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目的采用免疫亲和净化手段和高效液相色谱(HPLC)检测技术,建立富含色素类中药材中黄曲霉毒素(AFT)的测定方法,为中药材中AFT的检测和污染调查提供技术支撑。方法中药材样品经过高速粉碎均匀后,粉末状样品经80%甲醇水高速均质提取,提取液以磷酸盐缓冲液(PBS)稀释后,用免疫亲和柱(IAC)净化,以含吐温-20的PBS淋洗柱子后,用甲醇洗脱AFT,洗脱液经三氟乙酸衍生后用HPLC分析,C18反相色谱柱分离,用荧光检测器(FLD)测定,外标法定量。结果黄曲霉毒素G1、B1、G2和B2的最低检出限(LOD,S/N=3)分别为0.1、0.02、0.05和0.05μg/kg;以不含AFT、色素含量较高的枳实、绞股蓝、枇杷叶、鱼腥草和乌梅为基质,进行加标回收试验,加标水平为6.0~24μg/kg(以AFT总量计),AFT的平均回收率在62.3%~115%之间,RSD在0.92%~15.9%之间(n=3)。结论 IAC净化技术和HPLC-FLD相结合,用于4种AFT的检测可取得较为满意的结果,方法的准确度高、精密度好,各项指标均满足欧盟对食品中AFT检测的要求,方法可满足色素含量较高的中药材中4种黄曲霉毒素(AFT)的测定。
Objective To establish an HPLC method for the determination of aflatoxins (AFT) in Chinese herbal medicines rich in pigment, and to provide technical support for the investigation and pollution investigation of AFT in Chinese herbal medicines by immunoaffinity purification and high performance liquid chromatography (HPLC). Methods After the Chinese herbal medicines were homogenized at high speed, the powder samples were homogenized by high speed homogenization in 80% methanol water. The extracts were diluted with phosphate buffered saline (PBS) and purified by immunoaffinity column (IAC) After washing the column with -20 PBS, the AFT was eluted with methanol. The eluate was derivatized with trifluoroacetic acid and analyzed by HPLC. The residue was separated on a C18 reversed-phase column and quantified by a fluorescence detector (FLD). Results The detection limits (LOD, S / N = 3) of aflatoxins G1, B1, G2 and B2 were 0.1,0.02,0.05 and 0.05μg / kg, respectively. , Gynostemma pentaphyllum, loquat leaf, Houttuynia cordata Thunb.and black plum as matrix, the spiked level was 6.0 ~ 24μg / kg (total amount of AFT), the average recoveries of AFT ranged from 62.3% to 115% , RSD ranged from 0.92% to 15.9% (n = 3). Conclusion The combination of IAC purification technology and HPLC-FLD can achieve satisfactory results for the four AFT tests. The method has high accuracy and precision, and all the indexes meet the EU requirements for AFT detection in food. Methods To meet the high pigment content of Chinese herbal medicines four kinds of aflatoxin (AFT) determination.