CN109211818A - 一种基于铂纳米粒子比色法同时检测汞离子和银离子的方法 - Google Patents
一种基于铂纳米粒子比色法同时检测汞离子和银离子的方法 Download PDFInfo
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Abstract
在本发明中，制备了粒径均一的PVP包被的铂纳米粒子，并将其应用于同时检测Hg2+和Ag+。所制备的铂纳米粒子可以在过氧化氢(H2O2)存在时，显著催化氧化3,3,5,5‑四甲基联苯胺(TMB)变为蓝色，Hg2+和Ag+能大幅抑制PVP包被的Pt NPs的类过氧化物酶活性，导致吸光度下降。乙二胺四乙酸钠(EDTA)可以成功掩蔽Hg2+，对Ag+的影响可以忽略不计，由此实现选择性检测Ag+。经计算，Ag+检测限为9.75nM，Hg2+检测限为17.75nM。
Description
技术领域
本发明涉及汞离子和银离子的检测。尤其是一种基于铂纳米粒子比色法同时检测汞离子和银离子的方法，属于分析化学和纳米技术领域。
背景技术
汞离子和银离子是在自然环境中广泛存在的两种重金属离子。汞的化合物来源于燃煤工业、火山爆发、金矿开采和垃圾焚烧。汞离子会导致多种疾病，包括神经系统，免疫系统，生殖系统，肾脏，心脏甚至是基因遗传相关的疾病。银的化合物主要来源于摄影、电子和制镜工业的废料，可以引起生物酶失活，引起相关健康问题。银离子可在肝组织中累积，对过度使用含有银盐的药物的病人有负面作用。因此，建立一种快速、灵敏、选择性高的方法来检测汞离子和银离子是十分必要的。
汞离子和银离子经常在同一系统中共存。如今，已有很多方法应用于同时检测汞离子和银离子，比如电感耦合等离子质谱，原子吸收光谱。这些方法虽然非常灵敏，并且可以实现多重检测，但是所需仪器相当笨重，不便携带，因此不可用于现场检测。近几年，荧光法和电化学法在检测汞离子和银离子领域发展迅速。比色法由于可简单直接用裸眼观察而备受关注。近年来，通过把比色技术与其他方法结合，人们制备了多种纳米材料用于检测重金属离子。用吐温20、缩氨酸、巯基丙酸和腺一磷酸修饰金纳米粒子，加入汞离子和银离子可引起这些金纳米离子的聚合，可将此应用于检测高效汞离子和银离子。这类方法虽然简单但是特异性和灵敏度差。利用汞离子和银离子与DNA(脱氧核糖核酸)之间存在T(胸腺嘧啶)-Hg2+-T和C(胞嘧啶)-Ag+-C的化学作用，DNA修饰的金纳米粒子也可用于检测汞离子和银离子。然而，由于DNA非常不稳定且价格昂贵，这一技术在日常检测应用中非常受局限。金属纳米粒子如金纳米粒子、铂纳米粒子、钯纳米粒子具有独特的催化性质且这些性质已应用于生化检测。在这些金属纳米粒子中，铂纳米粒子是最重要的一种催化剂之一。它具有很多模拟酶性质，可催化超氧物歧化、过氧化、氧化。PVP(聚乙烯吡咯烷酮)包被的纳米粒子分散性好，形貌一致。在本方法中，制备了粒径均一的PVP包被的铂纳米粒子，并将其应用于同时检测Hg2+和Ag+。所制备的铂纳米粒子可以在过氧化氢(H2O2)存在时，显著催化氧化3,3,5,5-四甲基联苯胺(TMB)变为蓝色，Hg2+和Ag+能大幅抑制PVP包被的Pt NPs的类过氧化物酶活性，导致吸光度下降。乙二胺四乙酸钠(EDTA)可以成功掩蔽Hg2+，对Ag+的影响可以忽略不计，由此实现选择性检测Ag+。本检测方法灵敏度高、检测仪器简单、无需高技术人员。
发明内容
一种基于铂纳米粒子比色法同时检测汞离子和银离子的方法，包括以下步骤：
(1)合成PVP包被的铂纳米粒子
用H2PtCl6作前驱体，乙二醇既作为溶剂，同时也作为还原剂，PVP做包被剂，采用一锅法快速合成PVP包被的Pt NPs。
(2)建立测定汞离子的标准曲线
配制一定浓度梯度的汞离子标准溶液，加入pH 4.0的磷酸缓冲液中，再加入一定量的铂纳米粒子、TMB和H2O2，然后孵育20分钟，将所得溶液转移至石英比色皿，用紫外分光光度计测定其在652nm处的吸光度。以汞离子浓度为横坐标，所记录反应溶液吸光度与空白组吸光度之差为纵坐标，建立标准曲线。
(3)建立测定银离子的标准曲线
配制一定浓度梯度的银离子标准溶液，加入pH 4.0的磷酸缓冲液中，再加入一定量的铂纳米粒子、TMB和H2O2，然后孵育20分钟，将所得溶液转移至石英比色皿，用紫外分光光度计测定其在652nm处的吸光度。以银离子浓度为横坐标，所记录反应溶液吸光度与空白组吸光度之差为纵坐标，建立标准曲线。
(4)加入不同的金属离子，评价该检测方法的选择性。
(5)自来水样品中加标检测。
附图说明：
图1：合成铂纳米粒子的透射电子显微镜图。
图2：检测Ag+和Hg2+的线性校准曲线。
图3：不同金属离子对PVP包被的Pt NPs的类过氧化物酶活性的影响。
具体实施方式：
实施例1：
首先，在50mL烧瓶中加入4mL乙二醇，加热至110℃。将0.637mL 100mM的H2PtCl6和0.045g PVP分别溶解在2mL乙二醇中。然后，在1.5min内将两种溶液同时滴加到烧瓶中。在110℃下继续回流反应3小时后，获得深棕色均相Pt NPs。所得Pt NPs保存在室温下备用，使用时用超纯水稀释。
实施例2：
将100μL不同浓度Ag+或Hg2+(0，20nM，40nM，60nM，80nM和100nM)加入到1760μL pH4.0PBS溶液中。然后加入20μL 5.4nM PVP包被的Pt NPs，100μL 0.01M TMB和20μL10M H2O2并孵育20分钟。将混合物转移至石英比色皿中，记录UV-vis吸收光谱。
以汞离子浓度为横坐标，所记录反应溶液吸光度与空白组吸光度之差为纵坐标，建立标准曲线。该定量曲线在0-100nM的浓度范围内表现出良好的线性关系。曲线的相关系数分别r＝0.992，基于信噪比(S/N)＝3，该方法测定汞离子的检测限约为17.75nM。
以银离子浓度为横坐标，所记录反应溶液吸光度与空白组吸光度之差为纵坐标，建立标准曲线。该定量曲线在0-100nM的浓度范围内表现出良好的线性关系。曲线的相关系数分别r＝0.999，基于信噪比(S/N)＝3，该方法测定汞离子的检测限约9.75nM。
实施例3：
一定量的100μL0.1μM的金属离子，如Pb2+、Cd2+、Co2+、Fe2+、K+、Mg2+、Na+、Ni2+、Zn2+、Cu2+、Sr2+、Fe3+、Ca2+、Ba2+、Al3+和Cr3+，分别加入100μL1μMHg2+和Ag+，转移到1.84mL pH 4.0的PBS溶液中，然后加入20μL的5.4nM PVP封端的Pt NP，20μL的0.01M TMB和20μL的10M H2O2。孵育20分钟，将所得溶液转移至石英其特征在于记录了紫外-可见吸收光谱波长范围从500nm到800nm。从图2可以看出，只有汞离子和银离子对催化有抑制作用。
实施例4：
为了评估该方法在实际样品中应用，应用加标法检测自来水样品中的Hg2+和Ag+的浓度，首先，取100μL加入一系列浓度梯度Hg2+和Ag+(样品1，15nM和35nM；样品2，35nM和15nM；样品3，25nM和25nM；样品4，25nM和65nM；样品5，65nM和25nM；样品6，45nM和45nM)的自来水，分别加入1760μL pH 4.0PBS溶液中，再加入20μL 5.4nM的PVP包被的Pt NPs、100μL0.01M TMB和20μL 10M H2O2，然后孵育20分钟，将所得溶液转移至石英比色皿，用紫外分光光度计测定其在652nm处的吸光度。然后取含相同浓度的Hg2+和Ag+的加标自来水，加入EDTA(终浓度1mM),孵育20分钟，将所得溶液转移至石英比色皿，用紫外分光光度计测定其在652nm处的吸光度。
将所测得吸光度代入银离子标准曲线方程，由代数关系可得样品中银离子和汞离子的浓度。具体计算方式如下
结果如表1所示。样品中银离子的回收率在99.4–102.8％的范围内，RSD从1.7％到2.6％，汞离子的回收率在100.4–108.1％的范围内，RSD从0.8％到2.2％，证明了该检测方法能检测实际样品中汞离子和银离子的含量。
表1通过PVP包被的Pt NPs检测自来水样品中的Ag+和Hg2+
Claims (7)
1.一种基于铂纳米粒子比色法同时检测汞离子和银离子的方法，其特征在于包括以下步骤：
(1)用H2PtCl6作前驱体，乙二醇既作为溶剂，同时也作为还原剂，PVP做包被剂，采用一锅法快速合成PVP包被的Pt NPs。
(2)配制一定浓度梯度的银离子标准溶液，加入磷酸缓冲液中，再加入一定量的铂纳米粒子、TMB和H2O2，然后孵育20分钟，将所得溶液转移至石英比色皿，用紫外分光光度计测定其在652nm处的吸光度。以银离子浓度为横坐标，所记录反应溶液吸光度与空白组吸光度之差为纵坐标，建立标准曲线。
(3)测定Hg2+和Ag+共存的样品，加入EDTA掩蔽Hg2+，先测定Ag+引起的吸光度减少量，再通过扣除Ag+的影响来计算出Hg2+的浓度。
2.如权利要求书1所述的方法，其特征在于：所述磷酸缓冲液的pH为4.0，体积为1760μL。
3.如权利要求书1所述的方法，其特征在于：所述铂纳米粒子的粒径为5nm浓度为20μL，体积为5.4nM。
4.如权利要求书1所述的方法，其特征在于：所述TMB的浓度为0.01M，体积为100μL。
5.如权利要求书1所述的方法，其特征在于：所述H2O2的浓度为10M，体积为20μL。
6.如权利要求书1所述的方法，其特征在于：EDTA的中浓度为1mM。
7.如权利要求书1所述的方法，其特征在于：Ag+检测限为9.75nM，Hg2+检测限为17.75nM。
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111220610A (zh) * | 2020-02-05 | 2020-06-02 | 江苏大学 | 一种基于铁醇盐纳米酶的As5+比色检测法 |
| CN111239124A (zh) * | 2020-03-06 | 2020-06-05 | 中国药科大学 | 一种利用二硫化铂的过氧化氢比色检测法 |
| CN112881585A (zh) * | 2021-01-12 | 2021-06-01 | 赣南师范大学 | 一种基于纳米酶催化驱动的银源检测方法 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111220610A (zh) * | 2020-02-05 | 2020-06-02 | 江苏大学 | 一种基于铁醇盐纳米酶的As5+比色检测法 |
| CN111239124A (zh) * | 2020-03-06 | 2020-06-05 | 中国药科大学 | 一种利用二硫化铂的过氧化氢比色检测法 |
| CN112881585A (zh) * | 2021-01-12 | 2021-06-01 | 赣南师范大学 | 一种基于纳米酶催化驱动的银源检测方法 |
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