考马斯亮蓝法检测不同乳中乳清蛋白含量
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摘要
目的建立考马斯亮蓝法(coomassie brilliant blue method, CCB)检测乳品中蛋白质的方法,并对几种常见乳中总蛋白质含量及乳清蛋白含量进行检测比较。方法用考马斯亮蓝G-250显色,以牛血清蛋白为对照品,采用可见分光光度法在检测波长595nm处测定各种乳中蛋白质的含量。结果该方法在0.005～0.1 mg/mL范围内与吸光度呈现良好的线性关系(r=0.9995),回收率为99.45%、相对标准偏差(relative standard deviation,RSD)为2.03%;不同种类乳中蛋白质含量存在差别,乳清蛋白占比也有很大的不同。结论考马斯亮蓝法检测蛋白,显色灵敏,具有简便,快捷,成本低等特点。各种乳蛋白检测比较中,马乳的乳清蛋白占比最接近母乳,这为马乳的进一步开发提供了新的理论基础。
Objective To establish a method for determination of protein in dairy products by coomassie brilliant blue method(CCB), and to compare the total protein content and whey protein content of several common milks. Methods Coomassie brilliant blue G-250 was used for color development, and bovine serum albumin was used as a reference. The content of protein in various milk was determined by visible spectrophotometry at the detection wavelength of 595 nm. Results The method showed a good linear relationship with absorbance in the range of 0.005-0.1 mg/mL(r=0.9995), the recovery rate was 99.45%, and the relative standard deviation(RSD) was 2.03%. There were differences in the protein content of different types of milk, and the proportion of whey protein was also very different. Conclusion The coomassie brilliant blue method is sensitive to color, which is simple, fast and low cost. In the comparison of various milk protein tests, the proportion of whey protein in horse milk is the closest to breast milk, which provides a new theoretical basis for the further development of horse milk.
Objective To establish a method for determination of protein in dairy products by coomassie brilliant blue method(CCB), and to compare the total protein content and whey protein content of several common milks. Methods Coomassie brilliant blue G-250 was used for color development, and bovine serum albumin was used as a reference. The content of protein in various milk was determined by visible spectrophotometry at the detection wavelength of 595 nm. Results The method showed a good linear relationship with absorbance in the range of 0.005-0.1 mg/mL(r=0.9995), the recovery rate was 99.45%, and the relative standard deviation(RSD) was 2.03%. There were differences in the protein content of different types of milk, and the proportion of whey protein was also very different. Conclusion The coomassie brilliant blue method is sensitive to color, which is simple, fast and low cost. In the comparison of various milk protein tests, the proportion of whey protein in horse milk is the closest to breast milk, which provides a new theoretical basis for the further development of horse milk.
引文
[1]冯昕,王吉中,尧俊英,等.考马斯亮蓝法测定乳与乳制品中蛋白质含量[J].粮食与食品工业,2010,17(3):57-59.Feng X,Wang JZ,Rao JY,et al.Determination of protein content in milk and dairy products by coomassie brilliant blue method[J].Food Ind,2010,17(3):57-59.
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[4]陆东林,刘朋龙,王生俊,等.新疆马乳产业发展初探[J].中国乳业,2012,(11):16-19.Lu DL,Liu PL,Wang SJ,et al.A preliminary study on the development of Xinjiang horse milk industry[J].China Dair Ind,2012,(11):16-19.
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[6]孙士青,王少杰,李秋顺,等.考马斯亮蓝法快速测定乳品中蛋白质含量[J].山东科学,2011,24(6):53-55.Sun SQ,Wang SJ,Li QS,et al.Rapid determination of protein content in dairy products by coomassie brilliant blue method[J].Shandong Sci,2011,24(6):53-55.
[7]陈立红.未变性乳清蛋白的测定及其对产品特性的影响[D].天津:天津科技大学,2015.Chen LH.Determination of undenatured whey protein and its effect on product characteristics[D].Tianjin:Tianjin University of Science and Technology,2015.
[8]刘立新.牛乳蛋白质检测技术的研究[D].西安:陕西科技大学,2011.Liu LX.Research on protein detection technology of milk[D].Xi’an:Shaanxi University of Science and Technology,2011.
[9]蒋大程,高珊,高海伦,等.考马斯亮蓝法测定蛋白质含量中的细节问题[J].实验科学与技术,2018,88(3):123-126.Jiang DC,Gao S,Gao HL,et al.Details of the determination of protein content by coomassie brilliant blue method[J].Exper Sci Technol,2018,88(3):123-126.
[10]赵璐,何婷,丁文欢,等.考马斯亮兰法(Bradford法)测定驼乳中蛋白质的含量[J].应用化工,2016,(12):165-167,171.Zhao L,He T,Ding WH,et al.Determination of protein content in camel milk by coomassie brilliant blue method(Bradford method)[J].Appl Chem Ind,2016,(12):165-167,171.
[11]陆东林,刘朋龙,王生俊,等.马乳的营养价值和开发利用[J].新疆畜牧业,2012,(4):4-9.Lu DL,Liu PL,Wang SJ,et al.Nutritional value and development and utilization of horse milk[J].Xinjiang Anim Husbandr,2012,(4):4-9.
[12]许家喜.蛋白质的检测方法与乳制品中蛋白含量测定[J].大学化学,2009,24(1):66-69.Xu JX.Determination of protein and determination of protein content in dairy products[J].Univ Chem,2009,24(1):66-69.
[13]赖世云.婴儿配方奶粉中乳清蛋白测定方法的研究[D].杭州:浙江工业大学,2015.Lai SY.Study on the determination method of whey protein in infant formula[D].Hangzhou:Zhejiang University of Technology,2015.
[14]Inglingstad RA,Devold TG,Eriksen EK,et al.Comparison of the digestion of caseins and whey proteins in equine,bovine,caprine and human milks by human gastrointestinal enzymes[J].Dair Sci Technol,2010,90(5):549-563.
[15]陈渝.我国奶粉产业发展初探[J].中国乳业,2007,(11):10-13.Chen Y.Preliminary study on the development of milk powder industry in China[J].China Dair Ind,2007,(11):10-13.
[16]李春强.婴儿配方奶粉中乳清蛋白替代品的开发[D].长春:东北农业大学,2008.Li CQ.Development of whey protein substitutes in infant formula[D].Changchun:Northeast Agricultural University,2008.
[2]刘正,刘亚兵,张进,等.乳及乳制品中蛋白质含量的测定[J].农业科技与信息,2014,(7):24-25.Liu Z,Liu YB,Zhang J,et al.Determination of protein content in milk and dairy products[J].Agric Sci Technol Inf,2014,(7):24-25.
[3]杨梅.不同泌乳期人乳与牛乳中非酪蛋白差异蛋白质组学研究[D].沈阳:沈阳农业大学,2017.Yang M.Differential proteomics study of non-casein in human milk and milk during different lactation[D].Shenyang:Shenyang Agricultural University,2017.
[4]陆东林,刘朋龙,王生俊,等.新疆马乳产业发展初探[J].中国乳业,2012,(11):16-19.Lu DL,Liu PL,Wang SJ,et al.A preliminary study on the development of Xinjiang horse milk industry[J].China Dair Ind,2012,(11):16-19.
[5]GB/T 5009.5-2003食品中蛋白质的测定[S].GB/T 5009.5-2003 Determination of protein in food[S].
[6]孙士青,王少杰,李秋顺,等.考马斯亮蓝法快速测定乳品中蛋白质含量[J].山东科学,2011,24(6):53-55.Sun SQ,Wang SJ,Li QS,et al.Rapid determination of protein content in dairy products by coomassie brilliant blue method[J].Shandong Sci,2011,24(6):53-55.
[7]陈立红.未变性乳清蛋白的测定及其对产品特性的影响[D].天津:天津科技大学,2015.Chen LH.Determination of undenatured whey protein and its effect on product characteristics[D].Tianjin:Tianjin University of Science and Technology,2015.
[8]刘立新.牛乳蛋白质检测技术的研究[D].西安:陕西科技大学,2011.Liu LX.Research on protein detection technology of milk[D].Xi’an:Shaanxi University of Science and Technology,2011.
[9]蒋大程,高珊,高海伦,等.考马斯亮蓝法测定蛋白质含量中的细节问题[J].实验科学与技术,2018,88(3):123-126.Jiang DC,Gao S,Gao HL,et al.Details of the determination of protein content by coomassie brilliant blue method[J].Exper Sci Technol,2018,88(3):123-126.
[10]赵璐,何婷,丁文欢,等.考马斯亮兰法(Bradford法)测定驼乳中蛋白质的含量[J].应用化工,2016,(12):165-167,171.Zhao L,He T,Ding WH,et al.Determination of protein content in camel milk by coomassie brilliant blue method(Bradford method)[J].Appl Chem Ind,2016,(12):165-167,171.
[11]陆东林,刘朋龙,王生俊,等.马乳的营养价值和开发利用[J].新疆畜牧业,2012,(4):4-9.Lu DL,Liu PL,Wang SJ,et al.Nutritional value and development and utilization of horse milk[J].Xinjiang Anim Husbandr,2012,(4):4-9.
[12]许家喜.蛋白质的检测方法与乳制品中蛋白含量测定[J].大学化学,2009,24(1):66-69.Xu JX.Determination of protein and determination of protein content in dairy products[J].Univ Chem,2009,24(1):66-69.
[13]赖世云.婴儿配方奶粉中乳清蛋白测定方法的研究[D].杭州:浙江工业大学,2015.Lai SY.Study on the determination method of whey protein in infant formula[D].Hangzhou:Zhejiang University of Technology,2015.
[14]Inglingstad RA,Devold TG,Eriksen EK,et al.Comparison of the digestion of caseins and whey proteins in equine,bovine,caprine and human milks by human gastrointestinal enzymes[J].Dair Sci Technol,2010,90(5):549-563.
[15]陈渝.我国奶粉产业发展初探[J].中国乳业,2007,(11):10-13.Chen Y.Preliminary study on the development of milk powder industry in China[J].China Dair Ind,2007,(11):10-13.
[16]李春强.婴儿配方奶粉中乳清蛋白替代品的开发[D].长春:东北农业大学,2008.Li CQ.Development of whey protein substitutes in infant formula[D].Changchun:Northeast Agricultural University,2008.