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Colorimetric paper bioassay by horseradish peroxidase for the detection of catechol and resorcinol in aqueous samples.
Preparative Biochemistry & Biotechnology ( IF 2.9 ) Pub Date : 2020-05-07 , DOI: 10.1080/10826068.2020.1760883
Ajinkya Dabhade 1 , Sivaraman Jayaraman 2 , Balasubramanian Paramasivan 1
Affiliation  

Phenolic compounds such as catechol and resorcinol are toxic and persistent pollutants in the aqueous environment. Detection procedures such as chromatographic and spectrophotometric methods are time-consuming and require sophisticated instruments with skilled manpower. Development of a simple, cost effective, portable and disposable paper based biosensor could be a better alternative to the conventional methods. The present study attempted to develop a paper based biosensor by immobilizing horseradish peroxidase enzyme to detect catechol and resorcinol in aqueous samples. Horseradish peroxidase catalyzes the oxidation of phenolic compounds to semiquinones, which on reaction with a chromogen, 3-methyl 2-benzothiazolinone hydrazine (MBTH) gives faint pink to red color depending on the compound and its concentration in the sample is the basis for biosensing application. Different methods of enzyme immobilization on filter paper like physical adsorption, covalent coupling, and polysaccharide entrapment were executed. The performance of the various enzyme immobilization methods was evaluated by analyzing the developed color intensity using ImageJ software. Entrapment technique is the most effective method of immobilizing enzyme on the filter paper that produces the highest color intensity with better stability. The visible limit of detection (LoD) was observed as 0.45 mM (50 mg/L) for catechol and 0.09 mM (10 mg/L) for resorcinol in aqueous samples.



中文翻译:

辣根过氧化物酶比色纸生物测定法,用于检测水性样品中的儿茶酚和间苯二酚。

苯酚和间苯二酚等酚类化合物在水性环境中是有毒且持久的污染物。诸如色谱法和分光光度法之类的检测程序非常耗时,并且需要具有熟练人力的精密仪器。开发简单,成本有效,便携式和一次性纸质生物传感器可能是传统方法的更好替代方案。本研究试图通过固定辣根过氧化物酶来检测水性样品中的儿茶酚和间苯二酚,从而开发出一种基于纸张的生物传感器。辣根过氧化物酶催化酚类化合物氧化为半醌,然后与色原反应,3-甲基2-苯并噻唑啉酮肼(MBTH)取决于化合物,呈淡粉色至红色,其在样品中的浓度是生物传感应用的基础。执行了不同的酶固定在滤纸上的方法,如物理吸附,共价偶联和多糖截留。通过使用ImageJ软件分析显影的颜色强度来评估各种酶固定方法的性能。包埋技术是将酶固定在滤纸上的最有效方法,该方法可产生最高的颜色强度并具有更好的稳定性。在含水样品中,邻苯二酚的可见检测极限(LoD)为0.45 mM(50 mg / L),间苯二酚为0.09 mM(10 mg / L)。执行了不同的酶固定在滤纸上的方法,如物理吸附,共价偶联和多糖截留。通过使用ImageJ软件分析显影的颜色强度来评估各种酶固定方法的性能。包埋技术是将酶固定在滤纸上的最有效方法,该方法可产生最高的颜色强度并具有更好的稳定性。在含水样品中,邻苯二酚的可见检测极限(LoD)为0.45 mM(50 mg / L),间苯二酚为0.09 mM(10 mg / L)。执行了不同的酶固定在滤纸上的方法,如物理吸附,共价偶联和多糖截留。通过使用ImageJ软件分析显影的颜色强度来评估各种酶固定方法的性能。包埋技术是将酶固定在滤纸上的最有效方法,该方法可产生最高的颜色强度并具有更好的稳定性。在含水样品中,邻苯二酚的可见检测极限(LoD)为0.45 mM(50 mg / L),间苯二酚为0.09 mM(10 mg / L)。包埋技术是将酶固定在滤纸上的最有效方法,该方法可产生最高的颜色强度并具有更好的稳定性。在含水样品中,邻苯二酚的可见检测极限(LoD)为0.45 mM(50 mg / L),间苯二酚为0.09 mM(10 mg / L)。包埋技术是将酶固定在滤纸上的最有效方法,该方法可产生最高的颜色强度并具有更好的稳定性。在含水样品中,邻苯二酚的可见检测极限(LoD)为0.45 mM(50 mg / L),间苯二酚为0.09 mM(10 mg / L)。

更新日期:2020-05-07
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