Determination of 3 Cyanobacteria Toxins in Aquatic Products by High Performance Liquid Chromatography-High Resolution Mass Spectrometry with QuEChERS Extraction
摘 要
取水产品样品2.00 g与2 g无水硫酸钠混匀后加入提取剂10 mL甲醇,涡旋振荡5 min后超声提取5 min,再离心5 min,取其上清液。在上清液中加入500 mg中性氧化铝和15 mg石墨化碳黑(GCB)涡旋振荡1 min,离心5 min,取上清液,于40℃下吹氮至近干,用体积比为1:1的流动相A-流动相B混合溶液溶解残渣并定容至2.0 mL,经0.22 μm滤膜过滤,取其滤液按高效液相色谱-高分辨质谱法测定其中3种蓝藻毒素的含量。色谱分离中用Hypersile Gold C8色谱柱(150 mm×2.1 mm,3 μm)为固定相,用不同比例的流动相A和流动相B两溶液的混合液作为流动相进行程序梯度洗脱。质谱测定中采用电喷雾离子源,正、负离子切换模式。由于3种蓝藻毒素均产生基质减弱效应,制作工作曲线需用基质标准溶液以消除基质效应。结果表明:所测3种蓝藻毒素均在10~200 μg·L-1内与其峰面积之间呈线性关系,其检出限(3S/N)均为5 μg·kg-1。通过标准加入法进行回收试验,测得回收率为68.3%~104%,测定值的相对标准偏差(n=6)为7.7%~17%。
Abstract
A portion of sample of aquatic product (2.00 g) was extracted ultrasonically for 5 min with 10 mL of CH3OH after mixing with 2 g of anhydrous sodium sulfate, and centrifuged for 5 min. The supernatant was taken and purified by swirling for 1 min with 500 mg of neutral alumina and 15 mg of graphitized carbon black, and then centrifuged for 5 min. The supernatant was taken and evaporated to near-dryness by N2-blowing at 40℃. The residue was dissolved in and with its volume made up to 2.0 mL of a mixed solution of mobile phase A-mobile phase B with the volume ratio of 1:1. The solution was then filtered through 0.22 μm filtering membrane and the filtrate was used for determination of 3 cyanobacteria toxins by high performance liquid chromatography-high resolution mass spectrometry. In the liquid chromatography separation, Hypersile Gold C8 column (150 mm×2.1 mm, 3 μm) was used as stationary phase and mixtures of solution (A) and (B) in various ratios were used as mobile phases in the gradient elution. Electrospray ionization with mode of switching of cation and anion was adopted in mass spectrometry determination. Matrix matching was used in preparation of standard curves of the 3 cyanobacteria toxins to overcome matrix effects. Standard curves of the 3 cyanobacteria toxins were obtained in the same range of 10-200 μg·L-1, with their same detection limits (3S/N) of 5 μg·kg-1. Values of recovery found by test for recovery by standard addition method were in the range of 68.3%-104%, and values of RSDs (n=6) were found ranged from 7.7% to 17%.
中图分类号 O657.63 DOI 10.11973/lhjy-hx202001003
所属栏目 工作简报
基金项目 原国家质检总局科技计划项目(2017IK070);浙江省基础公益研究计划项目(LGN18B050002)
收稿日期 2019/5/29
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备注方科益,高级工程师,硕士,主要从事化学分析研究工作,30028788@QQ.com
引用该论文: FANG Keyi,CHEN Shubing,LI Shuang,WANG Yongjian,WANG Chunfang,CAO Guozhou. Determination of 3 Cyanobacteria Toxins in Aquatic Products by High Performance Liquid Chromatography-High Resolution Mass Spectrometry with QuEChERS Extraction[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2020, 56(1): 14~19
方科益,陈树兵,李双,王永健,王春芳,曹国洲. QuEChERS提取-高效液相色谱-高分辨质谱法测定水产品中3种蓝藻毒素的含量[J]. 理化检验-化学分册, 2020, 56(1): 14~19
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【2】KOSIBA J, KRZTON W, WILK-WOZNIAK E. Effect of microcystins on proto-and metazooplankton is more evident in artificial than in natural waterbodies[J]. Microbial Ecology, 2018,75(2):293-302.
【3】STOBO A, JEAN-PIERRE L, SCOTT A, et al. Liquid chromatography with mass spectrometry-detection of lipophilic shellfish toxins[J]. Journal of AOAC International, 2005,88(5):1371-1382.
【4】谢平.蓝藻水华及其次生危害[J].水生态学杂志, 2015,36(4):1-13.
【5】FEURSTEIN D, HOLST K, FISCHER A, et al. Oatp-associated uptake and toxicity of microcystins in primary murine whole brain cells[J]. Toxicology and Applied Pharmacology, 2009,234(2):247-255.
【6】CHRAPUSTA E, WEGRZYN M, ZABAGLO K, et al. Microcystins and anatoxin-a in Arctic biocrust cyanobacterial communities[J]. Toxicon, 2015,101:35-40.
【7】PONIEDZIAŁEK B, RZYMSKI P, KOKOCIHSKI M, et al. Toxic potencies of Metabolite(S) of non-cylindrospermopsin producing Cylindrospermopsis raciborskii isolated from temperate zone in human white cells[J]. Chemosphere, 2015,120:608-614.
【8】CHEN Y Y, HUANG X H, WANG J Z, et al. Effect of pure microcystin-LR on activity and transcript level of immune-related enzymes in the white shrimp (Litopenaeus vannamei)[J]. Ecotoxicology, 2017,26(5):702-710.
【9】PAULINO M G, ROSSI P A, VENTURINI F P, et al. Hepatotoxicity and metabolic effects of cellular extract of cyanobacterium Radiocystis fernandoi containing microcystins RR and YR on neotropical fish (Hoplias malabaricus)[J]. Chemosphere, 2017,175:431-439.
【10】范赛,赵榕,李兵,等.水产品中微囊藻毒素检测方法的研究进展[J].色谱, 2012,20(5):434-439.
【11】DE LA CRUZ A A, HISKIA A, KALOUDIS T, et al. A review on cylindrospermopsin: the global occurrence, detection, toxicity and degradation of a potent cyanotoxin[J]. Environmental Science: Processes and Impacts, 2013,15(11):1979-2003.
【12】AKCAALAN R, KÖKER L, OĜUZ A, et al. First report of cylindrospermopsin production by two cyanobacteria (dolichospermum mendotae and chrysosporum ovalisporum) in Lake iznik, turkey[J]. Toxins, 2014,6(11):3173-3186.
【13】黄会,刘慧慧,李佳蔚,等.水产品中微囊藻毒素检测方法及污染状况研究进展[J].中国渔业质量与标准, 2019,9(2):32-43.
【14】张维昊,金丽娜,徐小清.鱼肉中微囊藻毒素的高效液相色谱法分析[J].分析测试学报, 2011(2):117-119.
【15】KELLMANN M, MUENSTER H, ZOMER P, et al. Full scan MS in comprehensive qualitative and quantitative residue analysis in food and feed matrices: How much resolving power is required[J]. Journal of the American Society for Mass Spectrometry, 2009,20(8):1464-1476.
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