CN112903771A - 一种提高卟啉ecl体系发光强度的方法 - Google Patents
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Abstract
本发明公开了一种提高卟啉ECL体系发光强度的方法,将光敏剂四羧基苯基卟啉溶于水中,加入含有共反应剂K2S2O8的磷酸盐缓冲溶液中,再加入重水D2O,得到能够提高发光强度的四羧基苯基卟啉/D2O‑K2S2O8的阴极电化学发光体系。在相同条件下,由于原本溶剂H2O中的H被加入的重水中的D取代,使得卟啉产生的单线态氧的发光寿命延长,材料的稳定性良好;TCPP/D2O‑K2S2O8体系的ECL强度约为TCPP‑K2S2O8体系的3倍,而且材料制备过程简单;体系具有较好的稳定性,发光材料水溶性良好,体系易于构建,发光体用量少;该新的ECL传感器具有很好的灵敏度和高的选择性,以便用于后续应用。
Description
技术领域
本发明属于卟啉ECL技术领域,具体涉及一种提高卟啉ECL体系发光强度的方法。
背景技术
卟啉及其衍生化合物广泛存在于生物体内和能量转移相关的重要细胞器内,由于其电化学发光性能较低,限制了应用领域;之前的研究表明,重水可用于增强发光和延长发光寿命,提高量子产率。而单线态氧,即激发态氧分子,是分子氧的顺磁性状态的一种通称,属于活性氧,是普通氧的激发态。单线态氧的寿命与其所处环境密切相关,在气体环境室温下,可达到1小时以上(有报道为72分钟),而在不同溶液中寿命不同。之前有文献报道,单线态氧在重水中的寿命大约是普通水中的十倍。1O2在体内会不断生成与猝灭,且在多种生理及病理过程中起作用(包括好的和坏的)。例如,在染料光敏化氧化条件下,各种生物成分(蛋白质、氨基酸、核酸等)很容易与氧反应而使有机体损坏,如在动物和人体中会引起蛋白质光氧化疾病等。因此利用卟啉这一光敏剂通过增强单线态氧的发光而提高整个体系的电化学发光性能,继而通过单线态氧与碘离子的氧化还原反应来表明单线态氧的作用以及达到检测碘离子的目的。
发明内容
本发明的目的在于解决上述现有技术中存在的问题,提供一种提高卟啉ECL体系发光强度的方法,得到的体系稳定性较好,实验操作简单,应用范围广。
本发明的目的通过以下技术方案来具体实现:
一种提高卟啉ECL体系发光强度的方法,将光敏剂四羧基苯基卟啉溶于水中,加入含有共反应剂K2S2O8的磷酸盐缓冲溶液中,再加入重水D2O,得到能够提高发光强度的四羧基苯基卟啉/D2O-K2S2O8的阴极电化学发光体系。
进一步的,所述方法的步骤如下:
(1)将四羧基苯基卟啉溶于水中,以K2S2O8作为共反应剂,以磷酸盐缓冲溶液作为支持电解质,将四羧基苯基卟啉水溶液加入含有K2S2O8的磷酸盐缓冲溶液中,再加入重水D2O,得四羧基苯基卟啉/D2O-K2S2O8阴极电化学发光体系;
(2)采用传统的三电极体系:裸的玻碳电极作为工作电极,Ag/AgCl电极作为参比电极,铂柱电极作为对电极,测定发光体系的ECL强度。
更进一步的,步骤(1)中,所述含有K2S2O8的磷酸盐缓冲溶液的pH为7.0,K2S2O8浓度为0.1mol/L,所述发光体系中四羧基苯基卟啉的浓度为30μmol/L,重水D2O的浓度为0.32μmol/L。
更进一步的,步骤(2)中,测定发光体系ECL强度的扫描条件为pH值7.0、扫速0.15V/S、电位窗在0~-1.2V。
本发明公开了一种新的利用溶剂效应来增强发光材料的电化学发光性能的方法,选取的发光材料为四羧基苯基卟啉(TCPP),当其作为光敏剂时,会产生单线态氧,由于四羧基苯基卟啉是水溶性卟啉,当添加D2O后,溶剂H2O中的羟基的氢被氘取代后,由于O-D的振动频率比O-H低,使得O2的无辐射驰豫速率相对减小,因而单线态氧在重水中的寿命相对较长,因此会导致卟啉产生的单线态氧的发光寿命增长,从而提高ECL强度;利用简单的溶剂效应可以明显增强卟啉的电化学发光强度,其操作简单方便且灵敏度高;形成的体系具有良好的生物相容性,稳定、均匀,拓宽了水溶性卟啉在水相中的应用,同时解决了滴涂在电极上长时间进行检测时掉落的问题。
本发明具有以下有益效果:
本发明提供的提高卟啉ECL体系发光强度的方法,在相同条件下,由于原本溶剂H2O中的H被加入的重水中的D取代,使得卟啉产生的单线态氧的发光寿命延长,材料的稳定性良好;TCPP/D2O-K2S2O8体系的ECL强度约为TCPP-K2S2O8体系的3倍,而且材料制备过程简单;体系具有较好的稳定性,发光材料水溶性良好,体系易于构建,发光体用量少;该新的ECL传感器具有很好的灵敏度和高的选择性,以便用于后续应用。
附图说明
图1是TCPP材料的紫外及荧光表征图。
图2是不同材料的ECL强度对比图。
图3是TCPP/D2O-K2S2O8体系中所添加的重水浓度与相应的ECL强度图。
图4是体系扫描23圈的ECL稳定性。
具体实施方式
以下对本发明的优选实施例进行说明,应当理解,此处所描述的优选实施例仅用于说明和解释本发明,并不用于限定本发明。
实施例1
(1)TCPP材料的制备
圆底烧瓶中装入4-羧基苯甲醛(3.64g),磁子和丙酸(50mL),将反应混合物加热到80℃直到4-羧基苯甲醛完全溶解,将新蒸馏的吡咯(0.7mL)加入到反应液中并将所得溶液加热140℃回流2h,将反应混合物冷却至室温,然后将烧瓶置于-4℃下,过夜,诱导沉淀,过滤沉淀,用CH2Cl2(5×50mL)里面加一点丙酮(CH2Cl2:丙酮=10:1)洗涤,抽滤。将沉淀溶于乙醇50℃超声20min,旋干并真空干燥得到目标产物。
(2)TCPP材料的表征
通过紫外表征,发现步骤(1)制备的TCPP的水溶液在420~450nm的波长范围(Soret谱带)和500~700nm的可见波长范围(Q谱带)中,具有TCPP独特的光学性能。通过荧光表征,TCPP水溶液的发射峰在680nm和720nm处,也具有TCPP独特的光学性能。通过UV-vis和FL表明了发光材料四羧基苯基卟啉TCPP的成功制备。见图1。
(3)TCPP/D2O-K2S2O8发光体系的制备
将TCPP溶于超纯水(H2O)中得到TCPP水溶液,将TCPP水溶液加入含有K2S2O8的磷酸盐缓冲溶液中,TCPP质子化后,加入重水(D2O),摇匀,以使其H被D取代,从而得到所需发光体系。其中,含有K2S2O8的磷酸盐缓冲溶液的pH值为7.0,K2S2O8的浓度为0.1mol/L,得到的发光体系中TCPP浓度为30μmol/L,重水(D2O)浓度为0.32μmol/L。
比较例1
除了不加入重水(D2O)之外,其余操作与实施例1的步骤(3)相同。
TCPP发光材料的ECL信号测量
①实施例1步骤(1)制备的TCPP材料备用。
②将玻碳电极依次用0.3μm和0.05μm的三氧化二铝粉打磨抛光,然后用二次蒸馏水冲洗,待用。
③实验室采用传统的三电极体系:裸的玻碳电极(GCE,直径3.0mm)作为工作电极,Ag/AgCl电极(含饱和KCl溶液)作为参比电极,铂柱电极作为对电极。MPI-A型毛细管电泳-电化学发光分析仪(购买于西安瑞迈分析仪器有限责任公司)用于电化学以及电化学发光实验。紫外可见光谱仪Agilent-8453用于对TCPP材料进行定性分析。整个实验中:MPI-A型毛细管电泳-电化学发光分析仪的光电倍增管偏压设置为800V,TCPP作为光敏剂,磷酸盐缓冲溶液(PBS,pH值7.0)作为支持电解质,0.1mol/L的K2S2O8作为体系的共反应剂,用于整个体系中。
图2是实施例1步骤(3)的TCPP/D2O-K2S2O8体系、比较例1的TCPP-K2S2O8体系、纯K2S2O8体系(0.1mol/L的PBS溶液,pH值7.0)的ECL对比图。在相同条件下,添加重水后的TCPP材料和未添加重水的TCPP材料相比,由于溶剂中H被D所取代,使得TCPP所产生的单线态氧的发光寿命延长,从而此体系的ECL强度是之前体系的3倍。
固定发光体系中TCPP的浓度(30μmol/L),通过改变重水(D2O)的浓度,在上述三电极体系中测其ECL强度的变化,即对重水的浓度进行优化。图3是表示电化学发光强度(ECL)与重水(D2O)的关系图。
图4是表示实施例1的TCPP/D2O-K2S2O8体系连续扫描23圈之后的ECL强度,表明在三电极体系下,新型溶剂效应体系的ECL强度具有较好的ECL稳定性。
由以上可知,本发明构建的新型溶剂效应的ECL体系,实际操作简单,无毒无害,稳定性好,灵敏度高。
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,尽管参照前述实施例对本发明进行了详细的说明,对于本领域的技术人员来说,其依然可以对前述各实施例所记载的技术方案进行修改,或者对其中部分技术特征进行等同替换。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 (4)
1.一种提高卟啉ECL体系发光强度的方法,其特征在于,将光敏剂四羧基苯基卟啉溶于水中,加入含有共反应剂K2S2O8的磷酸盐缓冲溶液中,再加入重水D2O,得到能够提高发光强度的四羧基苯基卟啉/D2O-K2S2O8的阴极电化学发光体系。
2.根据权利要求1所述的提高卟啉ECL体系发光强度的方法,其特征在于,所述方法的步骤如下:
(1)将四羧基苯基卟啉溶于水中,以K2S2O8作为共反应剂,以磷酸盐缓冲溶液作为支持电解质,将四羧基苯基卟啉水溶液加入含有K2S2O8的磷酸盐缓冲溶液中,再加入重水D2O,得四羧基苯基卟啉/D2O-K2S2O8阴极电化学发光体系;
(2)采用传统的三电极体系:裸的玻碳电极作为工作电极,Ag/AgCl电极作为参比电极,铂柱电极作为对电极,测定发光体系的ECL强度。
3.根据权利要求2所述的提高卟啉ECL体系发光强度的方法,其特征在于,步骤(1)中,所述含有K2S2O8的磷酸盐缓冲溶液的pH为7.0,K2S2O8浓度为0.1mol/L,所述发光体系中四羧基苯基卟啉的浓度为30μmol/L,重水D2O的浓度为0.32μmol/L。
4.根据权利要求2所述的提高卟啉ECL体系发光强度的方法,其特征在于,步骤(2)中,测定发光体系ECL强度的扫描条件为pH值7.0、扫速0.15V/S、电位窗在0~-1.2V。
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