微囊藻毒素的提取与分析研究进展

本文简介了微囊藻毒素的形成、分子结构、危害及分析研究微囊藻毒素意义 ,然后从藻毒素样品的预处理、提取、分离和分析四个方面详细介绍了微囊藻毒素的提取与分析方法现状和最新进展 ,并客观评价了目前提取和分析藻毒素的各种方法 ,指出了其优缺点。在实验时 ,要根据实验的要求 ,进行不同方式的组合 ,进而达到满意的结果。以后随着性能更好的萃取头涂层材料的出现 ,SPME-HPLC联用技术在藻毒素分析检测领域将发挥更大的作用。

一种快速提取分析微囊藻毒素的方法

以五种群体和单细胞微囊藻及自然微囊藻水华为材料 ,在沸水浴中经不同时间处理 ,过滤后直接进行HPLC UV检测 ,发现 12min的沸水浴处理就足以达到抽提目的。研究中发现 ,去离子水比蒸馏水是更有效的抽提溶剂。传统的甲醇抽提结果与沸水浴处理的相对误差主要在 0 2 %— 16 5 9%之间。结果还显示 ,群体微囊藻需要比单细胞微囊藻抽提更长时间。本研究提供了一种经过改进的高效、廉价和快速的微囊藻毒素抽提分析方法。

多甲藻素在一种海生黄藻中的存在

采用聚酰胺薄层层析分离、分光光度法 ,对一种海生超微型黄藻品系PP983的色素组成进行了研究测定。结果表明 ,该黄藻中只含有叶绿素a而不含叶绿素c,检出 8种类胡萝卜素 ,除作为黄藻特征色素的Heteroxanthin和Vaucheriaxanthin两种外 ,还发现该藻含有一个吸收峰在 471nm ,呈现多甲藻素 (peridinin)特征的色素。用高效液相色谱 (HPLC) ,并以微小多甲藻 (Peridiniumpusillum)作对照 ,对其光合色素作进一步的分析 ,确证该黄藻中不含有叶绿素c而含

人工湿地系统去除藻毒素研究

以含水华的鱼塘水作为进水灌溉两套上行流 -下行流人工湿地系统 ,系统内种植芦苇等水生植物。灌溉一周后收集人工湿地进出水水样 ,用HPLC法检测其藻毒素含量 ,结合温度、电位、溶氧等理化指标和细菌、藻类数量的变化 ,分析人工湿地系统对藻毒素的去除效果及影响其效果的可能因素。结果表明 ,人工湿地系统对藻毒素有一定的去除作用 ,对藻毒素含量 0 .117g/L的进水 ,两套系统去除率分别为 6 8.5%和 34.6 % ,芦苇 -水葱组合的湿地系统去除效果优于茭白 -石菖蒲组合的湿地系统。在三种主要藻毒素RR、

铜绿微囊藻毒株M.8641的毒素

M.8641是从武汉东湖分离培养的一株有毒的铜绿微囊藻(Microcystis aeruginosa),它产生两种环状短肽肝毒素。经凝胶过滤及HPLC等过程纯化,Waters Pico Tag系统测定,其主毒素(毒素Ⅰ)的氨基酸组成为:Glu(1),β-Masp(1),Ala(Ⅰ),Arg(2),Mdha(1),FAB-MS和MS/MS测定分子量为m/z1038,元素组成为C_(49)H_(76)O_(12)N_(13)。毒素Ⅱ的氨基酸组成,除一分子Arg为Leu取代外,其余与毒素Ⅰ相同,分子量m/z 9

氯霉素在健康、沙门氏菌病和肝损害小型猪体内的药代动力学研究

本文研究了氯霉素单次静脉注射后,在健康、沙门氏菌病和肝损害小型猪体内的药代动力学,以观察疾病对氯霉素在动物体内变化过程的影响.从颈静脉导管采血.药物浓度用高效液相色谱法测定.结果表明,沙门氏菌感染和肝损伤后。氯霉素的动力学发生显著改变.在健康动物体内为一室开放模型.消除半衰期(l_(1/2)β)为1.1h.而在两种疾病动物体内则为二室开放模型,消除半衰期(l(1/2)β)分别为1.68h和1.63h。极显著延长(p<0.01).最后根据有关动力学参数制定了相应的给药方案.

东湖铜绿微囊藻毒素的分离与鉴定

1984—1985年从武昌东湖定期采集形成水华的铜绿微囊藻Microcystisaeruginosa,并进行了毒素的分离与鉴定。结果表明:(1)用反复冻融的藻细胞糊对20—25g小白鼠进行腹腔注射,其最低致死浓度为100mg/kg(LD_(100)=100mg/kg),致死时间为60—120min。(2)用匀浆、抽提、离子交换层析及高压液相层析(HPLC)纯化了毒素,纯化毒素对小白鼠的最低致死量约为lmg/kg,引起小白鼠中毒致死的特征表明它是一种与以前文献报道相似的肝毒素。(3)纯化毒素在230nm和2

The synthesis of astaxanthin esters, independent of the formation of cysts, highly correlates with the synthesis of fatty acids in Haematococcus pluvialis

The compositions and contents of astaxanthin esters and fatty acids in four types of Haematococcus pluvialis cells were studied by HPLC and GC-MS. Results showed that the synthesis and accumulation of astaxanthin was independent of the formation of cysts, but was highly correlated with the synthesis and accumulation of fatty acids, though it is an well known phenomenon that the accumulation of astaxanthin is usually accompanied by the formation of cyst. The red cysts contain more than 30% of fatty acids, with 81% of the unsaturated fatty acids. Taken together, besides a resource of astaxanthin, H. pluvialis would be a good resource of valuable fatty acids.

Non-microcystin producing Microcystis wesenbergii (Komarek) Komarek (Cyanobacteria) representing a main waterbloom-forming species in Chinese waters

It is well known that several morphospecies of Microcystis, such as Microcystis aeruginosa (Kutzing) Lemmermann and Microcystis viridis (A. Brown) Lemmermann can produce hepatotoxic microcystins. However, previous studies gave contradictory conclusions about microcystin production of Microcystis wesenbergii (Komarek) Komarek. In the present study, ten Microcystis morphospecies were identified in waterblooms of seven Chinese waterbodies, and Microcystis wesenbergii was shown as the dominant species in these waters. More than 250 single colonies of M. wesenbergii were chosen, under morphological identification, to examine whether M. wesenbergii produce hepatotoxic microcystin by using multiplex PCR for molecular detection of a region (mcyA) of microcystin synthesis genes, and chemical analyses of microcystin content by ELISA and HPLC for 21 isolated strains of M. wesenbergii from these waters were also performed. Both molecular and chemical methods demonstrated that M. wesenbergii from Chinese waters did not produce microcystin. (C) 2007 Elsevier Ltd. All rights reserved.

Cytotoxicity evaluation of three pairs of hexabromocyclododecane (HBCD) enantiomers on Hep G2 cell

Hexabromocyclododecanes (HBCDs) are additive brominated flame retardants mainly used in plastics and textiles. At the present time, these compounds are found in almost all environmental and human samples. In order to evaluate the environmental safety and health risk of HBCDs, the enantiomerically pure alpha-, beta-, and gamma-HBCD were prepared using high performance liquid chromatography (HPLC) on a PM-P-CD column and the cytotoxicities of their enantiomers were evaluated in Hep G2 cells. Results from the 3-(4,5-dimethylthioazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), resazurin reduction and lactate dehydrogenase (LDH) release assays showed a good agreement that the order of cytotoxicity was gamma-HBCD >= beta-HBCD > alpha-HBCD, and that significantly lower cell viability and higher LDH release were observed in all (+)-enantiomers ((+) alpha-, (+) beta- and (+) gamma-HBCD) than the corresponding (-)-forms ((-) alpha-, (-) beta- and (-) gamma-HBCD). Additionally, the formation of reactive oxygen species (ROS) induced by these HBCD enantiomers were detected. The positive correlation between the LDH release and ROS formation demonstrated that the toxic mechanism might be mediated by oxidative damage. These results suggest that environmental and human health risks of HBCDs must be evaluated at the level of individual enantiomers. (C) 2008 Published by Elsevier Ltd.

Proteomic analysis of the sarcosine-insoluble outer membrane fraction of Flavobacterium columnare

Outer membrane proteins (OMPs) of bacteria are key molecules interacting with the host environment. Flavobacterium columnare, a pathogen-causing columnaris disease of fish worldwide, was studied in order to understand the composition of its OMPs. The sarcosine-insoluble membrane fraction of the OMPs was analysed using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) in combination with reverse-phase high-performance liquid chromatography-tandem mass spectrometry (RP-HPLC MS/MS). Thirty-six proteins were identified, including proteins involved in cell wall/membrane biogenesis, specific transport of various nutrients and in essential metabolism. The present study is the first report on the OMPs of F. columnare, and may serve as the basis for understanding the pathogenesis of the bacterium.

Simultaneous determination of microcystin-LR and its glutathione conjugate in fish tissues by liquid chromatography-tandem mass spectrometry

A sensitive and selective liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantitative determination of microcystin-LR (MC-LR) and its glutathione conjugate (MC-LR-GSH) in fish tissues. The analytes were extracted from fish liver and kidney using 0.01 M EDTA-Na-2-5% acetic acid, followed by a solid-phase extraction (SPE) on Oasis HLB and silica cartridges. High-performance liquid chromatography (HPLC) with electrospray ionization mass spectrometry, operating in selected reaction monitoring (SRM) mode, was used to quantify MC-LR and its glutathione conjugate in fish liver and kidney. Recoveries of analytes were assessed at three concentrations (0.2, 1.0, and 5 mu g g(-1) dry weight [DW]) and ranged from 91 to 103% for MC-LR, and from 65.0 to 75.7% for MC-LR-GSH. The assay was linear within the range from 0.02 to 5.0 mu g g(-1) DW, with a limit of quantification (LOQ) of 0.02 mu g g(-1) DW. The limit of detection (LOD) of the method was 0.007 mu g g(-1) DW in both fish liver and kidney. The overall precision was determined on three different days. The values for within- and between-day precision in liver and kidney were within 15%. This method was applied to the identification and quantification of MC-LR and its glutathione conjugate in liver and kidney of fish with acute exposure of MC-LR. (c) 2007 Elsevier B.V. All rights reserved.

A laboratory study on risk assessment of microcystin-RR in cropland

The persistence time and risk of microcystin-RR (MC-RR) in cropland via irrigation were investigated under laboratory conditions. In order to evaluate the efficiency of the potential adsorption and biodegradation of MC-RR in cropland and the persistence time of MC-RR for crop irrigation, high performance liquid chromatography (HPLC) was used to quantify the amount of MC-RR in solutions. Our study indicated that MC-RR could be adsorbed and biodegraded in cropland soils. MC-RR at 6.5 mg/L could be completely degraded within 6 days with a lag phase of 1 - 2 days. In the presence of humic acid, the same amount of MC-RR could be degraded within 4 days without a lag phase. Accordingly, the persistence time of MC-RR in cropland soils should be about 6 days. This result also suggested the beneficial effects of the organic fertilizer utilization for the biodegradation of MC-RR in cropland soils. Our studies also demonstrated that MC-RR at low concentration (< 10 mu g/L) could accelerate the growth of plants, while high concentration of MC-RR (> 100 mu g/L) significantly inhibited the growth of plants. High sensitivity of the sprouting stage plants to MC-RR treatments as well as the strong inhibitory effects resulting from prolonged irrigation further indicated that this MC-RR growth-inhibition may vary with the duration of irrigation and life stage of the plants. (c) 2007 Published by Elsevier Ltd.

Sequential ultrastructural and biochemical changes induced in vivo by the hepatotoxic microcystins in liver of the phytoplanktivorous silver carp Hypophthalmichthys molitrix

Up to now, in vivo studies on the toxic effects of microcystins (MCs) on the ultrastructures of fish liver have been very limited. The phytoplanktivorous silver carp was injected i.p. with extracted hepatotoxic microcystins (mainly MC-RR and -LR) at a dose of 1000 mu g MC-LReq. kg(-1) body weight, showing a time-dependent ultrastructural change in liver as well as significant increases in enzyme activity of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH). We observed for the first time the occurrence of a large amount of activated secondary lysosomes, which might be an adaptive mechanism to eliminate or lessen cell damage caused by MCs through lysosome activation. Quantitative and qualitative determinations of MCs in the liver were conducted by HPLC and LC-MS2, respectively. MCs concentration in the liver reached the maximum (114.20 mu g g(-1) dry weight) after 3 h post-injection, and then rapidly dropped to 7.57 mu g g(-1) dry weight at 48 h, indicating a deputation of 99% accumulated MC-LReq. On the other hand, a decrease trend in glutathione (GSH) concentration was observed in the liver of silver carp while the activity of glutathione S-transferase (GST) increased significantly after injection. The high tolerance of silver carp to MCs might be due to the high basic GSH level in their liver, and/or an increased GSH synthesis. (C) 2007 Elsevier Inc. All rights reserved.

An improved method for the extraction of low molecular weight organic acids in variable charge soils

Due to specific adsorption to variable charge soils, low molecular weight organic acids (LMWOAs) have not been sufficiently extracted, even if common extractants, such as water and 0.1 M sodium hydroxide (NaOH), were employed. In this work, the method for extracting LMWOAs in soils with 0.1 M NaOH was improved for variable charge soils; e.g. 1.0 M potassium fluoride (KF) with pH 4.0 was applied as an extractant jointed with 0.1 M NaOH based on its stronger ability to change the electrochemical properties of variable charge soils by specific adsorption. With the proposed method, the recoveries of oxalic, tartaric, malic, citric and fumaric acids were increased from 83 4, 93 1, 22 2, 63 +/- 5 and 84 +/- 3% to 98 +/- 2, 100 +/- 2, 85 +/- 2, 90 +/- 2 and 89 +/- 2%, respectively, compared with NaOH alone. Simultaneously, the LMWOAs in Agri-Udic Ferrosol with field moisture were measured with a satisfactory result.