新興污染物鍶與有機酸結合動力及其螯合物之毒性研究

Studies on the Combination Dynamics and Toxicity of Emerging Contaminant Strontium with Organics Acids and Its Chelates

吳唯菱(Wei-Ling Wu)

指導教授 ： 黃武章

國立屏東科技大學/工學院/環境工程與科學系所/碩士(2013年)

https://doi.org/10.6346/NPUST.2013.00138

摘要

近年由於環境監測技術上的進步以及各種精密檢測儀器的開發，發現了以往未知但對人體有害之污染物，這些污染物未被各種管制標準所列管，但在環境中達到一定濃度時，可能會危害到環境生態及人體健康，其稱之為新興污染物。美國環保署公佈的「候選污染物名單3」，列出可能污染飲水水質但尚未列管的化學物質，其中極少數為無機性，例如鍶。由於鍶化合物的毒性資料不多，本研究特別選擇可能於環境中生成之三種鍶化合物(分別為硝酸鍶、氫氧化鍶、碳酸鍶)，進一步探討其細胞毒性與生物毒性。鍶化合物具不易溶解之特性，於是我們以三種不同溫度水(4 ℃、25 ℃、90 ℃)作為溶劑加以溶解，觀察其最大溶解度時的濃度與其飽合溶液之毒性變化。另外，考量人們飲用含鍶的水後可能對於哺乳類動物腎臟之負擔，我們將鍶水溶液與尿酸水溶液混合，觀察結合後溶液之分光光度計強度與測量毒性值之變化。本研究選擇溴化噻唑藍四氮唑之細胞存活率分析法作為非洲綠猴腎細胞(Vero cell) 之細胞毒性量測；混濁度分析作為大腸桿菌(E. coli) 之生態毒性量測。
本研究意外發現試驗所使用的純水對於所量測到的細胞毒性有嚴重干擾現象:超純水與去離子水對於細胞毒性分別會產生數值極高及極低之現象 (由超純水配製的樣品其細胞存活率平均落在80% 以上；以去離子水配製之樣品其數值呈現負值之情況)。我們將超純水及去離子水依不同配比混和後，0.01 mL混合純水樣品加入0.6 mL D2O中，測定純水樣之核磁共振氫譜，發現強度落在8以上者，以這類的混合純水進行細胞毒性實驗，所測得的量測細胞毒性值相較穩定(細胞存活率65 ~ 68 %)。
結果顯示，在以混濁度分析方法量測到的生態毒性方面，只有碳酸鍶對大腸桿菌有較高之毒性。在以溴化噻唑藍四氮唑之細胞存活率分析法量測細胞毒性方面，濃度為100 mg/L之各種鍶化合物水溶液之細胞存活率分別為: 硝酸鍶113.15±1.01 %；氫氧化鍶72.58±1.04 %；碳酸鍶87.98±3.46%。將100 mg/L之鍶化合物溶液與100 mg/mL之尿酸水溶液以不同比例混合後發現最高的量測毒性值分別出現在: 硝酸鍶：尿酸為7：3 時，存活率55.91±2.69%；氫氧化鍶：尿酸為8：2時，存活率68.24±0.49%；碳酸鍶：尿酸為1：9時，存活率72.14±0.21%。結果也顯示，在以混濁度分析方法量測到的100 mg/L之鍶化合物與100 mg/L尿酸之混合水溶液的生態毒性方面，大腸桿菌生長曲線經由波茲曼函數調和後所得之參數，隨著尿酸之混合比例增加，其參數值下降，顯示出毒性增強。顯示出三種鍶化合物與尿酸結合之後其螯合物對哺乳類動物之腎臟細胞及大腸桿菌會產生更高的毒性。

關鍵字

鍶 ； 細胞毒性 ； 生態毒性 ； 非洲綠猴腎細胞 ； 大腸桿菌 ； 尿酸

並列摘要

In recent years, due to the improvement of environmental monitoring techniques and the development of sophisticated detection equipments, people have found many un-explored contaminants in the past. These contaminants are not controlled by any regulatory standards, when their concentrations reach a certain level in the environment, they might be harmful for our environment, ecology and human health. These pollutants are called as Emerging Contaminants (ECs). U.S. EPA have announced " Contaminant Candidate List 3 (CCL3)". CCL 3 is a list of contaminants that are currently not subject to any proposed or promulgated national primary drinking water regulations. It is very few among those contaminants that are inorganic elements; one of them is Strontium (Sr). In the past, the toxicities of strontium compounds are lacked, therefore we have choice three strontium compounds (strontium nitride, strontium hydroxide and strontium carbonate) to investigate their cytotoxicities and ecotoxicities. Due to the extremely low solubility of those strontium compounds, we dissolved them in water under three temperatures (4 ℃、25 ℃、90 ℃) and observed the concentration and toxicity of these saturated strontium solutions. In addition, to understand the impact of those solutions on the human’s kidney cells, strontium solution was mixed with uric acid and to observe the changes in UV and experimental toxicity. In this study, the cytotoxicity of African green monkey kidney (Vero) cells is obtained by using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-
tetrazolium bromide as indicator in cell viability analysis; while the ecotoxicity is performed on the growth curve of E. coli. through the detection of turbidity. An unexpected finding is found in this study, ultra-pure water and deionized water will cause a too high (mostly cell survival over 80 %) or too low (mostly less than zero) measured toxicity value. We mixed ultra-pure water and deionized water by different ratios. After mixing, 0.01 mL of pure water mixture samples were added into 0.6 mL D2O and their proton intensity of Nuclear Magnetic Resonance (NMR) spectroscopy were performed. Results found that if ultrapure water / deionized water mixture has a high proton value ( > 8), such water can be used for the cytotoxicity test (cell survival near 65-68 %).
From the growth curve of E. Coli. analysis, only strontium carbonate possesses a significantly ecotoxicity toxicity. The Vero cell viability of 100 mg/L strontium nitrate is 113.15 ± 1.01%; strontium hydroxide is 72.58 ± 1.04%; strontium carbonate is 87.98 ± 3.46%. When 100 mg/L of strontium compounds combined with 100 mg/mL of uric acid, strontium nitrate: uric acid = 7 : 3 had the lowest cell vability 55.91±2.69%; strontium hydroxide: acid = 8 : 2 had the lowest cell vability 68.24±0.49%; strontium: uric acid = 1 : 9 had the lowest cell vability 72.14±0.21%. Results of growth curve of E. Coli. of strontium/uric acid mixtures show that as the mixed ratio of uric acid is increased, the fitted parameter of Bozeman function is decreased, that is indicated an increment of ecotoxicity.
Our results have illustrated that the chelates of strontium and uric acid cause more toxic for kidney cell and E. coli.

並列關鍵字

Strontium ； Cytotoxicity ； Ecotoxicity ； African green monkey kidney cells ； E. coli ； Uric acid

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