CN115057864A - 基于aie分子/葫芦脲7检测金刚烷胺的荧光探针及其检测方法 - Google Patents
基于aie分子/葫芦脲7检测金刚烷胺的荧光探针及其检测方法 Download PDFInfo
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Abstract
本发明涉及一种基于AIE分子/葫芦脲7检测金刚烷胺的荧光探针及其检测方法,所述荧光探针的分子式为C77H74IN29O14,由葫芦脲7(CB[7])与聚集诱导发光分子(TPM)制成,其中CB[7]与TPM的摩尔比为1:1。本发明的荧光探针能够实现对溶液中金刚烷胺进行高灵敏检测,并体现了探针的特异性和选择性,具有检测灵敏度高、成本低、检测效率高,且利于对复杂微观系统的分析。本发明与传统的仪器分析检测方法相比,样品处理简单,操作方便,可以实时检测,其检测成本大大降低。本发明探针能对溶液中的金刚烷胺进行检测,能实现单探针对氨基酸高度选择性的检测,能够达到测定快速以及实时检测的有益效果。
Description
技术领域
本发明涉及一种高灵敏性荧光探针,特别是一种用于高灵敏检测金刚烷胺的荧光探针及其应用。
背景技术
金刚烷胺最早作为一种人用的抗病毒药物,具有较好的抵抗流感病毒、治疗流感的功效。但是随着畜禽养殖业的发展,金刚烷胺逐渐移植并大量投入到畜禽类疾病的治疗中。由于对金刚烷胺使用计量的增大和使用频率的增加,使畜禽出现中毒现象,直接诱导病毒变异,产生较强的耐药性。同时人类在长期食用金刚烷胺药物过量的食品时,会使金刚烷胺药物进入人体并积累,产生心情烦躁、神经恍惚、身体疲劳、精神抑郁等一系列中枢神经系统疾病,还会使人产生下肢肿胀、眩晕、言语不清、疲劳乏力等一些心脑血管疾病,严重影响人类身体健康。
因此实现对金刚烷胺高灵敏检测显得尤为重要,而荧光检测方法作为一种新型的检测方法,具有灵敏度高、选择性好、检测成本低、操作方便以及实时检测的优点而备受人们的青睐。研究新型的用于检测溶液中金刚烷胺的荧光探针及荧光探针检测方法很有必要。
此外,具有聚集诱导发光(Aggregation-Induced Emission,AIE)特性的荧光探针在溶液或分散状态下不发光,而在聚集状态下发光强烈(限制分子内运动受限机理,RIM),利用这种特性研发了许多荧光检测器,为荧光分析检测提供了新的思路和方案。
发明内容
针对现有技术的不足,本发明的目的是提供一种用于高灵敏检测金刚烷胺的荧光探针及其应用。
为了实现上述目的,本发明所采用的技术方案是:
基于AIE分子/葫芦脲7检测金刚烷胺的荧光探针,所述荧光探针的分子式为C77H74IN29O14,结构式为:
所述荧光探针由葫芦脲7(CB[7])与聚集诱导发光分子(TPM)制成,其中CB[7]与TPM的摩尔比为1:1;CB[7]与TPM的结构式为:
所述的荧光探针的制备方法,包括以下步骤:
(1)取葫芦脲7加蒸馏水溶解,得溶液A;
(2)取聚集诱导发光分子加二甲基亚砜溶解,得溶液B;
(3)将溶液A和溶液B混合后,加蒸馏水稀释,即得。
所述溶液A中葫芦脲7的浓度为1.0×10-3mol/L。
所述溶液B中聚集诱导发光分子的浓度为1.0×10-3mol/L。
将溶液A和溶液B按照葫芦脲7与聚集诱导发光分子的摩尔比为1:1的比例进行混合,在常温下反应30s,加蒸馏水稀释至总浓度为1.0×10-5mol/L,即得。
所述的荧光探针在检测金刚烷胺中的应用。
所述的荧光探针在检测溶液中金刚烷胺的应用。
所述溶液中金刚烷胺的检测方法为:
(1)向制备的荧光探针溶液中加入不同浓度的金刚烷胺标准溶液,放置搅拌10s,得到标准体系,备用;
(2)向制备的荧光探针溶液中加入待检测溶液,放置搅拌10s,备用;
(3)以固定激发波长380nm,进行荧光发射光谱测定,分别测定加入待检测溶液的荧光探针溶液及标准体系,在600nm下的荧光强度;
(4)建立标准体系与荧光强度的线性关系,计算待测溶液中的金刚烷胺含量。
所述金刚烷胺标准溶液的浓度为0~5.0×10-6mol/L。
本发明有益效果:
1、本发明的荧光探针能够实现对溶液中金刚烷胺进行高灵敏检测,并体现了探针的特异性和选择性,具有检测灵敏度高、成本低、检测效率高,且利于对复杂微观系统的分析。
2、本发明是通过利用葫芦脲7(CB[7])与聚集诱导发光分子(TPM)形成超分子配合物,当检测到金刚烷胺时,金刚烷胺与荧光探针分子中的葫芦脲7(CB[7])络合,从而形成新的复合物,使聚集诱导发光分子(TPM)脱离,导致探针的荧光发生降低作用。因此,可以简单、快速、灵敏的对溶液中的金刚烷胺进行检测。
3、本发明与传统的仪器分析检测方法相比,样品处理简单,操作方便,可以实时检测,其检测成本大大降低。本发明探针能对溶液中的金刚烷胺进行检测,能实现单探针对氨基酸高度选择性的检测,能够达到测定快速以及实时检测的有益效果。
附图说明
图1实施例1中摩尔比法得到的紫外吸收光谱图。
其中,图A表示,不同摩尔比[CB[7]]/[TPM]下的波长与吸光度的关系;图B中,横坐标CB[7](μmol/L)表示,溶液中CB[7]的浓度;纵坐标中的λ380nm(au),表示380nm下的吸光度。
图2实施例1中等摩尔连续变换法得到的紫外吸收光谱图。
其中,图A表示,不同摩尔比[TPM]/([CB[7]]+[TPM])下的波长与吸光度的关系;图B中,横坐标X(TPM)表示,[TPM]/([CB[7]]+[TPM])的摩尔比;纵坐标中的△λ380nm*X(TPM)表示,380nm的吸光度与X(TPM)的乘积。
图3实施例2中测定不同浓度的金刚烷胺标准溶液的荧光光谱图。
图4实施例2中得到的金刚烷胺标准溶液与荧光强度的线性关系图。
其中,I0表示,荧光强度在600nm处,没有加入AM时的荧光强度;I表示,荧光强度在600nm处,加入AM后的荧光强度。
具体实施方式
以下结合实施例对本发明的具体实施方式作进一步详细说明。如无特别说明,实施例中所涉及的仪器设备均为常规仪器设备;涉及原料均为市售常规原料;涉及试验方法均为常规方法。
实施例中葫芦脲7(CB[7])简写为CB[7]、聚集诱导发光分子(TPM)简写为TPM。CB[7]溶液采用蒸馏水配置,TPM溶液采用二甲基亚砜(DMSO)(分析纯)溶液配置。
实施例1探究CB[7]与TPM所形成荧光探针的最佳摩尔比
一种荧光探针,分子式为C77H74IN29O14,由葫芦脲7(CB[7])与聚集诱导发光分子(TPM)制成,包括以下步骤:
(1)取葫芦脲7(CB[7])加蒸馏水溶解,得溶液A;
(2)取聚集诱导发光分子(TPM)加分析纯二甲基亚砜(DMSO)溶解,得溶液B;
(3)将溶液A和溶液B混合,在常温下反应30s,即得。
为了探究CB[7]与TPM所形成荧光探针的最佳摩尔比,我们采用紫外吸收光谱检测CB[7]与TPM之间的相互作用。
1)摩尔比法测定CB[7]与TPM之间的紫外吸收光谱数据,具体方法为:将CB[7]与TPM分别配制成1.0mmol/L水溶液和1.0mmol/L的DMSO溶液备用;固定TPM的浓度为0.01mmol/L,逐渐滴加改变CB[7]的浓度,形成[CB[7]]/[TPM]摩尔比为0,0.2,0.4,0.6,0.8,1.0,1.2,1.4,1.6,1.8,2.0,2.2,2.2,2.6,2.8,3.0的待测溶液,在室温下测定各待测溶液的紫外-可见吸收光谱(图1)。
2)随后采用等摩尔连续变换法(Job’s曲线)测试CB[7]与TPM之间的紫外吸收光谱数据:
制备浓度为0.01mmol/L的TPM溶液和浓度为0.01mmol/L的CB[7]溶液,备用。固定CB[7]与TPM的总浓度为0.01mmol/L,总体积为2mL,不断改变CB[7]与TPM之间的物质的量之比,配置一系列溶液,具体方法为:
分别准确移取0、0.2、0.4、0.6、0.8、1.0、1.2、1.4、1.6、1.8、2.0mL的TPM于离心管中,然后依次向离心管中加入2.0、1.8、1.6、1.4、1.2、1.0、0.8、0.6、0.4、0.2、0mL的CB[7],控制溶液总体积为2mL,配制出一系列不同摩尔比[TPM]/([CB[7]]+[TPM])=0,0.1,0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9,1.0的待测溶液混合均匀后,放置一段时间,在室温下测试各待测溶液的紫外-可见吸收光谱(图2)。
综上可知,本发明荧光探针的制备方法中:CB[7]与TPM所形成荧光探针的最佳摩尔比为1:1。
本实验选用:溶液A中葫芦脲7(CB[7])的浓度为1.0×10-3mol/L;溶液B中聚集诱导发光分子(TPM)的浓度为1.0×10-3mol/L;所得荧光探针溶液中,将溶液A和溶液B按照摩尔比1:1混合后,加蒸馏水稀释至总浓度为1.0×10-5mol/L,备用。
实施例2金刚烷胺的定量检测
向本发明的荧光探针总浓度为1.0×10-5mol/L的标准溶液(按实施例1进行制备)中,加入不同浓度的金刚烷胺(AM)标准溶液(金刚烷胺的浓度分别为0、0.5、1.0、1.5、2.0、2.5、3.0、3.5、4.0、4.5、5.0μmol/L),放置搅拌10s,以固定激发波长380nm,进行荧光发射光谱测定,收集的荧光波长范围为480~780nm。
检测结果如图3所示,可以看出,不同浓度的金刚烷胺可使荧光探针溶液发生不同程度的荧光降低(图中曲线从上到下,对应的AM浓度依次增加,荧光逐渐降低)。且在波长600nm时,对应的荧光强度最强。因此,以固定激发波长380nm、荧光波长600nm进行荧光发射光谱测定。根据不同浓度金刚烷胺与荧光强度的线性关系可知,金刚烷胺响应的线性范围为(0-5.0)×10-6mol/L,检出限为0.072×10-6mol/L,结果如图4所示。说明荧光变化与AM的浓度具有较好的线性关系,能够准确的进行检测。
利用荧光探针检测溶液中金刚烷胺的方法为:
(1)向总浓度为1.0×10-5mol/L的荧光探针溶液中加入待检测溶液或不同浓度的金刚烷胺标准溶液,放置搅拌10s,备用。
(2)以固定激发波长380nm,进行荧光发射光谱测定,分别测定待检测溶液及不同浓度金刚烷胺标准溶液,在600nm下的荧光强度。
(3)绘制金刚烷胺标准溶液与荧光强度的线性关系图,计算待测溶液中的金刚烷胺含量。
Claims (10)
3.如权利要求1所述的荧光探针的制备方法,其特征在于,包括以下步骤:
(1)取葫芦脲7加蒸馏水溶解,得溶液A;
(2)取聚集诱导发光分子加二甲基亚砜溶解,得溶液B;
(3)将溶液A和溶液B混合后,加蒸馏水稀释,即得。
4.如权利要求3所述的制备方法,其特征在于,所述溶液A中葫芦脲7的浓度为1.0×10- 3mol/L。
5.如权利要求3所述的制备方法,其特征在于,所述溶液B中聚集诱导发光分子的浓度为1.0×10-3mol/L。
6.如权利要求3所述的制备方法,其特征在于,将溶液A和溶液B按照葫芦脲7与聚集诱导发光分子的摩尔比为1:1的比例进行混合,在常温下反应30s,加蒸馏水稀释至总浓度为1.0×10-5mol/L,即得。
7.如权利要求1所述的荧光探针在检测金刚烷胺中的应用。
8.如权利要求1所述的荧光探针在检测溶液中金刚烷胺的应用。
9.如权利要求8所述的应用,其特征在于,所述溶液中金刚烷胺的检测方法为:
(1)向权利要求3制备的荧光探针溶液中加入不同浓度的金刚烷胺标准溶液,放置搅拌10s,得到标准体系,备用;
(2)向权利要求3制备的荧光探针溶液中加入待检测溶液,放置搅拌10s,备用;
(3)以固定激发波长380nm,进行荧光发射光谱测定,分别测定加入待检测溶液的荧光探针溶液及标准体系,在600nm下的荧光强度;
(4)建立标准体系与荧光强度的线性关系,计算待测溶液中的金刚烷胺含量。
10.如权利要求9所述的应用,其特征在于,所述金刚烷胺标准溶液的浓度为0~5.0×10-6mol/L。
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