ABSTRACT
This paper explored the specific peptides from Bubali Cornu by ultra-performance liquid chromatography-tandem mass spectrometry and based on mathematics set theory. Following the profile analysis of peptides from Bubali Cornu, Bovis Grunniens Cornu, Caprae Hircus Cornu, and Suis Cornu by nano LC-LTQ-Obitrap-MS after digestion with trypsin, the relationship of peptide composition among different samples was analyzed using the mathematics set theory. The ones that existed only in the Bubali Cornu set rather than in any other set were considered as the specific peptides of Bubali Cornu. The further bioinformatic analysis revealed four specific peptides from Bubali Cornu, whose specificity was verified by UPLC-QQQ-MS. The results showed that these four peptides could be used for distinguishing Bubali Cornu from Caprae Hircus Cornu and Suis Cornu. This study has provided a rapid and simple method for seeking the specific peptides in animal medicines, which can be utilized for quality evaluation of animal medicines, thus making them authenticable and traceable.
Subject(s)
Animals , Chromatography, Liquid , Cornus , Horns/chemistry , Peptides/chemistry , Tandem Mass SpectrometryABSTRACT
Objective: To establish a rapid on-site method for identifying Chinese medical material recombinase-mediated amplification (RAA) technology for the use of identifying medicinal Bubali Cornu from yak horn. Method: Based on the differences of mitochondrial genome sequences between Bubali Cornu and adulterants,the specific RAA primer (SNJ-1.F,SNJ-1.R) and fluorescence probe SNJ-1.probe were designed by variation sites. Alkaline lysis method was used to extract DNA from milled samples,and optimize RAA reaction system. The incubation was made at 37℃ for 15-20 min, the reaction results were monitored through gel electrophoresis and a mobile fluorescence amplification instrument. The RAA identification result was compared with COI DNA sequencing. Result: After incubation at 37℃ for 20 min,about 140 bp of bright and simple bands was amplified from DNA templates of Bubalus bubalis,whereas Bos mutus were negative. By the Real-time fluorescent RAA identify method,all reactions in DNAs from Bubali Cornu samples were amplified from 6.21 to 8.37 min,whereas DNAs from yak samples were amplified after 10.08 min,COI sequencing results conformed to the Real-time fluorescent RAA identification. Conclusion: Specific RAA could rapidly identify Bubali Cornu in 20 minutes, and thus be applied in medical material and its products. This method is simple,rapid and sophisticated instrument-free,with promises in on-site identification of traditional Chinese medicine.