基于纳米金催化的血清尿酸纸芯片的构建及应用

doi:10.7503/cjcu20160045

高等学校化学学报 ›› 2016, Vol. 37 ›› Issue (5): 829.doi: 10.7503/cjcu20160045

基于纳米金催化的血清尿酸纸芯片的构建及应用

赵甜甜¹, 陈雨晴¹, 张敏²(), 王月荣¹, 章弘扬¹, 胡坪¹()

1. 华东理工大学化学与分子工程学院, 上海市功能性材料化学重点实验室,上海市新药设计重点实验室, 上海 200237
2. 药学院, 上海市新药设计重点实验室, 上海 200237

收稿日期:2016-01-18 出版日期:2016-05-10 发布日期:2016-04-15
作者简介:联系人简介: 胡坪, 女, 博士, 教授, 博士生导师, 主要从事药物分析方面的研究. E-mail:huping@ecust.edu.cn
张敏, 男, 博士, 副教授, 主要从事药物分析方面的研究. E-mail: zhangm@ecust.edu.cn
基金资助:
国家“重大新药创制”科技重大专项课题(批准号: 2013ZX09507005)、国家自然科学基金(批准号: 81573397)和中央高校基本科研业务费探索项目资助

Fabrication of Paper-based Microfluidic Chips and the Application on the Determination of Uric Acid in Serum Based on Gold Nanoparticle-assisted Catalysis^†

ZHAO Tiantian¹, CHEN Yuqing¹, ZHANG Min^2,^*(), WANG Yuerong¹, ZHANG Hongyang¹, HU Ping^1,^*()

1. Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering,School of Pharmacy,East China University of Science and Technology, Shanghai 200237, China
2. Shanghai Key Laboratory of New Drug Design, School of Pharmacy,East China University of Science and Technology, Shanghai 200237, China

Received:2016-01-18 Online:2016-05-10 Published:2016-04-15
Contact: ZHANG Min,HU Ping E-mail:zhangm@ecust.edu.cn;huping@ecust.edu.cn
Supported by:
† Supported by the the National Science and Technology Major Project for Significant New Drugs Development, China(No.2013ZX09507005), the National Natural Science Foundation of China(No.81573397) and the Fundamental Research Funds for the Central Universities, China

摘要/Abstract

摘要：

建立了一种以纸芯片为平台, 利用纳米金(AuNPs)的过氧化物模拟酶特性对血清中尿酸(UA)含量进行快速检测的方法. 在改装的中性笔中灌注疏水性材料溶液, 直接在滤纸上绘制所需要的图案, 经干燥后形成纸芯片. 将纳米金、四甲基联苯胺(TMB)和H₂O₂的混合液依次滴加于纸芯片检测区域, 无色的TMB被氧化成蓝色, 然后将待测样品滴加于蓝色区域, 氧化态TMB被还原为无色, 根据手机相机记录的检测区域灰度值计算试样中尿酸的浓度. 实验优化了纳米金在纸芯片上的用量、反应时间和反应温度等参数, 在最优实验条件下, 检测尿酸的线性范围为10.6~125 mg/L, 检出限为4.64 mg/L, 加样回收率为94.8%~108.5%. 该方法选择性良好, 可用于测定血清样品中尿酸的含量.

关键词: 纸芯片, 纳米金, 尿酸

Abstract:

A simple and fast method for the determiantion of uric acid(UA) in serum was developed using paper-based microfluidic chips and gold nanoparticles(AuNPs) as peroxidase mimic. The paper-based microfluidic chips were fabricated by directly drawing on filter paper with a modified gel pen filled with hydrophobic solution and then dried. When the AuNPs and the mixed solution of TMB and H₂O₂ were droped on the detection zone of paper-based microfluidic chips, the colorless TMB was oxidated to blue substance. Then the sample was added onto the blue zone, the blue-oxidated TMB was reducted to colorless. Finally the picture of detection zone could be captured by a mobile phone camera and the concentration of UA could be caclulated according to the gray value of the detection zone. Parameters influencing the UA determination were investigated, including volume of AuNPs, reaction time and temperature. Under the optimized conditions, the detection limit of UA could be reached to 4.64 mg/L, the linear range was from 10.6 mg/L to 125 mg/L and the recovery was from 94.8% to 108.5%. The method could be applied to determining uric acid in serum.

Key words: Paper-based microfluidic chip, Gold nanoparticles, Uric acid

中图分类号:

O657

TrendMD:

赵甜甜, 陈雨晴, 张敏, 王月荣, 章弘扬, 胡坪. 基于纳米金催化的血清尿酸纸芯片的构建及应用. 高等学校化学学报, 2016, 37(5): 829.

ZHAO Tiantian, CHEN Yuqing, ZHANG Min, WANG Yuerong, ZHANG Hongyang, HU Ping. Fabrication of Paper-based Microfluidic Chips and the Application on the Determination of Uric Acid in Serum Based on Gold Nanoparticle-assisted Catalysis^†. Chem. J. Chinese Universities, 2016, 37(5): 829.

图/表 9

Fig.1 Front views(A, C) and back views(B, D) of two paper-based microfluidic chips fabricated by the modified pen with a nib width of 0.3 mm(A, B) and 0.4 mm(C, D)

Fig.2 Influence of without(A) and with(B) the addition of vacuum silicone on the resolution of paper-based microfluidic chips

Fig.3 Effects of (+)AuNPs volume on the TMB oxidation(A) and detection of UA(n=3)(B)

Fig.4 Imaging time for UA detection(n=3)a. TMB substrate; b. UA(25 mg/L); c. UA(50 mg/L); d. UA(75 mg/L).

Fig.5 Effect of temperature on the detection of UA(n=3)

Fig.6 Effects of different substances existing in serum on the UA detectiona. Blank; b.1.0 mg/L of inosine; c. 10 mg/L of creatinine; d. 7.0 mg/L of hypoxathine; e. 0.4 mg/L of xanthine; f. 1000 mg/L of glucose; g. 200 mg/L of vitamin C; h—l. 10 mg/L of NO2-, Ca2+, Na+, Cl- and Fe3+; m. 50 mg/L of UA; n. the mixture of all above(n=3).

Table 1 Recovery of UA in serum samples spiked with different concentration of UA standard solution(n=3)

c(UA)/(mg·L^-1)	Add/(mg·L^-1)	Found/(mg·L^-1)	Recovery(%)
39.6	20	58.8±4.7	96.0
39.6	40	77.5±5.1	94.8
39.6	60	104.7±7.7	108.5

Fig.7 Results of colorimetric assay of UA on paper-based microfluidic chips(A) and the calibration curve(B)#3 and #5 are samples number in Table 2.

Table 2 Results of determination of UA by paper-based microfluidic chips and HPLC(n=3)

No.	Paper-based chips	HPLC	δ(%)
#1	36.1	39.6	-8.9
#2	41.1	46.4	-11.4
#3	64.4	60.3	6.8
#4	43.4	40.3	7.7
#5	102.1	105.0	-2.8
#6	102.0	98.4	3.7
#7	51.6	57.6	-10.4
#8	96.0	100.0	-4.0

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基于纳米金催化的血清尿酸纸芯片的构建及应用

Fabrication of Paper-based Microfluidic Chips and the Application on the Determination of Uric Acid in Serum Based on Gold Nanoparticle-assisted Catalysis^†

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基于纳米金催化的血清尿酸纸芯片的构建及应用

Fabrication of Paper-based Microfluidic Chips and the Application on the Determination of Uric Acid in Serum Based on Gold Nanoparticle-assisted Catalysis†

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Fabrication of Paper-based Microfluidic Chips and the Application on the Determination of Uric Acid in Serum Based on Gold Nanoparticle-assisted Catalysis^†