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Objective To compare the detection results of aflatoxin B1 (AFB1) content in various matrices such as rice, wheat products, and peanut products using 2 kinds of different pretreatment methods: Fully automated immunomagentic bead purification and immunoaffinity column pretreatment method. Methods After extraction with methanol:water (70:30, V:V), a portion of the extract was purified using the immunoaffinity column method with an AFB1-specific immunoaffinity column, while another portion of the extract was processed by a fully automated immunomagnetic bead purification system paired with an immunomagnetic bead kit. Finally, both purified extracts were analyzed using high performance liquid chromatography coupled with post-column derivatization for detection. Results The efficacy of the immunomagetic bead purification kit and the column efficiency of the immunoaffinity column were both greater than 95%, which could meet the experimental requirements. Within mass concentration range of 1.00-50.00 ng/mL, the linear correlation coefficient (r) was 0.99993, indicating a good linear relationship. The 3 level spiked recovery experiments were conducted in rice matrix (n=6), and the recovery rates of the 2 kinds of pretreatment methods ranged from 98.2%-106.0%, with a relative standard deviation range of 1.0%-5.0%; single level spiked recovery experiments were conducted in wheat products (n=6), with a recovery rate exceeding 80% and a relative standard deviation of 3.7%-7.2%. Meanwhile, the 2 kinds of methods were used to detected AFB1 positive peanut products and the quality control sample of corn flour matrix, their results were subjected to t-test, which showed no significant difference. Further, Bland-Altman statistical analysis was carried out on the results obtained from the quality control samples of corn flour matrix after 2 kinds of pretreatment methods were used. The results showed that the 2 kinds of methods could replace each other in the detection of AFB1 in this matrix. The purification step of the immunoaffinity column method was time-consuming, 20 batches of samples take about 3-4 hours. And could only be manually operated, which generated about 1000 mL waste liquid; while the immunomagetic bead method used a fully automatic magnetic bead purifier, which automatically completed 20 batches of samples in 40 minutes at one time, with almost no waste liquid generated. Conclusion The fully automated immunomagnetic bead purification method and the immunoaffinity column method both can achieve good results in detecting AFB1 in various matrices. Also, the immunomagnetic bead purification method is more efficient, easy to handle, generates less waste liquid, and the cost can be reduced by about 50%.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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