超高效液相色谱-质谱联用法检测地理标志产品松江大米中8种真菌毒素的含量

Determination of 8 Mycotoxins in Product of Geographical Indication Songjiang Rice by Ultra-high Performance Liquid Chromatography-Mass Spectrometry

建立超高效液相色谱-三重四极杆质谱法同时检测大米基质中8种真菌毒素的方法,对地理标志产品松江大米中真菌毒素的污染情况进行监测。大米样品经提取液乙腈水混合液(84∶16,体积比)超声提取后,取部分提取液加入内标溶液,混匀后过多功能净化柱MycoSep229净化,采用Waters Acquity UPLC BEH C18色谱柱(2.1 mm×50 mm,1.7μm),以体积分数0.1%甲酸水溶液-乙腈作为流动相进行梯度洗脱,分离黄曲霉毒素B 1(Aflatoxin B 1,AFB 1)、黄曲霉毒素B 2(Aflatoxin B 2,AFB 2)、黄曲霉毒素G 1(Aflatoxin G 1,AFG 1)、黄曲霉毒素G 2(Aflatoxin G 2,AFG 2)、伏马毒素B 1(Fumonisins B 1,FB 1)、伏马毒素B 2(Fumonisins B 2,FB 2)、伏马毒素B 3(Fumonisins B 3,FB 3)、赭曲霉毒素A(Ochratoxin A,OTA)8种真菌毒素化合物,该方法采用电喷雾离子化正离子模式,以多反应监测方式进行。AFB 1、AFG 1、FB 1、FB 2、FB 3、OTA 6种真菌毒素在0.5~15.0μg/L范围内线性良好,AFB 2、AFG 2在0.125~3.750μg/L范围内线性良好,线性相关系数r均大于0.999;8种化合物定量限(limit of quantifications,LOQs)范围为0.01~0.25μg/kg,比国标中相同方法的定量限降低2~80倍不等;在低、中、高三水平加标,其回收率均在95%~120%范围内,相对标准偏差(relative standard deviations,RSDs)为1.5%~8.9%(N=6)。本方法前处理简单,方法利用率高,结果准确可靠,利用该方法检测的20批新上市松江大米中真菌毒素污染情况比较乐观,对人体的健康危害较低。

To establish a method for the determination of 8 mycotoxins in rice matrix by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UHPLC-MS/MS)and monitor the pollution levels of mycotoxins in product of geographical indication Songjiang rice.The rice products were processed with acetonitrile aqueous solution(84∶16,volume ratio)by ultrasonic extraction.Parts of the extract were added some internal standard solutions and purified by the Mcosep 229 multifunctional cleanup column.The 8 mycotoxin compounds are separated by Waters Acquity UPLC BEH C18 column(2.1 mm×50 mm,1.7μm)with volume fraction 0.1%formic acid aqueous solution-acetonitrile as mobile phases for gradient elution.They were detected by electrospray ion source in positive mode with multiple reaction monitoring(MRM)mode.The calibration curves showed great linearities in the range of 0.5-15.0μg/L,such as Aflatoxin B 1 and other 5 mycotoxins.The other calibration curves of Aflatoxin B 2 and Aflatoxin G 2 also showed good linearities in the range of 0.125-3.750μg/L.The correlation coefficient(r)of the calibration curves of all the compounds were above 0.999.The limits of quantifications(LOQs)of the 8 compounds were in the range of 0.01-0.25μg/kg,being 2-80 times lower than those of the same methods in the national standards.The recoveries of the 8 compounds at low,medium and high levels could reach 95%-120%,respectively,and the relative standard deviations(RSDs)were 1.5%-8.9%(N=6).In general,the method had the advantages of simple pretreatment,high utilization rate and accurate and reliable results.The pollution situation of mycotoxins in 20 batches of new Songjiang rice in the market determined by the above method was optimistic,and the risk to human health was at a lower level.