利用線上微透析取樣配合層析與原子吸收光譜法進行活體動物血中砷物種之分析研究

Arsenic Speciation in Blood of Living Animals using On-Line Microdialysis Sampling Coupled with HPLC and AAS

鄭冠文(Guan-Wen Cheng)

指導教授 ： 黃友利 副教授 ； 吳信隆 教授

高雄醫學大學/藥學院/藥學研究所/碩士(2003年)

摘要

摘 要
砷是一種具有毒性且分佈相當廣泛的微量元素之一，並且已被認定為是致癌物質之一。最近的研究發現三氧化二砷對於白血病的治療頗具療效。然而，目前對於三氧化二砷在活體血液中砷物種代謝變化的情形仍無法直接偵測與了解。因此，本實驗採用多種儀器系統連線方式建立可自動、連續與動態監測血中砷物種變化情形之分析方法。
本研究建立了連續與動態偵測給予三氧化二砷的活體動物血液中所包含AsIII，AsV，MMA和DMA 四種砷物種之連線分析方法。此連線分析系統是利用微透析取樣技術連接高效能液相層析儀與氫化物產生原子吸收光譜儀來進行分析。本分析系統進行AsIII，AsV，MMA和DMA 四種砷物種分析之偵測極限分別是2.0, 2.9, 2.4 與4.2 ng ml-1。對濃度各為10 ng ml-1 之四種砷物種之分析精密度分別為7.0% (AsIII), 7.0% (MMA), 5.1% (DMA) 和 8.7% (AsV)。以標準水溶液偵測四種砷物種之回收率皆在標準水溶液濃度±10%以內。標準水溶液製備AsIII，AsV，MMA和DMA濃度各為10 ng ml-1時，以in-vivo 分析方法對大白兔血液中AsIII，AsV，MMA和DMA之平均回收率分別為6.9±1.4%, 12.7±2.6%, 8.6±2.0% 與 11.6±2.4%。應用所建立之連線分析系統，每隔20分鐘即可偵測得知給予三氧化二砷之大白兔血中四種砷物種的連續與動態變化情形。

關鍵字

三氧化二砷 ； 微透析 ； 砷物種 ； 連線分析

並列摘要

Abstract
In the nature, arsenic is one of the wide distribution toxic trace element and carcinogen. Recently, the arsenic trioxide was found to be an effective drug for leukemia. However, the direct detection of arsenic species in blood of living organisms treated with arsenic trioxide is still a challenge task. In the present study we try to develop an on-line analytical system to automatic, continuous, and dynamic speciation of arsenic in blood of living animal.
An on-line and fully automatic method was developed for the continuous and dynamic in vivo monitoring of four arsenic species [arsenite (AsIII), arsenate (AsV), monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA)] in blood of living organisms treated with arsenic trioxide. Microdialysis sampling technique was employed to couple with high performance liquid chromatography (HPLC) and hydride generation atomic absorption spectrometry (HGAAS). Detection limits of 2.0, 2.9, 2.4 and 4.2 ng ml-1 were obtained for AsIII, MMA, DMA and AsV, respectively. Precision values of 7.0% (AsIII), 7.0% (MMA), 5.1% (DMA) and 8.7% (AsV) were obtained, respectively, at a 10 ng ml-1 level. Recoveries close to 100%, relative to an aqueous standard, were observed for each species. The average in vivo recoveries of AsIII, MMA, DMA and AsV in rabbit blood were 6.9 ± 1.4%, 12.7 ± 2.6%, 8.6 ± 2.0% and 11.6 ± 2.4%, respectively, at a 10 ng ml-1 level. The applicability of the proposed method permits the determination of four arsenic species in the blood of rabbits treated with arsenic trioxide with a temporal resolution of 20 min sampling.

並列關鍵字

As2O3 ； Microdialysis ； Arsenic specices ； On-line analysis

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