羅丹明B類和萘酰亞胺類熒光探針的合成及性能研究
發(fā)布時(shí)間：2018-08-16 12:33
【摘要】：熒光分析方法具有靈敏度高、選擇性好、分析速度快、操作簡(jiǎn)單和易于細(xì)胞原位成像等優(yōu)點(diǎn),該分析方法常見(jiàn)于生物化學(xué)、環(huán)境科學(xué)和醫(yī)療診斷等領(lǐng)域。同時(shí),基于羅丹明B類和萘酰亞胺類熒光染料具有較高的熒光量子產(chǎn)率、良好的光熱穩(wěn)定性、結(jié)構(gòu)容易修飾和價(jià)格低廉等性質(zhì),這兩類熒光染料被普及應(yīng)用。因此,本論文利用羅丹明B和萘酰亞胺為熒光母體設(shè)計(jì)合成了兩種熒光探針?lè)謩e應(yīng)用于檢測(cè)線粒體中的ATP和溶酶體中的H_2S。具體內(nèi)容如下:1.設(shè)計(jì)并合成了熒光探針?lè)肿覯ito-Rh和Lyso-Nh,并通過(guò)了飛行時(shí)間質(zhì)譜,核磁共振氫譜和核磁共振碳譜等表征手段對(duì)其結(jié)構(gòu)確認(rèn)。2.我們利用羅丹明B,二乙烯三胺和三苯基膦分別作為熒光探針?lè)肿覯ito-Rh的熒光團(tuán),識(shí)別基團(tuán)和定位基團(tuán)。熒光探針?lè)肿覯ito-Rh自身無(wú)色無(wú)熒光,而當(dāng)加入ATP后,ATP能與該探針?lè)肿覯ito-Rh發(fā)生相互作用,使熒光團(tuán)羅丹明B內(nèi)酰胺環(huán)被打開(kāi),導(dǎo)致探針?lè)肿覯ito-Rh在583 nm處發(fā)出強(qiáng)烈的紅色熒光。該探針?lè)肿覯ito-Rh對(duì)ATP具有較高的靈敏性,反應(yīng)前后熒光增強(qiáng)了81倍,檢測(cè)范圍為0.1-10 mM,與線粒體中的ATP含量相匹配。同時(shí),相比于其他生物性的陰離子,比如:ADP,AMP,GTP,CTP和UTP等,該探針對(duì)ATP具有特異的選擇性,主要?dú)w功于該探針對(duì)ATP具有雙識(shí)別位點(diǎn)—?dú)滏I相互作用和π-π相互堆積作用。此外,該探針能精準(zhǔn)地定位于線粒體中并成功應(yīng)用于實(shí)時(shí)監(jiān)控線粒體中ATP的含量變化。3.我們利用萘酰亞胺,硝基苯并二唑和嗎啉分別作為熒光探針?lè)肿覮yso-Nh的熒光團(tuán),識(shí)別基團(tuán)和定位基團(tuán)。該探針自身存在嗎啉基團(tuán)N原子的孤對(duì)電子和淬滅基團(tuán)(識(shí)別基團(tuán))產(chǎn)生的PET過(guò)程影響,該探針無(wú)熒光發(fā)射,在pH=5的條件下加入H_2S時(shí),識(shí)別基團(tuán)離去且N原子的孤對(duì)電子質(zhì)子化,在536 nm處產(chǎn)生綠色的萘酰亞胺特征熒光發(fā)射峰。反應(yīng)前后熒光增強(qiáng)倍數(shù)為72倍,表明該探針的靈敏性較高。同時(shí),該探針具有較好的選擇性,不受其他生物性相關(guān)的物質(zhì)干擾影響。此外,該探針可以檢測(cè)溶酶體中的H_2S含量的變化。
[Abstract]:Fluorescence analysis method has the advantages of high sensitivity, good selectivity, fast analysis speed, simple operation and easy in situ imaging of cells. It is widely used in the fields of biochemistry, environmental science and medical diagnosis. At the same time, Rhodamine B and naphthalimide fluorescent dyes are widely used because of their high fluorescence quantum yield, good photothermal stability, easy structure modification and low price. Therefore, in this thesis, Rhodamine B and naphthalimide were used to design and synthesize two fluorescent probes for the detection of ATP in mitochondria and H _ 2s in lysosome, respectively. The details are as follows: 1. The fluorescent probe molecules Mito-Rh and Lyso-Nhwere designed and synthesized, and their structures were confirmed by time-of-flight mass spectrometry (TF-MS), nuclear magnetic resonance (NMR) and nuclear magnetic resonance (NMR) carbon spectroscopy. Rhodamine B, diethylenetriamine and triphenylphosphine were used as fluorescence groups of Mito-Rh, respectively, to recognize and locate the groups. The fluorescence probe molecule Mito-Rh itself is colorless and fluorescence free, and when ATP is added, the ATP can interact with the probe molecule Mito-Rh, which makes the fluorescence group Rhodamine B lactam ring open, resulting in a strong red fluorescence of the probe molecule Mito-Rh at 583nm. The probe molecule Mito-Rh has a high sensitivity to ATP. The fluorescence increases 81 times before and after the reaction, and the detection range is 0.1-10 mm, which matches the content of ATP in mitochondria. At the same time, compared with other biological anions, such as UTP and UTP, the probe has a specific selectivity for ATP, which is mainly attributed to the double recognition site hydrogen bond interaction and 蟺-蟺 stacking of the probe to ATP. In addition, the probe can be accurately located in mitochondria and successfully used to monitor the changes of ATP content in mitochondria in real time. Naphthalene imide, nitrobenzodiazole and morpholine were used as fluorescence groups of Lyso-Nh, respectively, to recognize and locate the groups. The probe itself has the effect of solitary pair electrons of morpholine group N atom and the PET process produced by quenching group (recognition group). The probe has no fluorescence emission, and when H _ S _ 2S is added under pH=5 condition, The characteristic fluorescence emission peak of naphthalimide is produced at 536 nm by the recognition of the absence of the group and the protonation of the lone pair electrons of the N atom. The fluorescence enhancement was 72 times before and after the reaction, which indicated that the probe was sensitive. At the same time, the probe has good selectivity and is not affected by other biologically related substances. In addition, the probe can detect changes in H 2S content in lysosomes.
【學(xué)位授予單位】：湘潭大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O657.3
本文編號(hào)：2185995
[Abstract]:Fluorescence analysis method has the advantages of high sensitivity, good selectivity, fast analysis speed, simple operation and easy in situ imaging of cells. It is widely used in the fields of biochemistry, environmental science and medical diagnosis. At the same time, Rhodamine B and naphthalimide fluorescent dyes are widely used because of their high fluorescence quantum yield, good photothermal stability, easy structure modification and low price. Therefore, in this thesis, Rhodamine B and naphthalimide were used to design and synthesize two fluorescent probes for the detection of ATP in mitochondria and H _ 2s in lysosome, respectively. The details are as follows: 1. The fluorescent probe molecules Mito-Rh and Lyso-Nhwere designed and synthesized, and their structures were confirmed by time-of-flight mass spectrometry (TF-MS), nuclear magnetic resonance (NMR) and nuclear magnetic resonance (NMR) carbon spectroscopy. Rhodamine B, diethylenetriamine and triphenylphosphine were used as fluorescence groups of Mito-Rh, respectively, to recognize and locate the groups. The fluorescence probe molecule Mito-Rh itself is colorless and fluorescence free, and when ATP is added, the ATP can interact with the probe molecule Mito-Rh, which makes the fluorescence group Rhodamine B lactam ring open, resulting in a strong red fluorescence of the probe molecule Mito-Rh at 583nm. The probe molecule Mito-Rh has a high sensitivity to ATP. The fluorescence increases 81 times before and after the reaction, and the detection range is 0.1-10 mm, which matches the content of ATP in mitochondria. At the same time, compared with other biological anions, such as UTP and UTP, the probe has a specific selectivity for ATP, which is mainly attributed to the double recognition site hydrogen bond interaction and 蟺-蟺 stacking of the probe to ATP. In addition, the probe can be accurately located in mitochondria and successfully used to monitor the changes of ATP content in mitochondria in real time. Naphthalene imide, nitrobenzodiazole and morpholine were used as fluorescence groups of Lyso-Nh, respectively, to recognize and locate the groups. The probe itself has the effect of solitary pair electrons of morpholine group N atom and the PET process produced by quenching group (recognition group). The probe has no fluorescence emission, and when H _ S _ 2S is added under pH=5 condition, The characteristic fluorescence emission peak of naphthalimide is produced at 536 nm by the recognition of the absence of the group and the protonation of the lone pair electrons of the N atom. The fluorescence enhancement was 72 times before and after the reaction, which indicated that the probe was sensitive. At the same time, the probe has good selectivity and is not affected by other biologically related substances. In addition, the probe can detect changes in H 2S content in lysosomes.
【學(xué)位授予單位】：湘潭大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O657.3
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 曹成波;朱艷麗;于學(xué)麗;張長(zhǎng)橋;;萘酰亞胺類功能材料應(yīng)用研究進(jìn)展[J];精細(xì)與專用化學(xué)品;2007年Z1期
2 趙同豐,趙德豐,于華云,程侶柏;1,8-萘酰亞胺類熒光材料的進(jìn)展[J];染料工業(yè);1997年01期
，本文編號(hào)：2185995
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