Study On The Chemical Constituents And Quality Analysis Of Yinxing Mihuan Oral Solution

Yinxing Mihuan Oral Solution(YMOS)is a compound preparation made of ginkgo leaf extract and Armillariella mellea powder,which is scientifically formulated for treating coronary heart disease,angina pectoris,and ischemic cerebrovascular disease,with pharmacological activities such as anti-inflammatory,antioxidant,vasodilator,inhibition of platelet aggregation and activation,and promotion of blood vessel renewal.Because of the complex chemical composition of YMOS,further characterization of its substance basis is of great significance.However,the current standard of YMOS lacks the detection of Armillariella mellea powder,which cannot be used to comprehensively evaluate the overall quality,making an in-depth study of the quality control of YMOS inevitable.In this study,a research method of “the characterization of chemical composition,the quantitative analysis of multi-compounds,and the study of characteristic fingerprint” was established to address the key problems in the quality control of YMOS,and the stability of flavonol glycosides was explored based on the chemical kinetic theory,which enhanced the quality control method of YMOS and provided the scientific basis for its production,packaging,and storage.1.Based on UHPLC-Q-Orbitrap MS technology,the compounds in YMOS were systematically characterized.Combined with relevant data reported in literature and monographs,a total of 67 compounds in YMOS were identified,mainly including nucleosides(adenosine,guanosine,uridine,et al),organic acids(gallic acid,protocatechuic acid,caffeic acid,et al),terpene lactones(bilobalide,ginkgolide A,ginkgolide B,et al),flavonoids(quercetin,quercetin-3-O-β-D-glucoside,rutin,quercetin-3-O-[2’’-(6’’-p-coumaroyl)-β-Dglucosyl]-α-L-rhamnoside,et al),amino acids(proline,leucine,et al),and additives,among which 34 compounds were unambiguously identified in comparison with reference substances,which further clarified the chemical composition of YMOS.2.Based on the UPLC-PDA technique,a quantitative multi-compounds analytical method was established for YMOS using uridine,adenosine,guanosine,acesulfame,5-hydroxymethylfurfural,protocatechuic acid,4-hydroxybenzoic acid,sodium benzoate,quercetin-3-O-rutinoside/quercetin-3-O-β-D-glucoside,kaempferol-3-O-rutinoside,isorhamnetin-3-O-rutinoside,and quercetin-3-O-[2’’-(6’’-p-coumaroyl)-β-D-glucosyl]-α-Lrhamnoside as indicators.The established method was applied to the determination of the index compounds in different batches of unexpired and expired products,whose results showed that the unexpired products showed good lot-to-lot consistency;the dosages of two additives,acesulfame K and sodium benzoate,were in accordance with the World Health Organization(WHO)or the Chinese Pharmacopoeia 2020 edition;and the flavonol glycosides showed a significant decreasing trend in the expired YMOS.3.Based on the UPLC-PDA method,the characteristic fingerprints of YMOS were constructed,which was further employed to carry out the characteristic fingerprints of different batches of products.Different batches of YMOSs showed 19 characteristic fingerprint peaks with similar relative retention times for each compound,revealing good stability of each characteristic peak,followed by the identification of the characteristic fingerprint peaks by comparison with reference substances and LC-MS analysis.The similarity analysis of different batches of YMOS was performed by the Similarity Evaluation System for Chromatographic Fingerprint of TCM with results showing similarity of 22 batches of products greater than 0.9,indicating that the product quality was relatively stable.In conclusion,this UPLC characteristic fingerprinting method is scientific and feasible and can be used as the quality evaluation method of YMOS.Using the characteristic fingerprinting method,the correlation between the YMOS and its raw materials was investigated,whose results suggested that the preparation had good consistency in the component assignment.4.To further investigate the stability on flavonol glycosides in YMOS,this study systematically carried out the stability study of five flavonol glycosides including QGR,KGR,IGR,QRc G,and QG under different temperatures and p H according to the analysis of chemical kinetics.The degradation pattern of flavonol glycosides was systematically investigated,in which the results demonstrated that the presence of ortho-diphenolic hydroxyl group on B-ring and the type of saccharide connected to 3-hydroxyl on C-ring play a pivotal role in the stability of the tested compounds;and the flavonol glycosides have good stability at low temperatures and acidic conditions,providing a basis for the development,production,packaging,and storage of relevant preparations containing flavonol glycosides.