| Mebendazole (MEB), hardly soluble in water, is a broad-spectrum insecticide with the nature of low toxicity to mammals. Applied both in human and animals, EMB can usually be served as a kind of gastrointestinal drug for animals and fish. In addition, it can also be used to kill the phyla Annelida in fish.At present, the methods used for detecting mebendazole include titration, UV-spectrophotometry, colorimetry, phosphorescence-spectrophotometry, fluorescence-spectrophotometry, infrared-spectrophotometry, thin-layer chromatography and high performance liquid chromatography. However, most of these methods are used to detect the content of mebendazole contained in mebendazole troches or the amount of mebendazole in the blood, while few of them are used to detect the residue of mebendazole in aquatic products. And these methods have a high detection limit, which can hardly meet the standard——20μg/kg stipulated in the confirmative list established by Japan.This paper has systematically studied the detection methods of mebendazole in aquatic products with the aid of high performance liquid chromatography, and then the optimum detection condition was obtained according to the characteristic of HPLC and the nature of mebendazole.In this paper, the condition of chromatographic column, mobile phase, ratio of mobile phase, longest absorption wavelength and column temperature, which might have a significant effect on the results, were chosen as variable factors to determine the chromatographic condition of HPLC. Through test of precision, detecting limit, sample stability and spiked standard experiment, we have checked the methodology of the method.By means of experiments , the optimum chromatography condition—chromatography column using anti-phase Mightysil RP18 chromatography column(250mm×4.6 mm, 5μm); mobile phase, 0.3% phosphoric acid - acetonitrile mixed solution(65+35, 0.3% acid solution using ethylamine to adjust PH to 2.5 ); mobile speed, 1.0ml/min;detection wavelength,247nm;column temperature, 35℃. The results indicated that the mebendazole with concentration of between 0.1μg/mL～2.4μg/mL have a good linear relation(r=0. 9988).The detection limit of the method is 10μg/kg which is lower than 20μg/kg stipulated in the confirmative list established by Japan. Using previous method to perform experiments on the recovery rate of common aquatic products, and the recovery rate is between 86.42% and 96.82%, the average relative standard deviation is 1.58%.This paper also discussed the factors that affect uncertainty during the measurement in the HPLC, then give an analysis and evaluation on it, so as to gain a completely rationed description of the measurement result quality, and make the result traceable . Thus, make the newly-built analysis method more systematic and reliable. Through the uncertainty analysis of measurement, we can find out a standard curve that has the largest effect on the measurement result.Making use of certain experiment condition and according to actual needs, we have worked out a local standard in Fujian Province on the remain of mebendazole in aquatic products, and it has been announced and carried out in the province.The study was carried out by this subject on analyzing the residue of mebendazole by HPLC, the operation of which is easy, sensitive, reliable and has reference value in actual use. The method is suitable for analyzing and measuring the residue of mebendazole in aquatic products.
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