奈米粉體在分離及電化學感測上之應用研究

黃世宏

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奈米粉體在分離及電化學感測上之應用研究

Studies on application of nanopowders in separation and electrochemical detection

黃世宏 , Ph.D　　Advisor：陳東煌　　

繁體中文

電化學感測；氧化鐵；吸附；脂肪分解酵素；聚丙烯酸；磁性奈米吸附劑；重金屬離子；分離；氨基化；回收；碳管；正腎上腺素； recovery ； carbon nanotube ； Norepinephrine (NE) ； magnetic nano-adsorbent ； polyacrylic acid ； Lipase ； adsorption ； Amino-functionalization ； heavy metal ions ； electrochemical detection ； iron oxide ； separation

Abstract │ Reference (273) │ Times Cited (17) │ Altmetrics

Abstract 〈TOP〉

本論文係有關功能性磁性奈米吸附劑在分離及PAA被覆奈米碳管在電化學感測上的應用。首先探討PAA被覆磁性奈米粒子在脂肪分解酵素(lipase)分離上的應用；其次利用化學修飾的方法進一步將PAA被覆磁性奈米粒子，使表面具備氨基化，並應用在重金屬(陰/陽離子)的分離與回收；最後利用PAA分散奈米碳管，並將PAA被覆奈米碳管修飾在網印電極上，並在電化學感測上，用於偵測維生素C、正腎上腺素及尿酸。
   關於PAA被覆磁性奈米粒子在Candida rugosa lipase分離上的研究，吸附百分比取決於溶液的pH值，吸附量隨著pH值降低而增加，在pH 7-5.5從20%增加到90%，在pH 4.5-3.5吸附量達到98%，其吸附行為遵守Langmuir恆溫吸附模式，在2.5℃下0.03M、pH 3.5磷酸鹽緩衝溶液中，最大吸附容量(qm)和Langmuir平衡常數(K)分別為0.605 mg/mg與14.5 mL/mg。lipase吸附後，能夠在0.03M、pH 9磷酸鹽緩衝溶液中進行脫附，且經過吸附/脫附後，並沒有明顯的活性損失，根據pH影響、脫附和活性分析的研究，推測在使用的lipase，可能含有20%不純物或是不具活性的蛋白質，另外，lipase和PAA之間的靜電交互作用力，並沒有受到溫度(15~35℃)的影響。由於PAA被覆磁性奈米吸附劑無內物擴散阻力，故能在短時間內進行lipase的吸附/脫附實驗。基於上述結果，PAA被覆磁性奈米吸附劑可以有效率被用於並回收lipase。
   關於氨基化磁性奈米吸附劑在重金屬回收之研究，係先製備表面被覆PAA的氧化鐵奈米粒子，再藉由碳二醯胺的活化將二次乙基三胺(DETA)共價鍵結在氧化鐵奈米粒子上。穿透式電子顯微鏡(TEM)分析顯示，氨基化磁性奈米粒子具有分散性，平均粒徑為11.2±2.8 nm。由X射線繞射儀(XRD)分析得知，磁性奈米粒子為四氧化三鐵之尖晶石結構，且不因鍵結程序而改變其結構，此外，由磁性分析得知，所得磁性奈米粒子具有超順磁性且飽和磁化量為63.2 emu/g，另外由傅立葉轉換紅外線光譜儀(FTIR) 和界面電位分析儀測量其等電點為2.45之分析，可確認表面被覆PAA的氧化鐵奈米粒子已被氨基化，等電點從2.64偏移至4.59。關於氨基化磁性奈米吸附劑在水溶液對於吸附金屬陽/陰離子有相當高的吸附容量和效率，可以利用螯合或是離子交換機制去除金屬離子。氨基化的磁性奈米吸附劑吸附Cu2+ 和Cr(VI)皆依循Langmuir恆溫吸附模式，最大吸附容量(qm) 和Langmuir平衡常數(K) 對於Cu2+ 和Cr(VI)分別為12.43 mg/g 與 0.06 L/mg；11.24 mg/g 與 0.0165 L/mg。
    關於PAA被覆奈米碳管修飾在網印電極上，並在電化學感測上，用於偵測維生素C、正腎上腺素及尿酸的研究。係首先將碳管加入PAA水溶液中，在超音波下均勻混合形成PAA被覆奈米碳管複合物，並在網印電極表面上塗佈上一層PAA-MWNTs。經過PAA被覆奈米碳管修飾的網印電極，具有較大的表面積，而對於NE和UA分別可經由氫鍵或是靜電作用力的機制吸附在電極表面上。PAA被覆奈米碳管修飾的網印電極在磷酸鹽緩衝溶液(0.1 M、pH 7.5)中，偵測AA、 NE 和UA非常好的電化學催化活性，AA的氧化過電壓會降低，而NE 和UA氧化電流密度有明顯的增加。在使用微分脈衝伏安法偵測AA、 NE 和UA三成份混合溶液，氧化電位差NE-AA和UA-AA分別為228mV和112mV。經PAA被覆奈米碳管修飾的網印電極利用微分脈衝伏安法能夠將偵測三成份的訊號完全區分開來，也因此可以同時得到這三成份。而AA、NE 和UA濃度的線性範圍分別為100~1000μM、0~10μM 0~30μM;且偵側極限(S/N=3)分別為49.772μM 0.131μM和0.458μM。

Parallel Abstract 〈TOP〉

This dissertation concerns the application of functional magnetic nano-adsorbent in enzyme separation and heavy metal ions recovery and  PAA-coated multiwalled carbon nanotube in electrochemical detection. The adsorption of lipase from an aqueous solution by PAA-bound iron oxide magnetic nanoparticles was studied. Then the PAA-coated magnetic nanoparticles were further amino-functionalization using diethylenetriamine via carbodiimide activation and used for the recovery of heavy metal ions from aqueous solution. Finally, using PAA to dispersion carbon nanotube and PAA-coated multiwalled carbon nanotube composite modified Screen printed carbon electrode for the simultaneous determination ascorbic acid (AA)、Norepinephrine (NE) and Uric acid (UA).
    The feasibility of the polyacrylic acid (PAA)-bound magnetic nano-adsorbent for the recovery of Candida rugosa lipase from aqueous solutions was studied. The adsorption percentage was strongly dependent on the solution pH. With decreasing pH, the adsorption percentage increased rapidly from 20% to 90% at pH 7-5.5 and 98% adsorption could be achieved at pH 4.5-3.5. The adsorption behavior followed the Langmuir isotherm with a maximum adsorption amount of 0.605 mg mg-1 and a Langmuir adsorption equilibrium constant of 14.5 mL mg-1 in 0.03 M phosphate buffer at pH 3.5 and 25C. The adsorbed lipase could be desorbed in 0.03 M phosphate buffer at pH 9, and no significant activity loss was observed after adsorption/desorption. According to the investigations on the pH effect, desorption, and activity assay, it was suggested that the lipase used in this work might contain 20% impure or inert protein. In addition, the electrostatic interaction between lipase and PAA was not significantly affected by the temperature at 15-35C, and both the adsorption and desorption of lipase were quite fast due to the absence of internal diffusion resistance. The whole result demonstrated that the PAA-bound magnetic nano-adsorbent could be practically used for the efficient and fast recovery of lipase.
    A novel magnetic nano-adsorbent has been developed by the covalent binding of polyacrylic acid (PAA) on the surface of Fe3O4 nanoparticles and the followed amino-functionalization using diethylenetriamine (DETA) via carbodiimide activation. Transmission electron microscopy image showed that the amino-functionalized Fe3O4 nanoparticles were quite fine with a mean diameter of 11.2±2.8 nm. X-ray diffraction analysis indicated that the binding process did not result in the phase change of Fe3O4. Magnetic measurement revealed they were nearly superparamagnetic with a saturation magnetization of 63.2 emu/g Fe3O4. The binding of DETA on the PAA-coated Fe3O4 nanoparticles was demonstrated by the analyses of Fourier transform infrared (FTIR) spectroscopy and zeta potential. After amino-functionaliztion, the isoelectric point of PAA-coated Fe3O4 nanoparticles shifted from 2.64 to 4.59. The amino-functionalized magnetic nano-adsorbent shows a quite good capability for the rapid and efficient adsorption of metal cations and anions from aqueous solutions via the chelation or ion exchange mechanisms. The studies on the adsorption of Cu(II) and Cr(VI) ions revealed that both obeyed the Langmuir isotherm equation. The maximum adsorption capacities and Langmuir adsorption constants were 12.43 mg/g and 0.06 L/mg for Cu(II) ions and 11.24 mg/g and 0.0165 L/mg for Cr(VI) ions, respectively.
    The use of PAA-coated multiwalled carbon-nanotubes (PAA-MWNTs) composite modified Screen printed carbon electrode (SPE) for the simultaneous determination ascorbic acid (AA)、Norepinephrine (NE) and Uric acid (UA). PAA-MWNTs composite was prepared by mixing of MWNTs powers into PAA aqueous solution under sonication. SPE surface was modified with PAA-MWNTs film by casting. The PAA-MWNTs/SPE is of a high surface area and of affinity adsorption via ion exchange for NE  and hydrogen bonding mechanisms for UA, respectively. The PAA-MWNTs/SPE displayed excellent electrochemical catalytic activity towards AA、NE and UA, the oxidation overpotentials of AA was decreased and the enhanced oxidation peak currents significantly for NE and UA were observed at the PAA-MWNTs/SPE in phosphate buffer solution (0.1 M, pH 7.5). Differential pulse voltammetry was used for the simultaneous dertermination of AA, NE and UA in their ternary mixture. The peak separation between NE and AA, UA and NE was 228mV and 112mV, respectively. Therefore, the voltammetric responses of three compounds can be well resolved on the PAA-MWNTs/SPE, and simultaneous dertermination of these three compounds can be achieved. The calibration curves for AA, NE, UA were obtained in the range of 100~1000μM, 0~10μM, 0~30μM, respectively. The lowest detection limits (S/N=3) were 49.772μM, 0.131μM and 0.458μM for AA, NE and UA, respectively.

Reference ( 273 ) 〈TOP〉

8. Schmid, G.. (2001) Metals. In：Klabunde, K. J., ed. Nanoscale
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10. 廖敏宏 (2002) 磁性奈米載體在生物觸媒和生化分離之應用，國立
連結：
12. 張揚狀 (2005) 表面被覆幾丁聚醣之多功能磁性奈米載體的製備
連結：
13. Chen, D. H.; Chen, C. J. (2002) J. Mater. Chem., 12, 1557.
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15. Michaels, A. M.; Jiang, J.; Brus, L. (2000) J. Phys. Chem. B, 104, 11965.
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Times Cited (17) 〈TOP〉

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