目的 找到鹿角胶正品来源梅花鹿、马鹿中区别于易掺伪品种驯鹿、驼鹿的特征肽段并验证专属性。方法 使用鹿角胶样品,处理得到肽段混合物进入纳升液相色谱-高分辨质谱联用仪(nano HPLC-orbitrap fusion)分析,获得目的蛋白质全肽段质谱图。使用PEAKS8.5软件对质谱数据进行分析,进行肽段混合物的从头测序以及肽段序列的匹配,得到的数据通过比对找到特征离子对,确定特征肽段序列并合成,使用液相-三重四级杆质谱联用多反应检测(MRM)方法验证特征肽段专属性。结果 共找到梅花鹿、马鹿专属的2个特征离子对,对应2条特征肽段,专属性验证结果好。结论 找到的2条肽段可以作为鉴别梅花鹿与马鹿鹿角胶的特征肽段,并提供了一条特征肽段研究策略,为鹿角胶新的鉴别及含量测定方法奠定基础。
Abstract
OBJECTIVE To find the characteristic peptides of Cervi Cornus Colla and verify the specificity. METHODS To obtain the mass spectrum of the target protein, the peptide mixture was analyzed using nano ultra-high performance liquid chromatography-high resolution mass spectra (nano HPLC-orbitrap fusion). Use PEAKS 8.5 to analyze the mass spectral data in order to achieve denovo sequencing of peptide mixtures as well as prediction and identification of peptide sequences. The obtained data were compared to identify the characteristic ion pairs, after which the characteristic peptide sequences were located and synthesized. Triple quadrupole mass spectrometry was used to confirm the experimental results. RESULTS Two characteristic ion pairs which belong to Cervus nippon and Cervus elaphus were found, corresponding to two characteristic peptides, and the specificity verification results were satisfied. CONCLUSION The two peptides can be used as the characteristic peptides to identify Cervus nippon and Cervus elaphus, providing a strategy for the study of characteristic peptides. The results lay a foundation for the development of new identification and quantification methods of Cervi Cornus Colla.
关键词
鹿角胶 /
特征肽段 /
梅花鹿 /
马鹿 /
定性鉴别
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Key words
Cervi Cornus Colla /
characteristic peptide /
Cervus nippon /
Cervus elaphus /
identification
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中图分类号:
R282
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参考文献
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脚注
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基金
山东省重点研发计划重大科技创新工程项目资助(2021CXGC010511);山东省重点研发计划项目资助(2020RKB24001);山东省人文社会科学课题资助(2021-YYGL-44)
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