Internal And External Pollutants Investigation Of Sojae Semen Praeparatum And Massa Medicata Fermentata And Analysis Of Main Active Substances

Object:The present situation of microbial contamination,pesticide residue and mycotoxin content in different batches of SSP and MMF sold on the market were investigated,and dietary risk assessment was carried out for pesticide residues and mycotoxins contamination.To separate the dominant probiotics from SSP and MMF sold on the market,and use them for pure seed fermentation,then the pure fermented SSP was evaluated with active ingredients,when the pure fermented MMF was evaluated with activity of digestive enzymes.This study provides a basis for the establishment of safety standards for SSP and MMF.Methods:10 batches of SSPs and 5 batches of MMFs were checked for aerobic bacteria,molds and yeasts and control bacteria as per 1105 and 1106of Part IV,Ch P 2020.After being boiled for 30 min at 100℃,quantity of thermoduric bacteria was investigated and the dominant strain was identified by Biolog microbial identification system.The content of OTA and ZEN in different batches of SSP and MMF were determined by HPLC according to the first method of the 2351 general rule of Part IV,Ch P 2020.An UPLC-Q-TOF-MS was used to determine AFs and the first method of the 2341general rule of Part IV,Ch P 2020 was used to determine Residues of organochlorine pesticides.Exposure risk assessments of OTA、ZEN、pesticide residues for SSPs and MMFs are based on deterministic approach and to estimate the risk of liver cancer AFs.The strains of MMF sold on market were isolated by dilution coating method,when isolation and identification of the strains of SSP has been completed by our laboratory.We selected prebiotics for pure fermentation to obtain 2 batches of SSP and 6 batches of MMF.We evaluate the quality of pure fermented SSP by daidzein and genistein,and evaluate pure MMF by the activity of five digestive enzymes(amylase,saccharifying enzyme,protease,lipase,cellulase)as indexes.Results:1.Microbial contamination:In different batches of SSPs,quantities of aerobic bacteria ranged from 10~4to 10~8cfu/g and that of molds and yeasts from 10~3to 10~8cfu/g;in different batches of MMFs,quantities of aerobic bacteria,molds and yeasts all ranged from 10~3to 10~5cfu/g;control bacteria were detected in 10 batches of SSPs and 4 batches of MMFs.Both SSP and MMF contain a certain number of heat-resistant bacteria,among which the number of heat-resistant bacteria in SSP is high,between 10~3to 10~8cfu/g,and the number of heat-resistant bacteria in MMF is less than 10~2.Upon identification,thermoduric bacteria were entirely bacillus,dominated by bacillus subtilis.2.Exogenous Pollution and Health Risk Assessment:ZEN was only detected in MMF,and maximum pollution exceeding 2.23 times standard;OTA was detected in two kinds of decoction pieces,of which SSP had a higher rate of over-standard;aflatoxin only AFB1 was detected and exceeded the standard(GB2761—2017)in a batch of MMFs.No mycotoxins were detected in the samples of purebred fermented soybean.BHC-D detection rate is 100%and the maximum value exceeds the standard(GB 2763-2019);OP’DDT was detected in 1 batch of SSP and MMF respectively,and the content was within the standard range.All kinds of exogenous pollution evaluation results are still within the human acceptance level,the risk is acceptable,Child exposure risk greater than adult with a maximum TDI%of62.22%.Some samples are rich in pollution types and exceed the standard.3.Main components and enzyme activity test results:The total contents of daidzein and genistein in 2 of 10 batches of SSP were in accordance with the pharmacopoeia standard(≥400μg/g),which were 1216μg/g and 517.28μg/g respectively.The total content of daidzein and genistein was 276.42μg/g and 242.76μg/g respectively,which did not meet the standard of pharmacopoeia.Only 1 batch of 5 batches of commercially sold MMF showed high glycosylase and amylase activity,which were 137.09 U/g and 549.75 U/g respectively.The digestive enzyme activity of MMF of pure fermentation was lower than that of single strain fermented.Conclusion:TCM decoction pieces fermented by natural microbiota are seriously contaminated by endogenous and exogenous microorganisms.The contamination of mycotoxins and pesticides was detected in some samples,and the detection rate and content were relatively high,while pure bacterial species fermentation could effectively avoid contamination.The microbial community abundance exist large differences in different Fermented TCM decoction pieces(FTCMDP)and corresponding microbial limits should be formulated to ensure the safety and effectiveness of medicines.It is suggested to use a variety of pure microbial species to ferment,and to optimize the fermentation process of fermented black bean and six Shenqu pieces from various angles from fermentation conditions,microbial species or raw materials,so as to ensure the quality of FTCMDP.This study provides a scientific basis for the production,management,supervision and control of fermented Chinese medicine pieces.