College of Chemistry, Nanchang University, Nanchang, 330031, China.
Jiangxi Province Key Laboratory of Modern Analytical Science, Nanchang University, Nanchang, 330031, China.
Mikrochim Acta. 2021 Apr 6;188(5):155. doi: 10.1007/s00604-021-04799-w.
A colorimetric assay for acid phosphatase (ACP) was constructed that is based on in situ polymerization of aniline catalyzed by gold nanoparticles (AuNPs). Aniline can be polymerized by ammonium persulfate (APS) in acidic condition and form gold-polyaniline core-shell nanoparticles (Au@PANI NPs) in the presence of AuNPs with the assistance of sodium dodecyl sulfate (SDS). AuNPs were also found to accelerate the polymerization process of aniline and thus shorten the reaction time. Upon the introduction of ascorbic acid (AA), the oxidant APS was consumed via the redox reaction. That led to the suppression of the formation of PANI. Consequently, ACP activity can be supervised on the basis of hydrolysis of 2-phospho-L-ascorbic acid trisodium salt (AAP) catalyzed by ACP to release AA. With the increase of ACP activity, the intensity ratio of the absorbance at λ nm (A) and the absorbance at λ nm (A) gradually decreased and the color gradually changed from dark-green to light-green to blue-gray to purple and eventually to pink. This method for ACP determination worked in the range 0.40 to 2.00 U·L. The detection limit is 0.043 U·L. The assay was applied to determine ACP in human serum. The recovery ranged from 81.0 to 104.6%. Relative standard deviation was less than 5%. This suits the request for biological sample analysis. Graphical abstract Schematic presentation of the colorimetric determination of acid phosphatase activity and inhibitor screening based on in situ polymerization of aniline catalyzed by gold nanoparticles. : acid phosphatase (ACP); : gold nanoparticles (AuNPs); : gold-polyaniline core-shell nanoparticles (Au@PANI NPs); ascorbic acid (AA); 2-phospho-L-ascorbic acid trisodium salt (AAP).
一种基于金纳米粒子（AuNPs）催化苯胺原位聚合的酸性磷酸酶（ACP）比色测定法被构建。在酸性条件下，过硫酸铵（APS）可以使苯胺聚合，并在 AuNPs 的存在下形成金-聚邻苯二胺核-壳纳米粒子（Au@PANI NPs），同时借助十二烷基硫酸钠（SDS）的协助。AuNPs 还被发现可以加速苯胺的聚合过程，从而缩短反应时间。当引入抗坏血酸（AA）时，氧化还原反应会消耗氧化剂 APS。这导致 PANI 的形成受到抑制。因此，可以根据 ACP 催化 2-磷酸-L-抗坏血酸三钠盐（AAP）水解产生 AA 来监测 ACP 活性。随着 ACP 活性的增加，λnm 处的吸光度（A）与 λnm 处的吸光度（A）的强度比逐渐降低，颜色逐渐从深绿色变为浅绿色，再变为蓝灰色、紫色，最终变为粉红色。这种 ACP 测定方法的工作范围为 0.40 至 2.00 U·L。检测限为 0.043 U·L。该测定法用于测定人血清中的 ACP。回收率在 81.0%至 104.6%之间。相对标准偏差小于 5%。这符合生物样品分析的要求。 示意图 基于金纳米粒子催化苯胺原位聚合的酸性磷酸酶活性比色测定和抑制剂筛选。：酸性磷酸酶（ACP）；：金纳米粒子（AuNPs）；：金-聚邻苯二胺核-壳纳米粒子（Au@PANI NPs）；抗坏血酸（AA）；2-磷酸-L-抗坏血酸三钠盐（AAP）。
