Development Of Natural Anti-mold Agent And Its Application In Detection Of Polytoxin In Chinese Herbal Medicine

ObjectiveNatural plant essential oil monomer was used as research object to develop natural anti-fungal agent.The inhibitory effects on Aspergillus flavus,Aspergillus niger and Penicillium sp.,as well as the inhibition of mold growth and production of ginseng,gastrodia elata and lily in Chinese medicinal materials were studied.At the same time,the LC-MS/MS method was established to simultaneously qualitatively and quantitatively detect various mycotoxins in Chinese herbal medicine matrix,and applied it to detection the actual sample.MethodsFirstly,the"fumigation method"was used to select the best effect among the seven essential oil monomers.As the research object,and the minimum inhibitory concentration was determined,polyvinyl alcohol（PVA）was selected as the film-forming material to prepare the carvacrol.Mold proofing agent.Secondly,UFLC-MS/MS was established to simultaneously detect AFS and OTA in ginseng,gastrodia and lily.At the same time,a UPLC method for the determination of ginsenoside Rg₁,ginsenoside Rb and ginsenoside Re in ginseng was established,and the content of ginseng samples under the action of antifungal agents developed in this paper was determined.A UPLC method for the determination of gastrodin and p-hydroxybenzyl alcohol in Gastrodia elata was established.The content of the gastrodia elata sample under the action of the antifungal agent developed in this paper was also determined.Methodological investigations were carried out on the two methods.Finally,an external standard method was used to quantify the method of simultaneous determination of 19 mycotoxins in different batches of lotus seeds by ultra performance liquid chromatography-tandem mass spectrometry.Due to the complex matrix of lotus seeds,it is possible to contaminate many types of mycotoxins at the same time.In this study,the extraction solution and purification method were optimized.At the same time,the linearity,method specificity,matrix effect,recovery rate and precision of the method were investigated.ResultsCarvacrol mildew film can significantly inhibit the growth of three molds and the accumulation of aflatoxin（AFS）and ochratoxin（OTA）in ginseng,gastrodia elata and lily.It was found by UPLC method that it does not affect the active ingredients in Chinese herbal medicines.The UFLC-MS/MS method established in this thesis has strong specificity,high sensitivity,and simple pretreatment process.It is suitable for detecting a large number of mycotoxins in lotus seed samples at the same time.A total of 45 batches of lotus seeds were tested and the results showed that 26 batches（57.8%of the contamination ratio）were contaminated by more than one mycotoxin at different levels,in addition,a batch of samples were simultaneously contaminated with 7 different toxins.ConclusionsThe natural mildewproof film developed in this paper has a significant effect on inhibiting mold growth and toxicity.It does not affect the active ingredients in the three Chinese herbal medicines of ginseng,gastrodia and lily.The UFLC-MS/MS method established in this paper has strong specificity,high sensitivity and simple pretreatment process,which is suitable for detecting a large number of mycotoxins in Chinese herbal medicine samples at the same time.