Laboratório de Eletroforese Capilar, Departamento de Química, Universidade Federal de Santa Catarina, Trindade, CEP 88040-900, Florianópolis, SC, Brazil.
Laboratório de Estudo em Materiais Poliméricos, Departamento de Química, Universidade Federal de Santa Catarina, Trindade, CEP 88040-900, Florianópolis, SC, Brazil.
Anal Chim Acta. 2023 Sep 22;1275:341566. doi: 10.1016/j.aca.2023.341566. Epub 2023 Jun 27.
Enzyme assays can be performed with the capillary electrophoresis technique (CE) in many approaches, such as the immobilized enzyme micro-reactor. Acetylcholinesterase is a promising enzyme to be used when pursuing such a method, as it has already been explored in the proposal of similar methods of miniaturizing enzyme assays. The present work proposes a novel enzyme micro-reactor, based on the anchorage of the enzyme on magnetic nanoparticles of MnFeO with chitosan and glutaraldehyde as the cross-linker in the capillary by means of an arrange of neodymium magnets. The calculated K of the enzyme evaluated by this method was 1.12 mmol L, comparable to other studies in the literature that utilizes immobilized enzymes. Also, IC for neostigmine was assessed in 3 different micro-reactors, with an average of 29.42 ± 3.88 μmol L. In terms of the micro-reactor stability, it was possible to perform at least 25 experiments with assembled micro-reactor. The method was applied to hydroalcoholic extracts of 7 plant species. Plinia cauliflora had the best result, with 42.31 ± 6.81% of enzyme inhibition in a concentration of 100 mg L.
酶测定法可以通过毛细管电泳技术 (CE) 以多种方式进行，例如固定化酶微反应器。乙酰胆碱酯酶是一种很有前途的酶，可用于此类方法的研究，因为它已经在微型化酶测定方法的类似方法的探索中得到了研究。本工作提出了一种新型酶微反应器，该酶微反应器基于将酶锚定在 MnFeO 磁性纳米粒子上，壳聚糖和戊二醛作为交联剂，通过一系列钕磁铁固定在毛细管中。通过该方法评估的酶的计算 K 值为 1.12mmol L，与文献中利用固定化酶的其他研究相当。此外，还评估了 3 种不同微反应器中对新斯的明的 IC，平均为 29.42±3.88μmol L。就微反应器的稳定性而言，组装后的微反应器至少可以进行 25 次实验。该方法已应用于 7 种植物的水醇提取物。Plinia cauliflora 的抑制率最高，在 100mg L 的浓度下，酶抑制率为 42.31±6.81%。
