中国乳业 ›› 2022, Vol. 0 ›› Issue (9): 95-101.doi: 10.12377/1671-4393.22.09.19

• 乳品加工 • 上一篇    下一篇

乳及乳制品中牛乳铁蛋白含量测定方法比较研究

陈柔含, 韩奕奕, 马颖清, 刘洋, 丰东升*   

  1. 上海市农产品质量安全中心曁农业农村部食品质量监督检验测试中心（上海）,上海 201708
  • 出版日期:2022-09-25 发布日期:2022-09-30
  • 通讯作者: *丰东升（1966-）,男,江苏徐州人,硕士,高级农艺师,研究方向为农产品质量安全监管。
  • 作者简介:陈柔含（1996-）,女,浙江台州人,硕士,助理畜牧师,研究方向为食品营养品质检测技术研究;韩奕奕（1972-）,女,上海人,硕士,正高级畜牧师,研究方向为食品营养品质检测技术研究;马颖清（1984-）,女,上海人,硕士,畜牧师,研究方向为食品营养品质检测技术研究;刘洋（1994-）,女,河北承德人,硕士,助理畜牧师,研究方向为食品营养品质检测技术研究。
  • 基金资助:
    上海市科委项目“乳及乳制品中乳铁蛋白等活性物质检测技术研究及其应用”（19391902700）

Comparative Study on Determination Methods of Bovine Lactoferrin in Dairy Products

CHEN Rouhan, HAN Yiyi, MA Yingqing, LIU Yang, FENG Dongsheng*   

  1. Shanghai Center of Agri-Products Quality and Safety,Food Quality Supervision and Testing Center of the Ministry of Agriculture（Shanghai）,Shanghai 201708
  • Online:2022-09-25 Published:2022-09-30

PDF (PC)

183

摘要： 目的 探讨两种不同检测方法测定乳与乳制品中牛乳铁蛋白的含量及应用研究。方法 对比高效液相色谱（HPLC）法和超高效液相色谱串联质谱（UPLC- MS/MS）法测定生鲜乳、调制乳、超高温瞬时灭菌乳、含乳饮料和奶粉样品等乳及乳制品中牛乳铁蛋白的含量,并对测定结果进一步比较和分析原因。结果 两种测定方法均具有较高回收率和较优的重现性,但两者方法对实际样品的测定结果具有一定差异性。结论 HPLC法测定牛乳铁蛋白是通过肝素亲和柱纯化富集乳铁蛋白,测定的是非热变性牛乳铁蛋白的含量;而UPLC-MS/MS法测定的是热变性和非热变性牛乳铁蛋白的含量,不受热处理加工工艺的影响。实验室可针对不同检测目的选择不同方法进行定量研究。

关键词: 牛乳铁蛋白, 液相色谱, 质谱, 乳及乳制品

Abstract: [Objective] To investigate two different methods for the determination of lactoferrin in dairy products. [Method] The content of lactoferrin in raw milk modified milk,UHT milk,milk beverage and milk powder were determined by High performance liquid chromatography（HPLC）and Ultra high performance liquid chromatography tandem mass spectrometry（UPLC-MS/MS）,and further analyzed the reasons for the results. [Result] The two methods had high recovery and good reproducibility. However,there were some differences between the two methods. [Conclusion] The determination of lactoferrin by HPLC is to purify and enrich lactoferrin by heparin affinity column,and the content of non heat denatured lactoferrin is determined;UPLC-MS/MS method is used to determine the content of heat denatured and non heat denatured lactoferrin,which is not affected by the processing technology of heat treatment. Therefore,the laboratory can choose different methods for quantitative research for different testing purposes.

Key words: bovine lactoferrin, liquid chromatography, mass spectrometry, dairy products

[1] Caccavo D,Pellegrino N M,Altamura M,et al.Antimicrobial and immunoregulatory functions of lactoferrin and its potential therapeutic application[J]. Journal of Endotoxin Reshearch,2002,8(6):403-417.
[2] Rachman A B,Maheswari R R A,Bachroem M S . Composition and isolation of lactoferrin from colostrum and milk of various goat breeds[J]. Procedia Food Science,2015,3:200-210.
[3] Fernández-Menéndez S,Peixoto R R A,Fernández-Colomer B,et al. Effect of holder pasteurisation on total concentrations and iron-binding profiles of holo -lactoferrin used as fortifier in donor human milk[J]. International Dairy Journal,2020,100:104564.
[4] 吴春香,刘志楠,赵源,等. 酶联免疫吸附法快速检测婴儿配方奶粉中乳铁蛋白含量[J]. 食品安全质量检测学报,2018,9(4):803-807.
[5] 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.
[6] 朱丹丹,洪辉,吴红宇,等. 高效液相色谱法测定特殊医学用途婴幼儿配方食品中乳铁蛋白含量[J]. 现代食品,2020(18):190-193.
[7] Yao X,Bunt C,Cornish J,et al.Improved RP-HPLC method for determination of bovine lactoferrin and its proteolytic degradation in simulated gastrointestinal fluids[J]. Biomedical Chromatography:BMC,2013,27(2):197-202.
[8] Zhang Y,Lou F,Wu W,et al.Determination of Bovine Lactoferrin in Food by HPLC with a Heparin Affinity Column for Sample Preparation[J]. Journal of AOAC International,2017,100(1):133-138.
[9] Chen M X,Wen F,Zhang Y D,et al.Determination of native lactoferrin in milk by HPLC on HiTrapTM Heparin HP column[J]. Food Analysis Methods,2019,12:2518-2526.
[10] 邢志恩,王军,戴蕴青,等.SDS-PAGE-薄层扫描联用法测定3种不同来源的乳铁蛋白[J]. 食品科学,2011,16:274-278.
[11] Li J,Ding X,Chen Y,et al.Determination of bovine lactoferrin in infant formula by capillary electrophoresis with ultraviolet detection[J]. Journal f Chromatography A,2012,1244:178-183.
[12] 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.
[13] Zhu C,Lia L S,Yang G,et al.High-efficiency selection of aptamers for bovine lactoferrin by capillary electrophoresis and its aptasensor application in milk powder[J]. Talanta,2019,205:120088.
[14] 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.
[15] Zhang J S,Lai S Y,Cai Z X,et al.Determination of bovine lactoferrin in dairy products by ultra-high performance liquid chromatography-tandem mass spectrometry based on tryptic signature peptides employing an isotope-labeled winged peptide as internal standard[J]. Analytica Chimica Acta,2014,829:33-39.
[16] Yuan M M,Feng C,Wang S Y,et all. Selection of possible signature peptides for the detection of bovine lactoferrin in infant formulas by LC-MS/MS[J]. PLoS One. 2017,12(9):1-11.
[17] Chen H, Shi P, Fan F, et al.Complementation of UPLC-Q-TOF-MS and CESI-Q-TOF-MS on identification and determination of peptides from bovine lactoferrin[J]. Journal of Chromatography B,2018,1084:150-157.
[18] Ounis W B, Gauthier S F, Turgeon S L, et al.Separation of minor protein components from whey protein isolates by heparin affinity chromatography[J]. International Dairy Journal, 2008, 18(10-11): 1043-1050.
[19] 孙姗姗,罗娇依,李刚,等.不同地域来源乳粉及乳制品中乳铁蛋白含量分析[J].中国食物与营养,2019,25(8):20-25.
[20] Vreeke G J C,Lubbers W,Vincken J P,et al. A method to identify and quantify the complete peptide composition in protein hydrolysates[J]. Analytica Chimica Acta,2022,1201:339616.
[21] 郑仕剑,徐腾洋,焦玉芬,等. 高效液相色谱法测定婴幼儿乳粉中乳铁蛋白含量[J]. 现代食品,2021(19):190-194.
[22] Paiardi G,Richter S,Oreste P,et al.The binding of heparin to spike glycoprotein inhibits SARS-CoV-2 infection by three mechanisms[J]. Journal of Biological Chemistry,2022,298(2):101507.
[23] 章寅,高磊,郑隽,等. 乳制品中乳铁蛋白的测定[J]. 现代农业科技,2013(16):291-292.
[24] 楼飞,杨秀敏,章寅. 肝素亲和柱在乳铁蛋白检测中的应用优化[J]. 食品研究与开发,2019,40(6):161-165.
[25] García J M,Udenigwe C C,Duitama J,et al.Peptidomic analysis of whey protein hydrolysates and prediction of their antioxidant peptides[J]. Food Science and Human Wellness,2022,11(2):349-355.
[1] 何瑛, 曹学思, 纪坤发, 杨爱君, 陈欣, 陈春裕. 高效液相色谱法测定巴氏杀菌乳中糠氨酸含量的不确定度评定[J]. 中国乳业, 2022, 0(9): 71-80.
[2] 何瑛, 纪坤发, 杨美丰, 杨爱君, 罗少杰, 曹学思. 高效液相色谱法测定牛奶中糠氨酸含量[J]. 中国乳业, 2022, 0(8): 83-89.
[3] 徐旖, 王荣艳, 陈青俊, 任一平. 液相色谱-单四级杆法检测奶粉中泛酸的应用[J]. 中国乳业, 2022, 0(7): 59-64.
[4] 高爽. 高效液相色谱法在牛奶中磺胺类药物残留检测的应用[J]. 中国乳业, 2022, 0(6): 55-59.
[5] 陈锡龙, 赵贵, 黄鑫, 谢丽丽, 王庆红, 孙真峥, 杨强. 超高效液相色谱-串联质谱法测定牛奶中20 种常用兽药残留量的研究[J]. 中国乳业, 2022, 0(6): 60-67.
[6] 陈柏锡, 孙嘉昊, 刘威, 张文琪, 吴晓辉. 乳制品检验液相色谱分析方法的建立与优化[J]. 中国乳业, 2022, 0(4): 72-76.
[7] 李媛, 范芳芳, 张敏娟, 张利娟, 康婕. 半制备-高效液相色谱法测定乳粉中维生素D2和D3含量[J]. 中国乳业, 2022, 0(3): 66-70.
[8] 陈茂姣, 邓燕琴, 何娓娓, 兰宁, 张红梅. 流动相对液相色谱法测量乳制品中三聚氰胺影响的探究[J]. 中国乳业, 2022, 0(1): 69-72.
[9] 陈忱. 各阶段母乳中维生素A含量测定与比较[J]. 中国乳业, 2021, 0(5): 100-104.
[10] 陈增鑫, 潘芸芸, 闫泽文, 王恒. 气相色谱-质谱联用技术在乳制品检测中的应用[J]. 中国乳业, 2021, 0(3): 69-72.
[11] 霍路曼, 李艺, 郑百芹, 李明阳, 张淼怡, 郝立武, 张书宏. 液相色谱-飞行时间质谱法快速筛查生乳中29 种兽药残留的研究[J]. 中国乳业, 2021, 0(12): 100-104.
[12] 黄隽灏, 王珉道, 伍嘉耀, 纪坤发, 杨爱君, 余保宁. 生鲜乳中黄曲霉毒素M1检测方法研究[J]. 中国乳业, 2021, 0(1): 55-59.
[13] 谢婷婷, 黄子珍, 曾庆坤, 杨攀, 李玲, 黄丽. 液相色谱技术在特种乳制品中掺入其他乳源的研究现状[J]. 中国乳业, 2020, 0(3): 67-70.
[14] 王荣艳, 徐聪玲, 李海龙, 任一平. UPLC-MS/MS法测定乳基食品中谷氨酰胺含量研究[J]. 中国乳业, 2019, 0(2): 63-67.
[15] 房启波, 于春艳. 婴幼儿配方乳粉中添加低聚果糖的必要性及含量测定[J]. 中国乳业, 2018, 0(3): 60-62.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
ISSN 1671-4393
CN 11-4768/S
主管：中华人民共和国农业农村部
主办：中国农业科学院农业信息研究所
地址：北京市海淀区中关村南大街12号
邮编：100081
电话：010-82106772
E-mail：zhgry@caas.cn

版权所有 © 《中国乳业》编辑部