摘要
微囊藻毒素(Microeystins,MCs)是富营养化淡水水体中蓝藻的爆发性繁殖可能产生的最常见的藻毒素,它是一种单环七肽毒素,该毒素具有强烈的肝毒性和肿瘤促进作用。近年来作为福州市第二饮用水源地的山仔水库水体呈现不同程度的富营养化,监测饮用水水源地藻毒素的含量及污染状况对于控制藻毒素污染,减轻对人体的危害,保护人民的身体健康,具有重要的意义。
本研究对MC-LR、MC-RR、MC-YR三种微囊藻毒素的高效液相色谱检测及固相萃取前处理过程中的关键环节进行优化研究,并应用该方法对2007年山仔水库微囊藻毒素的污染水平及环境因子进行了为期一年的连续监测。通过本研究得出以下结论:
(1)微囊藻毒素分析的关键技术环节
HPLC进样分析时,以甲醇:0.06%TFA水溶液=60:40(V/V)为流动相进行等度洗脱,对MC-YR、MC-LR和MC-RR有较好的分离效果。在固相萃取前处理过程中,以0.1%TFA-甲醇为洗脱液对MC-YR、MC-LR和MC-RR三种毒素均具有较高的回收率,回收率分别可达到91%、89.8%,81%;对于环境样品,以20%甲醇为淋洗液,可以减少杂质峰对HPLC分析测试的影响,并保证回收率。
(2)山仔水库2007年微囊藻毒素的变化特征
2007年山仔水库的微囊藻毒素形态主要是MC-YR;在藻类的高发季节6～9月份均检出微囊藻毒素MC-YR,水中检出的MC-YR最大浓度为0.374μg/L,低于国家生活饮用水卫生规范的标准1.0μg/L。在水中加入冰乙酸破藻细胞后,水中微囊藻毒素含量成倍增加,最多增加了41.1倍。
(3)影响微囊藻毒素水平的水质条件
水温是微囊藻毒素浓度水平高低的关键影响因子,其次为TN/TP、TP,水体第一生产力—叶绿素a与藻毒素之间不存在显著的直接相关关系。在水温较高,水体中磷元素供应充足的条件下,水体藻类可能产生出更多的藻毒素,水质安全的关键环节在于控制每年高温期水体的TP水平。水质参数之间,TP—TN—叶绿素a之间呈现很强的相关性,叶绿素a其水平高低受到TN、TP水平高低、以及它们之间的交互作用的影响。
(4)水质评价结果与微囊藻毒素浓度水平
2007年山仔水库春季水体有从中营养向富营养化水平发展的趋势,夏季水体达到了富营养化水平,秋冬季节的水体尚处于中营养水平。水体的营养状态指数与微囊藻毒素的浓度水平呈显著正相关。
Microcystins (Microcystins, MCs) is the most popular algae toxin generated by the explosive propagation of cyanobacteria in the fresh water in the state of eutrophication. It is a kind of single-ring seven peptide toxin, the toxin has a strong liver toxicity and tumor promoting role. Shanzai reservoir as a the second drinking water resource for Fuzhou citizens showed somewhat eutrophication in recent years, Therefore it is urgently necessary to monitor the MC levels and understand its environmental behaviors in Shanzai reservoir, for controlling the harmful effects of MC to local citizens and keeping the health of citizens.
In this study, some key techniques for monitoring the three microcystin MC-LR, MC-RR, MC-YR by use of high-performance liquid chromatography (HPLC) analysis and solid-phase extraction for sample preparation were optimized, and with the monitoring technique, the microcystin pollution levels and environmental factors in Shanzai reservoir were monitored in the whole year of 2007. Some meaningful research conclusions were listed as following:
(1) The key techniques of monitoring MCs
It is a better separation for the MC-YR, MC-LR and MC-RR in HPLC sample analysis: the methanol: 0.06% TFA solution = 60:40 (V/V) as the mobile phase. It will have a higher recovery rate during the Solid Phase Extraction in pre-treatment process: using 0.1% TFA-methanol as the eluate for the three toxins MC-YR, MC-LR and MC-RR, and the recovery rate would reach 91%, 89.8%, 81% respectively. For environmental samples to use 20% methanol as eluent can reduce the impurity peaks impact on the HPLC analysis, and can ensure the recovery.
(2) The characteristics of MCs in Shanzai Reservoir in 2007
MC-YR is the main form of microcystins in Shanzai reservoir in 2007. In the season of June to September with high algae growth, MC-YR were detected out with the highest concentration 0.374μg/L in the water, lower than the national drinking water standard 1.0μg/L. Adding glacial acetic acid in the water to breaking algal cells, the content of microcystins increased up to 41.1 times of the original level.
(3) The relationship among MCs and water environmental indicators
The critical water environmental indicators which directly influence the MC levels were proved as Water temperature、TN/TP and TP. There is not significant correlation between the Water Primary Productivity - chlorophyll-a and microcystins. Under the conditions of higher water temperature and adequate supply of phosphorus, higher level of MC will exist in water. Therefore, it is regarded that the key problem of keeping the safety of drinking water is to reduce the TP level in June to Sept. with the higher temperature in whole year. For water quality parameters, TP-TN-chlorophyll-a showed a significant correlations. Chlorophyll-a level was strongly impacted by the level of TN and TP, as well as the interaction between them.
(4) Assessment of the water quality and relations between MCs and water quality
In 2007, the nutrition status of Shanzai reservoir is from mesotrophic to eutroph in Spring and is from light eutrophic to middle eutropher in Summer and mesotrophic in Autumn and Winter. There is significant correlation between the nutritional status index and the concentration levels of Microcystins.
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