Determination of aflatoxin by HPLC-post-column derivatization-fluorescence detection

Aflatoxin is a group of toxic metabolites produced by Aflatoxin during growth and reproduction. Aflatoxin mainly pollutes grain and oil crops, such as peanuts, corn, rice, cottonseeds, etc. Aflatoxin has a destructive effect on human and animal liver tissues. In severe cases, it can cause liver cancer or even death. Therefore, there are strict international requirements for aflatoxins in food.

In this paper, the post-column derivatization method of high-performance liquid fluorescence detector was used to determine aflatoxin. Through the selection of chromatographic column, optimization of mobile phase ratio and adjustment of detector conditions, the best separation effect of aflatoxin was obtained.

1. Experimental method

Cometro high performance liquid chromatography system;

Fluorescence detector, excitation wavelength 360nm, emission wavelength 450nm,

Cometro post-column derivatization system, post-column derivatization at 70 degrees, derivatizing agent: 0.03% methanol aqueous solution of iodine, flow rate 0.3ml / min;

Mobile phase: methanol-acetonitrile-water;

Flow rate: 0.8ml / min;

Chromatography column: octadecyl bonded silica gel column of various brands;

Sample: Aflatoxin stock solution;

2. Experimental process

1. Optimization of mobile phase ratio

Through experiments, select the Comatex column with the best separation effect, adjust the ratio of mobile phase to methanol-acetonitrile-water = 40:10:50, other experimental conditions remain unchanged.

Under this condition, a better separation effect can be obtained.

2. Adjustment of detector conditions

Optimize the detector conditions, use Comatex C18 column, optimize the mobile phase, and repeat the above experiment

The results show that baseline noise (0.2mv), sensitivity, signal-to-noise ratio (> 10), column efficiency (> 9000), and resolution between main peaks (> 1.5).

3. Conclusion

Through the choice of chromatographic column, the optimization of the mobile phase ratio, and the adjustment of the energy of the lamp, the resolution can be improved, the noise can be reduced, the sensitivity can be improved, and a good separation effect can be obtained. The post-column derivatization method of high-performance liquid fluorescence detector was used to determine aflatoxin, and the Comatex column was used; the mobile phase was methanol-acetonitrile-water = 40: 10: 50; the optimal separation conditions could be obtained by optimizing the detector conditions.

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