基于特征肽段的鹿茸及其混伪品种的专属性鉴别研究

Specificity Identification of Cervi Cornu Pantotrichum and Its Adulterant Based on Characteristic Peptides

目的:基于特征肽段建立鹿茸及其混伪品种的专属性鉴别方法。方法:针对鹿茸的主要成分蛋白质,采用胰蛋白酶对正品鹿茸(梅花鹿、马鹿)及伪品鹿茸(驯鹿)进行酶解,利用超高效液相色谱-四极杆飞行时间质谱法(UPLC-QTOF-MS)进行测定,结合数据分析软件,以主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)对3种鹿茸样品的液质数据进行分析,找出驯鹿的专属性特征肽。利用超高效液相色谱-三重四极杆质谱法(UPLC-QQQ/MS)进一步验证驯鹿特征肽段的专属性,同时基于特征肽段建立正品鹿茸的特征图谱。结果:确定了伪品鹿茸的2个特征肽段并建立了正品鹿茸的特征图谱。结论:伪品鹿茸的2个特征肽段可用于正品鹿茸中混有伪品鹿茸的专属性鉴别,建立的正品鹿茸的特征图谱方法重复性好、操作简便,可用于鹿茸及其混伪品种的鉴别。

Objective:To develop a method for the specificity identification of Cervi Cornu Pantotrichum(Cervus nippon Temminck or C.elaphus Linnaeus)and its adulterant(Rangifer tarandus Linnaeus)based on the characteristic peptides.Methods:According to the characteristics of the main protein components in Cervi Cornu Pantotrichum,trypsin was used to digest it and its adulterant.The obtained peptides were analyzed with the ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS),the data of which were then investigated depending on principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA).On this basis,the characteristic peptides of R.tarandus were screened out.Their specificity was further verified with the ultra performance liquid chromatography coupled with triple quadrupole mass spectrometry(UPLC-QQQ/MS).Moreover,the characteristic spectrum of Cervi Cornu Pantotrichum was established based on characteristic peptides.Results:Two characteristic peptides of the adulterant were identified and a characteristic spectrum of Cervi Cornu Pantotrichum was established.Conclusion:The two characteristic peptides of the adulterant can be used for the specificity identification of Cervi Cornu Pantotrichum mixed with the adulterant.The characteristic spectrum of Cervi Cornu Pantotrichum features good reproducibility and high convenience,which is of great significance to its quality control.