Application Of Digital Standard Materials In The Overall Quality Control Of Danshen Polyphenolic Acid Preparation For Injection

Salviae Miltiorrhizae Radix et Rhizoma is the dry root and rhizome of Salvia miltiorrhiza Bge.Its water-soluble components belong to phenolic acids,including Danshensu,protocatechuic aldehyde,salvianolic acid B,rosmarinic acid,salvianolic acid D,salvianolic acid A and salvianolic acid E,etc.Salvianolic acid for Injection,produced by Tianjin Tasly,is a single herb medicine,whose effective components are salvianolic acids.The existing quality standards controlled only 1 or 2 of the components,which is insufficient to embody its holistic quality.Besides,the determination method of total salvianolic acid in preparation standard using molecular weight ratio method to calculate the relative correction factor,while the response of UV detector used in the method does not depend solely on the molecular weight of the constituents.So the accuracy of the content of total salvianolic acid and the relative correction factor obtained by this method still needs further discussion.In addition,salvianolic acids have poor stability,so it is difficult to prepare monomer reference standards.New methods should be sought for the analysis and detection.This research mainly includes the following parts:1.Fingerprint method was established for salvianolic acids through optimizing chromatographic conditions.Similarity analysis for the fingerprints was carried out by Chempattern,with 11 common peaks are indicated including sodium Danshensu,protocatechuic aldehyde,caffeic acid,salvianolic acid D,peak X,peak M,rosmarinic acid,lithospermic acid,salvianolic acid B,peak N and salvianolic acid Y.These 11 peaks' content is relatively stable in each batch and can be used as quality control index of fingerprint.The method was validated with good stability,repeatability and precision.Similarity analysis and multivariate statistical analysis indicated the difference between the 10 batches of extracts was small.2.27 salvianolic acids were preliminarily identified in the extract through ion trap mass spectrometry and the presence of some of the 27 compounds were verified by Q-TOF mass spectrometry.The 11 chromatographic peaks in the aqueous extract of Salvia miltiorrhiza were identified by means of reference standard comparison method,including sodium Danshensu,protocatechuic aldehyde,caffeic acid,salvianolic acid D,salvianolic acid E(peak X),salvianolic acid H/I(peak M),rosmarinic acid,lithospermic acid,salvianolic acid B,salvianolic acid L(peak N),salvianolic acid Y.Based on the data of qualitative research,Digital Reference Standard software(DRS ORIGIN)is designed and developed.Combined with substitute reference standard method,spectrum and mass spectrometry assisted matching technology and big data concept,the retention time of chromatographic peaks can be predicted using this software on the basis of reducing the use cost of the reference standards.DRS ORIGIN contains several different substitute reference standard methods.In this study,the linear calibration using two reference standard(LCTRS)method and relative retention time(RR)method was established for salvianolic acids for the prediction of HPLC peaks using DRS ORIGIN.The results indicated that the prediction accuracy and column durability of LCTRS method are better than RR method.2.The assay method for salvianolic acid exact and preparation.8 peaks were determined including salvianolic acid D,salvianolic acid E(peak X),salvianolic acid H/I(peak M),rosmarinic acid,lithospermic acid,salvianolic acid B,salvianolic acid L(peak N),salvianolic acid Y.The quantitative control components accounted for about 70% of the extract content.The results of method validation meet the requirements.The relative correction factor determination method of salvianolic acids was studied.The correction factors were determined by the concentration averaging method and the influence of the key factors,such as the wavelength of inspection and the ratio of mobile phase,on the relative correction factor was investigated.The results show that the relative correction factors obtained are basically stable.The content of 10 batches of extracts and 2 batches of preparations was determined by external standard method and relative correction factor method.The results of the two methods are of little difference,which proves that the established relative correction factor method is reliable.Combined with digital substitute reference standard technology,the paper studied on the holistic quality control method of salvianolic acids.It provides a useful reference for the revision of the quality standard of Salvianolic Acid for Injection and provides a new model for the quality control of other traditional Chinese medicine extracts and preparations.