改良QuEChERS法结合UPLC-QTRAP-MS/MS技术测定玉米中的伏马毒素B_1、B_2
|
摘要
改进QuEChERS样品制备方法,为玉米中伏马毒素B1、B2的UPLC-QTRAP-MS/MS检测提供良好的分析结果。该方法包括将样品与酸化的乙腈水溶液混合振荡提取,然后使用MgSO4、NaCl进行盐析液体分配,通过用MgSO4、PSA、C18、石墨化碳黑和中性氧化铝分散固相萃取净化,C18色谱柱分离,以甲醇-0. 1%甲酸水溶液为流动相梯度洗脱,正模式扫描,采用多反应检测(MRM)-信息依赖采集(IDA)-增强子离子扫描(EPI)模式检测,在线EPI谱库定性分析,外标法定量。结果表明:在10～500μg/L的质量浓度范围内,2种伏马毒素的线性相关系数均大于0. 999,检出限分别为5μg/kg和3μg/kg;添加低、中、高3个水平,2种伏马毒素的平均回收率在89. 11%～104. 61%之间,相对标准偏差在2. 33%～9. 89%之间。与免疫亲和柱法比较发现,两种方法的回收率和特异性差别不大,但是改良的QuEChERS方法更具有时间和成本效益。
The sample preparation method of QuEChERS was improved to provide a good analytical result for the determination of fumonisins B_1 and B_2 in maize with UPLC-QTRAP-MS/MS. The samples were shakingly extracted with acidified acetonitrile aqueous solution,after liquid-liquid partition using MgSO_4 and NaCl,the supernatant was purified using Mg SO_4,PSA,C_(18),graphitized carbon black and neutral alumina as adsorbing material,then separation of fumonisins B_1 and B_2 was performed with C_(18) column.The mobile phase gradiently eluted with methanol and 0. 1% formic acid aqueous solution. The electrospray was operated in the positive ionization mode and two kinds of fumonisins were identified by multi-reaction detection( MRM)-information dependent acquisition( IDA)-enhanced production ion( EPI)scan mode. Qualitative analysis was performed by online EPI spectrum library search,and quantification was conducted by external standard method. The results showed that the calibration curves showed good linearity within the mass concentration of 10-500 μg/L with the correlation coefficients all above 0. 999.The detection limits for fumonisins B_1 and B_2 were 5 μg/kg and 3 μg/kg respectively. The recoveries of two kinds of fumonisins were 89. 11%-104. 61%at low,medium and high levels of addition,and the relative standard deviation were 2. 33%-9. 89%. Compared with immunoaffinity column method,the recoveries and specificity of the two methods were not much different,but the improved QuEChERS method had more advantages in analytical time and cost.
The sample preparation method of QuEChERS was improved to provide a good analytical result for the determination of fumonisins B_1 and B_2 in maize with UPLC-QTRAP-MS/MS. The samples were shakingly extracted with acidified acetonitrile aqueous solution,after liquid-liquid partition using MgSO_4 and NaCl,the supernatant was purified using Mg SO_4,PSA,C_(18),graphitized carbon black and neutral alumina as adsorbing material,then separation of fumonisins B_1 and B_2 was performed with C_(18) column.The mobile phase gradiently eluted with methanol and 0. 1% formic acid aqueous solution. The electrospray was operated in the positive ionization mode and two kinds of fumonisins were identified by multi-reaction detection( MRM)-information dependent acquisition( IDA)-enhanced production ion( EPI)scan mode. Qualitative analysis was performed by online EPI spectrum library search,and quantification was conducted by external standard method. The results showed that the calibration curves showed good linearity within the mass concentration of 10-500 μg/L with the correlation coefficients all above 0. 999.The detection limits for fumonisins B_1 and B_2 were 5 μg/kg and 3 μg/kg respectively. The recoveries of two kinds of fumonisins were 89. 11%-104. 61%at low,medium and high levels of addition,and the relative standard deviation were 2. 33%-9. 89%. Compared with immunoaffinity column method,the recoveries and specificity of the two methods were not much different,but the improved QuEChERS method had more advantages in analytical time and cost.
引文
[1]SAUVIATT M P,LAURENT D,KOHLER F,et al.Fumonisin,a toxin from the fungus Fusarium moniliforme sheld,blocks both the calcium current and the mechanical activity in frog atrial muscle[J].Toxicon Off J Int Soc Toxicol,1991,29(8):1025-1031.
[2]GELDERBLON W C,JASKIEWICZ K,MARASAS W F,et al.Fumonisins-novel mycotoxins with cancer-promoting activity produced by Fusarium moniliforme[J].Appl Environ Microbiol,1988,54(7):1806-1811.
[3]蒋艳成,陈志刚,许广人,等.伏马毒素对猪的毒性研究进展[J].中国饲料,2016(1):35-38.
[4]BRANDWAGT B F,MESBAH L A,TAKKEN F L W,et al.A longevity assurance gene homolog of tomato mediates resistance to Alternaria alternata f.sp.lycopersici toxins and fumonisin B1[J].Proc Nat Acad Sci USA,2000,97(9):4961-4966.
[5]刘中勇,刘津,张喆,等.粮谷类食品中伏马毒素限量标准和检测方法进展[J].粮食与饲料工业,2009(7):45-47.
[6]李成龙,米铁军,张会艳,等.荧光偏振免疫分析检测伏马毒素B1研究[C].郑州:中国畜牧兽医学会兽医药理毒理学分会第十二次学术讨论会,2013.
[7]管笛,潘灿平,王文,等.磁微粒酶联免疫吸附法测定玉米中的伏马毒素B1[J].食品科学,2014,35(8):208-211.
[8]王军淋,胡玲玲,蔡增轩,等.超高压液相色谱法同时检测玉米中的伏马毒素B1、B2、B3[J].食品安全质量检测学报,2013,4(1):215-223.
[9]李为喜,郑床木,武力,等.测定玉米中伏马毒素的免疫亲和层析净化高效液相色谱法[J].作物学报,2012,38(3):556-562.
[10]李永刚,张瑞,聂晓明,等.超高效液相色谱-串联质谱法测定玉米油中的伏马毒素[J].中国卫生检验杂志,2017,27(3):336-338,343.
[11]李正翔,陈小龙,曹赵云,等.液相色谱-串联质谱法测定粮谷中的伏马毒素[J].分析测试学报,2014,33(2):167-172.
[12]郑翠梅,张艳,王松雪,等.液相色谱-质谱联用同时检测粮食中多种真菌毒素的应用进展[J].粮食科技与经济,2012,37(1):45-49.
[13]魏云计,冯民,黄娟,等.固相萃取/高效液相色谱-串联质谱法快速测定饲料中的3种伏马毒素[J].分析测试学报,2018,37(6):729-733.
[14]薛秋艳,徐剑宏,祭芳,等.玉米中伏马毒素FB1液相色谱-电喷雾串联质谱测定方法的建立[J].江苏农业学报,2010,26(3):627-630.
[15]孙伟华,杨欢,曹赵云,等.基于分散固相萃取液相色谱-串联质谱法测定大米中8种真菌毒素[J].分析测试学报,2017,36(1):47-53.
[2]GELDERBLON W C,JASKIEWICZ K,MARASAS W F,et al.Fumonisins-novel mycotoxins with cancer-promoting activity produced by Fusarium moniliforme[J].Appl Environ Microbiol,1988,54(7):1806-1811.
[3]蒋艳成,陈志刚,许广人,等.伏马毒素对猪的毒性研究进展[J].中国饲料,2016(1):35-38.
[4]BRANDWAGT B F,MESBAH L A,TAKKEN F L W,et al.A longevity assurance gene homolog of tomato mediates resistance to Alternaria alternata f.sp.lycopersici toxins and fumonisin B1[J].Proc Nat Acad Sci USA,2000,97(9):4961-4966.
[5]刘中勇,刘津,张喆,等.粮谷类食品中伏马毒素限量标准和检测方法进展[J].粮食与饲料工业,2009(7):45-47.
[6]李成龙,米铁军,张会艳,等.荧光偏振免疫分析检测伏马毒素B1研究[C].郑州:中国畜牧兽医学会兽医药理毒理学分会第十二次学术讨论会,2013.
[7]管笛,潘灿平,王文,等.磁微粒酶联免疫吸附法测定玉米中的伏马毒素B1[J].食品科学,2014,35(8):208-211.
[8]王军淋,胡玲玲,蔡增轩,等.超高压液相色谱法同时检测玉米中的伏马毒素B1、B2、B3[J].食品安全质量检测学报,2013,4(1):215-223.
[9]李为喜,郑床木,武力,等.测定玉米中伏马毒素的免疫亲和层析净化高效液相色谱法[J].作物学报,2012,38(3):556-562.
[10]李永刚,张瑞,聂晓明,等.超高效液相色谱-串联质谱法测定玉米油中的伏马毒素[J].中国卫生检验杂志,2017,27(3):336-338,343.
[11]李正翔,陈小龙,曹赵云,等.液相色谱-串联质谱法测定粮谷中的伏马毒素[J].分析测试学报,2014,33(2):167-172.
[12]郑翠梅,张艳,王松雪,等.液相色谱-质谱联用同时检测粮食中多种真菌毒素的应用进展[J].粮食科技与经济,2012,37(1):45-49.
[13]魏云计,冯民,黄娟,等.固相萃取/高效液相色谱-串联质谱法快速测定饲料中的3种伏马毒素[J].分析测试学报,2018,37(6):729-733.
[14]薛秋艳,徐剑宏,祭芳,等.玉米中伏马毒素FB1液相色谱-电喷雾串联质谱测定方法的建立[J].江苏农业学报,2010,26(3):627-630.
[15]孙伟华,杨欢,曹赵云,等.基于分散固相萃取液相色谱-串联质谱法测定大米中8种真菌毒素[J].分析测试学报,2017,36(1):47-53.