液质联用多肽识别技术鉴别鳖甲胶的研究

Identification Study of Trionycis-Shell Glue by UPLC-MS/MS Marker Peptide

关键词:  鳖甲胶  特征离子  超高效液相色谱-串联质谱法  多肽

Key words:  trionycis-shell glue  characteristic ion  UPLC-MS/MS  peptide

摘要:

目的 找出鳖甲胶与龟甲胶、鹿角胶、阿胶、黄明胶、新阿胶、佛罗里达鳖甲胶、山瑞鳖甲胶的差异性特征肽段，从而建立专属性的鉴别方法。方法 用水提取样品，胰蛋白酶进行酶解，采用超高效液相色谱-四级杆飞行时间质谱和数据处理软件对酶解样品进行分析处理，查找鳖甲胶特征肽；采用超高效液相色谱-串联四级杆质谱的多反应检测建立专属性鉴别方法。结果 找出鳖甲胶的特征离子为m/z 784.90（双电荷），鉴定序列为GETGPVGVTGSVGPAGAR，为鳖的Ⅰ型a2链胶原蛋白中的一部分。利用特征肽段二级质谱图，确定了专属性检测离子对m/z 784.90（双电荷）→872.45，1 028.55，建立了鳖甲胶专属性鉴别方法。结论 确定的鳖甲胶特征肽专属性强，该方法的专属性、精密度和灵敏度符合分析检测的技术要求。

Abstract:

OBJECTIVE To establish a specific method for identification of trionycis-shell glue by finding the marker peptide different from glue of tortoise shell, deerhorn glue, donkey-hide gelatin, bovine-hide gelatin, pig-hide gelatin, Apalone ferox glue and Palea steindachneri glue. METHODS Samples were extracted with water, trypsin was used for enzymolysis. UPLC-Q-TOF/MS and data processing software were used to analyze the enzymolyzed samples to find the marker peptides of trionycis-Shell glue. Specific method was established by Multi-reaction detection of UPLC-Q-TOF/MS. RESULTS The marker peptide of trionycis-shell glue[m/z 784.90(n=2)] was found. This polypeptide sequence was GETGPVGVTGSVGPAGAR, one of fragmentation of collagen α-2(I) chain[Trionyx sinensis]. Used this characteristic ions, m/z 784.90(n=2)→872.45, 1 028.55, established an analysis method for identification of trionycis-shell glue. CONCLUSION The established present method was specific, precise and reliable. The research provided a possible approach for trionycis-shell glue.