基于MALDI-TOF MS肽指纹图谱技术的中药鹿血晶掺伪鉴别研究  

Identification of Adulterated Deer Blood Crystals in Traditional ChineseMedicine Based on MALDI-TOF MS Peptide Fingerprints Technique

作　　者：王慎兴 曹玲 黄青 马聪玉 潘建斌 赵浩东 施海蔚 狄斌[1] 陈洪渊[5] WANG Shen-xing;CAO Ling;HUANG Qing;MA Cong-yu;PAN Jian-bin;ZHAO Hao-dong;SHI Hai-wei;DI Bin;CHEN Hong-yuan(School of Pharmacy,China Pharmaceutical University,Nanjing 211112,China;Jiangsu Insitite for Food andDrug Control,Nanjing 210019,China;Jiangsu Provincial Drug Administration Audit and Inspection Center,Nanjing 210019,China;Institue of Materia Medica Chinese Academy of Medical Science,Beijing 100050,China;School of Chemistry and Chemical Engineering,Nanjing University,Nanjing 210046,China)

机构地区：[1]中国药科大学药学院,江苏南京211112 [2]江苏省食品药品监督检验研究院,江苏南京210019 [3]江苏省药品监督管理局审核查验中心,江苏南京210019 [4]中国医学科学院药物研究所,北京100050 [5]南京大学化学化工学院,江苏南京210046

出　　处：《质谱学报》2025年第1期115-122,共8页Journal of Chinese Mass Spectrometry Society

基　　金：江苏省药品监督管理局2023年药品监管科学科研计划项目(202302)。

摘　　要：本研究应用基质辅助激光解吸电离-飞行时间质谱(MALDI-TOF MS)技术,构建了具有物种特异性的白蛋白和血红蛋白特征肽指纹图谱,用于中药鹿血晶的掺伪鉴别。以鹿血晶真品(梅花鹿、马鹿)和常见掺伪品(猪、牛、羊、驴)共6个物种的血晶制品作为研究对象,通过理论肽段预测并结合高分辨质谱,筛选出1组能够反映物种差异的特征肽段。采集不同物种来源血晶制品以及不同来源市售中药鹿血晶的特征肽指纹图谱,并结合化学计量学分析,实现了不同来源血晶制品的快速准确区分。该方法能够实现复杂基质样品中目标特征肽段的灵敏、特异检测,具有高专属性、快速、便捷的优势,为鹿血晶及血晶类中药制品的掺伪鉴别提供了准确的分析手段,也为其他复杂蛋白类制品的快速鉴别提供了有价值的参考。This study was focused on analyzing blood crystal products from six species of Chinesemedicine deer blood crystals(sika deer,red deer)and commonly adulterated products(pig,cow,sheep,donkey).A method based on matrix-assisted laser desorption ionization-time of flight massspectrometry(MALDI-TOF MS)combined with chemometric blood analysis was established toidentify adulteration in deer blood crystals used in traditional Chinese medicine.In this study,the bottom-up mass spectrometric proteomics approach was applied to acquire the peptide fingerprints ofthe highly abundant blood proteins,hemoglobin and albumin in samples.The blood crystal samplesunderwent pretreatments including reduction,alkylation,trypsinolysis,and desalting.Firstly,Skylinesoftware was used to predict candidate peptides by identifying the theoretical characteristic peptidesfor each species,and the mass spectrometry data of the tryptic digests were collected using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometer(UPLC-Q-TOFMS).Then,the mass spectrometry data were combined with UNIFI software and Uniprot database toaccurately select a set of characteristic peptides that can reflect the differences between the targetproteins,which was achieved by examining the presence of candidate characteristic peptides in thesamples,as well as considering parameters such as the response,missed cut rate,and detection rate.Thereafter,a one-way controlled variable experiment was conducted to determine the optimal matrixfor MALDI-TOF MS analysis.The laser energy was set to 90,and the spotting was performed usingthe mixing method.α-Cyano-4-hydroxycinnamic acid(CHCA)was identified as the optimal matrixfor the blood crystal samples.The optimized MALDI-TOF MS method was employed to obtain thecharacteristic peptide fingerprints of 10 batches of blood crystals for each of the six species.Thesefingerprints were then combined with principal component analysis(PCA)in order to achieve rapidand accurate differentiation of blood crystals from

关 键 词：基质辅助激光解吸电离-飞行时间质谱(MALDI-TOF MS) 中药鹿血晶 肽指纹图谱 掺伪鉴别 

分 类 号：O657.63[理学—分析化学]