitosterol and the other 4 kinds of sterols were 0.05-50.00 mg/L and 0.05-20.00 mg/L, respectively. The correlation coefficients were all greater than 0.999. The spiked recoveries ranged from 84.7%-101.6% with relative standard deviations of 1.4%-4.1%. The limits of detection and limits of quantitation for the 5 kinds of phytosterols were 3.0and 10.0 mg/kg, respectively. Conclusion The method developed in this study is simple and convenient for the rapid determination of phytosterols in vegetable oil.
作者
李洁
李玲
王艳丽
李海霞
鞠香
刘艳明
胡梅
崔玉花
张卉
LI Jie;LI Ling;WANG Yan-Li;LI Hai-Xia;JU Xiang;LIU Yan-Ming;HU Mei;CUI Yu-Hua;ZHANG Hui(Shandong Institute for Food and Drug Control,Key Laboratory of Supervising Technology for Meat and Meat Products for State Market Regulation,Shandong Research Center of Engineering and Technology for Quality Control of Food for Special Medical Purposes,Shandong Research Center of Engineering and Technology for Safety Inspection of Food and Drug,Jinan 250101,China)
出处
《食品安全质量检测学报》
CAS
北大核心
2023年第4期24-31,共8页
Journal of Food Safety and Quality
基金
山东省食品药品检验研究院院级科研课题项目(SDIFDC-KY-2021013)。
关键词
气相色谱-串联质谱法
甾醇
酶解
皂化
植物油
gas chromatography-tandem mass spectrometry
phytosterols
enzymolysis
saponification
vegetable oils
