Based On The Quality Of Markers T2?T5 Ophiocordyceps Sinensis Quality Evaluation Research

As dry complex,the Cordyceps sinensis(Ophiocordyceps sinensis)are formed from Cordyceps sinensis which is parasitic in the live or dead larvae of Lepidoptera.They are mainly distributed in the Qinghai-Tibet Plateau.The alpine meadows and shrubs with altitude above 2800 m are the principal distribution area for Cordyceps sinensis.They have the effect of immune regulation,lipid-lowering,anti-tumor,anti-bacterial and anti-fatigue,et al.Currently,Cordyceps are confused with much products which are the sibling species to Cordyceps in the market.The confused Cordyceps have high similarity with the right so that the existing distinguishing standard Chinese Pharmacopoeia is not the effective method and seriously affecting the safety of clinical medication.Based on the above matter,it is necessary to explore out the specific components in Cordyceps sinensis and develop a more scientific and reasonable quality control standard for Cordyceps sinensis.In this paper,the specific components T2 and T5 in Cordyceps sinensis were isolated and purified by preparative chromatography,and the stability of T2 was investigated.The preparation conditions of T2 were determined as follows: column of Ultimate T5B-C18(250 mm × 10 mm,5 ?m),detection wavelength of 270 nm and detection time of 30 min.T5 was prepared as Ultimate T5B-C18 column(250 mm × 10 mm,5?m),the detection wavelength of 240 nm,the detection time is 30 min.T2 was partially degraded to T3 and T4 under the influence of temperature and air in the stability test to T2.The optimum drying method of T2 and T5 was freeze-dried.The preparative T2 and T5 sample were carried on Q-TOF / MS and found that T2,T5 may be small molecules of the peptide,then the prepared T2 and T5 were sequenced by LC-MS / MS.The liquid chromatograph is Venusil × BPC,C18,5 ?m,150 A,and the column is homemade injection needle one column,The flow rate is 0.4 ?l / min,the sample volume is 10 ?l,the gradient elution program is 0 ~ 6 min,5% ~ 8% B;6 ~ 40 Min,8% to 30% B;0 ~ 45 min,30% ~ 60B;45 ~ 48 min,60% ~ 80 B;48 ~ 56 min,80% B;56 ~ 58 min,80% 5% B;58 ~ 65 min,5% B;Mass Spectrometer is Q Exactive,Trap column is Acclaim PePmap 100,75 ?m × 2 cm,nanoviper,C18,3 ?m,100 A,ion mode is positive ion mode,The results were compared with the local database and NCBI database by MASCOT,the peptide sequence derived from Cordyceps sinensis and its protein belonged to Ophiocordyceps unilateralis was obtained by comparing the mass information with the local database and NCBI database.The next work is to sort out the highest scores which several peptides SVTDPFHPGGPGLIR,IILLILK,ETGNLLR,LLLALVK from the sequencing results,and compare verify with the original samples by chromatography and mass spectrometry to determine the structure of T2 and T5?In this paper,the new methods of quality control of Cordyceps sinensis based on T5 were established,and the activities of T2 and T5 were studied in order to contribute to the quality control and efficacy of Cordyceps sinensis.