纸芯片显色法用于L-半胱氨酸的快速测定
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摘要
该文以5,5'-二硫硝基苯甲酸(DTNB)为含巯基化合物的衍生化试剂,建立了其纸芯片快速检测新方法。显色剂DTNB和样品在显色区相遇并显色,用手机拍照记录,采用Photoshop软件分析显色强度,进行比色检测,并以L-半胱氨酸(L-Cys)为例,优化得到纸芯片检测L-Cys的最佳显色条件。结果显示,L-Cys浓度在0.01～0.1 mmol/L范围内与显色强度呈良好的线性关系(r=0.999 8),检出限达0.001 mmol/L。多种氨基酸和干扰离子对L-Cys在纸芯片上的测定干扰不超过±5%。采用该方法测得牛血清样品中L-Cys含量为0.013 mmol/L,加标回收率为99.1%～103%,该方法检测L-Cys具有操作简单、耗样量少、检测快速、重现性好等优点。
A new colorimetric method based on paper chip was established for the rapid detection of L-Cysteine(L-Cys) using 5,5ʼ-dithiobis-(2-nitrobenzoic acid)(DTNB) as derivatizing agent.A normal phone was used to record the color intensity.The color intensity was analyzed by Photoshop software and used to calculate the concentration of thiol-contained molecules in samples.The optimum coloration conditions for the detection were investigated.Results showed that there was a good linear relationship between color intensity and L-Cys concentration in range of 0.01-0.1 mmol/L with the linear correlation coefficient of 0.999 8 and the detection limit of 0.001 mmol/L.In the range of ±5%,the interference of several amino acids and ions on the determination of L-Cyse was small.The method was applied in the detection of L-Cys content in bovine serum sample with a result of 0.013 mmol/L and a spiked recovery of 99.1%-103%.Results showed that this method has high selectivity and reproducibility.The colorimetric method based on paper-based microfluidic chip has several advantages such as simplicity,low-reagent consumption,rapidness and repeatability.
A new colorimetric method based on paper chip was established for the rapid detection of L-Cysteine(L-Cys) using 5,5ʼ-dithiobis-(2-nitrobenzoic acid)(DTNB) as derivatizing agent.A normal phone was used to record the color intensity.The color intensity was analyzed by Photoshop software and used to calculate the concentration of thiol-contained molecules in samples.The optimum coloration conditions for the detection were investigated.Results showed that there was a good linear relationship between color intensity and L-Cys concentration in range of 0.01-0.1 mmol/L with the linear correlation coefficient of 0.999 8 and the detection limit of 0.001 mmol/L.In the range of ±5%,the interference of several amino acids and ions on the determination of L-Cyse was small.The method was applied in the detection of L-Cys content in bovine serum sample with a result of 0.013 mmol/L and a spiked recovery of 99.1%-103%.Results showed that this method has high selectivity and reproducibility.The colorimetric method based on paper-based microfluidic chip has several advantages such as simplicity,low-reagent consumption,rapidness and repeatability.
引文
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[27] Arena A,Donato N,Saitta G,Bonavita A,Rizzo G,Neri G.Sens.Actuators B,2010,145(1):488-494.