固相萃取-液相色谱-串联质谱法测定动植物油脂中胆固醇
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摘要
采用固相萃取-液相色谱-串联质谱法测定动植物油脂中胆固醇的含量。样品在无水乙醇和氢氧化钾溶液中皂化,用石油醚-乙醚(1+1)混合液提取,经硅胶固相萃取柱净化。以Zorbox SB C_(18)色谱柱为分离柱,以不同体积比的5mmol·L~(-1)乙酸铵溶液和乙腈-甲醇(1+1)溶液的混合液为流动相进行梯度洗脱,采用电喷雾正离子源和多反应监测模式检测。采用内标法定量,胆固醇的质量浓度在0.1~5.0mg·L~(-1)内与其对应的峰面积呈线性关系,胆固醇在植物油脂和动物油脂中的检出限(3S/N)分别为0.02,0.2mg·kg~(-1)。在0.5,1.0,5.0mg·kg~(-1)等3个浓度水平进行加标回收试验,回收率为85.2%~92.0%,测定值的相对标准偏差(n=6)为3.6%~9.4%。
LC-MS/MS was applied to the determination of cholesterol in fats or oils of animals or vegetables with solid phase extraction.The sample was saponified by ethanol and potassium hydroxide,and extracted with mixture of petroleum ether and ethyl ether(1+1),and then purified by silica gel solid phase extraction column.Zorbox SB C_(18) chromatographic column was used as stationary phase,and the mixture of 5 mmol·L~(-1) ammonium acetate solution and acetonitrile-methanol(1+1)solution mixed in different ratios was used as mobile phase in gradient elution.ESI+and multi-reactions monitor mode were adopted in MS/MS.Internal standard method was used for quantitative.Linear relationship between values of peak area and mass concentration of cholesterol was kept in the range of 0.1-5.0 mg·L~(-1),with detection limits(3 S/N)of cholesterol in fats or oils of vegetables or fats or oils of animals of 0.02,0.2 mg·kg~(-1),respectively.Tests for recovery were made by standard addition method at the concentration levels of 0.5,1.0,5.0 mg·kg~(-1),giving values of recovery and RSD′s(n=6)in the ranges of85.2%-92.0% and 3.6%-9.4% respectively.
LC-MS/MS was applied to the determination of cholesterol in fats or oils of animals or vegetables with solid phase extraction.The sample was saponified by ethanol and potassium hydroxide,and extracted with mixture of petroleum ether and ethyl ether(1+1),and then purified by silica gel solid phase extraction column.Zorbox SB C_(18) chromatographic column was used as stationary phase,and the mixture of 5 mmol·L~(-1) ammonium acetate solution and acetonitrile-methanol(1+1)solution mixed in different ratios was used as mobile phase in gradient elution.ESI+and multi-reactions monitor mode were adopted in MS/MS.Internal standard method was used for quantitative.Linear relationship between values of peak area and mass concentration of cholesterol was kept in the range of 0.1-5.0 mg·L~(-1),with detection limits(3 S/N)of cholesterol in fats or oils of vegetables or fats or oils of animals of 0.02,0.2 mg·kg~(-1),respectively.Tests for recovery were made by standard addition method at the concentration levels of 0.5,1.0,5.0 mg·kg~(-1),giving values of recovery and RSD′s(n=6)in the ranges of85.2%-92.0% and 3.6%-9.4% respectively.
引文
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[13]彭科怀,廖应萍,赵年华.直接皂化测定猪肝中胆固醇含量[J].预防医学情报杂志,2008,24(8):663-664.
[14]陈树东,林晓佳,吴钟玲,等.固相萃取-气相色谱-质谱联用法测定植物油中的胆固醇和4种植物甾醇[J].中国油脂,2016,41(7):94-98.
[15]陈果.凝胶渗透色谱-高效液相色谱法测定食用油中胆固醇含量[J].食品与药品,2016,18(5):361-363.
[2]周永生,罗士平,孔泳.固相萃取-气相色谱-液质联用检测地沟油中胆固醇[J].色谱,2012,30(2):207-210.
[3]郭涛,杜蕾蕾,万辉,等.高效液相色谱法测定胆固醇含量鉴别地沟油[J].食品科学,2009,30(22):286-289.
[4]何文绚,杜方润,李艳霞,等.快速测定植物油胆固醇含量在地沟油筛查中的应用[J].分析化学,2015,43(3):394-398.
[5]DB34/T 1765-2012动物油脂中胆固醇含量的测定液相色谱串联质谱法[S].
[6]陈红,杨梅,朱蓉,等.超高效液相色谱三重四极杆质谱法测定火锅油、潲水油及地沟油中胆固醇含量[J].中国食品卫生杂志,2011,23(5):429-432.
[7]许秀丽,任荷玲,李娜,等.氘代同位素内标-气相色谱-质谱法测定胆固醇含量鉴别地沟油的研究[J].检验检疫科学,2012,22(2):16-23.
[8]朱蓉,陈红,杨梅,等.UPLC-MS/MS法测定常用植物油料中胆固醇含量[J].中国卫生检验杂志,2012,22(1):14-18.
[9]张丽,汤佳佳,徐国兵,等.高效液相色谱法测定牛奶中的胆固醇[J].食品科学,2012,33(2):216-218.
[10]郭玮,陈方俊,潘柏中.胆固醇检测方法的发展历史及标准化[J].检验医学,2013,28(11):970-973.
[11]盛灵慧,王晶,武利庆,等.液相色谱-质谱联用测定蛋粉中胆固醇含量[J].食品科学,2008,29(12):632-634.
[12]余雯静,郑利,沈崇玉.地沟油中胆固醇的LC/MS/MS定性定量检测[J].重庆理工大学学报(自然科学),2011,25(12):41-45.
[13]彭科怀,廖应萍,赵年华.直接皂化测定猪肝中胆固醇含量[J].预防医学情报杂志,2008,24(8):663-664.
[14]陈树东,林晓佳,吴钟玲,等.固相萃取-气相色谱-质谱联用法测定植物油中的胆固醇和4种植物甾醇[J].中国油脂,2016,41(7):94-98.
[15]陈果.凝胶渗透色谱-高效液相色谱法测定食用油中胆固醇含量[J].食品与药品,2016,18(5):361-363.