Contamination And Health Risk Assessment Of Aflatoxins In Wild Jujube Seeds And Multiple Mycotoxins In Body Samples

As one of the main exogenous pollutants affecting the quality and safety of traditional Chinese medicines,mycotoxins not only affect the quality of traditional Chinese medicines,but also pose potential health hazards to users.Due to its high pollution rate in the natural environment and the difficulty for natural degradation,it will accumulate in the host once it is produced,and then be accumulated and transmitted in animals and humans through the food chain.Long-term accumulation of mycotoxins in the body can lead to serious toxic damages such as liver and kidney dysfunction,carcinogenesis,and teratogenicity,which seriously affect human health.Therefore,the establishment of highly sensitive,high-throughput and rapid detection methods to monitor the contamination of mycotoxin in vitro and in vivo has important practical significance for the quality assurance of Chinese medicine and the safety of medication;it also helps to promote the prevention and control of mycotoxin contamination to achieve the ultimate goal of reducing the risk of exposure to the population.This paper is based on the needs of in vitro and in vivo exposure assessment of mycotoxins.The main research contents are as follows:1.Established aflatoxin residue analysis and health risk assessment methods for Semen Ziziphi Spinosae.Through analyzing the residues of aflatoxins（AFs）in 72 batches of Semen Ziziphi Spinosae from different regions,the risk of AFB₁ intake by the Semen Ziziphi Spinosae was evaluated.The contamination of AFs in 72 batches of Semen Ziziphi Spinosae was determined by IAC-HPLC-FLD method.An exposure assessment model based on Monte Carlo simulation combined with pollution data was established,and the risk of exposure（MOE）was used to assess the health risk of aflatoxin contamination in Semen Ziziphi Spinosae.The results showed that 44.4%of the Semen Ziziphi Spinosae samples were found to contain AFB₁.The average and maximum contamination level in the samples were 5.42μg/kg and 55.09μg/kg,respectively.There was no significant regional difference in the contamination level distribution.The average daily exposure,97.5%（90%confidence interval）of AFB₁ intake by Semen Ziziphi Spinosae was calculated to be 0.0086（0.0081⁰.0092）and 0.0573（0.0532-0.0614）μg/（kg·d）,respectively.The results of health risk assessment showed that the health risk of AFB₁ exposure was lower by taking Semen Ziziphi Spinosae,but the risk of high intake was higher than that of regular intake,which needs attention.This study can provide reference for the clinical evaluation of Semen Ziziphi Spinosae.2.Established UFLC-MS/MS determination method for 10 mycotoxins in plasmaA highly sensitive UPLC-MS/MS method for simultaneous determination of aflatoxin B₁（AFB₁）,aflatoxin B₂（AFB₂）,aflatoxin G₁（AFG₁）,aflatoxin G₂（AFG₂）,aflatoxin M₁（AFM₁）,ochratoxin A（OTA）,Fumonisin B₁（FB₁）,Fumonisin B₂（FB₂）,Deoxynivalenol（DON）,Zearalenone（ZEN）contamination levels in human plasma samples was established by optimizing extraction solvent,chromatography and mass spectrometry conditions.The results showed that the linear relationships of the 10 mycotoxins in the linear range were good,the correlation coefficient R² was greater than 0.998,the detection limit was 0.03 ng·mL^-10.2ng·mL^-1,the limit of quantification was 0.06 ng·mL^-10.4 ng·mL^-1,and the average recovery was 76.50%¹14.00%.The method involves less sample volume（100μL）,simple pretreatment operation,low cost,and short detection time,and can provide a reference for high-throughput detection of various mycotoxins in other in vivo samples.3.Assessment of internal exposure of mycotoxin biomarkers in different populationsThe content of 10 mycotoxin biomarkers in 500 human plasma samples was detected based on the established high-throughput UFLC-MS/MS detection method.The results showed that OTA was detected in three groups of healthy pregnant women,liver cancer patients and patients taking Chinese medicine.The other nine mycotoxin biomarkers were not detected.The average exposure levels of the three groups were 0.57μg·L^-1,0.30μg·L^-11 and 0.36μg·L^-1,respectively.The detection rate of males under 60 years old taking Chinese medicine was about47.83%,and the detection rate of pregnant women is the lowest（9.7%）.There was no significant correlation between the differences in exposure levels between the population and gender,age,and population classification（P>0.05）.The Provisional Daily Intake（PDI）exposures of patients taking Chinese medicine,healthy pregnant women and liver cancer patients ranged from 0.017 to0.325μg/kg·bw/day,and the maximum appeared to be 0.325μg/kg·bw/day in healthy pregnant women;the OTA content in about 3.0%pregnant women body exceeded Provisional Maximum Tolerable Daily Intake（PMTDI）,and about 1.54%of liver cancer patients exceeded PMTDI;the PDI values of the patients taking Chinese medicine did not exceed PMTDI.It should be noted that the pregnant women are high-risk groups in the three groups of people.