化学氧化法去除微囊藻毒素的研究进展

有害藻类水华污染已经成为世界范围性的环境难题。微囊藻毒素是藻细胞释放的一种毒性物质,具有极强的肝毒性和致癌性。当水源水受到藻类污染时,高效的微囊藻毒素去除技术十分必要。氧化法包括氯化、臭氧、高锰酸盐等化学氧化以及芬顿和基于紫外线的高级氧化,已经成功应用于微囊藻毒素的降解去除。本文综述了各种化学氧化法去除藻毒素的动力学、影响因素、降解机制和产物毒性等方面的研究进展,比较分析了各种氧化法的优、劣以及适用范围,并对未来藻毒素去除技术研究进行了展望,为微囊藻毒素的有效去除提供参考。     

水环境中微囊藻毒素分析技术新进展

微囊藻毒素是常见的蓝藻毒素,具有很强的肝脏神经系统和肾脏毒性. 由于水的富营养化,蓝藻会爆发产生大量的微囊藻毒素,进而对水生生物和食用它们的人类构成巨大威胁. 随着浓缩、富集、分离方法和仪器技术的进步,定量分析微囊藻毒素的方法也在不断进步,且应用越来越广泛. 综述了水、沉积物和生物中微囊藻毒素的富集和检测方法,结果显示:目前常见的采样方法是主动采样法,开发简便可靠和实用的被动采样方法是急需的研究方向之一. 衍生化方法可以降低基质效应,有利于使用不同的检测方法和试验观察,因此开发一种高效、灵敏的衍生化方法检测微囊藻毒素将是重要的研究方向之一.     

蓝藻水华衍生的微囊藻毒素污染及其对水生生物的生态毒理学研究

富营养化导致的蓝藻水华频发,引发各种衍生物污染,严重时造成重大生态灾害事件,甚至危及人类健康。其中微囊藻毒素以其毒性大、分布广和结构稳定的特点,成为水环境中常见的潜在危害物质,它主要由微囊藻产生,是一类具有多种异构体的环状七肽物质。本文根据微囊藻毒素污染现状及其水生生态毒理学研究的最新研究进展,介绍了微囊藻毒素的理化性质及其产生、迁移和转化,在我国天然水体、水库源水和饮用水中的污染现状以及部分水产品中的微囊藻毒素累积情况,较全面地评述了微囊藻毒素的分子致毒机理以及对水生生态系统的重要组成成分--常见水生植物和鱼类的生态毒理学效应,并提出了该领域未来研究的主要方向。     

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

通过超声提取、固相萃取纯化、超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF-MS)联用技术快速测定水产品中的微囊藻毒素-RR、-YR、-LR和节球藻毒素.分别采用选择离子监测质荷比(m/z)为519.84、1045.66、995.67、825.54分子离子峰进行定量分析.该法检出限为5.0～10.0μg/kg,在浓度0.02～5mg/kg的范围内,峰面积与样品浓度呈良好线性关系;4种藻毒素的回收率为76.2％～93.7％,相对标准偏差为2.0％～7.1％.采用上述方法对45个太湖水产品样品进行测定,发现有少量水产品中存在藻毒素污染,其中微囊藻毒素-RR最高含量为15.2μg/kg,微囊藻毒素-LR最高含量为0.84μg/kg,MC-YR、节球藻毒素均未检出.此方法可作为监测水产品体内蓄积藻毒素的分析方法.     

一种水产品中微囊藻毒素含量的检测方法

<正>申请公布号:CN104155372A申请公布日:2014.11.19申请人:深圳市格瑞斯特环保技术有限公司摘要:本发明公开了一种水产品中微囊藻毒素含量的检测方法,其包括如下步骤:(1)分别配制若干梯度浓度的微囊藻毒素的标准使用液,对其分别进行色谱分析,获得峰面积,并分别以所述梯度浓度为横纵、峰面积为坐标绘制成标准曲     

基因工程抗体在微囊藻毒素检测分析中的应用研究

微囊藻毒素是蓝藻水华过程中产生的一类小分子多肽生物毒素,能对人和动物的肝脏造成严重损伤,属于2B类致癌物质。由于全球性工业和生活废水的大量排放以及气候变暖等现实因素,蓝藻水华现象日趋频繁和加剧,使得微囊藻毒素在水体和土壤中大量蓄积,严重威胁生态环境和农产品质量安全。基于"抗体-抗原"识别原理的免疫学检测方法是当前生物毒素快速检测研究的热点,近年抗体创制技术突飞猛进,现已从传统的多克隆抗体和单克隆抗体逐渐发展到了新型的基因工程人工抗体创制阶段。该文系统阐述了微囊藻毒素产生的内外因素及其对生态系统、人和动物的主要危害风险;全面梳理了基因工程抗体创制技术及其在微囊藻毒素检测应用研究上的最新进展;并结合作者及所在科研团队最新研究成果,深入探讨了基因工程抗体在微囊藻毒素检测应用上的发展趋势、存在的问题以及拟解决的对策。     

芯片液相色谱-质谱法高灵敏度检测水产品中微囊藻毒素

建立了一种芯片液相色谱-高分辨质谱测定3种微囊藻毒素（MC-RR、MC-YR和MC-LR）的分析方法.在最优条件下,3种藻毒素在16 min内实现基线分离,并选择电喷雾源正离子模式进行质谱测定.方法在2.5~2 000 ng/mL范围内线性良好,线性相关系数大于0.996 5,检测限介于0.5~2.0 ng/mL之间.鱼肉和虾肉样品经固相萃取法净化处理后未测得上述毒素残留,加标回收率为82.8%~120.3%.方法具有样品用量少、定性快速准确、灵敏度高等优点,适合用于水产品等生物样品中藻毒素的痕量监测.     

甲基丙烯酸酯毛细管整体柱分离微囊藻毒素的研究

以甲基丙烯酸丁酯为单体,乙二醇二甲基丙烯酸酯为交联剂,在致孔剂存在的条件下原位聚合制备了甲基丙烯酸丁酯毛细管整体柱(150 μm i.d.)。实验中优化了用此整体柱分离3种微囊藻毒素(MC-LR,-YR和-RR)的色谱条件(流动相种类、缓冲溶液浓度、pH、流动相流速),建立了微囊藻毒素的整体柱毛细管液相色谱分离方法,该法可以在9 min之内实现3种微囊藻毒素的基线分离。将该方法应用于实际水样中微囊藻毒素的分析,成功实现了培养水样和巢湖水样中微囊藻毒素的快速分离,两种样品中均检测到MC-LR。结果表明,所制备的甲基丙烯酸酯毛细管整体柱具有良好的重现性、渗透性,在微囊藻毒素的常规检测中具有很好的应用前景。     

液相色谱–质谱联用技术在水产品兽药残留检测中的应用

液相色谱–质谱联用技术逐渐成为水产品中兽药残留检测的重要手段。从技术原理、样品处理方法的改进和新方法的应用,以及多兽药残留同时分析等方面论述了液相色谱–质谱联用技术在水产品兽药残留检测中的应用与进展。     

液相色谱-离子阱质谱对水华蓝藻提取物中微囊藻毒素的定性研究

本文研究了野外蓝藻水华样品甲醇提取物的液相色谱和电喷雾质谱特征图谱,利用电喷雾离子化-串联质谱(ESI-MSn)离子阱技术对特征图谱中主要微囊藻毒素的分子离子峰进行了二级质谱分析,获得相应的子离子质谱图,并对其进行了结构解析,确定了野外蓝藻水华样品中微囊藻毒素种类,为准确鉴定和分析水华蓝藻中微囊藻毒素提供了基础.     

Optimal strategies for determination of free/extractable and total microcystins in lake sediment

The optimization of analytical procedures for the quantification of free and total microcystins (MCs) in natural sediments was systematically examined based on solvent extraction and Lemieux oxidation. In this optimized analytical procedure, a sequential solvent extraction using 50% (v/v) methanol and EDTA-sodium pyrophosphate was selected as the optimal extraction solvent for free MCs analysis, after which the purified extracts and sediment residuals were applied to the optimized Lemieux oxidation for determination of total MCs in lake sediments. The optimized procedures were shown to be efficient and reliable for the routine analysis of both free and total MCs in lake sediment samples, as indicated by the minimal adverse impact of sediment organic matter on the recovery of free MCs and yield of MMPB (2-methyl-3-methoxy-4-phenylbutyric acid). Finally, the developed procedures were applied to field sediment samples collected from Lake Dianchi during a bloom season and seven of thirty samples showed positive results.     

Determination of microcystins in natural blooms and cyanobacterial strain cultures by matrix solid-phase dispersion and liquid chromatography–mass spectrometry

An analytical procedure based on matrix solid-phase dispersion (MSPD) and liquid chromatography–mass spectrometry (LC-MS) was developed for determining three microcystins (MCs) in natural water blooms and cyanobacteria strain cultures. The procedure involves sample homogenization with C₁₈, washed with dichloromethane to eliminate interfering compounds, and elution with acidic methanol. Results were compared to those achieved by using an organic solvent standard method. Mean recoveries of MCs with MSPD were 85–92% with intra-day relative standard deviation (RSDs) of 9–19%, whereas organic solvent extraction resulted in recovery rates of 92–105% with intra-day RSDs ranging from 8 to 18%. Limits of quantification (LOQs) were 1 g g^–1 dry weight for the MCs either by MSPD or organic solvent extraction. The two analytical methods tested were specific and sensitive to the extraction of MCs and were applied to the detection of MCs in water blooms and culture strains. The concentration of MCs varied from 7 to 3,330 g g^–1 of lyophilized cells with MC-LR always showing the highest concentration. MCs levels were higher in culture strains than in water blooms, except for MC-LR, whose concentration in blooms was slightly superior to that determined in culture strains.     

A Fungus Capable of Degrading Microcystin-LR in the Algal Culture of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> PCC7806

Microcystins (MCs) are a family of natural toxins produced by cyanobacteria (blue-green algae). Microbial degradation is considered an efficient method for eliminating cyanobacteria and MCs in environmental conditions. This study examines the ability of Trichaptum abietinum 1302BG, a white rot fungus, to degrade microcystin-LR in the harmful algal culture of Microcystis aeruginosa PCC7806. Results showed that microcystin-LR could not be detected by high-performance liquid chromatography after 12 h in algal culture incubated with the fungus. There were also high activities of catalase and peroxidase in algal culture incubated with the fungus. However, similar to the control, they decreased to normal levels after 72 h. Meanwhile, the micronucleus test in the toxicity studies revealed that the degraded algal culture had low toxicity.     

全自动在线固相萃取-高效液相色谱法测定水体中痕量微囊藻毒素

建立了全自动化在线固相萃取-高效液相色谱法快速、准确测定水体中3种痕量微囊藻毒素的新方法.样品用自动进样器连续注入固相萃取小柱完成富集后,通过双梯度高效液相系统中的一个泵(上样泵)实现净化,再通过阀切换将固相萃取小柱切换至分析流路中,用另一个泵(分析泵)将待测物冲洗至分析柱进行测定.方法使用Acclaim PA柱芯(1...     

固相萃取-超高效液相色谱-电喷雾串联质谱法同时测定地表水中9种微囊藻毒素

建立了固相萃取-超高效液相色谱-电喷雾串联三重四极杆质谱(SPE-UPLC-ESI-MS/MS)联用技术分析水中9种微囊藻毒素的方法。样品经SPE提取和净化后,以Waters ACQUITY UPLCTM BEH C18色谱柱为分离柱,以含0.1%甲酸乙腈和含0.1%甲酸水作为流动相进行梯度洗脱,电喷雾离子源电离、正离子多反应监测模式质谱进行定性和定量分析。9种微囊藻毒素在0.1～50 μg/L或0.5～100 μg/L质量浓度范围内线性良好,相关系数为0.9990～0.9998,方法的检出限(以3倍信噪比计)为0.1～0.5 ng/L;高、中、低3个添加水平的回收率为75.8%～109%,相对标准偏差为0.49%～10.0%。结果表明,该方法灵敏、准确,检测范围广,分析速度快。应用该方法检测了杭州市两处水库水样中的微囊藻毒素,分别检出了3种和8种微囊藻毒素。     

Evaluation of extraction approaches linked to ELISA and HPLC for analyses of microcystin-LR, -RR and -YR in freshwater sediments with different organic material contents

The efficiencies of conventional extraction techniques and analytical methods (HPLC–DAD and ELISA) were investigated for analyses of microcystins (MCs) in sediments. Our results showed several limitations. First, the extraction efficiency strongly depends on the extraction solvent, and extraction with 5% acetic acid in 0.2% trifluoroacetic acid (TFA)–methanol was confirmed as being the most appropriate for three different sediments (recovery: 33.1–44.9% of total MCs according to HPLC analyses). Second, the recovery of MCs was affected by the type of sediment but did not clearly correlate with the content of organic carbon. These results suggest that the sorption of MCs onto inorganic materials such as clay minerals is probably a more important process than interactions of the MCs with organic sediment matter. Third, the structure of the MCs is another crucial factor that affects the sorption of MCs and their recovery from sediments. Hydrophilic MC-RR gave much lower recoveries (20.0–38.8%) than MC-YR (44.1–59.5%) or MC-LR (55.3–77.8%) from three different types of spiked sediments. Recovery results analysed with HPLC–DAD correlated well with ELISA analyses. Further, extraction with 5% acetic acid in 0.2% TFA–methanol was used for analyses of MCs in 34 natural sediment samples collected from Brno reservoir (Czech Republic) from April to October 2005. Concentrations of MCs in sediments ranged from 0.003 to 0.380 μg/g sediment d.m. (ELISA results) or 0.016–0.474 μg/g d.m. (HPLC results). These values are equivalent to 0.63–96.47 μg/L of sediment (ELISA) or 4.67–108.68 μg/L (HPLC), respectively. Concentrations of sediment MCs showed both temporal and spatial variability, with the highest MC contents observed in the spring (April and May) and the lowest concentrations in July and August. Our results demonstrate the suitability of the methods described here for studying the occurrence, fate and ecological role of MCs in the aquatic environment.      

Rapid quantification of total microcystins in cyanobacterial samples by periodate-permanganate oxidation and reversed-phase liquid chromatography

Microcystins (MCs) comprise a family of more than 80 related cyclic hepatotoxic heptapeptides. Oxidation of MCs causes cleavage of the chemically unique C₂₀ β-amino acid (2S, 3S, 8S, 9S)-3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid (Adda) amino to form 2-methyl-3-methoxy-4-phenylbutanoic acid (MMPB), which has been exploited to enable analysis of the entire family. In the present study, the reaction conditions (e.g. concentration of the reactants, temperature and pH) used in the production of MMPB by oxidation of cyanobacterial samples with permanganate-periodate were optimized through a series of well-controlled batch experiments. The oxidation product (MMPB) was then directly analyzed by high-performance liquid chromatography with diode array detection. The results of this study provided insight into the influence of reaction conditions on the yield of MMPB. Specifically, the optimal conditions, including a high dose of permanganate (≥50 mM) in saturated periodate solution at ambient temperature under alkaline conditions (pH ∼9) over 1-4 h were proposed, as indicated by a MMPB yield of greater than 85%. The technique developed here was applied to determine the total concentration of MCs in cyanobacterial bloom samples, and indicated that the MMPB technique was a highly sensitive and accurate method of quantifying total MCs. Additionally, these results will aid in development of a highly effective analytical method for detection of MMPB as an oxidation product for evaluation of total MCs in a wide range of environmental sample matrices, including natural waters, soils (sediments) and animal tissues.     

Identification of several Tonalide® transformation products in the environment

This work reports on the survey carried out to determine the presence of microcystins (MCs) by using ELISA tests and HPLC-UV for such determination in different sample sites along the Spanish course of the Guadiana River. The most important cyanobacteria species identified were, Microcystis aeruginosa and Oscillatoria spp. The highest total microcystin content recorded was 6.40?µg?L^?1 in 2002. The main toxins found were microcystins RR and LR, with microcystin YR present at trace levels. Improvements in sample clean up were carried out by using Immunoaffinity solid phase extraction (SPE) and its advantages regarding to conventional SPE were clearly demonstrated. The confirmation of MCs presence in the evaluated water reservoirs, underlines the necessity of monitoring programs as well as the improvement of analytical methodologies to efficiently prevent the human health risks as a consequence of MCs contamination.