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壳聚糖不同分子量作为葡萄糖生物传感器固定基质的研究。

Study on different molecular weights of chitosan as an immobilization matrix for a glucose biosensor.

机构信息

School of Pharmaceutical Sciences, Universiti Sains Malaysia, Minden, Penang, Malaysia.

出版信息

PLoS One. 2013 Aug 5;8(8):e70597. doi: 10.1371/journal.pone.0070597. Print 2013.

Abstract

Two chitosan samples (medium molecular weight (MMCHI) and low molecular weight (LMCHI)) were investigated as an enzyme immobilization matrix for the fabrication of a glucose biosensor. Chitosan membranes prepared from acetic acid were flexible, transparent, smooth and quick-drying. The FTIR spectra showed the existence of intermolecular interactions between chitosan and glucose oxidase (GOD). Higher catalytic activities were observed on for GOD-MMCHI than GOD-LMCHI and for those crosslinked with glutaraldehyde than using the adsorption technique. Enzyme loading greater than 0.6 mg decreased the activity. Under optimum conditions (pH 6.0, 35°C and applied potential of 0.6 V) response times of 85 s and 65 s were observed for medium molecular weight chitosan glucose biosensor (GOD-MMCHI/PT) and low molecular weight chitosan glucose biosensor (GOD-LMCHI/PT), respectively. The apparent Michaelis-Menten constant ([Formula: see text]) was found to be 12.737 mM for GOD-MMCHI/PT and 17.692 mM for GOD-LMCHI/PT. This indicated that GOD-MMCHI/PT had greater affinity for the enzyme. Moreover, GOD-MMCHI/PT showed higher sensitivity (52.3666 nA/mM glucose) when compared with GOD-LMCHI/PT (9.8579 nA/mM glucose) at S/N>3. Better repeatability and reproducibility were achieved with GOD-MMCHI/PT than GOD-LMCHI/PT regarding glucose measurement. GOD-MMCHI/PT was found to give the highest enzymatic activity among the electrodes under investigation. The extent of interference encountered by GOD-MMCHI/PT and GOD-LMCHI/PT was not significantly different. Although the Nafion coated biosensor significantly reduced the signal due to the interferents under study, it also significantly reduced the response to glucose. The performance of the biosensors in the determination of glucose in rat serum was evaluated. Comparatively better accuracy and recovery results were obtained for GOD-MMCHI/PT. Hence, GOD-MMCHI/PT showed a better performance when compared with GOD-LMCHI/PT. In conclusion, chitosan membranes shave the potential to be a suitable matrix for the development of glucose biosensors.

摘要

两种壳聚糖样品(中分子量(MMCHI)和低分子量(LMCHI))被研究作为酶固定化基质,用于制备葡萄糖生物传感器。从乙酸制备的壳聚糖膜具有柔韧性、透明性、光滑性和快速干燥性。FTIR 光谱表明壳聚糖和葡萄糖氧化酶(GOD)之间存在分子间相互作用。与使用吸附技术相比,GOD-MMCHI 的催化活性更高,而 GOD-LMCHI 则与戊二醛交联的催化活性更高。酶负载大于 0.6mg 会降低活性。在最佳条件下(pH6.0、35°C 和施加电位 0.6V),观察到中分子量壳聚糖葡萄糖生物传感器(GOD-MMCHI/PT)和低分子量壳聚糖葡萄糖生物传感器(GOD-LMCHI/PT)的响应时间分别为 85s 和 65s。发现 GOD-MMCHI/PT 的表观米氏常数([Formula: see text])为 12.737mM,而 GOD-LMCHI/PT 的表观米氏常数为 17.692mM。这表明 GOD-MMCHI/PT 对酶具有更大的亲和力。此外,与 GOD-LMCHI/PT(9.8579nA/mM 葡萄糖)相比,GOD-MMCHI/PT 具有更高的灵敏度(52.3666nA/mM 葡萄糖),当 S/N>3 时。与 GOD-LMCHI/PT 相比,GOD-MMCHI/PT 在葡萄糖测量方面具有更好的重复性和再现性。在研究的电极中,GOD-MMCHI/PT 表现出最高的酶活性。GOD-MMCHI/PT 和 GOD-LMCHI/PT 遇到的干扰程度没有显著差异。尽管涂有 Nafion 的生物传感器由于研究中的干扰物而显著降低了信号,但它也显著降低了对葡萄糖的响应。评估了生物传感器在大鼠血清中葡萄糖测定中的性能。与 GOD-LMCHI/PT 相比,GOD-MMCHI/PT 获得了更好的准确性和回收率结果。因此,与 GOD-LMCHI/PT 相比,GOD-MMCHI/PT 表现出更好的性能。总之,壳聚糖膜具有作为葡萄糖生物传感器开发的合适基质的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac52/3734260/ea248b215da8/pone.0070597.g001.jpg

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