【摘要】：近年來,將熒光探針與熒光成像技術(shù)進(jìn)行結(jié)合并運(yùn)用到對一些特定物質(zhì)的檢測已成為極其重要的科學(xué)命題,同時(shí)因其操作方便快捷,靈敏度和選擇性都較高,也被普遍應(yīng)用到食品、環(huán)境等各個(gè)領(lǐng)域。氟硼吡咯(BODIPY)類熒光染料因具有高的摩爾消光系數(shù)、良好的光穩(wěn)定性、高的熒光量子產(chǎn)率等優(yōu)點(diǎn)可作為熒光分子探針的理想染料,因此,本論文以BODIPY為熒光母體,設(shè)計(jì)合成了BODIPY類衍生物B1和B2,檢測試驗(yàn)結(jié)果顯示該探針不僅能特異性識別半胱氨酸(Cys)、同型半胱氨酸(Hcy),而且能進(jìn)一步通過反應(yīng)動(dòng)力學(xué)上的差異,又可以將Cys與Hcy區(qū)別開來。具體工作內(nèi)容如下:第一章:介紹了本論文的研究背景。介紹了熒光探針的組成及其識別機(jī)理,根據(jù)探針與生物硫醇的反應(yīng)機(jī)理進(jìn)行簡單歸納分類,為本論文中生物硫醇小分子探針的設(shè)計(jì)、合成與應(yīng)用奠定文獻(xiàn)基礎(chǔ)。第二章:設(shè)計(jì)并合成了兩個(gè)BODIPY類衍生物B1和B2,并且分別用飛行時(shí)間質(zhì)譜(TOP-MS)、氫譜(1H-NMR)和碳譜(13C-NMR)和元素分析方法,對其結(jié)構(gòu)進(jìn)行了表征。第三章:重點(diǎn)介紹了用于檢測實(shí)際樣品中Cys的“關(guān)-開型”BODIPY類熒光分子探針B1。此探針表現(xiàn)出對Cys高度選擇性,并不受Hcy和谷胱甘肽(GSH)的干擾。此分子探針與Cys在0.2-30μM保持良好的線性關(guān)系,檢測限為0.05μM。在pH=6.0-8.0的中性條件下,B1對Cys的響應(yīng)不受pH的影響,并且B1對Cys響應(yīng)迅速,響應(yīng)時(shí)間在5 min之內(nèi)。再者,分子探針B1對Cys選擇性響應(yīng)機(jī)理(PET機(jī)理)通過質(zhì)譜,核磁和高斯密度泛函理論(DFT)計(jì)算進(jìn)行了充分的驗(yàn)證。最后,將此分子探針應(yīng)用到活細(xì)胞成像試驗(yàn)中,并得到了滿意的結(jié)果。第四章:重點(diǎn)介紹了用于檢測實(shí)際樣品中Cys的“關(guān)-開型”近紅外BODIPY類熒光分子探針B2。此探針的發(fā)射在708 nm處,應(yīng)用在近紅外區(qū)域?qū)ys有高度的選擇性。此分子探針與Cys在0.2-30μM保持良好的線性關(guān)系,檢測限為0.05μM。實(shí)驗(yàn)結(jié)果顯示,探針B2對Cys的檢測在中性條件(pH=6.0-9.0)下基本不受pH的干擾且響應(yīng)迅速(少于5 min)。再者,分子探針B2對Cys選擇性響應(yīng)機(jī)理(PET機(jī)理)通過質(zhì)譜,核磁和高斯密度泛函理論(DFT)計(jì)算進(jìn)行了充分的驗(yàn)證。最后,將此分子探針應(yīng)用到活細(xì)胞成像和組織試驗(yàn)中,得到了令人滿意的結(jié)果。
[Abstract]:In recent years, it has become an extremely important scientific proposition to combine fluorescence probe with fluorescence imaging technology and apply it to the detection of some specific substances. At the same time, because of its convenient and rapid operation, high sensitivity and selectivity. It is also widely used in food, environment and other fields. Fluoroboron polypyrrole (BODIPY) fluorescent dyes can be used as ideal dyes for fluorescent molecular probes because of their high molar extinction coefficient, good photostability and high fluorescence quantum yield. Therefore, BODIPY is used as fluorescent parent in this paper. BODIPY derivatives B1 and B2 were designed and synthesized. the results showed that the probe could not only specifically recognize cysteine (Cys), homocysteine (Hcy), but also further pass the difference in reaction kinetics. You can also distinguish Cys from Hcy. The specific work is as follows: the first chapter: introduces the research background of this paper. The composition and recognition mechanism of fluorescent probe are introduced. According to the reaction mechanism between probe and biomercaptan, the composition and recognition mechanism of fluorescent probe are briefly summarized and classified, which lays a literature foundation for the design, synthesis and application of biomercaptan small molecular probe in this paper. In chapter 2, two BODIPY derivatives B1 and B2 were designed and synthesized, and their structures were characterized by time-of-flight mass spectrometry (TOP-MS), hydrogen spectroscopy (1H-NMR), carbon spectrum (13C-NMR) and elemental analysis, respectively. In chapter 3, the "off-open" BODIPY fluorescent molecular probe B1, which is used to detect Cys in practical samples, is introduced in detail. The probe showed high selectivity to Cys and was not interfered by Hcy and Hcy. The molecular probe has a good linear relationship with Cys at 0.2-30 渭 M, and the detection limit is 0.05 渭 M. Under the neutral condition of pH=6.0-8.0, the response of B1 to Cys is not affected by pH, and B1 responds quickly to Cys, and the response time is within 5 min. Furthermore, the selective response mechanism of molecular probe B1 to Cys (PET mechanism) was fully verified by mass spectrometry, nuclear magnetic field and Gao Si density functional theory (DFT) calculation. Finally, the molecular probe was applied to living cell imaging test, and satisfactory results were obtained. Chapter 4: the "off-open" near-infrared BODIPY fluorescent molecular probe B2, which is used to detect Cys in practical samples, is introduced in detail. The probe emits at 708 nm and is highly selective to Cys in the near infrared region. The molecular probe has a good linear relationship with Cys at 0.2-30 渭 M, and the detection limit is 0.05 渭 M. The experimental results show that the detection of Cys by probe B2 is basically undisturbed by pH under neutral condition (pH=6.0-9.0) and has a rapid response (less than 5 min). Furthermore, the selective response mechanism of molecular probe B2 to Cys (PET mechanism) was fully verified by mass spectrometry, nuclear magnetic field and Gao Si density functional theory (DFT) calculation. Finally, the molecular probe was applied to living cell imaging and tissue test, and satisfactory results were obtained.
【學(xué)位授予單位】：湘潭大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O657.3

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