利用線上微透析技術配合同時多元素電熱式原子吸收光譜連線分析系統進行生理鹽水及細胞懸浮液中銅、錳、鎳與硒之分析研究

Continuous Monitoring of Cu, Mn, Ni and Se in Saline and Cell Suspension Using On-line Microdialysis Coupled with Simultaneous Multielement Electrothermal Atomic Absorption Spectrometry

陳柏華(Po-Hua Chen)

指導教授 ： 黃友利

高雄醫學大學/醫學院/醫學研究所/碩士(2005年)

摘要

本研究目的在於發展微透析配合原子吸收光譜儀之同時多元素分析連線系統，用以測定在生物系統中參與調控氧化壓力相關機轉的銅(Cu)、錳(Mn)、鎳(Ni)與硒(Se)。本研究內容包括兩部分，第一部份為建立同時多元素原子吸收光譜儀測定0.9% NaCl中的銅、錳、鎳與硒之儀器參數，包括溫度程式及基質修飾劑等。本實驗使用3 mg Pd以及10 mg Mg(NO3)2當作基質修飾劑，並配合1200℃裂解溫度，使得基質中氯化鈉的干擾能順利去除，且保留待測元素避免在裂解過程中散失，並經由分析方法的驗證而確認本方法之精密度及準確度。第二部分則使用0.9% NaCl為灌流液，以1.0 mL/min的灌流速度來建立微透析結合原子吸收光譜儀之連線系統，並應用於含銅、錳、鎳與硒標準品之生理鹽水進行透析作用。本連線方法分析銅、錳、鎳與硒之探針回收率分別為：57.9 ± 1.4%、65.0 ± 1.0%、65.5 ± 4.1%、67.9 ± 4.6%。此外，本連線系統的長時間穩定度、反應性及添加回收率等方法驗證之結果，顯示本連線系統之分析可靠性。最後將所建立的連線方法應用於細胞懸浮液添加銅、錳、鎳與硒之測定，其結果顯示，本連線方法確實能原位、動態且連續的監測細胞懸浮液中的銅、錳、鎳與硒之濃度變化情形。

關鍵字

線上 ； 鎳 ； 硒 ； 錳 ； 銅 ； 細胞懸浮液 ； 生理鹽水 ； 同時多元素電熱式原子吸收光譜 ； 微透析技術

並列摘要

Microdialysis sampling coupled on-line with simultaneous electrothermal atomic absorption spectrometry (SIMAAS) for continuous monitoring of copper (Cu), manganese (Mn), nickel (Ni) and selenium (Se) in cell cultured suspension has been developed. The trace elements investigated in this study are considered to be markedly associated with oxidative stress in biomedicine. Two major works were carried out in this study. Firstly, the analytical conditions including temperature program and matrix modifiers have been developed for the determination of Cu, Mn, Ni and Se in saline solution. The results showed that 3 mg of Pd and 10 mg of Mg(NO3)2 were successfully used as the matrix modifiers to prevent target elements from evaporation and to eliminate the interference resulting from NaCl in 1200℃ of the pyrolysis step. The analytical performances such as linearity, accuracy, and precision were validated to confirm the reliability of the proposed method. Secondly, the utrapure saline solution (0.9% NaCl) was used as the perfusion solution at the flow rate of 1 mL/min through the microdialysis probe to develop the on-line microdialysis-SIMAAS system. The optimized conditions of on-line microdialysis- SIMAAS system were performed for continuous measurements of four trace elements in microdialysate. The probe recovery of on-line microdialysis-SIMAAS system measured by a saline solution contained Cu, Mn, Ni and Se was 57.9 ± 1.4%, 65.0 ± 1.0%, 65.5 ± 4.1%, and 67.9 ± 4.6%, respectively. The analytical performances for microdialysis system such as long-term stability, response, and spiked recovery were validated to confirm the reliability of on-line system. The use of an on-line microdialysis-SIMAAS system permitted the in situ, dynamic and continuous monitoring of extracellular diffusible Cu, Mn, Ni and Se in cell suspension after treated with trace elements.

並列關鍵字

SIMAAS ； Microdialysis ； On-line ； Saline ； Se ； Cu ； Mn ； Cell Suspension ； Ni

參考文獻

[1]B. Halliwell, and J. M. C. Gutteridge, Biochem. J., 1984, 219, 1.

[2]D. Wu, and A. I. Cederbaum, Alcohol Res. Health., 2003, 27, 277.

[3]J. M. C. Gutteridge, Clin. Chem., 1995, 41, 1819.

[4]S. M. Beak, Y. S. Lee, and J. A. Kim, Biochimie, 2004, 86, 425.

[5]H. J. Chae, S. W. Chae, J. S. Kang, B. G. Bang, J. I. Han, S. R. Moon, R. K. Park, H. S. So, K. S. Jee, H. M. Kim and H. R. Kim, J. Radiat. Res., 1999, 40, 323.