CN108822070A - 一种用于检测同型半胱氨酸结构荧光探针对位化合物及其应用 - Google Patents

一种用于检测同型半胱氨酸结构荧光探针对位化合物及其应用 Download PDF

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CN108822070A
CN108822070A CN201810606849.1A CN201810606849A CN108822070A CN 108822070 A CN108822070 A CN 108822070A CN 201810606849 A CN201810606849 A CN 201810606849A CN 108822070 A CN108822070 A CN 108822070A
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homocysteine
compound
fluorescence intensity
application
fluorescence probe
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朱彧
胡静仪
韩媛
么秀华
信欣
刘怡
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Tianjin Huanhu Hospital (tianjin Department Of Neurosurgery Research Institute Tianjin Brain Department Central Hospital)
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Tianjin Huanhu Hospital (tianjin Department Of Neurosurgery Research Institute Tianjin Brain Department Central Hospital)
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    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
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    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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    • C09K2211/1025—Heterocyclic compounds characterised by ligands
    • C09K2211/1088—Heterocyclic compounds characterised by ligands containing oxygen as the only heteroatom

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Abstract

本发明涉及临床医疗领域，尤其涉及一种用于检测同型半胱氨酸结构荧光探针对位化合物及其应用，具有如下的结构：通过对荧光强度的测定可有效看出本探针可以和同型半胱氨酸结构中的氨基和琉基基团特异性的发生亲核加成反应，共辄体的空间结构发生变化，引起体系中荧光强度增强，可选择性检测同型半胱氨酸，与半胱氨酸和其它氨基酸不响应。

Description

一种用于检测同型半胱氨酸结构荧光探针对位化合物及其 应用
技术领域
本发明涉及临床医疗领域，尤其涉及一种用于检测同型半胱氨酸结构荧光探针对位化合物及其应用。
背景技术
同型半胱氨酸(Homocysteine,HCY)又称高半胱氨酸，是一种含硫氨基酸且不属于组成蛋白质的20种氨基酸，体内不能合成，只能来源于蛋氨酸(甲硫氨酸)的分解代谢。正常情况下，血HCY含量甚微，一般空腹总HCY水平为5umol/L-15umol/L；轻度、中度、重度高HCY血症患者浓度分别为16umol/L-30umol/L、31umol/L-100umol/L、大于100umol/L。研究证实：高浓度的HCY是冠状动脉疾病、脑血管疾病、外周血管疾病独立的危险因子，危险度随着浓度的升高而增加。因此监测和降低同型半胱氨酸的浓度，具有十分重要的临床意义。
发明内容
本发明所要解决的技术问题是，克服现有技术中存在的不足，提供一种检测同型半胱氨酸结构荧光探针对位化合物及其应用。
一种用于检测同型半胱氨酸结构荧光探针对位化合物，具有如下的结构：
一种用于检测同型半胱氨酸结构荧光探针对位化合物应用，
a.化合物的稀释
称取上述化合物0.1g于1.5mlEP管中,加入1ml蒸馏水,溶解，混匀，梯度稀释至300倍；
b.化合物的荧光测定
取96孔黑色微孔板，加入上述化合物各100μl，激发光280nm,发射光590nm,测定其荧光强度；
c.同型半胱氨酸的荧光测定
取96孔黑色微孔板，加入同型半胱氨酸0μmol、1μmol、2μmol、5μmol、10μmol、20μmol、50μmol、和100μmol的同型半胱氨酸各100μl加入上述化合物各100μl，37度孵育10min，激发光280nm,发射光590nm,测定其荧光强度。
本发明的有益效果是：
通过对荧光强度的测定可有效看出本探针可以和同型半胱氨酸结构中的氨基和琉基基团特异性的发生亲核加成反应，共辄体的空间结构发生变化，引起体系中荧光强度增强，可选择性检测同型半胱氨酸，与半胱氨酸和其它氨基酸不响应。
附图说明
图1是探针与同型半胱氨酸反应的线性图。
具体实施方式
为了使本技术领域的技术人员更好地理解本发明的技术方案，下面结合附图和最佳实施例对本发明作进一步的详细说明。
本发明一种用于检测同型半胱氨酸结构荧光探针对位化合物，具有如下的结构：
一种用于检测同型半胱氨酸结构荧光探针对位化合物应用，
a.化合物的稀释
称取上述化合物0.1g于1.5mlEP管中,加入1ml蒸馏水,溶解，混匀，梯度稀释至300倍；
b.化合物的荧光测定
取96孔黑色微孔板，加入上述化合物各100μl，激发光280nm,发射光590nm,测定其荧光强度；
c.同型半胱氨酸的荧光测定
取96孔黑色微孔板，加入同型半胱氨酸0μmol、1μmol、2μmol、5μmol、10μmol、20μmol、50μmol、和100μmol的同型半胱氨酸各100μl加入上述化合物各100μl，37度孵育10min，激发光280nm,发射光590nm,测定其荧光强度。
本例中实验材料与设备为细胞成像微孔板分析仪(C5MVDG)BioTek,美国。
探针与同型半胱氨酸反应后，扣除探针荧光自发荧光，可得到探针与同型半胱氨酸反应的线性图(图1)。(相关系数R＝0.9985)
对其探针与同型半胱氨酸反应后荧光强度做重复性试验得到结果：
可得到试验结果稳定性高，实验结果可靠。
通过对荧光强度的测定可有效看出本探针可以和同型半胱氨酸结构中的氨基和琉基基团特异性的发生亲核加成反应，共辄体的空间结构发生变化，引起体系中荧光强度增强，可选择性检测同型半胱氨酸，与半胱氨酸和其它氨基酸不响应。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。

Claims (2)

1.一种用于检测同型半胱氨酸结构荧光探针对位化合物，其特征在于，具有如下的结构：
2.一种用于检测同型半胱氨酸结构荧光探针对位化合物应用，其特征在于：
a.化合物的稀释
称取上述化合物0.1g于1.5mlEP管中,加入1ml蒸馏水,溶解，混匀，梯度稀释至300倍；
b.化合物的荧光测定
取96孔黑色微孔板，加入上述化合物各100μl，激发光280nm,发射光590nm,测定其荧光强度；
c.同型半胱氨酸的荧光测定
取96孔黑色微孔板，加入同型半胱氨酸0μmol、1μmol、2μmol、5μmol、10μmol、20μmol、50μmol、和100μmol的同型半胱氨酸各100μl加入上述化合物各100μl，37度孵育10min，激发光280nm,发射光590nm,测定其荧光强度。
CN201810606849.1A 2018-06-13 2018-06-13 一种用于检测同型半胱氨酸结构荧光探针对位化合物及其应用 Pending CN108822070A (zh)

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105339438A (zh) * 2013-07-05 2016-02-17 富士胶片株式会社 色素多聚体、着色组合物、硬化膜、彩色滤光片、彩色滤光片的制造方法、固体摄像元件及图像显示装置

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105339438A (zh) * 2013-07-05 2016-02-17 富士胶片株式会社 色素多聚体、着色组合物、硬化膜、彩色滤光片、彩色滤光片的制造方法、固体摄像元件及图像显示装置

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
SAMUEL SAS ET AL.: "Highly luminescent hybrid materials based on smectites with polyethylene glycol modified with rhodamine fluorophore", 《APPLIED CLAY SCIENCE》 *

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