微囊藻毒素的检测和脱除方法综述

近年来,由于富营养化,全球有害蓝藻水华的发生率持续上升,形成水华的蓝藻会释放出多种毒素,微囊藻毒素是有害蓝藻产生的常见毒素,具有肝毒性、肾毒性、神经毒性、生殖毒性等多种毒性,严重威胁人类和生态系统健康。微囊藻毒素在水中非常稳定,难以通过传统水处理工艺去除。因此,寻求经济有效的微囊藻毒素检测和脱除方法至关重要。本文综述了定量检测和去除微囊藻毒素的方法,包括物理法、化学法和生物法,并分析总结了这三类方法用于检测和脱除微囊藻毒素的优势与局限性,总结了不同方法脱除微囊藻毒素的机理,重点介绍了绿色高效的光催化降解微囊藻毒素的脱除方法,最后,基于当前的研究结果,对未来微囊藻毒素脱除研究方向进行了展望,为解决环境中微囊藻毒素的污染问题提供思路。     

表面等离子体共振免疫传感器用于快速检测微囊藻毒素的研究

针对现有微囊藻毒素(MC-LR)检测方法操作复杂、因标记而污染环境、仪器贵重,不利于现场快速检测等问题,将自行研制的表面等离子体共振(SPR)生物芯片检测仪应用于微囊藻毒素的检测,提出抑制型SPR生物芯片快速检测痕量微囊藻毒素的方法。采用该方法分别对浓度为3.5、2.5、1.5、1、0μg/L的MC-LR样品进行了检测。结果表明:该方法检测限小于1μg/L,可满足世界卫生组织(WHO)对于饮用水和我国地表水环境质量标准中MC-LR最低含量检测的需求。该方法完成一个样品检测耗时约8 min,相比于高效液相色谱法(HPLC)和酶联免疫吸附法(ELISA)等传统检测方法,快速定量是其最大的优势,可用于食品质量监控和现场实时检测。     

全自动固相萃取-高效液相色谱法同时测定饮用水中3种微囊藻毒素

建立全自动固相萃取-高效液相色谱法同时测定饮用水中3种微囊藻毒素的方法。采用全自动固相萃取仪代替传统的前处理方式,对大体积水样进行富集,以KH2PO4(p H=3.0)-甲醇(43∶57,v∶v)为流动相,采用二极管阵列检测器进行检测,结果 3种微囊藻毒素在0.1~10.0μg/m L范围内线性关系良好,相关系数r2≥0.999,3种不同微囊藻毒素的检出限均可达到0.05μg/L(S/N=3),加标回收率在86.3%~99.7%范围内,标准偏差为1.70%~2.72%(n=6),该方法用于测定饮用水中3种微囊藻毒素具有操作简单、快速,精密度高、准确可靠等特点。     

太湖蓝藻培养单细胞蛋白的可行性

以蓝藻原料为氮源发酵生产酵母单细胞蛋白,考察了碳源、维生素、微量元素、矿物质元素、培养温度、接种量、初始pH、通气量等对酵母产量的影响,优化了发酵条件,1 g/dL(干重)的蓝藻培养基最终使酵母产量达到6.2 g/L,并利用高效液相色谱法检测酵母产物胞内微囊藻毒素.结果表明,酵母中微囊藻毒素含量为390 μg/kg.     

固相萃取-高效液相色谱法测定鲫鱼肉中微囊藻毒素

建立固相萃取-高效液相色谱法(SPE-HPLC)测定鲫鱼肉中微囊藻毒素的方法.鲫鱼肉经90%甲醇水溶液提取,反相硅胶萃取柱净化后,采用高效液相色谱法进行测定.以乙腈-水-三氟乙酸(体积比35:65:0.05)为流动相,经Zorbax Eclipse C18柱(4.6mm×250mm i.d.5μm)分离,外标法定性、定量分析,结果表明微囊藻毒素LR的线性定量范围0.1～10.0μg/mL,鲫鱼肉中检出限25 ng/g.微囊藻毒素RR的线性定量范围0.1～10.0 μg/mL,在鲫鱼肉中的检出限20ng/g.此方法准确,灵敏度高,专属性好,可作为监测淡水水产品微囊藻毒素污染的分析方法.     

微囊藻毒素致毒机理及防治方法研究进展

微囊藻毒素是水体污染的主要来源之一。本文对微囊藻毒素致毒机理及防治方法研究进展进行了阐述。     

固相萃取-反相高效液相色谱法检测太湖蛳螺中微囊藻毒素

建立一种高效液相色谱法测定太湖蛳螺中微囊藻毒素RR和LR的方法。该方法采用固相萃取法进行纯化富集,用2极管阵列检测器检测,比较不同提取溶剂、提取时间和淋洗液等因素对样品中微囊藻毒素提取效率的影响。色谱条件：色谱柱300SB Zorbax C18柱（250mm×4．6mm i．d．,5μm）,流动相为乙腈：水（V：V）=35：65（含0．1％甲酸）,检测波长为238nm。在2．5～100ng的范围内,进样量与峰面积呈良好的线性关系。该方法灵敏度高,简便快捷,可作为水生物中藻毒素风险评价和监测水生物体内蓄积的藻毒素较可靠的分析方法。     

高效液相色谱-质谱法测定蓝藻中的微囊藻毒素

微囊藻毒素是由淡水蓝绿藻产生的一类最常见的环七肽缩肝毒素,对动物及人类健康具有较大的危害性。本文以实验室培养铜绿微囊藻为原料,建立了固相萃取富集微囊藻毒素,液相色谱-电喷雾电离质谱测定藻样中的微囊藻毒素MC-LR的方法。该法绝对检出限为25pg,回收率为70%以上,方法灵敏度高,实用性强。在此方法的基础上,对实际藻样中的微囊藻毒素进行了定性分析。结果表明,该藻样含有MC-RR及可能为[Dha]-MC-HtyR、MC-YR、[D-Asp3]-MC-HtyR或[D-Asp3,Dhb7]-MC-HtyR的单体。     

三通道自动固相萃取-高效液相色谱四极杆串联质谱法检测水中8种痕量微囊藻毒素

目的 建立三通道自动固相萃取-高效液相色谱-四极杆串联质谱法检测水中痕量的8种微囊藻毒素的分析方法.方法 采用高自动化的前处理设备,采用C18固相萃取小柱富集浓缩,应用液相色谱-四极杆串联质谱法定性定量地检测8种微囊藻毒素.结果 该方法检测水中的8种微囊藻毒素检出限为0.01～0.6μg/L,在0.25～62.5μg/...     

高效液相色谱-串联质谱法测定水产品中7种微囊藻毒素

建立了高效液相色谇串联质谱法同时测定水产品中7种微囊藻毒素（微囊藻毒素-RR、微囊藻毒素-YR、微囊藻毒素-LR、微囊藻毒素-LA、微囊藻毒素-LW、微囊藻毒素-LF、微囊藻毒素．LY）的分析方法。样品用90%醇水（P／g）溶液提取,经离心、固相萃取柱净化后,用C18柱,以0．1％甲酸乙腈．0．1％甲酸溶液为流动相,梯度洗悦分离,使用三重四级杆质谱检测器捡测,以保留时间及特征离子对定性,外标法定量。结果表明,7种微囊藻毒素在线性范围内线性关系良好,相关系数均不低于0．9990,方法的定量限（以信噪比为10计）为0．2-0．5μg／kg,添加水平为1．0-20μg／kg时,回收率为71．5-106％,相对标准偏差（n=6）为312～9．1％。该法前处理简单、回收率高、精密度好,适用于水产品中7种微囊藻毒素的测定。     

Detection of the hepatotoxic microcystins in 36 kinds of cyanobacteria Spirulina food products in China

Gel filtration chromatography, ultra-filtration, and solid-phase extraction silica gel clean-up were evaluated for their ability to remove microcystins selectively from extracts of cyanobacteria Spirulina samples after using the reversed-phase octadecylsilyl ODS cartridge for subsequent analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The reversed-phase ODS cartridge/silica gel combination were effective and the optimal wash and elution conditions were: H(2)O (wash), 20% methanol in water (wash), and 90% methanol in water (elution) for the reversed-phase ODS cartridge, followed by 80% methanol in water elution in the silica gel cartridge. The presence of microcystins in 36 kinds of cyanobacteria Spirulina health food samples obtained from various retail outlets in China were detected by LC-MS/MS, and 34 samples (94%) contained microcystins ranging from 2 to 163 ng g(-1) (mean = 14 +/- 27 ng g(-1)), which were significantly lower than microcystins present in blue green alga products previously reported. MC-RR - which contains two molecules of arginine (R) - (in 94.4% samples) was the predominant microcystin, followed by MC-LR - where L is leucine - (30.6%) and MC-YR - where Y is tyrose - (27.8%). The possible potential health risks from chronic exposure to microcystins from contaminated cyanobacteria Spirulina health food should not be ignored, even if the toxin concentrations were low. The method presented herein is proposed to detect microcystins present in commercial cyanobacteria Spirulina samples.     

Detection of the hepatotoxic microcystins in 36 kinds of cyanobacteria Spirulina food products in China

Gel filtration chromatography, ultra-filtration, and solid-phase extraction silica gel clean-up were evaluated for their ability to remove microcystins selectively from extracts of cyanobacteria Spirulina samples after using the reversed-phase octadecylsilyl ODS cartridge for subsequent analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The reversed-phase ODS cartridge/silica gel combination were effective and the optimal wash and elution conditions were: H₂O (wash), 20% methanol in water (wash), and 90% methanol in water (elution) for the reversed-phase ODS cartridge, followed by 80% methanol in water elution in the silica gel cartridge. The presence of microcystins in 36 kinds of cyanobacteria Spirulina health food samples obtained from various retail outlets in China were detected by LC-MS/MS, and 34 samples (94%) contained microcystins ranging from 2 to 163 ng g^?1 (mean?=?14?±?27 ng g^?1), which were significantly lower than microcystins present in blue green alga products previously reported. MC-RR?–?which contains two molecules of arginine (R)?–?(in 94.4% samples) was the predominant microcystin, followed by MC-LR?–?where L is leucine?–?(30.6%) and MC-YR?–?where Y is tyrose?–?(27.8%). The possible potential health risks from chronic exposure to microcystins from contaminated cyanobacteria Spirulina health food should not be ignored, even if the toxin concentrations were low. The method presented herein is proposed to detect microcystins present in commercial cyanobacteria Spirulina samples.     

Development of an ultrarapid one-step fluorescence immunochromatographic assay system for the quantification of microcystins

Microcystins are a family of potent toxic oligopeptides produced by freshwater cyanobacteria genera and have been a great threat to the welfare of humans and animals. There has been a great demand for developing a fast and convenient analytical method to detect microcystins. Recently, direct competition ELISA using monoclonal or polyclonal antibody has become the prevailing method for detecting microcystins. In this study, we report rapid quantification methods of microcystins using fluorescence for a detection signal and a lateral-flow-type immunochromatography as a separation system. The assay systems consist of a test strip housed in a disposable cartridge and a portable laser-fluorescence scanner. The components of a test strip are as follows: a nitrocellulose membrane, a sample pad, an absorption pad, and a backing card. A fluorescence scanner was designed to fit the cartridge and to quantify the distribution of the fluorescence intensity along the strip. When the calibration curve for an antibody-immobilized system was determined, a good linearity was displayed in the range from 125 to 2000 pg/mL of microcystin-LR. The linear-regression coefficient (R) was 0.938 between relative fluorescence intensity and the microcystin concentration. The limit of detection was determined to be 95.38 pg/mL. We then designed another biosensor system by changing an experimental format from the competition type to the inhibition type. When compared to the antibody-immobilized system, the antigen-immobilized assay detected a lower level of microcystin but did not discern microcystin-LR above 1000 pg/mL. The detection of limit for the antigen-immobilized system was 47.23 pg/mL. The linear regression coefficient (R) in the antigen-immobilized system equaled to 0.927. The reproducibility in the antigen-immobilized system was good through the entire range. The reproducibility in the antibody-immobilized system was relatively poor when compared to a MC-immobilized system. However, it still registered in the acceptable range of 7.32-9.91% except for the extreme ends of the MC concentration. Finally, surface water was tested to check for potential matrix interference. The calibration curve displayed a similar pattern as did those for other matrixes, including PBS and tap water, although its sensitivity was a little less due to the interference with certain components in the surface water. Overall, either of the biosensor systems can be used as a useful on-site detection tool for checking drinking water or surface water for microcystins. The laser-fluorescence scanner we developed is relatively small, transportable, and easy to use. Thus, the samples can be analyzed for microcystins at the test site using a real-time base within 15 min without having to bring the samples back to the laboratory.     

Oxidation of the cyanobacterial hepatotoxin microcystin-LR by chlorine dioxide: reaction kinetics, characterization, and toxicity of reaction products

Cyanobacteria are known producers of cytotoxins, hepatotoxins, and neurotoxins. The main toxins are microcystins, cyclic heptapeptide hepatotoxins, produced by strains of several cyanobacterial genera frequently found in eutrophied freshwaters. Due to the acute and chronic toxicity of microcystins, successful removal of these toxins in drinking water treatment processes is of increasing concern. In the present work the kinetics of microcystin-LR (MC-LR) oxidation by chlorine dioxide (ClO2) was studied with UV-spectrometry and high performance liquid chromatography (HPLC). Characterization of reaction products was performed with mass spectrometric (MS) analysis, while the toxicity of reaction products was tested with a protein phosphatase inhibition assay (PPIA). The main reaction products formed, dihydroxy isomers of MC-LR as identified by MS, were nontoxic according to the PPIA. The overall rate constant k for the reaction between MC-LR and ClO2 at 293 K and pH 5.65 was modest, k = 1.24 M(-1) s(-1), suggesting that ClO2 is not a suitable oxidant for the degradation of microcystins in drinking water treatment processes.     

(类)Fenton试剂氧化降解微污染水中微囊藻毒素的进展研究

作为一种有效的高级氧化技术,(类)Fenton试剂在氧化降解微污染水中的微囊藻毒素方面具有独特的优势,是一种很有应用潜力的水处理技术。概述(类)Fenton试剂的类型和特点,并对其应用进展进行研究。     

Cloud-point extraction and preconcentration of cyanobacterial toxins (microcystins) from natural waters using a cationic surfactant

A new cloud-point extraction and preconcentration method using a cationic surfactant, Aliquat-336 (tricaprylylmethylammonium chloride), has been developed for the determination of cyanobacterial toxins, microcystins, in natural waters. Sodium sulfate was used to induce phase separation at 25 degrees C. The phase behavior of Aliquat-336 with respect to concentration of Na2SO4 was studied. The cloud-point system revealed a very high phase volume ratio compared to other established systems of nonionic, anionic, and cationic surfactants. At pH 6-7, it showed an outstanding selectivity in analyte extraction for anionic species. Only MC-LR and MC-YR, which are known to be predominantly anionic, were extracted (with averaged recoveries of 113.9 +/- 9% and 87.1 +/- 7%, respectively). MC-RR, which is likely to be amphoteric at the above pH range, was not detectable in the extract. Coupled to HPLC/UV separation and detection, the cloud-point extraction method (with 2.5 mM Aliquat-336 and 75 mM Na2SO4 at 25 degrees C) offered detection limits of 150 +/- 7 and 470 +/- 72 pg/mL for MC-LR and MC-YR, respectively, in 25 mL of deionized water. Repeatability of the method was 7.6% for MC-LR and 7.3% for MC-YR. The cloud-point extraction process can be completed within 10-15 min with no cleanup steps required. Applicability of the new method to the determination of microcystins in real samples was demonstrated using natural surface waters collected from a local river and a local duck pond spiked with realistic concentrations of microcystins. Effects of salinity and organic matter (TOC) content in the water sample on the extraction efficiency were also studied.     

Monitoring of shrimp and farmed fish sold in Canada for cyanobacterial toxins

Sixty-one samples of shrimp and 32 samples of farmed fish collected from retail markets across Canada were analyzed for cyanobacterial toxins, including microcystins, paralytic shellfish poisons (saxitoxins), cylindrospermopsin, and β-N-methylamino-L-alanine, using established methods of analysis. None of these toxins were detected in any of the samples. Some shrimp samples screened for paralytic shellfish poisons showed the presence of unknown peaks in the chromatogram after periodate oxidation.