多壁碳纳米管分散固相萃取净化在测定水产品中3种微囊藻毒素的应用及与固相萃取法的比较
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摘要
建立多壁碳纳米管净化-超高效液相色谱-串联质谱法对水产品中3种微囊藻毒素残留量的测定方法。水产品用80%甲醇溶液提取,经多壁碳纳米管净化后,采用电喷雾正离子多反应监测模式进行分析。对不同性质碳纳米管进行考察,并对净化条件进行优化。将优化后的分散固相萃取条件与采用HLB柱固相萃取的净化方式进行比较,两者在1~50 μg/L范围内呈现良好的线性,线性相关系数均大于0.99,3种微囊藻毒素的检出限为0.1~0.2 μg/kg。在不同浓度水平下进行加标实验,平均回收率在88.7%~95.5%之间,相对标准偏差不大于4.2%。该方法适用于水产品中微囊藻毒素的痕量检测。
An ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method was developed for the determination of 3 microcystins(MCs) in aquatic products. The samples were extracted with methanolwater(8:2, V/V) and the extract was then cleaned up by multi-walled carbon nanotubes(MWCNTs). The target analytes were detected by UPLC-MS/MS using positive electrospray ionization(ESI+) in the multiple reaction monitoring(MRM) mode. Different MWCNTs were investigated for their suitability as sorbents for dispersive solid phase extraction(DSPE) and the purification conditions were optimized. The efficiency of the optimized DSPE conditions was compared to that of solid phase extraction using HLB column. Both purification methods showed a good linearity(R2 ≥ 0.99) over the concentration range from 1 to 50 μg/L and their limits of quantification were in the range of 0.1–0.2 μg/kg. The average recoveries of three MCs at different spiked levels were 88.7%–95.5% with relative standard deviations(RSDs) equal to or lower than 4.2%. This DSPE method can be suitable for the rapid detection of trace microcystins in aquatic products.
An ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method was developed for the determination of 3 microcystins(MCs) in aquatic products. The samples were extracted with methanolwater(8:2, V/V) and the extract was then cleaned up by multi-walled carbon nanotubes(MWCNTs). The target analytes were detected by UPLC-MS/MS using positive electrospray ionization(ESI+) in the multiple reaction monitoring(MRM) mode. Different MWCNTs were investigated for their suitability as sorbents for dispersive solid phase extraction(DSPE) and the purification conditions were optimized. The efficiency of the optimized DSPE conditions was compared to that of solid phase extraction using HLB column. Both purification methods showed a good linearity(R2 ≥ 0.99) over the concentration range from 1 to 50 μg/L and their limits of quantification were in the range of 0.1–0.2 μg/kg. The average recoveries of three MCs at different spiked levels were 88.7%–95.5% with relative standard deviations(RSDs) equal to or lower than 4.2%. This DSPE method can be suitable for the rapid detection of trace microcystins in aquatic products.
引文
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[13]杨振宇,周瑶.液相色谱-串联质谱法检测动物源性水产品中7种微囊藻毒素[J].质谱学报, 2014, 35(5): 447-453. DOI:10.7538/zpxb.youxian.2014.0035.
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[15]杨晓红,蒲朝文,张仁平,等.水体微囊藻毒素污染对人群的非致癌健康风险[J].中国环境科学, 2013, 33(1): 181-185. DOI:10.3969/j.issn.1000-6923.2013.01.026.
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[17]黄艺,张郅灏.微囊藻毒素的致毒机理和人体健康风险评价研究进展[J].生态环境学报, 2013, 22(2): 357-364. DOI:10.16258/j.cnki.1674-5906.2013.02.002.
[18] ZERVOU S K, CHRISTOPHORIDES C, KALOUDIS T, et al. New SPE-LC-MS/MS method for simultaneous determination of multi-class cyanobacterial and algal toxins[J]. Journal of Hazardous Materials, 2017, 323(Part A): 56-66. DOI:10.1016/j.jhazmat.2016.07.020.
[19]沈强,李嗣新,胡俊.微囊藻毒素HPLC快速检测方法研究[J].环境科学与技术, 2017(2): 103-106.
[20]王阳,徐明芳,曾晓琮,等.CE和HPLC测定水源水体中微囊藻毒素方法比较[J].食品科学, 2016, 37(22): 210-215. DOI:10.7506/spkx1002-6630-201622032.
[21]刘永波,王迪迪,薛瑞芳.超快速高效液相色谱-三重四级杆串联质谱法测定水中微囊藻毒素的研究[J].化学研究与应用, 2016, 28(8): 1145-1149. DOI:10.3969/j.issn.1004-1656.2016.08.017.
[22]沈斐,许燕娟,姜晟,等.超高效液相色谱-串联质谱法测定水华样品中微囊藻毒素及其基质效应的消除[J].色谱, 2017, 35(7): 772-777.DOI:10.3724/SP.J.1123.2017.03046.
[23] LU Z W, WANG Q, WANG M L, et al. Using UHPLC Q-Trap/MS as complementary technique to in-depth mine UPLC Q-TOF/MS data for identifying modified nucleosides in urine[J]. Journal of Chromatography B, 2017, 1015: 108-117. DOI:10.1016/j.jchromb.2017.03.002.
[24]虞锐鹏,陶冠军,杨健,等.超高效液相色谱-四极杆-飞行时间质谱法快速测定水产品中微囊藻毒素和节球藻毒素[J].分析试验室,2012, 31(1): 80-83. DOI:10.3969/j.issn.1000-0720.2012.01.019.
[25]陈立坚,何敏恒,李秀英,等.高效液相色谱-串联质谱法测定水产品中7种微囊藻毒素[J].现代食品科技, 2012(9): 1243-1248.DOI:10.13982/j.mfst.1673-9078.2012.09.019.
[26] ZHANG L, PING X, YANG Z. Determination of microcystin-LR in surface water using high-performance liquid chromatography/tandem electrospray ionization mass detector[J]. Talanta, 2014, 62(1): 191-198. DOI:10.1016/j.talanta.2003.07.001.
[27]张玲玲,虞锐鹏,陶冠军,等.加速溶剂萃取-超高效液相色谱串联质谱法测定水产品中微囊藻毒素[J].分析实验室, 2016, 35(5): 604-607. DOI:10.13595/j.cnki.issn1000-0720.2016.0140.
[28]郑西强,刘群,陈云峰.活性炭纤维对水中微囊藻毒素的吸附性能[J].环境科学, 2013, 7(10): 3802-3806.
[29]李安峰,潘涛,杨冲,等.水体富营养化治理与控制技术综述[J].安徽农业科学, 2012, 40(16): 9041-9044. DOI:10.3969/j.issn.0517-6611.2012.16.095.
[30]徐潇颖,罗金文,陈万勤,等.磁性多壁碳纳米管固相萃取/高效液相色谱-串联质谱法测定蜂蜜中多组分兽药残留[J].分析测试学报,2017(1): 61-66. DOI:10.3969/j.issn.1004-4957.2017.01.010.
[2] WANG X, SUN M, XIE M, et al. Differences in microcystin production and genotype composition among Microcystis colonies of different sizes in Lake Taihu[J]. Water Research, 2013, 47: 5659-5669.DOI:10.1016/j.watres.2013.06.040.
[3]王莎飞,郭彩荣,徐向阳,等.环境水体藻毒素生物处理技术研究进展[J].应用生态学报, 2016, 27(5): 1683-1692. DOI:10.13287/j.1001-9332.201605.026.
[4] FAYAD P B, ROY-LACHAPELLE A, DUY S V, et al. On-line solidphase extraction coupled to liquid chromatography tandem mass spectrometry for the analysis of cyanotoxins in algal blooms[J]. Toxicon Official Journal of the International Society on Toxinology, 2015(108): 167-175. DOI:10.1016/j.toxicon.2015.10.010.
[5]甘南琴,魏念,宋立荣.微囊藻毒素生物学功能研究进展[J].湖泊科学, 2017, 29(1): 1-8. DOI:10.18307/2017.0101.
[6]VICHI S,BURATTI F M,TESTAI E.Microcystins:toxicological profile[M].Springer Netherlands,2016:1-16.DOI:10.1007/978-94-007-6419-4_24.
[7]熊珊珊,宋天宇,蒋辉.鱼腥藻毒素与微囊藻毒素的免疫毒性研究[J].公共卫生与预防医疗, 2017, 28(3): 1-5.
[8] KEHR J, ZILLIGES A. A mannan binding lectin is involved in cell-cell attachment in a toxic strain of Microcystis aeruginosa[J].Molecular Microbiology, 2006, 59(3): 893-906. DOI:10.1111/j.1365-2958.2005.05001.x.
[9]VAN APELDOORN M E,VAN EGMOND H P,SPEIJERS G J,et al.Toxins of cyanobacteria[J]. Molecular Nutrition & Food Research, 2007, 51(5): 7-60. DOI:10.1002/mnfr.200600185.
[10] SALVADOR D, CHURRO C, VALRION E J. Evaluating the influence of light intensity in mcy A gene expression and microcystin production in toxic strains of Planktothrix agardhii and Microcystis aeruginosa[J]. Microbiol Methods, 2016, 123: 4-12. DOI:10.1016/j.mimet.2016.02.002.
[11]BLáHOVá L, ORAVEC M, MAR?áLEK B, et al.The first occurrence of the cyanobacterial alkaloid toxin cylindrospermopsin in the Czech Republic as determined by immunochemical and LC/MS methods[J]. Toxicon, 2009, 53(5): 519-524. DOI:10.1016/j.toxicon.2009.01.014.
[12] MENG G, SUN Y, FU W, et al. Microcystin-LR induces cytoskeleton system reorganization through hyperphosphorylation of tau and HSP27 via PP2A inhibition and subsequent activation of the p38 MAPK signaling pathway in neuroendocrine (PC12) cells[J]. Toxicology, 2011,290(2/3): 218-229. DOI:10.1016/j.tox.2011.09.085.
[13]杨振宇,周瑶.液相色谱-串联质谱法检测动物源性水产品中7种微囊藻毒素[J].质谱学报, 2014, 35(5): 447-453. DOI:10.7538/zpxb.youxian.2014.0035.
[14] DIETRICH D, HOEGER S. Guidance values for microcystins in water and cyanobacterial supplement products (blue-green algal supplements): a reasonable or misguided approach?[J]. Toxicology and Applied Pharmacology, 2005, 203(3): 273-289. DOI:10.1016/j.taap.2004.09.005.
[15]杨晓红,蒲朝文,张仁平,等.水体微囊藻毒素污染对人群的非致癌健康风险[J].中国环境科学, 2013, 33(1): 181-185. DOI:10.3969/j.issn.1000-6923.2013.01.026.
[16]王伟琴,金永堂,吴斌.水源水中微囊藻毒素的遗传毒性与健康风险评价[J].中国环境科学, 2010, 30(4): 468-476.
[17]黄艺,张郅灏.微囊藻毒素的致毒机理和人体健康风险评价研究进展[J].生态环境学报, 2013, 22(2): 357-364. DOI:10.16258/j.cnki.1674-5906.2013.02.002.
[18] ZERVOU S K, CHRISTOPHORIDES C, KALOUDIS T, et al. New SPE-LC-MS/MS method for simultaneous determination of multi-class cyanobacterial and algal toxins[J]. Journal of Hazardous Materials, 2017, 323(Part A): 56-66. DOI:10.1016/j.jhazmat.2016.07.020.
[19]沈强,李嗣新,胡俊.微囊藻毒素HPLC快速检测方法研究[J].环境科学与技术, 2017(2): 103-106.
[20]王阳,徐明芳,曾晓琮,等.CE和HPLC测定水源水体中微囊藻毒素方法比较[J].食品科学, 2016, 37(22): 210-215. DOI:10.7506/spkx1002-6630-201622032.
[21]刘永波,王迪迪,薛瑞芳.超快速高效液相色谱-三重四级杆串联质谱法测定水中微囊藻毒素的研究[J].化学研究与应用, 2016, 28(8): 1145-1149. DOI:10.3969/j.issn.1004-1656.2016.08.017.
[22]沈斐,许燕娟,姜晟,等.超高效液相色谱-串联质谱法测定水华样品中微囊藻毒素及其基质效应的消除[J].色谱, 2017, 35(7): 772-777.DOI:10.3724/SP.J.1123.2017.03046.
[23] LU Z W, WANG Q, WANG M L, et al. Using UHPLC Q-Trap/MS as complementary technique to in-depth mine UPLC Q-TOF/MS data for identifying modified nucleosides in urine[J]. Journal of Chromatography B, 2017, 1015: 108-117. DOI:10.1016/j.jchromb.2017.03.002.
[24]虞锐鹏,陶冠军,杨健,等.超高效液相色谱-四极杆-飞行时间质谱法快速测定水产品中微囊藻毒素和节球藻毒素[J].分析试验室,2012, 31(1): 80-83. DOI:10.3969/j.issn.1000-0720.2012.01.019.
[25]陈立坚,何敏恒,李秀英,等.高效液相色谱-串联质谱法测定水产品中7种微囊藻毒素[J].现代食品科技, 2012(9): 1243-1248.DOI:10.13982/j.mfst.1673-9078.2012.09.019.
[26] ZHANG L, PING X, YANG Z. Determination of microcystin-LR in surface water using high-performance liquid chromatography/tandem electrospray ionization mass detector[J]. Talanta, 2014, 62(1): 191-198. DOI:10.1016/j.talanta.2003.07.001.
[27]张玲玲,虞锐鹏,陶冠军,等.加速溶剂萃取-超高效液相色谱串联质谱法测定水产品中微囊藻毒素[J].分析实验室, 2016, 35(5): 604-607. DOI:10.13595/j.cnki.issn1000-0720.2016.0140.
[28]郑西强,刘群,陈云峰.活性炭纤维对水中微囊藻毒素的吸附性能[J].环境科学, 2013, 7(10): 3802-3806.
[29]李安峰,潘涛,杨冲,等.水体富营养化治理与控制技术综述[J].安徽农业科学, 2012, 40(16): 9041-9044. DOI:10.3969/j.issn.0517-6611.2012.16.095.
[30]徐潇颖,罗金文,陈万勤,等.磁性多壁碳纳米管固相萃取/高效液相色谱-串联质谱法测定蜂蜜中多组分兽药残留[J].分析测试学报,2017(1): 61-66. DOI:10.3969/j.issn.1004-4957.2017.01.010.