Colorimetric determination of acid phosphatase activity and inhibitor screening based on in situ polymerization of aniline catalyzed by gold nanoparticles
Colorimetric determination of acid phosphatase activity and inhibitor screening based on in situ polymerization of aniline catalyzed by gold nanoparticles
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纳米金催化苯胺原位聚合比色法测定酸性磷酸酶活性及抑制剂筛选
DOI：
10.1007/s00604-021-04799-w
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发表时间：
2021-05-01
影响因子：
5.7
通讯作者：
Ma, Lihua
中科院分区：
文献类型：
--
作者：
Liu, Hui;Huang, Pengcheng;Ma, Lihua
基于纳米金催化苯胺的原位聚合，建立了酸性磷酸酶(ACP)的比色分析方法。在十二烷基硫酸钠(十二烷基硫酸钠)的辅助下，过硫酸铵(APS)可以在酸性条件下使苯胺聚合，形成金-聚苯胺核壳纳米粒子(Au@PANI NPs)。Au纳米粒子还能加速苯胺的聚合过程，从而缩短反应时间。在引入抗坏血酸(AA)后，氧化剂APS通过氧化还原反应消耗。这导致了对PANI形成的抑制。因此，ACP的活性可以根据ACP催化2-磷酸-L-抗坏血酸三钠(AAP)的水解释放AA来监测。随着酸性磷酸酶活力的增加，λ705 nm处的吸光度(A705)和λ530 nm处的吸光度(A530)的强度比逐渐减小，颜色由深绿色到浅绿色再到蓝灰色到紫色，最后变成粉红色。方法的线性范围为0.043~2.00U·L−1，检出限为0.40U·L−1，用于人血清中酸性磷酸酶的测定。回收率在81.0%~104.6%之间。相对标准偏差小于5%。这适合于生物样品分析的要求。图形摘要基于纳米金催化苯胺原位聚合的酸性磷酸酶活性比色法测定和抑制剂筛选的示意图。酸性磷酸酶(ACP)；金纳米粒子(AuNPs)；金-聚苯胺核壳纳米粒子(Au@PANI NPs)；抗坏血酸(AA)；2-磷酸-L-抗坏血酸三钠盐(AAP)。
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 atλ705nm (A705) and the absorbance atλ530nm (A530) 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−1. The detection limit is 0.043 U·L−1. 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 abstractSchematic 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).