Analysis Of Aflatoxins And Ochratoxin A In Traditional Chinese Medicine,Bruceae Fuctus And Zingiberis Rhizoma Recens

Mycotoxins are secondary toxic metabolites produced by fungi. AFB1, AFB2, AFG1, AFG2and OTA are not only toxic but also a wide range of pollution. Chinese herbal medicines are vulnerable to be contaminated by mycotoxins during planting, harvesting and storage and the harm could be more serious without any control. Therefore, it is necessary to establish accurate, sensitive, reliable and easy detection methods to monitor the contamination of mycotoxins in traditional Chinese medicine (TCM) for the purpose of drug safety and providing the basis for the development of appropriate limits. Through a literature review and preliminary experiments, aflatoxins and ochratoxin A detection methods have been established in this study and the main contents are as follows:1. An IAC-HPLC-PCD-FLD method has been developed for the simultaneous determination of AFB1, AFB2, AFG1and AFG2in Bruceae Fuctus.The samples were extracted with methanol/water solution, followed by an IAC clean-up step and then the eluates were analyzed by high performance liquid chromatography and on-line post-column photochemical derivatization-fluorescence detection (IAC-HPLC-PCD-FLD) and the positive samples were further confirmed by HPLC-MS/MS. Aflatoxin B1,G1showed a good linear relationship in the range of0.50～50.00ng/mL and aflatoxin B2, G2at a range of0.15～15.00ng/mL. The limits of detection (LODs) of aflatoxin B1, B2, G1and G2were0.08、0.015、0.06、0.02μg/kg, respectively. The recoveries were74.3%～100.8%with relative standard deviations (RSD) of all below8.7%. The levels of aflatoxin B1ranged from0.26to22.89μg/kg and the total content of aflatoxin B1, B2, G1and G2were between0.31and27.52μg/kg in the positive samples. The first established method was simple, accurate, reproducible and reliable for the determination of aflatoxins in Bruceae Fuctus2. An IAC-UPLC-FLD method was developed for the simultaneous determination of AFB1, AFB2, AFG1, AFG2and OTA in Zingiberis Rhizoma Recens. AFs and OTA were extracted from samples with methanol/water solution, followed by an IAC clean-up step and then analyzed by ultra-performance liquid chromatography coupled with fluorescence detection (IAC-UPLC-FLD). Positive samples were further confirmed by HPLC-MS/MS. The recoveries of aflatoxin B1, B2, G1, G2and OTA were in the range of75.2%～99.7%and the LODs were0.04,0.04,0.10,0.07and0.25μg/kg, respectively. The max levels of aflatoxin B1and OTA were6.50and20.66μg/kg. The proposed method was simple, sensitive and reproducible for the rapid determination of AFs and OTA in Zingiberis Rhizoma Recens.3. A MIP-SPE-UPLC-FLD method was proposed for the rapid determination of OTA in Zingiberis Rhizoma RecensThe samples were firstly extracted and then cleaned up with an AFFINIMIP(?) SPE OTA column for UPLC-FLD analysis. The limit of detection (LOD) and limit of quantification (LOQ) for OTA were0.09and0.30ng/mL, respectively. The recoveries of OTA spiked at5,15and25μg/kg ranged from87.6to94.5%. In addition, after a simple regenerated procedure, the MIP-based SPE column could be reused at least forty-one times to obtain more than80%recoveries of OTA in this study. Under the optimized UPLC conditions, the analysis took around3.3min. The established method was simple, rapid and low-cost for the determination of OTA in Zingiberis Rhizoma Recens.