高效液相色谱-光化学衍生法测定湖南产莲子中黄曲霉毒素G1、G2、B1、B2

Determination of aflatoxin G,G,B,and B in Nelumbinis Semen from Hunan Province by HPLC-photochemical derivatization

目的对不同来源的湖南产莲子中黄曲霉毒素G1、G2、B2、B1进行高效液相色谱-光化学衍生法测定。方法采用高效液相色谱–光化学衍生法,采用岛津GL Inertsil ODS-3色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇–乙腈–水(35∶13∶52),柱温35℃;光化学衍生器(254 nm)激发波长λex=360 nm,发射波长λex=450 nm。结果黄曲霉毒素G1、G2、B2、B1分别在6.7~33.3、11.4~56.8、10.3~51.5、4.1~20.3 pg线性关系良好;平均回收率分别为98.07%、97.72%、96.11%、99.52%,RSD值分别为1.69%、1.40%、2.72%、1.34%(n=6)。9批莲子样品中,有6批未检出黄曲霉毒素,来自于农贸市场农户自存的2批次检出黄曲霉毒素B1,实验室塑料袋包装储存1年的1批检出黄曲霉毒素B1、B2,但质量分数均低于法定标准限量。结论不同来源湖南产莲子中黄曲霉毒素质量分数均低于《中国药典》2020年版规定限量。该法测定结果准确、重复性良好,可为完善莲子安全性控制和质量标准提升提供实验依据。

Objective To establish an HPLC-photochemical derivatization method for determination of aflatoxin G1,G2,B2,and B1 in Nelumbinis Semen from different sources in Hunan Province.Methods The content of aflatoxins were determined by HPLC-photochemical derivatization method.Shimadzu GL Inertsil ODS-3 chromatographic column(250 mm×4.6 mm,5μm)was used with column temperature of 35℃,and methanol-acetonitrile-water(35∶13∶52)was used.Fluorescence detector:excitation wavelengthλex=360 nm and emission wavelengthλem=450 nm.Results Aflatoxin G2,G1,B2,and B1 showed good linear relationships in the ranges of 6.7—33.3,11.4—56.8,10.3—51.5,and 4.1—20.3 pg,and their average recoveries were 98.07%,97.72%,96.11%,and 99.52%with the RSD values were 1.69%,1.40%,2.72%,and 1.34%,respectively.Among 9 batches of Nelumbinis Semen samples,aflatoxin was not detected in 6 batches,and aflatoxin B1 was detected in 2 batches from farmers’market,and aflatoxin B1and B2 were detected in 1 batch of sample from laboratory plastic bags storage for one year.But the aflatoxin content was much lower than the Chinese Pharmacopoeia 2020 Edition legal standard limit.Conclusion Aflatoxin in Nelumbinis Semen samples of different sources in Hunan Province was lower than the limit stipulated in the Chinese Pharmacopoeia 2020 Edition.The method is accurate and reproducible,which can provide experimental basis for improving the safety control and quality standard of Nelumbinis Semen.