Safety Evaluation Of Codonopsis Pilosula And Study On The Regulation Of Fructans From Codonopsis Pilosula On Intestinal Microbiota In Vitro

Objective:1.Codonopsis pilosula is a Chinese medicinal material commonly used as medicine and food,and its quality is the premise of safety and effectiveness in clinical application.In this study,exogenous harmful substances from Codonopsis pilosula will be detected,including 33 pesticides in The Chinese Pharmacopoeia(2020edition),which are prohibited in the planting process of Traditional Chinese medicinal materials,heavy metals(lead,cadmium,arsenic,mercury,copper)and sulfur dioxide residues.This study was to provide data support for the quality evaluation of Codonopsis pilosula.2.Codonopsis pilosula contains a variety of chemical components.Among them,Codonopsis pilosula polysaccharide is one of the main effective components.Currently,fructans isolated are known to have a positive effect on the regulation of intestinal microbiota.Fructans are metabolized into short-chain fatty acids by intestinal microbiota in the intestine,which changes the microbial diversity of intestinal microbiota.This study was intended to further study whether CPA and CPB from Codonopsis pilosula could regulate intestinal microorganisms on the basis of our previous studies,so as to provide some theoretical basis for further development and application of Codonopsis pilosula.Methods:1.10 batches of Codonopsis pilosula from different origins were collected,and33 banned pesticides in 10 batches of Codonopsis pilosula were detected by the fifth method in the Determination of pesticide residues in The Four General Rules 2341 of Chinese Pharmacopoeia(2020 edition).The contents of heavy metals in 10 batches from Codonopsis pilosula were determined by the method 2321 of The Fourth General Rule of Chinese Pharmacopoeia(2020 edition),and the contents of arsenic and mercury were determined by atomic fluorescence method.The sulfur dioxide residues in 10 batches of Codonopsis pilosula were determined by the first method2331 in the fourth general rule of Chinese Pharmacopoeia(2020 edition).Finally,the pollution degree of heavy metals and sulfur dioxide residues in 10 batches of Codonopsis pilosula was evaluated by single factor pollution index method.2.The prebiotic effects of fructan CPA and CPB were investigated by fermentation of intestinal microorganisms in vitro.Human feces were fermented with different concentrations of CPA and CPB in vitro.The pH of fermentation broth was measured.Fecal DNA was extracted.16SV3V4 region was amplified and sequenced.The sequencing data were combined after removing low quality,and then further filtered by removing chimerism.We obtained appropriate reads for flora difference analysis,including species composition analysis,α diversity analysis,β diversity analysis,principal component analysis,difference analysis between species groups,correlation analysis between flora and sample pH.Result:1.None of the 10 batches of Codonopsis pilosula contained 33 banned pesticides specified in Chinese Pharmacopoeia(2020 Edition).Five heavy metals and sulfur dioxide residues were detected,but their residues were far below the limit standard of Pharmacopoeia.The single-factor pollution index of heavy metals and sulfur dioxide residues was less than 1.2.Broth of CPA and CPB with different concentrations could reduce the pH of fecal fermentation.The diversity of flora in the high concentration treatment group was higher than that in the low concentration treatment group.Both of them could promote the growth of beneficial bacteria such as Faecalibacterium,Prevotella,Bifidobacterium,Megamonas,Odoribacter,Butyricimonas,and inhibit the growth of harmful bacteria such as Fusobacterium,Flavonifractor,Colidextribacter,Oscillibacter,etc.In addition,fructan CPB from Codonopsis pilosula can also promote the growth of Phascolarctobacterium.Conclusion:1.The overall quality of Codonopsis pilosula was good.There were no 33 pesticides banned by Chinese Pharmacopoeia(2020 edition),and no excessive residues of heavy metals(lead,cadmium,arsenic,mercury,copper)and sulfur dioxide.2.Fructan CPA and CPB from Codonopsis pilosula could regulate the growth of human intestinal microorganisms and are potential prebiotics.