摘要
为对掺假蜂蜜识别提供新思路和新方法,本研究应用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱联用的方法鉴定4种未掺假蜂蜜及2种掺假蜂蜜中肽类物质的组成和结构,并探讨未掺假蜂蜜和掺假蜂蜜多肽的差异。以20%的三氯乙酸溶液沉淀蜂蜜蛋白,所得蜂蜜蛋白以胰蛋白酶酶解,然后通过C18固相萃取柱净化酶解肽段。采用0.1%甲酸和乙腈作为流动相进行梯度洗脱,以高分辨质谱(Q-Exactive)的Full MS/ddMS2模式对胰蛋白酶酶解后的蜂蜜多肽进行鉴定,MaxQuant软件分析质谱结果,分析所得肽段在Uniprot上进行Blast序列对比。结果显示,4种未掺假蜂蜜中均存在来自蜂王浆主要蛋白(MRJPs)的多肽,2种掺假蜂蜜中未检测出MRJPs,且4种未掺假蜂蜜中所鉴定出的多肽80%以上来自于蜜蜂(Apis cerana cerana、Apis cerana、Apis mellifera)。通过比对6种蜂蜜酶解肽段序列发现,肽段EYILVLSNK在4种未掺假蜂蜜中均有检测到,在2种掺假蜂蜜中未检出。因此,来自于Apis mellifera的MRJP1酶解产生的肽段EYILVLSNK或可作为辨别真伪蜂蜜的潜在肽段。
In order to provide a new idea and method for identification of adulterated honey,the composition and structure of peptides in four kinds of unauthorized honey and two kinds of adulterated honey were identified by ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry,and the differences between unauthorized honey and adulterated honey peptides were discussed.Honey protein was precipitated with 20%trichloroacetic acid.The obtained honey protein was enzymatically hydrolyzed by trypsin and then purified by C18 solid phase extraction column.Then the digested sample by trypsin was separated by gradient elution with 0.1%formic acid and acetonitrile as mobile phase and determined by high resolution mass spectrometry(Q-Exactive)in Full MS/ddMS2 mode.MaxQuant software was used to analyze the results of mass spectrometry.The analyzed peptides was comparised on Uniprot by Blast.The results showed that Major Royal Jelly Protein(MRJPs)existed in all four kinds of unauthorized honey,and were not detected in two kinds of adulterated honey,and more than 80%of the peptides identified in four kinds of unadulterated honey come from honeybees(Apis cerana cerana,Apis cerana,Apis mellifera).By comparing the enzymatic hydrolysis sequence of six kinds of honey,it was found that EYILVLSNK was detected in four kinds of unadulterated honey and not in two kinds of adulterated honey.Therefore,EYILVLSNK,a peptide digested by MRJP1 of Apis mellifera,might be a potential peptidest for distinguishing true and false honey.
作者
谢博
傅红
杨方
XIE Bo;FU Hong;YANG Fang(Inspection and Quarantine Technology Center,Fujian Entry-Exit Inspection and Quarantine Bureau,Fuzhou 350001,China;College of Biological Sciences and Engineering,Fuzhou University,Fuzhou 350108,China;Fujian Provincial Key Laboratory of Marine Enzyme Engineering,Fuzhou University,Fuzhou 350108,China)
出处
《食品工业科技》
CAS
北大核心
2020年第2期244-251,共8页
Science and Technology of Food Industry
基金
国家重点研发项目(2017YFF0211001)
关键词
多肽
掺假蜂蜜
超高效液相色谱-四极杆-静电场轨道阱高分辨质谱
peptides
adulterated honey
ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Exactive)
