高效液相色谱法测定乳制品中纳他霉素分析方法的改进
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摘要
目的优化高效液相色谱法(high performance liquid chromatography,HPLC)测定乳制品中纳他霉素含量的分析方法。方法对测定纳他霉素GB/T 21915-2008《食品中纳他霉素的测定》的高效液相色谱法中样品的提取溶剂、蛋白沉淀剂、色谱流动相及流速进行优化,并对优化后的方法进行方法学验证。结果选择10%的冰乙酸水溶液作为样品提取溶剂;乙酸锌和亚铁氰化钾作为蛋白沉淀剂;甲醇-水-冰乙酸(60:40:2,V:V:V)作为流动相;流速为1.0 mL/min。可以在7 min内完成对目标物的分离,线性方程为Y=43315X-1751,r~2=0.99997,方法平均回收率在98.5%~106.3%之间,相对标准偏差(relative standard deviation,RSD)均小于2.2%,方法检出限为0.02 mg/L。结论该方法前处理简单,回收率及灵敏度高,准确度和稳定性好,适用于对纯牛奶、乳酸菌饮料、奶粉等乳制品中纳他霉素的分离和定量检测。
Objective To optimize the method for determination of natamycin content in dairy products by high performance liquid chromatography(HPLC). Methods According to GB/T 21915-2008 Determination of namycin in food-High performance liquid chromatography, the extraction solvent, protein precipitant, chromatographic mobile phase and flow rate of the sample were optimized, and the optimized method was validated by methodology. Results The 10% glacial acetic acid aqueous solution was selected as extraction solvent of sample, methanol-water-acetic acid(60:40:2, V:V:V) was selected as the chromatographic mobile phase with zinc acetate and potassium ferricyanide as protein precipitants, and the flow rate was 1.0 mL/min. The separation of the target could complete within 7 min. The linear equation of natamycin was Y=43315 X-1751 with r~2 of 0.99997. The average recoveries were 98.5%-106.3% with the relative standard deviations less than 2.2%, and the limit of detection was 0.02 mg/L. Conclusion The proposed method is simple, accurate and precise, which is suitable for the separation and quantitative detection of natamycin content in pure milk, lactic acid bacteria beverage, powdered milk and other dairy products.
Objective To optimize the method for determination of natamycin content in dairy products by high performance liquid chromatography(HPLC). Methods According to GB/T 21915-2008 Determination of namycin in food-High performance liquid chromatography, the extraction solvent, protein precipitant, chromatographic mobile phase and flow rate of the sample were optimized, and the optimized method was validated by methodology. Results The 10% glacial acetic acid aqueous solution was selected as extraction solvent of sample, methanol-water-acetic acid(60:40:2, V:V:V) was selected as the chromatographic mobile phase with zinc acetate and potassium ferricyanide as protein precipitants, and the flow rate was 1.0 mL/min. The separation of the target could complete within 7 min. The linear equation of natamycin was Y=43315 X-1751 with r~2 of 0.99997. The average recoveries were 98.5%-106.3% with the relative standard deviations less than 2.2%, and the limit of detection was 0.02 mg/L. Conclusion The proposed method is simple, accurate and precise, which is suitable for the separation and quantitative detection of natamycin content in pure milk, lactic acid bacteria beverage, powdered milk and other dairy products.
引文
[1]范蓓,哈益明,王峰.食品中纳他霉素的国内外标准研究[J].食品工业科技,2011,(12):62-64.Fan B,Ha YM,Wang F.Studies on standards at home and abroad on of namycin in food[J].Sci Technol Food Ind,2011,(12):62-64.
[2]胡海洋,乔春明,葛菁萍,等.纳他霉素的特性和生产研究状况[J].中国现代药物应用,2009,3(2):200-201.Hu HY,Qiao CM,Ge QP,et al.Characteristics of namycin and its production[J].Chin J Mod Drug Appl,2009,3(2):200-201.
[3]高玉荣,王雪平.天然抗真菌食品生物防腐剂:纳他霉素的性质及在食品中的应用[J].农产品加工,2009,(5):7-13.Gao YR,Wang XP.Natural antifungal food preservative:the nature and application of namycin in food[J].Acad Period Farm Prod Process,2009,(5):7-13.
[4]贺家亮,张敏,赵胜娟.纳他霉素的研究现状[J].中国食品添加剂,2005,(3):30-33.He JL,Zhang,M,Zhao SJ.The research status of namycin[J].China Food Addit,2005,(3):30-33.
[5]刘红梅.纳他霉素在食品工业中的应用[J].宁夏农林科技,2012,53(11):96-97.Liu HM.Application of namycin in food industry[J].Ningxia J Agric,2012,53(11):96-97.
[6]GB 2760-2014食品安全国家标准食品添加剂使用标准[S].GB 2760-2014 National food safety standard-Standard for uses of food additives[S].
[7]GB/T 21915-2008食品中纳他霉素的测定-液相色谱法[S].GB/T 21915-2008 Determination of namycin in food-High performance liquid chromatography[S].
[8]王志强,胡国媛,李志勇,等.微生物抑制法快速检测鲜奶中多种抗生素残留[J].中国食品卫生杂志,2008,20(2):139-141.Wang ZQ,Hu GY,Li ZY,et al.Rapid detection of multiple antibiotic residues in fresh milk by microbial inhibition[J].Chin J Food Hyg,2008,20(2):139-141.
[9]周玉虎,常硕.超声萃取-紫外分光光度法测定酸奶中的纳他霉素[J].食品安全质量检测,2009,28(7):78-79.Zhou YH,Chang S.Ultrasonic extraction and ultraviolet spectrophotometry for the determination of namycin[J].J Food Saf Qual2009,28(7):78-79.
[10]Harry B.Analytical profiles of drug substances[M].New York:Academic Press,1994.
[11]周莉莉,祝建华,张卉,等.乳制品中那他霉素的超高效液相色谱-串联质谱法快速测定[J].分析测试学报,2010,(1):83-85.Zhou LL,Zhu JH,Zhang H,et al.Rapid determination of namycin in dairy products by UPLC-MS/MS[J].J Instrum Anal,2010,(1):83-85.
[12]Roberts DPT,Scotter MJ,Godula M,et al.Development of an RP-HPLC method for the simultaneous determination of benzoic acid sorbic acid,namycin and lysozyme in hard and pasta filata cheese[J].Food Chem,2011,127(3):1294-1299.
[13]朱聿元,李红梅,周露.高效液相色谱-紫外检测法测定糕点中纳他霉素的含量[J].绿色科技,2017,(20):196-197.Zhu YY,Li HM,Zhou L.High performance liquid chromatography with UV detection was used to determine the content of namycin in cakes[J].J Green Sci Tech,2017,(20):196-197.
[14]王亚君.高效液相色谱法检测运动食品中的纳他霉素[J].食品研究与开发,2017,38(9):164-167.Wang YJ.Detection of namycin in sports food by high performance liquid chromatography[J].Food Res Dev,2017,38(9):164-167.
[15]GB/T 27404-2008实验室控制规范食品理化检验[S].GB/T 27404-2008 Laboratory control specification-Food physical and chemical test[S].
[16]魏宝东,孟宪军.天然生物性食品防腐剂纳他霉素的特性及其应用[J].辽宁农业科学,2004,(2):24-25.Wei BD,Meng XJ.Characteristics and application of natural biotic food preservative,namycin[J].Liaoning Agric Sci,2004,(2):24-25.
[17]骆健美,金志华,岑沛霖,等,纳他霉素在不同溶剂中溶解度的测定与关联[J].高校化学工程学报,2008,22(5):733-737.Luo JM,Jin ZH,Cen PL,et al.Determination and correlation of solubility of namycin in different solvents[J].J Chem Eng Chin Univ,2008,22(5):733-737
[18]王丽丽.高效液相色谱法测定乳品中的纳他霉素残留[J].食品安全质量检测学报,2016,7(11):4688-4689.Wang LL.Determination of namycin residues in dairy products by high performance liquid chromatography[J].J Food Saf Qual,2016,7(11):4688-4689.
[19]闰磊,刘宁,孙亚范,等.乳制品中纳他霉素的液相色谱检测方法[J].食品研究与开发,2010,31(4):111.Run L,Liu N,Sun YF,et al.Determination of namycin in dairy products by liquid chromatography[J].Food Res Dev,2010,31(4):111.
[2]胡海洋,乔春明,葛菁萍,等.纳他霉素的特性和生产研究状况[J].中国现代药物应用,2009,3(2):200-201.Hu HY,Qiao CM,Ge QP,et al.Characteristics of namycin and its production[J].Chin J Mod Drug Appl,2009,3(2):200-201.
[3]高玉荣,王雪平.天然抗真菌食品生物防腐剂:纳他霉素的性质及在食品中的应用[J].农产品加工,2009,(5):7-13.Gao YR,Wang XP.Natural antifungal food preservative:the nature and application of namycin in food[J].Acad Period Farm Prod Process,2009,(5):7-13.
[4]贺家亮,张敏,赵胜娟.纳他霉素的研究现状[J].中国食品添加剂,2005,(3):30-33.He JL,Zhang,M,Zhao SJ.The research status of namycin[J].China Food Addit,2005,(3):30-33.
[5]刘红梅.纳他霉素在食品工业中的应用[J].宁夏农林科技,2012,53(11):96-97.Liu HM.Application of namycin in food industry[J].Ningxia J Agric,2012,53(11):96-97.
[6]GB 2760-2014食品安全国家标准食品添加剂使用标准[S].GB 2760-2014 National food safety standard-Standard for uses of food additives[S].
[7]GB/T 21915-2008食品中纳他霉素的测定-液相色谱法[S].GB/T 21915-2008 Determination of namycin in food-High performance liquid chromatography[S].
[8]王志强,胡国媛,李志勇,等.微生物抑制法快速检测鲜奶中多种抗生素残留[J].中国食品卫生杂志,2008,20(2):139-141.Wang ZQ,Hu GY,Li ZY,et al.Rapid detection of multiple antibiotic residues in fresh milk by microbial inhibition[J].Chin J Food Hyg,2008,20(2):139-141.
[9]周玉虎,常硕.超声萃取-紫外分光光度法测定酸奶中的纳他霉素[J].食品安全质量检测,2009,28(7):78-79.Zhou YH,Chang S.Ultrasonic extraction and ultraviolet spectrophotometry for the determination of namycin[J].J Food Saf Qual2009,28(7):78-79.
[10]Harry B.Analytical profiles of drug substances[M].New York:Academic Press,1994.
[11]周莉莉,祝建华,张卉,等.乳制品中那他霉素的超高效液相色谱-串联质谱法快速测定[J].分析测试学报,2010,(1):83-85.Zhou LL,Zhu JH,Zhang H,et al.Rapid determination of namycin in dairy products by UPLC-MS/MS[J].J Instrum Anal,2010,(1):83-85.
[12]Roberts DPT,Scotter MJ,Godula M,et al.Development of an RP-HPLC method for the simultaneous determination of benzoic acid sorbic acid,namycin and lysozyme in hard and pasta filata cheese[J].Food Chem,2011,127(3):1294-1299.
[13]朱聿元,李红梅,周露.高效液相色谱-紫外检测法测定糕点中纳他霉素的含量[J].绿色科技,2017,(20):196-197.Zhu YY,Li HM,Zhou L.High performance liquid chromatography with UV detection was used to determine the content of namycin in cakes[J].J Green Sci Tech,2017,(20):196-197.
[14]王亚君.高效液相色谱法检测运动食品中的纳他霉素[J].食品研究与开发,2017,38(9):164-167.Wang YJ.Detection of namycin in sports food by high performance liquid chromatography[J].Food Res Dev,2017,38(9):164-167.
[15]GB/T 27404-2008实验室控制规范食品理化检验[S].GB/T 27404-2008 Laboratory control specification-Food physical and chemical test[S].
[16]魏宝东,孟宪军.天然生物性食品防腐剂纳他霉素的特性及其应用[J].辽宁农业科学,2004,(2):24-25.Wei BD,Meng XJ.Characteristics and application of natural biotic food preservative,namycin[J].Liaoning Agric Sci,2004,(2):24-25.
[17]骆健美,金志华,岑沛霖,等,纳他霉素在不同溶剂中溶解度的测定与关联[J].高校化学工程学报,2008,22(5):733-737.Luo JM,Jin ZH,Cen PL,et al.Determination and correlation of solubility of namycin in different solvents[J].J Chem Eng Chin Univ,2008,22(5):733-737
[18]王丽丽.高效液相色谱法测定乳品中的纳他霉素残留[J].食品安全质量检测学报,2016,7(11):4688-4689.Wang LL.Determination of namycin residues in dairy products by high performance liquid chromatography[J].J Food Saf Qual,2016,7(11):4688-4689.
[19]闰磊,刘宁,孙亚范,等.乳制品中纳他霉素的液相色谱检测方法[J].食品研究与开发,2010,31(4):111.Run L,Liu N,Sun YF,et al.Determination of namycin in dairy products by liquid chromatography[J].Food Res Dev,2010,31(4):111.