基于UPLC-QTOF/MS和UNIFI二元比较鉴别巴西龟龟甲胶和草龟龟甲胶（英文）

Differentiating Trachemys scripta elegans Shell Glue from Chinemys reevesii Shell Glue by UPLC-QTOF/MS Coupled with Binary Compare Tool of UNIFI

目的本研究旨在建立一种区别巴西龟龟甲胶和中华草龟龟甲胶的方法。方法采用胰蛋白酶对龟甲胶样品进行酶解,利用超高效液相色谱-四极杆飞行时间质谱法分别对以巴西龟为来源的龟甲胶和以中华草龟为来源的龟甲胶的酶解样品进行测定,利用UNIFI中二元比较工具对二者酶解肽段进行比对分析,找出二者之间各自的特征肽段,然后用PEAKS■Studio软件对其进行鉴别,用于作为区别二者的根据。结果研究显示巴西龟龟甲胶酶解样品中含有的特征肽m/z 641.3和中华草龟龟甲胶酶解样晶的特征肽m/z 649.3可作为区别二者的根据,其氨基酸序列分别为GEAGPSGPAGPTGAR和GESGPSGPAGPTGAR。结论该方法可区别巴西龟龟甲胶和中华草龟龟甲胶,可用于龟甲胶的质量控制。

Objective This study was conducted to develop an method for identification of Trachemys scripta elegans(Ba Xi Gui,巴西龟)shell glue from Chinemys reevesii(Zhong Hua Cao Gui,中华草龟)shell glue.Methods In this research,an ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS)method coupled with Binary Compare tool of UNIFI software was validated to differentiate Trachemys scripta elegans(Ba Xi Gui,巴西龟)shell glue from Chinemys reevesii(Zhong Hua Cao Gui,中华草龟)shell glue.The gelatins were digested enzymatically into peptides using trypsin.The UPLC-QTOF/MS data of the trypsin digested samples was subjected to Binary Compare of UNIFI software in order to screen out the unique peptides.PEAKS■Studio software was utilized to identify the sequence of the marker peptides.Results It was found that m/z 641.3 was the marker peptide in samples of Trachemys scripta elegans(Ba Xi Gui,巴西龟)shell glue,and m/z 649.3 was the marker peptide in samples of Chinemys reevesii(Zhong Hua Cao Gui,中华草龟)shell glue,with possible amino acid sequence of GEAGPSGPAGPTGAR and GESGPSGPAGPTGAR respectively.Conclusions The results will be helpful for the differentiating between these 2 gelatins,and it can be also used for quality control of Tortoise shell glue(Gui Jia Jiao,龟甲胶).