CN111122531B - 一种可视化识别多种有机溶剂的比率荧光传感方法 - Google Patents

一种可视化识别多种有机溶剂的比率荧光传感方法 Download PDF

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CN111122531B
CN111122531B CN202010010250.9A CN202010010250A CN111122531B CN 111122531 B CN111122531 B CN 111122531B CN 202010010250 A CN202010010250 A CN 202010010250A CN 111122531 B CN111122531 B CN 111122531B
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杨伟强
张桂云
李飞明
倪建聪
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Minnan Normal University
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Abstract

本发明公开了一种可视化识别多种有机溶剂的比率荧光传感方法，包括以下步骤：(1)碳点的制备；(2)金簇的制备；(3)荧光探针比例的调制和液相检测；(4)荧光试纸的制备及可视化检测。本发明以荧光碳点和金簇作为双荧光探针来构建多色彩的比率荧光的传感模式，由于不同类型的有机溶剂对两种探针的荧光强度改变不同，试纸的荧光可呈现差异较大的不同颜色，易于为肉眼所区分，进而实现甲醇、乙腈、丙酮、乙酸乙酯、四氢呋喃五种有机溶剂的可视化识别检测。

Description

一种可视化识别多种有机溶剂的比率荧光传感方法
【技术领域】
本发明属于比率荧光传感技术领域，具体涉及一种可视化识别多种有机溶剂的比率荧光传感方法。
【背景技术】
无论在工业生产还是实验室中，有机溶剂的使用都不可或缺。然而这些常见的有机溶剂通常都是无色透明的液体，仅用感官难以进行有效的区分，一旦失去标签则造成处置的困难和资源的浪费。若鉴别其成分还需气相色谱等专用的仪器设备，费时费力，陈本高昂。为此，需要发展一种便携式识别常见有机溶剂的检测技术。
荧光光谱法具有灵敏度高、响应快速、简单经济的特点，在便携式检测中往往只需要特定的荧光探针以及合适的激发光源(如365nm的荧光笔)。相较于甄别单色荧光强度的变化，正常人眼对荧光颜色的敏感程度大大优于对荧光强度的分辨能力(人的肉眼可分辨大约一千万种颜色)。以荧光颜色变化作为检测依据，更易于实现可视化检测。比率荧光分析方法通过引入两个荧光探针来响应目标物，其荧光发射峰通常处于不同的波段，即具有不同的荧光颜色。当两种荧光探针的响应强度发生变化时，二者的混合荧光颜色也随之改变。根据荧光颜色的不同，可以用肉眼来识别性检测目标物，适用于有限条件下的便捷式检测。
碳点和金簇是近年来发展较快的荧光纳米材料，具有制备方法简单、光学稳定性好、成本经济、环境友好等优势。研究发现碳点的荧光发射强度和发射波长也受溶剂的影响。Huang等以鞣酸为碳源合成一种疏水性的碳点，其在纯水中的荧光强度很弱；而四氢呋喃的加入可使该溶液的荧光强度增强，据此可以检测水中四氢呋喃的含量(Chem.Commun.,2016,52,2063--2066)。Chao等以邻苯二胺为碳源水热合成碳点，其在四氢呋喃、丙酮、二甲基甲酰胺、甲醇、乙二醇几种溶剂中的最大荧光波长分别为512、520、525、550、560nm，呈现出一定的荧光颜色区分度(J.Mater.Chem.C,2018,6,7527--7532)。然而，这类方法均为单荧光探针模式，存在分析对象单一、可视化区分度不足等问题，无法满足多种有机溶剂的快速识别检测需要。
【发明内容】
本发明提供一种可视化识别多种有机溶剂的比率荧光传感方法，以解决如何便携式识别常见几种有机溶剂的问题。
为了解决以上技术问题，本发明采用以下技术方案：
一种可视化识别多种有机溶剂的比率荧光传感方法，包括以下步骤：
(1)碳点的制备；
(2)金簇的制备；
(3)荧光探针比例的调制和液相检测；
(4)荧光试纸的制备及可视化检测。
进一步地，步骤(1)中所述碳点的制备，包括以下步骤：称取柠檬酸和聚乙烯亚胺，溶于水中并振荡混匀，于水热反应釜中反应；所得溶液经透析袋透析，冷冻干燥并用水重新分散，得到碳点储备液。
进一步地，步骤(2)中所述金簇的制备，包括以下步骤：配制氯金酸和牛血清白蛋白溶液，将氯金酸溶液和牛血清白蛋白溶液在磁力搅拌器上搅拌混匀，逐滴加入NaOH溶液，继续搅拌反应，得到金簇储备液。
进一步地，步骤(3)中所述荧光探针比例的调制和液相检测，包括以下步骤：将碳点和金簇以质量浓度之比分别为1:4、1:5、1:6、1:7、1:8的比例配制成双荧光探针溶液；取50μL该溶液分别加入到450μL的甲醇、乙腈、丙酮、乙酸乙酯、四氢呋喃纯溶剂中混匀；用荧光光谱仪测试，采集激发波长为365nm时荧光发射在400-700nm之间的变化光谱图；以445nm和645nm处的荧光峰强度之比作为判定依据，选择碳点和金簇以1:6的比例配制成的双荧光探针溶液进行后续实验。
进一步地，步骤步骤(4)中所述荧光试纸的制备及可视化检测，包括以下步骤：将滤纸用打孔器裁成直径为5-6mm的圆形，浸泡在碳点/金簇为1:6的双荧光探针溶液，浸泡时间为1h，取出自然晾干，保存备用；取0.5mL甲醇、乙腈、丙酮、乙酸乙酯、四氢呋喃分别滴于试纸上，置于365nm紫外灯下，观察其荧光颜色并用相机拍摄照片作为标准比色卡(含RGB值)；对于未知有机溶剂，取0.5mL待测样品滴加于试纸上，置于365nm紫外灯下，观察其荧光颜色并与标准比色卡进行对比，判断该有机溶剂是哪一种。
本发明的技术原理：采用双荧光探针的模式。碳点和金簇的荧光发射峰分别位于445nm和645nm，且不随溶剂的改变而发生波长移动。其对不同有机溶剂的响应体现在荧光强度的升高或降低，即二者的强度比值变化，并表现为混合荧光颜色的变化。由于可呈现出的荧光颜色更多，图像可视化的区分度更高(RGB值的差异性)，因而可区分的有机溶剂种类也可以更多。本发明以荧光碳点和金簇作为双荧光探针来构建多色彩的比率荧光的传感模式，由于不同有机溶剂对两种荧光探针的荧光强度改变不同，试纸的荧光可呈现不同的颜色，易于为肉眼所区分和识别，进而实现甲醇、乙腈、丙酮、乙酸乙酯、四氢呋喃的可视化识别。
本发明具有以下有益效果：
(1)试验中碳点和金簇的最佳质量浓度比例为1:6，可制备荧光试纸。且其对不同类型的有机溶剂可呈现肉眼易于分辨的荧光色彩，RGB值差异性较大。
(2)在单荧光探针模式中，荧光纳米材料对不同有机溶剂所呈现的强度变化或者波长移动的响应微弱，并不易于为肉眼所区分，需要相应的分析仪器。而本发明采用了碳点和金簇两种荧光探针，其荧光发射波长差大，可呈现的荧光颜色更多，可视化区分度更高。
(3)目前报道的比率荧光方法都是用于分析某一种有机溶剂中水分的含量，均没有识别多种有机溶剂的能力。而本发明可用于多种有机溶剂(甲醇、乙腈、丙酮、乙酸乙酯、四氢呋喃)的识别性检测。
【附图说明】
图1为双荧光探针溶液(a)对甲醇(b)、乙腈(c)、丙酮(d)、乙酸乙酯(e)、四氢呋喃(f)的荧光响应光谱图，插图为分别的荧光颜色照片图及RGB值。
【具体实施方式】
为便于更好地理解本发明，通过以下实例加以说明，这些实例属于本发明的保护范围，但不限制本发明的保护范围。
一种可视化识别多种有机溶剂的比率荧光传感方法，包括以下步骤：
(1)碳点(CDs)的制备。称取0.5g柠檬酸和0.5g聚乙烯亚胺，溶于20mL超纯水中并振荡混匀，于水热反应釜中，在180℃下反应6h，所得溶液经透析袋(分子量1000)透析，冷冻干燥并用超纯水重新分散，得到1mg/mL的碳点储备液。
(2)金簇(AuNCs@BSA)的制备。配制10mM的氯金酸，50mg/mL的BSA(牛血清白蛋白)溶液各5mL；将氯金酸溶液和BSA溶液在37℃磁力搅拌器上搅拌混匀20min，逐滴加入1MNaOH溶液0.5mL，继续搅拌反应12h，得到的1mg/mL金簇储备液。
(3)荧光探针比例的调制和液相检测。将碳点和金簇以质量浓度之比分别为1:4、1:5、1:6、1:7、1:8的比例配制成双荧光探针溶液；取50μL该溶液分别加入到450μL的甲醇、乙腈、丙酮、乙酸乙酯、四氢呋喃纯溶剂中混匀；用荧光光谱仪测试，采集激发波长为365nm时荧光发射在400-700nm之间的变化光谱图；以445nm和645nm处的荧光峰强度之比作为判定依据，优选差值最大的比例进行后续实验(如1:6)。
(4)荧光试纸的制备及可视化检测。将滤纸用打孔器裁成直径为5-6mm的圆形，浸泡在碳点/金簇为1:6的双荧光探针溶液，浸泡时间为1h，取出自然晾干(24h以上)，保存备用；取0.5mL甲醇、乙腈、丙酮、乙酸乙酯、四氢呋喃分别滴于试纸上，置于365nm紫外灯下，观察其荧光颜色并用相机拍摄照片作为标准比色卡，其RGB值分别如图1所示；对于未知有机溶剂，取0.5mL待测样品滴加于试纸上，置于365nm紫外灯下，观察其荧光颜色并与标准比色卡进行对比，判断该有机溶剂是哪一种。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。

Claims (1)

1.一种可视化识别多种有机溶剂的比率荧光传感方法，其特征在于，包括以下步骤：
(1)碳点的制备；
(2)金簇的制备；
(3)荧光探针比例的调制和液相检测；
(4)荧光试纸的制备及可视化检测；
步骤(1)中所述碳点的制备，包括以下步骤：称取柠檬酸和聚乙烯亚胺，溶于水中并振荡混匀，于水热反应釜中反应；所得溶液经透析袋透析，冷冻干燥并用水重新分散，得到碳点储备液；
步骤(2)中所述金簇的制备，包括以下步骤：配制氯金酸和牛血清白蛋白溶液，将氯金酸溶液和牛血清白蛋白溶液在磁力搅拌器上搅拌混匀，逐滴加入NaOH溶液，继续搅拌反应，得到金簇储备液；
步骤(3)中所述荧光探针比例的调制和液相检测，包括以下步骤：将碳点和金簇以质量浓度之比分别为1:4、1:5、1:6、1:7、1:8的比例配制成双荧光探针溶液；取50μL该溶液分别加入到450μL的甲醇、乙腈、丙酮、乙酸乙酯、四氢呋喃纯溶剂中混匀；用荧光光谱仪测试，采集激发波长为365nm时荧光发射在400-700nm之间的变化光谱图；以445nm和645nm处的荧光峰强度之比作为判定依据，选择碳点和金簇以1:6的比例配制成的双荧光探针溶液进行后续实验；
步骤(4)中所述荧光试纸的制备及可视化检测，包括以下步骤：将滤纸用打孔器裁成直径为5-6mm的圆形，浸泡在碳点/金簇为1:6的双荧光探针溶液，浸泡时间为1h，取出自然晾干，保存备用；取0.5mL甲醇、乙腈、丙酮、乙酸乙酯、四氢呋喃分别滴于试纸上，置于365nm紫外灯下，观察其荧光颜色并用相机拍摄照片作为标准比色卡，其中含RGB值；对于未知有机溶剂，取0.5mL待测样品滴加于试纸上，置于365nm紫外灯下，观察其荧光颜色并与标准比色卡进行对比，判断该有机溶剂是哪一种。
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