特征肽技术用于龟甲(浙龟甲)中掺巴西龟的检查研究

Detection of Brazilian Tortoise in Tortoise Shell(Zhejia Tortoise Shell) by Characteristic Peptide Technique

关键词:  乌龟  黄缘闭壳龟  黄喉拟水龟  巴西龟  掺伪  特征肽  高效液相色谱-串联质谱法

Key words:  Chinemys reevesii (Gray)  Cyclemys flavomarginata Gray  Clemmys mutica(Cantor)  Trachemys scripta elegans (Wied.)  adulteration  characteristic peptide  UPLC-MS/MS

摘要:

目的 找出巴西龟特征肽段，建立龟甲（浙龟甲）中掺入巴西龟的专属性检查方法。方法 样品以水提法提取，胰蛋白酶酶解，利用超高效液相色谱-四级杆飞行时间质谱和数据处理软件查找巴西龟特征肽，并利用蛋白质搜库技术初步测序；利用找到的特征肽段，建立超高效液相色谱-串联四级杆质谱的多反应监测的巴西龟掺伪专属性检查方法。结果 找出巴西龟特征性肽段m/z 400.23（双电荷），并初步测定序列，通过分析该特征肽段二级质谱图，确定了专属性检测离子对m/z 400.23（双电荷）→374.05，142.90，并建立了龟甲（浙龟甲）中掺入巴西龟的专属性检查方法。20批样品中，11批样品检出巴西龟成分。结论 该方法的专属性强，符合分析检测的方法技术要求，可用于龟甲（浙龟甲）掺入巴西龟的检查。

Abstract:

OBJECTIVE To find out the characteristic peptide of Trachemys scripta elegans (Wied.) and establish a specific method for adulteration of tortoise-shell(Zhejiang tortoise shell). METHODS The samples were extracted by water extraction method, hydrolyzed by trypsin, searched for the characteristic peptide of Trachemys scripta elegans (Wied.) by ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) and data processing software, and sequenced by protein library searching technology. Ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-QQQ-MS) was used to establish the specific method based on the marker peptide of Trachemys scripta elegans (Wied.). RESULTS The marker peptides[m/z 400.23(n=2)] of Trachemys scripta elegans (Wied.) was found. Using this characteristic ions, m/z 400.23(n=2)→374.05, 142.90, established an analysis method for identification of Trachemys scripta elegans (Wied.). In 20 samples, 11 samples detected Trachemys scripta elegans (Wied.) characteristic peptide. CONCLUSION This method has strong specificity and meets the technical requirements of analysis. This method can be used for the inspection of tortoise shell (Zhejiang tortoise shell) mixed with Trachemys scripta elegans (Wied.).