液相色谱法和液质联用法检测玉米中的脱氧雪腐镰刀菌烯醇的比较
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摘要
为了选择实验室准确检测脱氧雪腐镰刀菌烯醇的快速经济的方法,比较免疫亲和柱-高效液相色谱(HPLC)法和免疫亲和柱-液质联用(LC-MS)法检测脱氧雪腐镰刀菌烯醇。结果表明,两种方法的线性关系均良好,R~2=0.999 9;HPLC法、LC-MS外标法及内标法精密度试验结果相对标准偏差(RSD)分别为1.31%、1.20%及0.37%,回收率分别在80.77%～91.92%、108.71%～105.56%及113.48%～119.27%,精密度、回收率及重复性试验结果RSD均<5%;HPLC法的定量限和检出限分别为76.82μg/kg、23.05μg/kg,LC-MS法的定量限和检出限分别为1.67μg/kg、0.502μg/kg,均低于国标给定标准。LC-MS法灵敏度高,检出限低,但在检测由本底样品时,容易造成假阳性,HPLC法的提取溶剂是水,LC-MS法提取溶剂是乙腈。综合分析,在均能满足实验要求情况下,HPLC法更简便、经济、环保,可作为实验室大批量检测的首选。
In order to select a rapid and economical method for the accurate detection of deoxynivalenol in laboratory, the deoxynivalenol was determined by immunoaffinity column HPLC and immunoaffinity column LC-MS. The results showed that the linear relationship of the two methods were both good, R~2=0.999 9. The relative standard deviations(RSD) of the precision tests results of HPLC method, external standard method and internal standard method of LC-MS were 1.31%, 1.20% and 0.37%, respectively, the recovering rates were 80.77%-91.92%, 108.71%-105.56%, and 113.48%-119.27%, respectively. The RSD of precision, recovering rate and repeatability tests results were all less than 5%. The quantitative limit and detection limit of HPLC method were 76.82 μg/kg and 23.05 μg/kg, respectively. The quantitative limit and detection limit of LC-MS method were 1.67 μg/kg and 0.502 μg/kg, respectively, which were lower than the national standard. But the sensitivity of LC-MS was high and the detection limit was low. It was easy to cause false positive in the detection of background samples, the extraction solvent of HPLC was water, and the extraction solvent of LC-MS was acetonitrile. As comprehensive analysis, under the condition that all could meet the experimental requirements, HPLC method was more convenient, economic and environmentally friendly, and was the first choice for large-scale laboratory detection.
In order to select a rapid and economical method for the accurate detection of deoxynivalenol in laboratory, the deoxynivalenol was determined by immunoaffinity column HPLC and immunoaffinity column LC-MS. The results showed that the linear relationship of the two methods were both good, R~2=0.999 9. The relative standard deviations(RSD) of the precision tests results of HPLC method, external standard method and internal standard method of LC-MS were 1.31%, 1.20% and 0.37%, respectively, the recovering rates were 80.77%-91.92%, 108.71%-105.56%, and 113.48%-119.27%, respectively. The RSD of precision, recovering rate and repeatability tests results were all less than 5%. The quantitative limit and detection limit of HPLC method were 76.82 μg/kg and 23.05 μg/kg, respectively. The quantitative limit and detection limit of LC-MS method were 1.67 μg/kg and 0.502 μg/kg, respectively, which were lower than the national standard. But the sensitivity of LC-MS was high and the detection limit was low. It was easy to cause false positive in the detection of background samples, the extraction solvent of HPLC was water, and the extraction solvent of LC-MS was acetonitrile. As comprehensive analysis, under the condition that all could meet the experimental requirements, HPLC method was more convenient, economic and environmentally friendly, and was the first choice for large-scale laboratory detection.
引文
[1] JAJIC I, KRSTOVIC S, KOS J, et al. Incidence of deoxynivalenol in Serbian wheat and barley[J]. J Food Protect, 2014, 77(5):853-858.
[2] BENNETT J W, KLICH M. Mycotoxins[J]. Clin Microbiol Rev, 2003,16(3):497.
[3]陆晶晶,杨大进. 2013年中国小麦粉中脱氧雪腐镰刀菌烯醇污染调查[J].卫生研究,2015,44(4):658-660.
[4] T譈RKER L, G譈M譈S S. A theoretical study on vomitoxin and its tautomers[J]. J Hazard Mat, 2009, 163(1):285-294.
[5] PESTKA J J, SMOLINSKI A T. Deoxynivalenol:toxicology and potential effects on humans[J]. J Toxicol Environ Health Part B, 2005, 8(1):39.
[6]万小乐.高效液相色谱法测定粮食中呕吐毒素的方法研究[J].粮食加工,2016,41(4):74-76.
[7]霍星华,赵宝玉,万学攀,等.脱氧雪腐镰刀菌烯醇的毒性研究进展[J].毒理学杂志,2008,22(2):151-154.
[8]李群伟,侯海峰,李亚鲁,等.脱氧雪腐镰刀菌烯醇致家兔多脏器损伤的实验观察[J].中国地方病防治杂志,2007,22(1):19-22.
[9]何樊,闫冬阁,陈渠玲,等.免疫亲和高效液相色谱法测定小麦粉品中脱氧雪腐镰刀菌烯醇的方法改进[J].现代食品科技,2018,34(1):1-6.
[10] KARAMI-OSBOO R, MAHAM M, MIRI R, et al. Evaluation of dispersive liquid-liquid microextraction-HPLC-UV for determination of deoxynivalenol(DON)in wheat flour[J]. Food Anal Meth, 2013, 6(1):176-180.
[11]魏润蕴,王竹天.谷物中脱氧雪腐镰刀菌烯醇和雪腐镰刀菌烯醇的气相色谱测定[J].卫生研究,1996,25(4):243-245.
[12] SHAR Z H, SUMBAL G A, SHERAZI S T H, et al. Determination of deoxynivalenol in poultry feed by three gas chromatographic detection techniques[J]. Chromatographia, 2014, 77(3-4):337-346.
[13]朱群英,索莉莉,胡美华.液质联用同位素内标法同时测定小麦粉中7种真菌毒素[J].现代预防医学,2014,41(23):4362-4365.
[14] KLINGLMAYR C, NOBAUER K, RAZZAZI-FAZELI E, et al. Determination of deoxynivalenol in organic and conventional food and feed by sol-gel immunoaffinity chromatography and HPLC-UV detection[J].J Chromatogr B, 2010, 878(2):187-193.
[15] SOLEIMANY F, JINAP S, FARIDAH A, et al. A UPLC-MS/MS for simultaneous determination of aflatoxin A, zearalenone, DON, fumonisins,T-2 toxin and HT-2 toxin, in cereals[J]. Food Control, 2012, 25(2):647-653.
[16]熊齐荣,金涌,刑仕歌,等.胶体金试纸条法快速筛查小麦和玉米中脱氧雪腐镰刀菌烯醇的研究[J].食品科技,2014,39(2):292-296.
[17]中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会. GB 5009.111—2016食品安全国家标准食品中脱氧雪腐镰刀菌烯醇及其乙酰化衍生物的测定[D].北京:中国标准出版社,2016.
[18]王培,徐广超,尹丽梅,等.免疫亲和柱净化-高效液相色谱法同时检测小麦中脱氧雪腐镰刀菌烯醇及其衍生物[J].粮油食品科技,2015,23(1):63-66.
[19]何攀,闫东阁,陈渠玲,等.免疫亲和高效液相色谱法测定小麦粉品中脱氧雪腐镰刀菌烯醇的方法改进[J].现代食品科技,2018,34(1):1-6.
[20]免疫亲和层析净化高效液相色谱法测定小麦粉中脱氧雪腐镰刀菌烯醇方法的研究[J].食品科学,2018,39(2):48-50.
[21]吴振兴,鲍蕾,静平,等.小麦和玉米中脱氧雪腐镰刀菌烯醇与雪腐镰刀菌烯醇的免疫亲和净化-高效液相色谱方法研究[J].食品安全质量检测学报,2014,5(12):3848-3852.
[22]许娇娇,黄百芬,蔡增轩,等.面粉中脱氧雪腐镰刀菌烯醇标准物质的研制[J].中国食品卫生杂志,2016,28(3):291-294.
[2] BENNETT J W, KLICH M. Mycotoxins[J]. Clin Microbiol Rev, 2003,16(3):497.
[3]陆晶晶,杨大进. 2013年中国小麦粉中脱氧雪腐镰刀菌烯醇污染调查[J].卫生研究,2015,44(4):658-660.
[4] T譈RKER L, G譈M譈S S. A theoretical study on vomitoxin and its tautomers[J]. J Hazard Mat, 2009, 163(1):285-294.
[5] PESTKA J J, SMOLINSKI A T. Deoxynivalenol:toxicology and potential effects on humans[J]. J Toxicol Environ Health Part B, 2005, 8(1):39.
[6]万小乐.高效液相色谱法测定粮食中呕吐毒素的方法研究[J].粮食加工,2016,41(4):74-76.
[7]霍星华,赵宝玉,万学攀,等.脱氧雪腐镰刀菌烯醇的毒性研究进展[J].毒理学杂志,2008,22(2):151-154.
[8]李群伟,侯海峰,李亚鲁,等.脱氧雪腐镰刀菌烯醇致家兔多脏器损伤的实验观察[J].中国地方病防治杂志,2007,22(1):19-22.
[9]何樊,闫冬阁,陈渠玲,等.免疫亲和高效液相色谱法测定小麦粉品中脱氧雪腐镰刀菌烯醇的方法改进[J].现代食品科技,2018,34(1):1-6.
[10] KARAMI-OSBOO R, MAHAM M, MIRI R, et al. Evaluation of dispersive liquid-liquid microextraction-HPLC-UV for determination of deoxynivalenol(DON)in wheat flour[J]. Food Anal Meth, 2013, 6(1):176-180.
[11]魏润蕴,王竹天.谷物中脱氧雪腐镰刀菌烯醇和雪腐镰刀菌烯醇的气相色谱测定[J].卫生研究,1996,25(4):243-245.
[12] SHAR Z H, SUMBAL G A, SHERAZI S T H, et al. Determination of deoxynivalenol in poultry feed by three gas chromatographic detection techniques[J]. Chromatographia, 2014, 77(3-4):337-346.
[13]朱群英,索莉莉,胡美华.液质联用同位素内标法同时测定小麦粉中7种真菌毒素[J].现代预防医学,2014,41(23):4362-4365.
[14] KLINGLMAYR C, NOBAUER K, RAZZAZI-FAZELI E, et al. Determination of deoxynivalenol in organic and conventional food and feed by sol-gel immunoaffinity chromatography and HPLC-UV detection[J].J Chromatogr B, 2010, 878(2):187-193.
[15] SOLEIMANY F, JINAP S, FARIDAH A, et al. A UPLC-MS/MS for simultaneous determination of aflatoxin A, zearalenone, DON, fumonisins,T-2 toxin and HT-2 toxin, in cereals[J]. Food Control, 2012, 25(2):647-653.
[16]熊齐荣,金涌,刑仕歌,等.胶体金试纸条法快速筛查小麦和玉米中脱氧雪腐镰刀菌烯醇的研究[J].食品科技,2014,39(2):292-296.
[17]中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会. GB 5009.111—2016食品安全国家标准食品中脱氧雪腐镰刀菌烯醇及其乙酰化衍生物的测定[D].北京:中国标准出版社,2016.
[18]王培,徐广超,尹丽梅,等.免疫亲和柱净化-高效液相色谱法同时检测小麦中脱氧雪腐镰刀菌烯醇及其衍生物[J].粮油食品科技,2015,23(1):63-66.
[19]何攀,闫东阁,陈渠玲,等.免疫亲和高效液相色谱法测定小麦粉品中脱氧雪腐镰刀菌烯醇的方法改进[J].现代食品科技,2018,34(1):1-6.
[20]免疫亲和层析净化高效液相色谱法测定小麦粉中脱氧雪腐镰刀菌烯醇方法的研究[J].食品科学,2018,39(2):48-50.
[21]吴振兴,鲍蕾,静平,等.小麦和玉米中脱氧雪腐镰刀菌烯醇与雪腐镰刀菌烯醇的免疫亲和净化-高效液相色谱方法研究[J].食品安全质量检测学报,2014,5(12):3848-3852.
[22]许娇娇,黄百芬,蔡增轩,等.面粉中脱氧雪腐镰刀菌烯醇标准物质的研制[J].中国食品卫生杂志,2016,28(3):291-294.