Quality Control Method And Batch Consistency Evaluation Of Fermented Cordyceps Sinensis Silitaris Solid Preparations

Fermented Cordyceps sinensis is a Chinese medicine preration which extracted and separated from Cordyceps sinensis by artifical fermentation.The study showed that the chemical composition of fermented Cordyceps sinensis was similar to Cordyceps sinensis.It mainly contained nucleosides,sterols,polysaccharides,Amino acids,alcohols,sphingolipds and other chemical components,but the content of the components was different.There are many kinds of fermented Cordyceps sinensis products on the market,but there is no uniform quality control standard,resulting in uneven quality of products.The quality standards of Jinshuibao capsules(tablets)and Bailing capsules have been included in the 2015 edition of Chinese Pharmacopoeia.The quality standards of Jinshuibao capsules(tablets)were as follows: TLC identification of three nucleosides,amino acids and mannitol;HPLC identification of five nucleosides;and HPLC determination of adenosine and ergosterol.The quality standards of Bailing capsules were as follows: TLC identification of ergosterol;HPLC identification of 6 nucleosides and 3 amino acids;amino acid analyzer determination of total amino acids;titration determination of mannitol;and HPLC determination of adenosine.Sterols and Amino acids are the main chemical components and pharmacodynamic substances of Fermented Cordyceps sinensis powder,but there was no systematic quality control.As a result,the existing quality standards could not fully reflect the quality of fermented Cordyceps sinensis,and the product process consistency has not been fully developed.The quality control of polysaccharides and nucleosides of fermented Cordyceps sinensis has been completed in the early stage of the project.In order to further improved the quality control standard of fermented Cordyceps sinensis and carried out the process consistency evaluation research,this study adopted the fingerprint combined with QAMS to control the quality of sterols in fermented Cordyceps sinensis,established the Amino acid fingerprint of fermented Cordyceps sinensis,and compared the determination methods of Amino acid content in fermented Cordyceps sinensis.To study the Terahertz Absorption Spectra of fermented Cordyceps sinensis.The quality of fermented Cordyceps sinensis products was evaluated by terahertz time-domain spectroscopy,frequency-domain spectroscopy and absorption coefficient curve spectroscopy.It provided some experimental basis for quality control and process consistency evaluation of fermented Cordyceps sinensis.1.Study on Fingerprint of sterols in fermentation Cordyceps sinensisJinshuibao capsules,jinshuibao tablets and Bailing capsules were taken as the sample,the qualitative analysis was carried out on the products of cordyceps sinensis by using HPLC system combine with similarity analysis,principal component analysis and cluster analysis.The results showed that the similarity between Jinshuibao capsules,Jinshuibao tablets and Bailing capsules were more than 0.9,there is high consistency and stability between batches of jinshuibao capsule,jinshuibao tablet and Bailing capsule.Jinshuibao capsules and Jinshuibao tablets identified four sterol substances,namely Ergosterol,?-sitosterol,Stigmasterol and Daucosterol.Bailing Capsule only contains Ergosterol and Stigmasterol.Principal component analysis and cluster analysis showed that Jinshuibao capsules and Jinshuibao tablets produced by the same strain,the same manufacturer were classified as one group,Bailing capsules were classified as another group,which was consistent with the actual situation.2.Determination of sterols content in fermented Cordyceps sinensis by QAMSErgosterol is used as an internal marker to determinate the content of ?-sitosterol,Stigmasterol and Daucosterol The influence of the factors such as instrument,velocity and column type on relative correction factor(RCF)was investigated.and The content measured by QAMS method was compared with that measured by the external standard method.The results shows that there was no significant difference between the two methods,It is suggested that QAMS method can be used as an efficient and convenient method for simultaneous determination of sterols substances in fermented Cordyceps sinensis.3.Study on Fingerprint of Amino acid in fermentation Cordyceps sinensisAmino acid fingerprints of fermented Cordyceps sinensis were established by high performance liquid chromatography.The similarity analysis,principal component analysis and cluster analysis were used to qualitatively analyze the Amino acid of the fermented Cordyceps powder.The results showed that the similarity between jinshuibao capsule,jinshuibao tablet and Bailing capsule were more than 0.9,there is high consistency and stability between batches of jinshuibao capsule,jinshuibao tablet and Bailing capsule.All three products identified 20 common peaks,of which 1,2,3,4,5,6,7,8,9,11,12,13,14,16,17,19,20 were identified.Aspartic acid,Glutamic acid,Hydroxyproline,Serine,Glycine,Histidine,Arginine,Threonine,Alanine,Valine,Tyrosine,Proline,Methylthio Acid,Isoleucine,Leucine,Phenylalanine and Lysine.Principal component analysis showed that the same strain,Jinshuibao capsules and Jinshuibao tablets by the same manufacturer were classified into one category,and Bailing capsules were further classified into one category.The results of cluster analysis showed that the samples were divided into three categories: Jinshuibao capsules were grouped into one category,Jinshuibao tablets were grouped into one category,and Bailing capsules were grouped into one group;Jinshuibao capsules and Bailing capsules were clustered and analyzed.The results showed that the samples were clustered into two categories,Jinshuibao capsules were grouped together,and Bailing capsules were grouped together,which was consistent with the actual situation.4.Comparative study on determination of Amino Acid Content in fermented Cordyceps sinensisThe content of amino acids in fermented Cordyceps powder was determined by RP-HPLC and AAA methods.The results of two amino acid determinations were compared and analyzed.There were some differences in the determination of individual amino acid content.Using glycine as an internal standard,quantitative analysis of 16 amino acids by QAMS.The influence of the factors such as instrument,column temperature,velocity and column type on relative correction factor(RCF)was investigated.and The content measured by QAMS method was compared with that measured by the external standard method.The results showed that the RSD was less than 5% under different conditions of instrument,flow rate,column temperature,different instruments and column types.The results of t test showed that the results measured by QAMS method and the external standard method were more than 0.05.There was no significant difference between the two groups of data,indicating that the QAMS method could be used as an efficient and convenient method for simultaneous determination of 16 amino acid substances in fermented Cordyceps sinensis.5.Study on terahertz wave spectrum of fermented Cordyceps sinensisUsing terahertz spectroscopy,the terahertz time-domain spectral information of the fermented Cordyceps sinensis products Jinshuibao capsules and Bailing capsules were measured respectively,and the frequency domain spectrum,the refractive index spectrum and the absorption curve spectrum were obtained by Fourier transform.The spectral information was combined with principal component analysis to evaluate the quality of the fermented Cordyceps sinensis product.The results showed that there was no significant difference in the refractive index profile of terahertz between Jinshuibao capsules and Bailing capsules.There were some differences in the terahertz time domain spectrum,frequency domain spectrum and absorption coefficient curve spectrum,Two kinds of products could be clearly distinguished by three spectral lines.Principal component analysis was performed after the absorption curve spectra of all samples were processed by element scattering correction,special point baseline method and standard normal variables.The results showed that Jinshuibao capsules were clustered into one group,and the clustering effect was better.Bailing capsules were clustered into one group,and the clustering effect was slightly worse.The clustering results reflected the consistency of Jinshuibao capsules over Bailing capsules to some extent.