婴幼儿配方奶粉中酪蛋白磷酸肽2种检测方法的比较
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摘要
目的比较婴幼儿配方奶粉中酪蛋白磷酸肽(casein phosphopeptides,CPPs)的2种主要检测方法。方法分别采用高效液相色谱法(high performance liquid chromatography,HPLC)和高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometry, HPLC-MS/MS)检测婴幼儿配方奶粉中酪蛋白磷酸肽的含量。结果 HPLC操作简便,检出限为7.5mg/100g,日内精密度≤2.7%,满足婴幼儿配方奶粉中酪蛋白磷酸肽的检测要求;而HPLC-MS/MS检出限为1mg/100 g,日内精密度≤8.8%,较HPLC具有更好的灵敏度。结论 HPLC高效准确,实验操作简单,适合于CPPs含量较高样品的检测分析;HPLC-MS/MS灵敏度高、特异性强,适用于CPPs含量较低样品的检测。
Objective To compare 2 main methods for detecting casein phosphopeptides(CPPs) in infant formula. Methods High performance liquid chromatography(HPLC) and high performance liquid chromatographytandem mass spectrometry(HPLC-MS/MS) were used to detect casein phosphopeptides in infant formula. Results High performance liquid chromatography was easy to operate, the detection limit was 7.5 mg/100 g, and the intra-day precision was less than 2.7%, which met the requirements for the detection of casein phosphopeptides in infant formula. The detection limit of high performance liquid chromatography tandem-mass spectrometry was 1 mg/100 g, and the intra-day precision was less than 8.8%, which had better sensitivity than high performance liquid chromatography. Conclusion HPLC method is effective and accurate, which is suitable for determination of the higher-content sample of CPPs. HPLC-MS/MS method has high sensitivity and specificity, which is suitable for the determination of the lower-content samples of CPPs.
Objective To compare 2 main methods for detecting casein phosphopeptides(CPPs) in infant formula. Methods High performance liquid chromatography(HPLC) and high performance liquid chromatographytandem mass spectrometry(HPLC-MS/MS) were used to detect casein phosphopeptides in infant formula. Results High performance liquid chromatography was easy to operate, the detection limit was 7.5 mg/100 g, and the intra-day precision was less than 2.7%, which met the requirements for the detection of casein phosphopeptides in infant formula. The detection limit of high performance liquid chromatography tandem-mass spectrometry was 1 mg/100 g, and the intra-day precision was less than 8.8%, which had better sensitivity than high performance liquid chromatography. Conclusion HPLC method is effective and accurate, which is suitable for determination of the higher-content sample of CPPs. HPLC-MS/MS method has high sensitivity and specificity, which is suitable for the determination of the lower-content samples of CPPs.
引文
[1]庞广昌,朱文欣,陈庆森,等.酪蛋白磷酸肽测定方法的研究[J].食品科学,2001,22(5):49-51.Panf GC,Zhu WX,Chen QS,et al.Study on the determination method of casein phosphopeptide[J].Food Sci,2001,22(5):49-51.
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[3]Sato R,Noguchil T,Naito H,et al.Casein phosphopeptide(CPP)enhances calcium absorption from the ligated segment of rat small intestine[J].J Nutr Sci Vitaminol,1986,32(1):67-76.
[4]Scholz-Ahrens KE,Schrezenmeir J.Effects of bioactive substances in milk on mineral and trace element metabolism with special reference to casein phosphopeptides[J].Br J Nut,2000,84(1):147-153.
[5]高然,陈俊德,易瑞灶,等.酪蛋白磷酸肽金属矿物质螯合物的研究进展[J].中国食品添加剂,2015,2015(12):167-171.Gao R,Chen JD,Yi RZ,et al.Research progress of casein phosphopeptide metal mineral chelate[J].Chin Food Addit,2015,2015(12):167-171.
[6]Matsui T,Yano H,Awano T,et al.The influences of casein phosphopeptides on metabolism of ectopic bone induced by decalcified bone matrix implantation in rats[J].Nutr Sci Vitaminol,1994,40(2):137-145.
[7]Zhou C,Zhang DL,Bai YX,et al.Casein phosphopeptide-amorphous calcium phosphate remineralization of primary teeth early enamel lesions[J].J Dentistr,2014,24(1):21-29.
[8]喻明玲,林居红,胡赟,等.酪蛋白磷酸肽-无定形磷酸钙对酸蚀牙釉质显微硬度的影响[J].第三军医大学学报,2013,35(2):138-141.Yu ML,Lin JH,Hu Y,et al.Effect of casein phosphopeptide-amorphous calcium phosphate on microhardness of acid etched enamel[J].Acta Acad Med Mil Tert,2013,35(2):138-141.
[9]Nagai T,Takenake T,Mori T,et al.Effects of caffeine and casein phosphopeptides on fertilization in vitro of pig oocytes matured in culture[J].Mol Reprod Dev,1994,37(4):452-456.
[10]Otani H,Kihara Y,Park M,et al.The immunoenhancing property of a dietary casein phosphopeptide preparation in mice[J].Food Agric Immunol,2000,12(2):165-173.
[11]Peregoa S,Zabeo A,Marasco E,et al.Casein phosphopeptides modulate calcium uptake and apoptosis in Caco2 cells through their interaction with the TRPV6 calcium channel[J].J Func Food,2013,5(2):847-857.
[12]GB 14880-2012食品安全国家标准-食品营养强化剂使用标准[S].GB 14880-2012 National standards for food safety-Standards for the use of nutritive fortifiers in food[S].
[13]梁艳,王艳萍,任新志,等.一种乳制品中酪蛋白磷酸肽含量的检测方法,中国:103792298[P].2014-05-14.Liang Y,Wang YP,Ren XZ,et al.A method for detecting casein phosphopeptide content in dairy products,China:103792298[P].2014-05-14.
[14]李则均.高效液相色谱法测定婴幼儿配方羊奶粉中酪蛋白磷酸肽含量[J].中国乳业,2017,182(2):66-70.Li ZJ.Determination of casein phosphopeptides in infant formula milk powder by high performance liquid chromatography[J].Chin Dairy,2017,182(2):66-70.
[15]冯凤琴,陈闽军,杜永盛,等.高效液相色谱法测定奶粉中酪蛋白磷酸肽(CPP)含量[J].中国食品学报,2006,6(3):128-132.Feng FQ,Chen MJ,Du YS,et al.Determination of casein phosphopeptide(CPP)in milk powder by high performance liquid chromatography[J].JChin Inst Food Sci Technol 2006,6(3):128-132.
[16]赖奕坚,陈闵军,刘玉敏,等.LC-MS法测定钙片中酪蛋白磷酸肽含量[J].食品科学,2006,27(12):672-674.Lai YJ,Chen MJ,Liu YM,et al.Determination of casein phosphopeptide content in calcium tablets by LC-MS[J].Food Sci,2006,27(12):672-674.
[17]胡蓓,张京顺,柯星,等.高效液相色谱串联质谱法测定婴幼儿配方奶粉中酪蛋白磷酸肽含量[J].食品科学,http://www.spkx.net.cn/CN/Y/V/I/0.Hu B,Zhang JS,Ke X,et al.Determination of casein phosphopeptides in infant formula by LC-MS/MS[J].Food Sci,http://www.spkx.net.cn/CN/Y/V/I/0.
[18]Dupas C,Adt I,Cottaz A,et al.A chromatographic procedure for semi-quantitative evaluation of caseinphosphopeptides in cheese[J].Dairy Sci Technol,2009,89(6):519-529
[2]Hansen M,Sandstrom B,Lonnerdal B,et al.The effect of casein phosphopeptide on zinc and calcium absorption from high phytate infant diets assessed in rat pups and Caco2 cells[J].Int Pediatr Res Found,1996,40(4):547-552.
[3]Sato R,Noguchil T,Naito H,et al.Casein phosphopeptide(CPP)enhances calcium absorption from the ligated segment of rat small intestine[J].J Nutr Sci Vitaminol,1986,32(1):67-76.
[4]Scholz-Ahrens KE,Schrezenmeir J.Effects of bioactive substances in milk on mineral and trace element metabolism with special reference to casein phosphopeptides[J].Br J Nut,2000,84(1):147-153.
[5]高然,陈俊德,易瑞灶,等.酪蛋白磷酸肽金属矿物质螯合物的研究进展[J].中国食品添加剂,2015,2015(12):167-171.Gao R,Chen JD,Yi RZ,et al.Research progress of casein phosphopeptide metal mineral chelate[J].Chin Food Addit,2015,2015(12):167-171.
[6]Matsui T,Yano H,Awano T,et al.The influences of casein phosphopeptides on metabolism of ectopic bone induced by decalcified bone matrix implantation in rats[J].Nutr Sci Vitaminol,1994,40(2):137-145.
[7]Zhou C,Zhang DL,Bai YX,et al.Casein phosphopeptide-amorphous calcium phosphate remineralization of primary teeth early enamel lesions[J].J Dentistr,2014,24(1):21-29.
[8]喻明玲,林居红,胡赟,等.酪蛋白磷酸肽-无定形磷酸钙对酸蚀牙釉质显微硬度的影响[J].第三军医大学学报,2013,35(2):138-141.Yu ML,Lin JH,Hu Y,et al.Effect of casein phosphopeptide-amorphous calcium phosphate on microhardness of acid etched enamel[J].Acta Acad Med Mil Tert,2013,35(2):138-141.
[9]Nagai T,Takenake T,Mori T,et al.Effects of caffeine and casein phosphopeptides on fertilization in vitro of pig oocytes matured in culture[J].Mol Reprod Dev,1994,37(4):452-456.
[10]Otani H,Kihara Y,Park M,et al.The immunoenhancing property of a dietary casein phosphopeptide preparation in mice[J].Food Agric Immunol,2000,12(2):165-173.
[11]Peregoa S,Zabeo A,Marasco E,et al.Casein phosphopeptides modulate calcium uptake and apoptosis in Caco2 cells through their interaction with the TRPV6 calcium channel[J].J Func Food,2013,5(2):847-857.
[12]GB 14880-2012食品安全国家标准-食品营养强化剂使用标准[S].GB 14880-2012 National standards for food safety-Standards for the use of nutritive fortifiers in food[S].
[13]梁艳,王艳萍,任新志,等.一种乳制品中酪蛋白磷酸肽含量的检测方法,中国:103792298[P].2014-05-14.Liang Y,Wang YP,Ren XZ,et al.A method for detecting casein phosphopeptide content in dairy products,China:103792298[P].2014-05-14.
[14]李则均.高效液相色谱法测定婴幼儿配方羊奶粉中酪蛋白磷酸肽含量[J].中国乳业,2017,182(2):66-70.Li ZJ.Determination of casein phosphopeptides in infant formula milk powder by high performance liquid chromatography[J].Chin Dairy,2017,182(2):66-70.
[15]冯凤琴,陈闽军,杜永盛,等.高效液相色谱法测定奶粉中酪蛋白磷酸肽(CPP)含量[J].中国食品学报,2006,6(3):128-132.Feng FQ,Chen MJ,Du YS,et al.Determination of casein phosphopeptide(CPP)in milk powder by high performance liquid chromatography[J].JChin Inst Food Sci Technol 2006,6(3):128-132.
[16]赖奕坚,陈闵军,刘玉敏,等.LC-MS法测定钙片中酪蛋白磷酸肽含量[J].食品科学,2006,27(12):672-674.Lai YJ,Chen MJ,Liu YM,et al.Determination of casein phosphopeptide content in calcium tablets by LC-MS[J].Food Sci,2006,27(12):672-674.
[17]胡蓓,张京顺,柯星,等.高效液相色谱串联质谱法测定婴幼儿配方奶粉中酪蛋白磷酸肽含量[J].食品科学,http://www.spkx.net.cn/CN/Y/V/I/0.Hu B,Zhang JS,Ke X,et al.Determination of casein phosphopeptides in infant formula by LC-MS/MS[J].Food Sci,http://www.spkx.net.cn/CN/Y/V/I/0.
[18]Dupas C,Adt I,Cottaz A,et al.A chromatographic procedure for semi-quantitative evaluation of caseinphosphopeptides in cheese[J].Dairy Sci Technol,2009,89(6):519-529