CN113980672B - 检测铜离子的荧光探针及其制备方法、检测方法和应用 - Google Patents
检测铜离子的荧光探针及其制备方法、检测方法和应用 Download PDFInfo
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Abstract
本发明公开了一种检测铜离子的荧光探针及其制备方法、检测方法和应用。该荧光探针通过将聚乙烯亚胺和1,1,2,2‑四‑(4‑羧基‑(1,1‑联苯))乙烯在超声条件下混匀形成。本发明利用聚乙烯亚胺的性质,一方面作为增强1,1,2,2‑四‑(4‑羧基‑(1,1‑联苯))乙烯荧光的络合剂,使其荧光量子产率由1.13%提升到39.66%;同时另外一方面也作为特异性检测Cu2+的螯合剂。此外,本发明所合成的荧光探针制备条件温和,步骤简便,在水中溶解性好,能适合水溶液检测的需求,且光稳定性强,荧光受环境影响小,目视荧光变化效果好,且对Cu2+具有较高选择性和灵敏度,在环境监测系统中具有广阔的应用前景。
Description
技术领域
本发明属于化学分析和环境安全检测领域,具体涉及一种检测铜离子的荧光探针及其制备方法、检测方法和应用。
背景技术
重金属离子的毒性和存在形式是危害人们身体健康的主要问题,许多国家都遭受重金属离子的污染。铜离子是一种重金属离子,常常在废水中通过食物链而进入餐桌。虽然铜离子是人体必需的微量元素,但是过多的摄取铜离子,会增加肝脏和肾脏的负担,从而导致威尔逊病、阿尔茨海默病等疾病。
目前对于废水和食品中的铜离子的特异性检测大多采用的还是原子吸收光谱法和比色法,这些方法灵敏度低、操作复杂且费用较高。荧光分子探针技术作为新型检测技术得到了更多的关注,但普通荧光分子探针在水溶液中对于金属离子的选择性较差,且荧光量子产率不足。
发明内容
本发明的目的是解决现有技术的不足,提供一种检测铜离子的荧光探针,具体采用以下的技术方案:
一种检测铜离子的荧光探针的制备方法,包括以下步骤:
将聚乙烯亚胺和1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯在超声条件下混匀,形成所述荧光探针。
优选地,聚乙烯亚胺和1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯的质量比为10:1。聚乙烯亚胺与1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯可以通过聚乙烯亚胺的氨基与1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯的羧基发生共价结合,先在磁力搅拌的条件下,将以上两种物质按10:1的质量比在磁力搅拌下混合均匀,并在超声的条件下,复合物会形成直径在10nm以下的荧光材料,将复合物透析3天,除去未反应的1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯,放置在4℃备用。
根据上述方法得到的荧光探针可以应用在废水检测和食品检测中,其对Cu2+浓度的检测范围在10-250nM,检测限为1.1nM。基于聚乙烯亚胺对Cu2+的螯合作用,可以特异的捕获Cu2+而达到检测的目的。上述荧光探针可用于水中Cu2+的检测分析(检测食品时可取样后进行提取溶解);其具体检测方法包括以下步骤:将荧光探针溶于水加入待检测液中,根据荧光强度和标准曲线得到待检测液中铜离子的浓度;所述标准曲线根据以下步骤制得:配制5-500nM的一系列铜离子浓度标准溶液和0nM铜离子浓度的空白溶液,将荧光探针加入到上述各个标准溶液和空白溶液中,混匀后测定各个标准溶液和空白溶液的荧光强度,以各个标准溶液的荧光强度与空白溶液的荧光强度的差值为纵坐标,铜离子浓度为横坐标制成标准曲线。
本发明的有益效果为:
(1)本发明巧妙的利用聚乙烯亚胺的性质,一方面作为增强1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯荧光的络合剂,使其荧光量子产率由1.13%提升到39.66%;同时另外一方面也作为特异性检测Cu2+的螯合剂。
(2)本发明所合成的荧光探针制备条件温和,步骤简便,该荧光探针中具有多个氨基,在水中溶解性好,能适合水溶液检测的需求,可用于检测水溶液中的Cu2+,同时,本发明的荧光探针光稳定性强,荧光受环境影响小,目视荧光变化效果好,且对Cu2+具有较高选择性和灵敏度,在环境监测系统中具有广阔的应用前景。
附图说明
图1所示为荧光探针的合成过程图;
图2所示为荧光探针的紫外吸收图谱和荧光发射光谱图;
图3所示为不同Cu2+浓度下荧光探针和1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯的发射光谱图;
图4所示为不同Cu2+浓度下荧光探针的荧光响应图谱和标准曲线;
图5所示为不同金属离子条件下荧光探针的荧光信号强度。
具体实施方式
以下将结合实施例和附图对本发明的构思及产生的技术效果进行清楚、完整的描述,以充分地理解本发明的目的、方案和效果。
实施例1:荧光探针的制备
一、配制1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯溶液
称取40mg的1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯溶解在40mL0.01M NaOH中,反复超声涡旋直至完全溶解,配制成1mg/mL的溶液。
二、配制聚乙烯亚胺溶液
称取500mg的聚乙烯亚胺溶解在5mL超纯水中,反复涡旋直至完全溶解,配制成100mg/mL的聚乙烯亚胺浓度,并用超纯水稀释至一系列浓度。
三、合成荧光探针
其反应过程如图1所示,具体为:向0.9mL超纯水中加入1mL的1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯溶液,混合均匀后,向上述溶液中加入0.1mL的聚乙烯亚胺溶液,在室温下搅拌1h,将体系超声30min后,4℃透析3天,放置4℃保存备用。图2所示为室温下,本发明的荧光探针的紫外吸收图谱和荧光发射光谱,图中纵坐标为吸光度和荧光强度,横坐标为波长,其中左插图(实物照片)为可见光下荧光探针的颜色,呈现浅黄色;右插图(实物照片)为紫外灯下荧光探针的荧光,荧光颜色为深绿色。
选取硫酸奎宁作为量子产率标准物质,通过计算其量子产率为55%。然后将硫酸奎宁、1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯和荧光探针分别稀释一系列浓度,同时测定不同浓度的吸光度和荧光光谱,通过荧光光谱在软件中计算出相应的荧光积分面积,选取吸光度为横坐标,荧光积分面积作为纵坐标,拟合出标准曲线,并通过下面公式计算得到各自量子产率。
是待测物质的量子产率,/>是硫酸奎宁的量子产率,kx、kQ分别是待测物质和硫酸奎宁拟合出来的标准曲线的斜率,nx、nQ分别是待测物质和硫酸奎宁的折射率(硫酸奎宁在0.01M H2SO4条件下折射率为1.33,待测物在超纯水条件下折射率为1.33)。
结果如表1所示,由表1可知,荧光探针中1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯荧光的络合剂的荧光量子产率由1.13%提升到了39.66%。
表1 1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯和荧光探针的相对量子产率
实施例2:测定荧光探针及1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯在不同Cu2+浓度中的荧光发射光谱
配制三个浓度的Cu2+溶液(0、100和1000nM),将实施例1配制的20μL荧光探针和1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯溶液分别加入300μL三种Cu2+浓度的溶液,激发光为360nm,发射在400-800nm。结果如图3所示,室温下,在不同Cu2+浓度下,左图为荧光探针的发射光谱,可以看到在不同浓度下的Cu2+浓度,荧光探针的荧光强度有明显的响应,插图(实物照片)也可以明显看出随着Cu2+浓度的增加,酶标孔中的荧光出现显著淬灭;右图为1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯的发射光谱,在不同浓度的Cu2+浓度下,荧光强度几乎不受Cu2+浓度的影响,插图(实物照片)也表明Cu2+浓度不会影响荧光信号;证明只有聚乙烯亚胺存在才能对Cu2+响应。
实施例3:应用荧光探针绘制标准曲线用于测定Cu2+
配制好一系列浓度的Cu2+溶液(0、5、10、25、50、100、250和500nM),取实施例1配制的20μL荧光探针加入300μL不同Cu2+浓度,通过测定荧光信号,制得荧光响应图谱(如图4左图所示)和标准曲线(如图4右图所示),图中纵坐标为荧光强度的差值,横坐标为Cu2+浓度,插图为实物图;可以发现,随着Cu2+浓度的增加,酶标孔中荧光信号逐渐减弱。
实施例4:荧光探针对Cu2+的特异性识别
配制好1μM浓度的Cu2+溶液,以及10μM浓度的Mn2+、Ca2+、K+、NH+ 4、Ag+、Na+、Cd2+、Mg2+、Zn2+、Fe3+溶液。取实施例1配制的20μL荧光探针加入300μL不同离子浓度,通过测定荧光信号,制得荧光响应图谱,如图5所示,图中纵坐标为荧光强度,横坐标为不同阳离子种类。可以发现,荧光信号在加入1μM浓度的Cu2+后出现显著下降,而除Fe3+略有影响外,其他10μM浓度的阳离子对荧光信号强度影响不大。
以上所述,只是本发明的较佳实施例而已,本发明并不局限于上述实施方式,只要其以相同的手段达到本发明的技术效果,都应属于本发明的保护范围。在本发明的保护范围内其技术方案和/或实施方式可以有各种不同的修改和变化。
Claims (5)
1.一种检测铜离子的荧光探针的制备方法,其特征在于,包括以下步骤:
将聚乙烯亚胺和1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯在超声条件下混匀,形成所述荧光探针,具体为:
称取40 mg的1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯溶解在40 mL 0.01M NaOH中,反复超声涡旋直至完全溶解,配制成1 mg/mL的溶液;
称取500 mg的聚乙烯亚胺溶解在5 mL超纯水中,反复涡旋直至完全溶解,配制成100mg/mL的聚乙烯亚胺浓液;
向0.9 mL超纯水中加入1 mL的1,1,2,2-四-(4-羧基-(1,1-联苯))乙烯溶液,混合均匀后,向上述混合溶液中加入0.1 mL的聚乙烯亚胺溶液,在室温下搅拌1 h,将体系超声30min后,4℃透析3天,放置4℃保存备用。
2.一种检测铜离子的荧光探针,其特征在于,由权利要求1所述的制备方法制得。
3.根据权利要求2所述的荧光探针,其特征在于,所述荧光探针对铜离子的检测范围为10-250 nM,检测限为1.1 nM。
4.一种检测铜离子的方法,其特征在于,包括以下步骤:将权利要求3所述的荧光探针加入待检测液中,根据荧光强度和标准曲线得到待检测液中铜离子的浓度;所述标准曲线根据以下步骤制得:配制5-500nM的一系列铜离子浓度标准溶液和0nM铜离子浓度的空白溶液,将荧光探针加入到上述各个标准溶液和空白溶液中,混匀后测定各个标准溶液和空白溶液的荧光强度,以各个标准溶液的荧光强度与空白溶液的荧光强度的差值为纵坐标,铜离子浓度为横坐标制成标准曲线。
5.权利要求2或3所述的荧光探针在废水检测和食品检测中检测铜离子的应用。
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