高效液相色谱法测定鳙鱼肉中三种微囊藻毒素

目的:建立能同时检测鳙鱼肉中三种微囊藻毒素(MC-RR、MC-YR、MC-LR)的高效液相色谱分析法。方法:鱼样经90%的甲醇提取、离心后经固相萃取柱富集、净化,甲醇洗脱,氮吹浓缩,以乙腈+水(含0.05%三氟乙酸)38+62为流动相,经C18柱分离2,38 nm波长检测。结果:三种微囊藻毒素在0.25 mg/L～2.50 mg/L均有良好的线性,其相关系数均大于0.9995;回收率在67%～83%;RSD均小于5.0%。结论:该法灵敏度高、操作简便、准确,适用于淡水鱼中微囊藻毒素的检测。     

高效液相色谱法同时测定水中三种微囊藻毒素

目的:建立能同时检测水中三种微囊藻毒素(MC-RR、MC-YR、MC-LR)的高效液相色谱分析法。方法:水样经固相萃取柱富集、净化,甲醇洗脱,氮吹浓缩,以乙腈-水(含0.06%三氟乙酸)38∶62为流动相,经C18柱分离,238 nm波长检测。结果:三种藻毒素在0.25 mg/L~2.50 mg/L均有良好的线性,其相关系数均大于0.9995;回收率在83.0%~89.1%;RSD均小于5.0%。结论:该法灵敏度高、操作简便、准确,适用于水中微囊藻毒素的检测。     

绍兴市重要水域微囊藻毒素污染状况调查

目的了解绍兴市重要水域微囊藻毒素污染情况,为水质安全风险评估和治理提供依据。方法于2016年9月—2017年9月在绍兴市三大水域设置15个点进行水样采集,采用超高效液相色谱-串联质谱法对6种微囊藻毒素(MC-LR、MC-RR、MC-YR、MC-LA、MC-LY、MC-LF)进行检测,比较不同时间、地点采集水样的微囊藻毒素检出情况。结果共采集水样135份,检出微囊藻毒素22份,检出率为16.3%。萧绍河网、浦阳江流域和曹娥江流域采集水样检出率分别为7.9%、18.5%和26.7%;7月、8月和9月采集水样检出率分别为13.3%、40.0%和46.7%,其他月份均未检出。22份阳性样品中,6种微囊藻毒素检出率由高到低为MC-LR(100.0%)、MC-RR(100.0%)、MC-YR(22.7%)、MC-LY(18.2%)、MC-LF(13.6%)和MC-LA(0.0%)。结论绍兴市三大水域夏秋季均存在微囊藻毒素污染,以MC-LR和MC-RR为主。     

液相色谱串联质谱法分析饮用水中微囊藻毒素

微囊藻毒素是一类具生物活性的单环七肽,目前所检测到的微囊藻毒素异构体已超过50多种,其中主要的有MC-LR、MC-YR和MC-RR三种.     

茂名市水源水及出厂水中藻毒素污染调查

目的了解茂名市出厂水及水源水中的微囊藻毒素的污染情况。方法于2010年6月至2011年5月,采用超快速液相色谱-质谱方法对茂名市水源水(良德水库、石骨水库出水口和石骨水库入水口)、茂名市山阁调蓄池水及茂名市河东水厂出厂水中的3种微囊藻毒素MC-RR、MC-LR、MC-YR含量进行动态监测。结果水源水水样中MC-RR、MC-LR、MC-YR的检出率分别为66.7%(24/36),58.3%(21/36),8.3%(3/36),检测范围分别为0.004～0.994、0.004～0.134、0.030～0.055μg/L,水源水中MC-RR在10月份出现高峰值(0.994μg/L)。4—9月调蓄池水中MC-LR的含量均高于水库水;而出厂水中MC-RR、MC-LR、MC-YR的含量均低于水库水,且仅检出MC-RR(0.008～0.012μg/L)。结论茂名市水源水主要以MC-RR、MC-LR的持续污染为主,出厂水受到低浓度MC-RR污染,饮用水安全存在一定的隐患。     

超高效液相色谱-串联质谱法同时检测生活饮用水中7种微囊藻毒素

目的建立检测生活饮用水中7种微囊藻毒素(MC)异构体MC-LR、MC-RR、MC-YR、MC-LA、MC-LF、MC-LY、MC-LW的方法。方法生活饮用水无需任何处理,过滤后直接上机测定;水源水经煮沸处理30 min后,用C18固相萃取小柱萃取净化,5 m L 0.1%甲酸甲醇溶液洗脱,氮气吹干后用甲醇定容至1 m L,过0.22μm滤膜后上机测定。结果在0~20μg/L范围内7种微囊藻毒素的线性关系良好,相关系数r≥0.998,最低检出限介于0.2~0.5μg/L间;在0.5、1.0、5.0μg/L3个浓度下加标回收,平均回收率在76.9%~113.0%之间,RSD在1.1%~11.3%之间。结论该法能快速、准确地检测生活饮用水中7种微囊藻毒素。     

郑州市主要生活饮用水源微囊藻细胞毒素特征分析

目的分析郑州市主要生活饮用水源微囊藻细胞毒素特征。方法以西流湖和黄河花园口段某调蓄池作为调查现场,采用96孔板结和极限稀释法对采集藻细胞进行分离纯化;应用全细胞PCR方法检测所分离微囊藻细胞株藻青蛋白基因中间序列(PC-IGS)和微囊藻毒素多肽合成酶基因B(mcyB),并对提取毒素应用固相萃取(SPE)高效液相色谱(HPLC)法进行检测。结果自西流湖分离的XLH细胞株和黄河花园口调蓄池分离的2株微囊藻细胞BM1、BM2,PC-IGS、mcyB基因扩增均为阳性;mcyB基因序列测定结果与Genbank中报道的mcy基因同源性达99%;HPLC检测3株藻细胞所含毒素异构体主要为MC-LR,占毒素总量的质量百分比分别为97.9%、98.6%和99.3%。结论自郑州市主要生活饮用水源分离的3株微囊藻细胞均为产毒株,产生毒素异构体主要为毒性较大的MC-LR。     

鱼、鸭样品中微囊藻毒素含量测定方法研究

目的建立鱼、鸭肉和肝脏样品中微囊藻毒素(MC-LR)含量的测定方法。方法取鱼、鸭肉和肝脏经组织捣碎机捣碎,甲醇提取,离心,Sep-pakCl8(ODS)小柱富集、净化,0.1%三氟乙酸(TFA)甲醇梯度解析,酶联免疫吸附试验(ELISA)法测定MC-LR含量。结果方法最小检出浓度为0.1ng/g,相对标准偏差为5.34%,加标回收率为90%～92%。结论该方法预处理样品分离效果好,对测定无干扰,所建立的方法经实际应用取得了满意的效果。     

微囊藻毒素检测方法与福州市售水产品污染调查

目的报道微囊藻毒素(MCs)检测方法,调查福州市售水产品MCs污染情况。方法于2015年夏季采集福州市售8种水产品44份,用固相萃取-高效液相色谱法,检测水产品中微囊藻毒素MC-LR、MC-RR、MC-YR含量。结果检测的44份水产品样品中,MCs检出率为27.3%(12/44);在花蛤、贻贝、田螺、鲫鱼、鲢鱼、草鱼样品中均有检出,以贝类污染率较高;MCs污染检出率9月(41.7%)高于7月(5.0%)。结论福州市售水产品存在微囊藻毒素污染,应加强对养殖水源污染的防治。     

水中微囊藻毒素的超高效液相色谱-四极杆-静电场轨道阱高分辨质谱测定法

目的建立水中微囊藻毒素(MC)煮沸浓缩-超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UPLCquadrupole-Orbitrap MS)的测定方法。方法水样在电热板上加热浓缩后,以0.22μm滤膜过滤,采用BEH C₁₈色谱柱(50mm×2.0 mm,2.1μm),以甲醇-2‰甲酸水溶液作为流动相,梯度洗脱,采用电喷雾电离源正离子模式(HESI+)在t-SIM模式下分析检测,通过测得的精确质量数与理论精确质量数吻合度、目标化合物的元素组成、保留时间进行定性。结果MC-LR、MC-YR、MC-RR、MC-LA、MC-LF、MC-LY在0.2～5μg/L,MC-WR在1～25μg/L之间线性良好,相关系数均>0.998;MC-RR、MC-YR、MC-LR、MC-WR、MC-LA、MC-LY、MC-LF的检出限分别为0.18、0.26、0.40、1.31、0.29、0.29、0.29ng/L;高、中、低浓度加标的RSD为4.11%～10.8%,平均回收率为61.8%～103%。结论该方法简便快捷,选择性好,灵敏度高,重现性好,可满足水中微囊藻毒素检测的要求。     

Bioaccumulation of the hepatotoxic microcystins in various organs of a freshwater snail from a subtropical Chinese lake, Taihu Lake, with dense toxic Microcystis blooms

In this paper, we describe the seasonal dynamics of three common microcystins (MCs; MC-RR, MC-YR, and MC-LR) in the whole body, hepatopancreas, intestine, gonad, foot, remaining tissue, and offspring of a freshwater snail, Bellamya aeruginosa, from Gonghu Bay of Lake Taihu, China, where dense toxic Microcystis blooms occur in the warm seasons. Microcystins were determined by liquid chromatography electrospray ionization mass spectrum. Microcystin (MC-RR + MC-YR + MC-LR) content of the offspring and gonad showed high positive correlation, indicating that microcystins could transfer from adult females to their young with physiological connection. This study is the first to report the presence of microcystins in the offspring of the adult snail. The majority of the toxins were present in the intestine (53.6%) and hepatopancreas (29.9%), whereas other tissues contained only 16.5%. If intestines are excluded, up to 64.3% of the toxin burden was allocated in the hepatopancreas. The microcystin content in the intestine, hepatopancreas, and gonad were correlated with the biomass of Microcystis and intracellular and extracellular toxins. Of the analyzed foot samples, 18.2% were above the tolerable daily microcystin intake recommended by the World Health Organization (WHO) for human consumption. This result indicates that public health warnings regarding human ingestion of snails from Taihu Lake are warranted. In addition, further studies are needed to evaluate the occurrence by Microcystis in relation to spatial and temporal changes in water quality.     

水体中微囊藻毒素和节球藻毒素的UPLC-MS~n分析方法研究

[目的]针对水体中3种代表性的微囊藻毒素（microcystin-RR,MC-RR;microcystin-YR,MC-YR;microcystin-LR,MC-LR）和节球藻毒素（nodularin,NOD）,建立一种快速、准确、可靠的痕量分析方法。[方法]样品经C18固相萃取柱净化富集后,采用超高效液相色谱-电喷雾串联三重四级杆质谱法（ultra performance liquid chromatography/tande mmass spectrometry,UPLC-MS/MS）,在多反应监测（Multi reaction monitoring,MRM）模式下进行检测,采用基质匹配标准曲线法消除基质效应。[结果]该方法加标回收率为95.7%~115.0%,精密度（RSD）范围为2.43%~6.04%,检测限可达0.05~0.20ng/L。将该方法应用于实际水样分析,水体中3种微囊藻毒素和节球藻毒素含量低于检测限。[结论]建立了水体中3种痕量微囊藻毒素和节球藻毒素的UPLC-MS/MS分析方法,该方法可用于实际水样的检测。     

高效液相色谱-串联质谱联用法测定饮用水中5种微囊藻毒素

目的建立液相色谱-串联质谱法同时测定饮用水中5种微囊藻毒素(MC-LR、MC-LW、MC-RR、MC-LF、MC-YR)检测的方法。方法采用直接进样方式,用高效液相色谱-串联质谱(HPLC/MS/MS)测定饮用水和水源水中的微囊藻毒素。水样经0.22μm微孔滤膜过滤,采用HPLC/MS/MS电喷雾电离阳离子(ESI+),多反应监测(MRM)模式检测,外标法定量。结果方法的线性范围为0.5～50.0μg/L,线性相关系数0.9994～1.0000,检出限0.06～0.08μg/L;高低两个水平的平均加标回收率分别为91.2%～102%和93.0%～99.0%;相对标准偏差(RSD)2.11%～3.264%;日内重复取样测定的RSD≤3.26%,日间重复取样测定的RSD≤4.36%。结论该方法可用于测定水中5种微囊藻毒素,方法操作简单、干扰少、快速、准确可靠,检测方法的检出限、精密度和加标回收率符合国家生活饮用水标准检验方法对质量控制的要求。     

江南某城市饮用水中微囊藻毒素调查及初步健康风险评价

[目的]调查江南某城市饮用水中微囊藻毒素[MCs（MC．LR、MC-RR、MC-YR）]浓度,并进行初步健康风险评估。[方法]2010年6月一2011年5月,按月采集各城区饮用水样品,经Cts固相萃取柱净化富集后,采用超高效液相色谱一电喷雾三重四级杆串联质谱对3种典型的MCs进行定量分析,并应用水环境健康风险评价模型对监测结果进行初步评价。[结果]从季节性变化看,MCs全年均有检出,浓度在ND～16．3ng／L,以MC-LR为主,其中8月达最高值;部分城区饮用水中多个月份中的MC．LR浓度超过10．0ng／L;由于供水差异,各区浓度差异较大,浓度最低的城区年平均值为0．2n班,最高者年平均值为6．7ng／L。个人年健康风险值范围为（0．15×10-6^（-5）．62×10^（-6）／a。各城区饮用水中MCs所导致的健康风险低于国际辐射防护委员会（ICRP）和美国环境保护署（USEPA）的推荐值[（5×10^（-5））／a和（1x10^（-4）／a]。[结论]供水水质不均衡,饮用水中藻毒素浓度差异较大,大部分城区中饮用水存在一定的健康风险。     

HPLC-PDA法同时测定水源水中主要3种微囊藻毒素方法的研究

[目的]建立水中微囊藻毒素(MC)含量的高效液相色谱初筛、光谱定性、质谱确认分析方法。[方法]以Waters Atlantis dC18(3μ2.1×100 mm),色谱质谱柱为分析柱,流动相:20%甲醇溶液,Surveyor二极管阵列检测器(PDA)和Finnigan LCQ Deca XP MAX质谱仪为检测器。[结果]回归方程和相关系数为:MC-RR y=7.401998×1025 x+5.028972×102,r=0.998;MC-LR y=5.689824×105x+5.925834×102,r=0.9996;MC-YR y=4.816086×105x+6.145898×102,r=0.9994。平均回收率为=95.1%~105.0%,RSD为3.00%~4.99%。[结论]用HPLC-PDA法检测水源水中的微囊藻毒素方法简便,重现性好,灵敏度高,定性准确,定量精确。     

Apoptotic effect of cyanobacterial blooms collected from Polish water reservoirs

Recently in many countries, including Poland, the problem of toxicity of cyanobacterial blooms has been of great importance. In many cases it is connected with the increase of microcystins (MCYSTs) concentration in fresh water. This problem is caused by excessive eutrophication of drinking and recreational water bodies. In humans, the most frequent symptoms of the MCYST effect are: cutaneous rash, fever, vomiting, diarrhoea, gastroenteritis and acute damage of the liver. The aim of this work was to estimate apoptotic effects of five different cyanobacterial hepatotoxic extracts containing MC-LR and other variants of MCYSTs (MC-RR, MC-YR, and MC-WR). These effects were analysed in rat hepatocytes--primary target of cyanobacterial hepatotoxins. Morphological changes in hepatocytes were examined by means of fluorescence and differential interference contrast microscopy with the DNA-specific dye, Hoechst 33342. The hepatocytes were treated with each cyanobacterial extracts containing MC-LR in the range between 100 nM-2000 nM for 30 min, 60 min and 120 min. The first characteristic apoptotic changes: shrinking and budding of cells were seen after 30 min, MC-LR = 100 nM. During the next 30 min the percentage of apoptotic cells increased by over 50%, MC-LR at concentrations ranging from 100 to 250 nM (the value dependent on a bloom sample). Highly condensed chromatin and apoptotic bodies were observed in 85-90% of hepatocytes after 120 min of treatment with MC-LR in concentration of 1000 nM. The apoptotic changes in rat hepatocytes confirm the high cytotoxic potential of cyanobacterial bloom samples collected during different months and years from reservoirs of drinking and recreational water in central Poland.     

Observation of Sward Destruction Caused by Irrigation with Toxic <Emphasis Type="Italic">Microcystis</Emphasis> Morphospecies Containing Water in Southern Hungary

In the summer of 2006 bloom-like phenomenon occurred in a garden pond in Szeged, Southern Hungary. After regular watering of a sward with pond water containing the algal mass, destruction of garden grass occurred. Microcystis aeruginosa, Microcystis viridis, Microcystis ichthyoblabe, and Microcystis wesenbergii were identified by light microscopy in the water sample; microcystin-FR, -LR, -RR and -YR were determined by matrix-assisted laser desorption/ionization—time-of-flight analysis. There was an 80% decrease in the green mass (87% in chlorophyll-content) of the grass in a 1 m² area of the garden irrigated with pond water.     

First reported case of turtle deaths during a toxic Microcystis spp. bloom in Lake Oubeira, Algeria

Microcystins analysis was conducted in field cyanobacterial bloom samples and dead terrapin tissues from Lake Oubeira (Algeria) with an aim of studying the cause of the mortality of the freshwater terrapin species Emys orbicularis and Mauremys leprosa during October 2005. The deaths of these two terrapin species were observed during a bloom of Microcystis spp. The total microcystin content per phytoplankton biomass evaluated with the methanol extraction-protein phosphatase methodology was 1.12 mg MCYST-LR equivalents/g dried bloom material. The analysis of this bloom extract by the LC/MS technique demonstrated the presence of three microcystin variants: microcystin-LR (MCYST-LR), microcystin-YR (MCYST-YR), and microcystin-RR (MCYST-RR). Microcystins were also detected in fresh carcasses of terrapin liver, viscera and muscle tissues using the GC/MS after Lemieux oxidation method and the PP2A inhibition assay. The high level of microcystins detected using the Lemieux oxidation-GC/MS method in the liver tissue (1192.8 microg MCYST-LR equivalent/g dw) and in the viscera tissue (37.19 microg MCYST-LR equivalent/g dw) of the species M. leprosa and E. orbicularis, respectively, and the liver crumbling observed after the necropsy examination of the fresh carcass of M. leprosa support the possibility that cyanobacterial microcystins contribute to the turtle mortalities.     

Direct FAME synthesis for rapid total lipid analysis from fish oil and cod liver oil

A direct method for preparation of fatty acid methyl ester (FAME) via transesterification of fatty acids from fish oil and cod liver oil using methanolic sulfuric acid and chloroform as co-solvents was optimized. The synthesized FAME were subsequently analysed by gas chromatography (GC) to identify and quantify the individual fatty acid in the oils. Temperature and heating times had a significant effect on the total fatty acid (TFA) recovery. In general, temperatures of 90°C and 100°C and reaction times of 90 and 30 min, respectively, resulted in maximum recovery of TFA from the oils. TFA recoveries of more than 95% suggest that this method is suitable for quantitative analysis of individual fatty acids. In comparison, a conventional method which involved a separate extraction procedure using chloroform and methanol before FAME synthesis yielded approximately 30% lower TFA recovery from the oils. It can be concluded that the direct FAME synthesis technique is superior to the current conventional method, not only because of its simplicity, speed and economical use of solvents, but also its precision.     

HPLC法测定花锚中落干酸和龙胆苦苷的含量

目的:建立反高效液相色谱法测定花锚中落干酸和龙胆苦苷的含量。方法:采用kromasil—C18(250mm×4.6mm,5μm)色谱柱,以流动相甲醇和水(含0.04%磷酸)的比例在0~15min内由23:77至30:70线性梯度洗脱,流速1 ml/min,检测波长238 nm,柱温35℃。结果:两种成分均达到基线分离,落干酸和龙胆苦苷的线性范围分别为0.198~5.544μg(r=0.9999),0.0262~0.7336μg(r=0.9999);回收率为96.12%(RSD=1.73%)、97.67%(RSD=1.91%)。结论:方法测定快速,结果准确、可靠。