Determination of Imidazole in Cigarette Additives by High Performance Liquid Chromatography with Pre-column Derivatization
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KeyWord:pre-column derivatization  high-performance liquid chromatography(HPLC)  cigarette additives  imidazole  dansyl chloride
CHENG Li-xia,HE Dong-xiang,YANG Jin-long,L Chang-ping 甘肃烟草工业有限责任公司技术研发中心
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      An effective method of high-performance liquid chromatography(HPLC)was developed for the determination of imidazole,the harmful ingredient in cigarette additives based on pre-column derivatization with dansyl chloride(Dns-Cl).The additive samples firstly underwent the treatment of a buffer of carbonate solution,followed by the pre-column derivatization with dansyl chloride,then the extraction with ethyl acetate,dryness,evaporation,dilution with acetonitrile and filteration,separation on a C18 column,elution with acetonitrile-water by gradient elation,and finally the detection with a fluorescence detector.The optimum derivatising conditions of imidazole with dansyl chloride are as follows:concentration of dansyl chloride:1 000 mg/L;temperature of reaction:45 ℃;pH of buffer solution:10.0;time of reaction:40 min. The imidazole derivative was quantified by the external standard method.There was a good linearity for the analyte in the range of 0.5-500 mg/L with a correlation coefficient of 0.999 5.The limits of detection of the method for imidazole derivative was 0.24 mg/kg,and the limits of quantitation was 0.80 mg/kg.The recoveries at three spiked levels ranged from 89.1% to 97.2% with RSDs not more than 7.5%.Results showed that the method was accurate and reliable,and was applicable to the determination of imidazole content in cigarette additives.