为了推动奶业振兴,2016年国家奶业科技创新联盟发起实施了推动奶业可持续发展的专项研究项目——优质乳工程,它涵盖了创建优质乳标识制度、推动奶牛养殖技术升级,全面实施乳制品加工工艺标准化监管3 个方面。其中优质乳标识制度提出了以活性酶类、活性蛋白和糠氨酸为核心的优质乳制品品质的评价方法。目前,在糠氨酸和乳铁蛋白检测中常用的检测方法有高效液相色谱法、液相色谱质谱联用法、毛细管电泳质谱联用法等。高效液相色谱法是最常用的检测方法,但前处理时间长且过程复杂,对样品纯度要求较高;液相色谱质谱联用法灵敏度高,但设备昂贵,对操作人员要求较高;毛细管电泳质谱法对生物大分子具有较好分离检测效果,但是毛细管电泳和质谱接口还存在一定缺陷,维护成本较高,检测重复性差。本文阐述了优质乳中糠氨酸和乳铁蛋白的检测方法,比较了方法的优缺点,旨在为进一步开发优质乳中糠氨酸和乳铁蛋白检测新方法提供参考。
In order to promote the revitalization of the dairy industry,the National Dairy Industry Science and Technology Innovation Alliance initiated and implemented a special research project to promote the sustainable development of the dairy industry -- high-quality milk Project in 2016,which covers three aspects: establishing a high quality milk labeling system,promoting the upgrading of dairy farming technology,and comprehensively implementing the standardization and supervision of dairy processing technology. The quality evaluation method of high quality dairy products based on active enzymes,active proteins and furosine is proposed in the high quality milk labeling system. At present,the commonly used detection methods for furosine and lactoferrin include high performance liquid chromatography,liquid chromatography mass spectrometry,capillary electrophoresis mass spectrometry,etc. High performance liquid chromatography is the most commonly used detection method,but the pretreatment time is long and the process is complex,and the sample purity is high. Liquid chromatography-mass spectrometry (LC-MS) is sensitive,but the equipment is expensive and the requirement for operators is high. Capillary electrophoresis mass spectrometry has a good separation and detection effect on biological macromolecules,but the interface between capillary electrophoresis and mass spectrometry still has some defects,high maintenance cost,poor detection repeatability. This paper described the detection methods of furosine and lactoferrin in high-quality milk,compared the advantages and disadvantages of the methods. The aim is to provide reference for further developing a new method for the detection of furosine and lactoferrin in high quality milk.
[1] Adeola K F,Adunni O O.Effect of storage temperature on microbial quality of infant milk[J]. Journal of Tropical Pediatrics,1998,44(1):54-55.
[2] 孙宇霞. 完善检测标准保障乳制品质量安全[J]. 中国乳业,2012(6):68-69.
[3] Burton H.Reviews of the progress of dairy science the bacterio-logical,chemical, biochemical and physical changes that occur in milk at temperatures of 100 150°C[J]. Journal of Dairy Research,1984,51(2):341-363.
[4] Seiquer I,Delgado-Andrade C,Haro A, et al.Assessing the effects of severe heat treatment of milk on calcium bioavailability in vitro and in vivo studies[J]. Journal of Dairy Science,2010, 93(12):5635-5643.
[5] ChoY H,Hong S M,Kim C H. Determination of lactulose and furosine formation in Heated milk as a milk quality indicator[J].Korean Journal for Food Science of Animal Resources,2012,32(5):540-544.
[6] Ruiz-Matute A I,Corzo-Martinez M,Montilla A,et al. Presence of mono-,di- and galactooligosaccharides in commercial lactose-free UHT dairy products[J]. Journal of Food Composition and Analysis,2012,28(2):164-169.
[7] Ho Y K,Zakrzewski S F,Mead L H.Hydrophobic interactions between the 5-alkyl group of 2,4-diamino-6-methylpyrimidines and dihydrofolate reductase[J]. Biochemistry-Us,1973,12(5):1003-1005.
[8] Suzuki T,Akiyama M,Matsui H, et al.Changes in physicochemical properties and sensory characteristics of UHT milk pasteurized by indirect and direct heating methods during storage at 10 degrees C[J]. Journal of the Japanese Society for Food Science and Technology-Nippon Shokuhin Kagaku Kogaku Kaishi,2014,61(5):199-205.
[9] Lorenzen P C,Clawin-Raedecker I,Einhoff K,et al.A survey of the quality of extended shelf life (ESL) milk in relation to HTST and UHT milk[J]. International Journal of Dairy Technology, 2011,64(2):166-178.
[10] Mayer H K,Raba B,Meier J,et al.RP-HPLC analysis of furosine and acid-soluble beta-lactoglobulin to assess the heat load of extended shelf life milk samples in Austria[J]. Dairy Science & Technology,2010,90(4):413-428.
[11] Pellegrino L,Cattaneo S,Masotti F, et al.Detection of milk powder and caseinates in Halloumi cheese[J]. Journal of Dairy Science,2010,93(8):3453-3460.
[12] 何瑛,纪坤发,杨美丰,等.高效液相色谱法测定牛奶中糠氨酸含量[J].中国乳业,2022(8):83-89.
[13] 黄文强,苏玉芳,张彩霞.乳制品中糠氨酸检测方法改进[J].食品安全导刊,2021,11(18):112-113.
[14] Troise A D,Fiore A,Wiltafsky M,et al.Quantification of Nepsilon- (2- Furoylmethyl)- L- lysine (furosine),Nepsilon- (Carboxymethyl)-L-lysine (CML), Nepsilon-(Carboxyethyl)-L-lysine(CEL) and total lysine through stable isotope dilution assay and tandem mass spectrometry[J]. Food Chemistry,2015,188:357-364.
[15] Bignardi C,Cavazza A,Corradini C,et al.Determination of furosine in food products by capillary zone electrophoresis-tandem mass spectrometry[J]. Electrophoresis,2012,33(15): 2382-2389.
[16] 龚广予,巫庆华,吴正钧,等.乳铁蛋白的检测方法——随机免疫扩散法[J].上海奶牛,2000(3):19-21.
[17] 丰东升,马颖清,陈柔含,等.乳铁蛋白双抗体夹心ELISA检测方法的建立[J].食品工业,2022,43(11):295-299.
[18] 张英华,迟玉杰,董平.酶联免疫法测定牛初乳中乳铁蛋白含量[J].中国乳品工业,1999(6):19-20,31.
[19] 郑云鹏,赵红杰,郭靖宇,等.酶联免疫法检测奶粉样本中的乳铁蛋白及其均匀度的探索[J].中国乳品工业,2019,47(6):41-44.
[20] 刘楚新. 转基因牛奶中人乳铁蛋白准确、快速检测方法的建立[D].武汉:华中农业大学,2011.
[21] 廖菁菁,谢志榕,李雅玫,等.高效液相色谱法测定婴幼儿配方奶粉中乳铁蛋白[J].中国口岸科学技术,2022,4(3):90-96.
[22] 梁政洋,黄丽,李玲,等.水牛乳中蛋白质基于RP-HPLC指纹图谱的建立及在乳源分析中的应用[J].食品科技,2019,44(8):329-334.
[23] 张敏,王瑶,苏运聪,等.不同乳制品中α-乳白蛋白、β-乳球蛋白和乳铁蛋白的测定[J].乳业科学与技术,2020,43(5):17-21.
[24] 刘宇,陈伟,史玉东,等.高效毛细管电泳法快速测定乳铁蛋白原料的纯度[J].中国乳品工业,2016,44(2):43-46.
[25] 顾媛,程利花,姜金斗,等.毛细管电泳法检测婴幼儿奶粉中乳铁蛋白的质量分数[J].中国乳品工业,2011,39(5):54-56.
[26] 孙娜娜,刘金虎,杨孟迪,等.乳及乳制品中乳铁蛋白的全自动高通量毛细管凝胶电泳检测方法研究[J].中国乳品工业,2021,49(9):52-56.