Study On The Quality Standard Of Polysaccharides In Ginkgo Armillaria Oral Solution

Aiming at the problem that the original standard WS1-XG-004-2001 for Ginkgo Armillaria Oral Solution and its original medicinal material,Armillaria mellea,the identification method for polysaccharides in Ginkgo Armillaria Oral Solution lacks specificity and its content determination error is large,the improvement Phenol sulfate method equipped with microplate reader assay and PMP-pre-column derivatization high performance liquid chromatography were used to study the quality control of polysaccharides in Ginkgo Armillaria Oral Solution,which provide ideas and solutions for the improvement of the standard of Ginkgo Armillaria Oral Solution and the quality control of polysaccharide drugs.In response to the shortcomings of traditional sulfuric acid-phenol method for the determination of total sugar content,such as cumbersome operation,poor repeatability and comparability,an improved sulfuric acid-phenol method with microplate reader was proposed,and the method was used to determine the total sugar content of the commercially available Ginkgo Armillaria Oral Solution,self-made oral solution.and its raw medicinal material Ginkgo biloba leaf extract（5 different manufacturers）,Armillaria mellea（2 different manufacturers）,stevia,and sodium benzoate.The results showed that the sugar content in the oral solution was derived from the three parts of Ginkgo biloba leaf extract,Armillaria mellea and the supplementary material steviose.In addition,PMP-pre-column derivatization HPLC was used to determine the monosaccharide composition of Ginkgo Armillaria Oral Solution and its ingredients,and the corresponding fingerprints were constructed by combination with traditional Chinese medicine fingerprinting similarity software as follows:the monosaccharide composition of the oral solutions was mannose,rhamnose,glucose,galactose,and arabinose;E manufacturer has a big difference in the monosaccharide composition of ginkgo biloba extract from the other 4 manufacturers;The monosaccharide composition of Armillaria mellea from G manufacturer is inconsistent with oral liquid.The polysaccharides from Ginkgo Armillaria Oral Solution and the original medicinal materials were further extracted,after which quality control was performed in three aspects:discrimination（controlled from four aspects:relative molecular weight distribution determination,monosaccharide composition,infrared discrimination,glycosidic bond analysis）,inspection,and content determination.respectively,to construct a draft quality control for the polysaccharides from the Ginkgo Armillaria Oral Solution.After identification,the polysaccharides from Ginkgo Armillaria Oral Solution and Armillaria mellea were consistent in their monosaccharides composition,which are composed of mannose,galactose,glucose,rhamnose and arabinose,and the molar ratio is different;The infrared pattern showed that the characteristic absorption peaks of Polysaccharides from oral solutions and Armillaria mellea were 3300cm^-1,2900cm^-1,1600cm^-1,1400cm^-1,1100cm^-1,1000cm^-1,550cm^-1;The glycosidic bond analysis showed:The main chain of polysaccharide glycosidic bond from oral liquid is→1)Glc（4→and→1）Gal（6→connection,contains a glucose branch at the 6 position of glucose,the end group is glucose,and the main chain of Armillaria mellea polysaccharide glycosidic bond is→1）Glc(4→,contains a glucose branch at the 6 position of glucose,and the end group is glucose.Therefore,the original medicinal materials of different sources will cause differences in the molar ratio,relative molecular weight,and glycosidic bond connection of the polysaccharides of the commercial oral liquid and the self-made oral liquid polysaccharide monosaccharide.In accordance with the current quality standard-the 2020 version of the Chinese Pharmacopoeia,the original medicinal materials stevioside,sodium benzoate,and Ginkgo biloba extract（5 different manufacturers）in the ginkgo honey ring oral solution were studied for quality control.Contains medicinal excipients stevia and preservative sodium benzoate,the content of which is 3g/L;only the Ginkgo biloba extract from E manufacturer does not meet the pharmacopoeia standards,corresponding to the results of monosaccharide composition of Ginkgo biloba leaf extract in the above mentioned raw medicinal materials.In summary,through the quality control of the raw medicinal materials of Ginkgo Armillaria Oral Solution,it further shows that the self-made oral solution is consistent with the commercially available oral solution,and the results obtained are accurate and reliable.The established modified sulfuric acid phenol method equipped with a microplate reader is applicable according to the production needs of enterprises;the proposed pre-column derivatization high performance liquid chromatography method can be used as a supplement for the identification of Ginkgo Armillaria Oral Solution and its original medicinal materials,and Armillaria mellea and Ginkgo biloba extract.It provides ideas and methods for the quality control of Ginkgo Armillaria Oral Solution.