基于罗丹明衍生物近红外染料的合成及性能研究

发布时间：2018-05-27 14:15

本文选题：荧光分子探针 + 近红外　；参考：《湘潭大学》2017年硕士论文

【摘要】：目前,荧光分子探针已成为分析化学的一个研究热点。由于具有分析速度快、操作简单、选择性好、灵敏度高等优点,荧光分析技术已被广泛应用于各个领域,如化学化工、材料化学、环境科学以及生物医学等领域。罗丹明染料具有大的刚性共轭结构,荧光量子产率高,是一类优良的荧光探针染料。但罗丹明类的探针发射波长比较短(650 nm),这限制了其在生物成像中的应用。近红外荧光分子探针的吸收和发射波长都位于650-900 nm,因此具有背景荧光低、强的穿透生物组织的能力、对生物样品的光损伤少的优点。近年来,越来越多的人研究近红外荧光分子探针。本论文利用费舍尔氏醛来增大荧光母体的共轭结构,将探针的波长延长到近红外区,设计合成了两个新型的罗丹明类近红外荧光分子探针,并分别应用于对谷胱甘肽(glutathione,GSH)和三价铝离子的检测。具体内容如下:1.设计并合成了荧光分子探针NIR-Rh1、NIR-Rh2,并通过质谱、核磁共振氢谱以及碳谱对两个分子的结构进行了确认及表征。2.将合成的近红外荧光分子探针NIR-Rh1用于对GSH的检测。在加入GSH后,该探针展现高的量子产率(Φ=0.43),当与2.5当量的GSH作用时,荧光强度增加了75倍。而且,该探针能高选择性的检测GSH,不受其它生物硫醇(半胱氨酸Cys、同型半胱氨酸Hcy)以及氨基酸的干扰。另外,GSH诱导的探针内酰胺开环的机理也通过质谱、核磁共振进行了验证。更重要的是,该荧光探针成功的被应用到生物样品中如细胞和组织中选择性的检测GSH。3.将合成的近红外荧光分子探针NIR-Rh2应用于铝离子的检测。据我们所知,这是第一个用于测铝并将其应用于细胞和组织中的近红外探针。在未加Al~(3+)时,探针溶液显浅黄色并且没有荧光。当加入Al~(3+)后,Al~(3+)与该探针NIR-Rh2发生络合作用,导致NIR-Rh2的内酰胺结构开环,在743 nm处出现一个强发射峰,同时溶液的颜色由浅黄色变成绿色。当加入2当量的Al~(3+)时,荧光强度增强至空白的77倍。探针溶液在743 nm处的荧光强度与1.0×10~(-7)-2.0×10~(-5)M范围内的Al~(3+)浓度呈良好的线性关系,检测限为3×10~(-8) M。另外,在pH值为6.0-8.0时,不会干扰探针NIR-Rh2检测Al~(3+),而且对Al~(3+)的选择性高。同时,我们还利用质谱、核磁以及DFT理论计算对荧光分子探针NIR-Rh2与Al~(3+)的响应原理进行了验证,并成功的将该探针应用到了活细胞和组织中荧光成像。
[Abstract]:At present, fluorescent molecular probes have become a hot spot in analytical chemistry. Because of its advantages of fast analysis, simple operation, good selectivity and high sensitivity, fluorescence analysis technology has been widely used in various fields, such as chemistry, material chemistry, environmental science and biomedicine. Rhodamine dyes have large rigid conjugated structure and high fluorescence quantum yield, so they are a kind of excellent fluorescent probe dyes. However, Rhodamine probe has a shorter emission wavelength of 650 nm, which limits its application in biological imaging. The absorption and emission wavelengths of near-infrared fluorescence probes are located at 650-900 nm, so they have the advantages of low background fluorescence, strong ability to penetrate biological tissues and less light damage to biological samples. In recent years, more and more people have studied near infrared fluorescent molecular probes. In this paper, Fischer's aldehyde was used to enlarge the conjugate structure of the fluorescence matrix, and the wavelength of the probe was extended to near infrared region. Two novel Rhodamine near infrared fluorescent probes were designed and synthesized. It was applied to the detection of glutathione (GSH) and trivalent aluminum ions, respectively. The details are as follows: 1. The fluorescent molecular probe NIR-Rh1 (NIR-Rh2) was designed and synthesized. The structures of the two molecules were confirmed and characterized by mass spectrometry, nuclear magnetic resonance (NMR) and carbon spectroscopy. The synthesized near infrared fluorescent probe NIR-Rh1 was used to detect GSH. After the addition of GSH, the probe exhibited a high quantum yield (桅 _ (0.43), and the fluorescence intensity increased 75 times when the probe was treated with 2.5 equivalent GSH. Moreover, the probe can detect GSH with high selectivity without interference from other biological mercaptan (cysteine cysts, homocysteine) and amino acids. In addition, the mechanism of GSH-induced ring opening of the probe lactam was also verified by mass spectrometry (MS) and nuclear magnetic resonance (NMR). More importantly, the fluorescent probe was successfully applied to the selective detection of GSH.3 in biological samples such as cells and tissues. The synthesized near infrared fluorescence probe NIR-Rh2 was applied to the detection of aluminum ions. To our knowledge, this is the first near-infrared probe to be used for the determination of aluminum and its application in cells and tissues. When Al~(3 is not added, the probe solution is yellowish and has no fluorescence. After the addition of Al~(3), the Al~(3) complex with the probe NIR-Rh2 resulted in a ring opening of the lactam structure of NIR-Rh2, and a strong emission peak at 743nm, and the color of the solution changed from light yellow to green. When 2 equivalent Al~(3 was added, the fluorescence intensity increased to 77 times of the blank. The fluorescence intensity of the probe solution at 743 nm showed a good linear relationship with the concentration of Al~(3 in the range of 1.0 脳 10 ~ (-7) ~ (-7) -2.0 脳 10 ~ (10) ~ (-5) M, and the detection limit was 3 脳 10 ~ (-8) M. In addition, when pH was 6.0-8.0, the probe NIR-Rh2 could not interfere with the detection of Al~(3, and the selectivity to Al~(3 was high. At the same time, the response principle of fluorescent molecular probe NIR-Rh2 and Al~(3 was verified by mass spectrometry, nuclear magnetic field and DFT theory, and the probe was successfully applied to fluorescent imaging in living cells and tissues.
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3

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