高位阻卟啉類(lèi)熒光探針的設(shè)計(jì)合成及其熒光性能的研究
發(fā)布時(shí)間：2019-05-19 13:22
【摘要】：卟啉是一種性能優(yōu)良的近紅外熒光發(fā)色團(tuán),以其為母體設(shè)計(jì)合成的熒光探針,在近紅外光區(qū)進(jìn)行檢測(cè)時(shí)可以有效減小復(fù)雜生物樣品中背景熒光的干擾,提高檢測(cè)的準(zhǔn)確性和分辨率。目前基于卟啉結(jié)構(gòu)設(shè)計(jì)合成的熒光探針以陽(yáng)離子探針居多,而中性分子及陰離子探針鮮少報(bào)道。中性分子和陰離子在探針識(shí)別過(guò)程中的相互作用較弱,且大多數(shù)的中性分子及陰離子探針是通過(guò)與分析物之間發(fā)生化學(xué)反應(yīng)導(dǎo)致體系熒光強(qiáng)度變化來(lái)達(dá)到識(shí)別目的。因此,設(shè)計(jì)合成卟啉類(lèi)中性分子和陰離子探針較陽(yáng)離子探針更具有挑戰(zhàn)性。同時(shí),研究表明,簡(jiǎn)單地改變熒光團(tuán)中取代基的位置可以很好地提高熒光探針?lè)肿拥倪x擇性。選擇性的改變探針會(huì)更好地避免生物體及環(huán)境中干擾離子的影響,使探針對(duì)目標(biāo)化合物的檢測(cè)更為精準(zhǔn)。本文以卟啉為母體,根據(jù)PET機(jī)理設(shè)計(jì)合成了高位阻可識(shí)別硫離子、苯硫酚及巰基氨基酸的熒光探針。因?yàn)檫@些探針受光激發(fā)后發(fā)生PET過(guò)程,并且擁有高的位阻,所以探針本身的空白熒光很弱。而當(dāng)加入目標(biāo)化合物后,分子內(nèi)PET過(guò)程被抑制,體系熒光得到明顯的提高。主要內(nèi)容如下:首先,我們以不同取代位置的單羥基苯基卟啉為母體,2,4-二硝基苯磺酰氯為識(shí)別部分合成得到了兩種硫離子熒光探針M1、M2,利用核磁、質(zhì)譜等分析手段進(jìn)行了結(jié)構(gòu)表征。優(yōu)化探針M1、M2的測(cè)試條件后,得知兩個(gè)探針對(duì)硫離子均有較好的響應(yīng)。但將兩個(gè)探針的選擇性比較之后發(fā)現(xiàn):探針M1較M2更具有選擇的專一性。經(jīng)分析,探針M1的位阻較高,在一定程度上抑制了巰基化合物的進(jìn)攻,而促使體積較小的硫離子進(jìn)入發(fā)生親核反應(yīng)。因此,增大探針的空間位阻有利于探針選擇性的提高。其中,探針M1的線性范圍為3×10-6 mol/L~9×10-6 mol/L,檢出限為40 nM,探針M2的檢出限為62nM,線性范圍為1×10-6 mol/L~8×10-6 mol/L。其次,基于上述結(jié)論,將2-單羥基卟啉和NBD-Cl為原料合成了一種高位阻的苯硫酚熒光探針M3。該探針在加入苯硫酚前后熒光發(fā)生變化,并且在1×10-6 mol/L~7×10-6mol/L的濃度范圍內(nèi)呈現(xiàn)良好的線性關(guān)系,檢出限為54 nM。最后,我們?cè)?-單氨基卟啉接入馬來(lái)酰亞胺基團(tuán),得到了一種熒光增強(qiáng)型的可識(shí)別巰基氨基酸的探針M4。當(dāng)加入巰基氨基酸(GSH、Cys、Hcy)后,與探針之間發(fā)生環(huán)合反應(yīng),抑制了PET過(guò)程,體系的熒光強(qiáng)度得到增強(qiáng)。同時(shí),還檢測(cè)了6種非巰基氨基酸,結(jié)果發(fā)現(xiàn)體系的熒光光譜并沒(méi)有發(fā)生變化,表明探針M4可以用來(lái)區(qū)分巰基氨基酸和非巰基氨基酸。
[Abstract]:Porphyrin is a kind of near infrared fluorescence chromophore with excellent properties. The fluorescence probe designed and synthesized with it as the parent can effectively reduce the interference of background fluorescence in complex biological samples when it is detected in the near infrared region. Improve the accuracy and resolution of detection. At present, cationic probes are the most common fluorescent probes based on porphyrin structure design, while neutral molecules and anion probes are rarely reported. The interaction between neutral molecules and anions in the process of probe recognition is weak, and most neutral molecules and anion probes achieve the recognition purpose by chemical reaction with analytes to cause the change of fluorescence intensity of the system. Therefore, it is more challenging to design and synthesize porphyrin neutral molecules and anion probes than cationic probes. At the same time, it is shown that simply changing the position of the substituted group in the fluorescent cluster can improve the selectivity of the fluorescent probe molecule. Selective change of probe will better avoid the influence of interference ions in organism and environment, and make the detection of target compounds more accurate. In this paper, a fluorescence probe with high resistance to recognize sulfur ions, phlorophenol and sulfhydryl amino acids was designed and synthesized according to the mechanism of PET. Because the PET process occurs after these probes are excited by light and has high steric resistance, the blank fluorescence of the probes itself is very weak. When the target compound was added, the intramolecular PET process was suppressed and the fluorescence of the system was improved obviously. The main contents are as follows: first, we synthesized two kinds of sulfur ion fluorescence probes M1, M2by using monohydroxyphenyl porphyrins with different substitution positions as parent and 2, 4-dinitrobenzenesulfonyl chloride as recognition part. The structure was characterized by mass spectrometry and other analytical methods. After optimizing the test conditions of probe M _ 1 and M _ 2, it was found that the two probes had good responses to sulfur ions. However, after comparing the selectivity of the two probes, it is found that probe M1 is more selective and specific than M2. The results show that the steric hindrance of probe M1 is high, which suppresses the attack of sulfhydryl compounds to a certain extent, and causes the smaller sulfur ions to enter the nucleophilic reaction. Therefore, increasing the steric hindrance of the probe is beneficial to the improvement of the selectivity of the probe. The linear range of probe M1 is 3 脳 10 鈮，
本文編號(hào)：2480763
[Abstract]:Porphyrin is a kind of near infrared fluorescence chromophore with excellent properties. The fluorescence probe designed and synthesized with it as the parent can effectively reduce the interference of background fluorescence in complex biological samples when it is detected in the near infrared region. Improve the accuracy and resolution of detection. At present, cationic probes are the most common fluorescent probes based on porphyrin structure design, while neutral molecules and anion probes are rarely reported. The interaction between neutral molecules and anions in the process of probe recognition is weak, and most neutral molecules and anion probes achieve the recognition purpose by chemical reaction with analytes to cause the change of fluorescence intensity of the system. Therefore, it is more challenging to design and synthesize porphyrin neutral molecules and anion probes than cationic probes. At the same time, it is shown that simply changing the position of the substituted group in the fluorescent cluster can improve the selectivity of the fluorescent probe molecule. Selective change of probe will better avoid the influence of interference ions in organism and environment, and make the detection of target compounds more accurate. In this paper, a fluorescence probe with high resistance to recognize sulfur ions, phlorophenol and sulfhydryl amino acids was designed and synthesized according to the mechanism of PET. Because the PET process occurs after these probes are excited by light and has high steric resistance, the blank fluorescence of the probes itself is very weak. When the target compound was added, the intramolecular PET process was suppressed and the fluorescence of the system was improved obviously. The main contents are as follows: first, we synthesized two kinds of sulfur ion fluorescence probes M1, M2by using monohydroxyphenyl porphyrins with different substitution positions as parent and 2, 4-dinitrobenzenesulfonyl chloride as recognition part. The structure was characterized by mass spectrometry and other analytical methods. After optimizing the test conditions of probe M _ 1 and M _ 2, it was found that the two probes had good responses to sulfur ions. However, after comparing the selectivity of the two probes, it is found that probe M1 is more selective and specific than M2. The results show that the steric hindrance of probe M1 is high, which suppresses the attack of sulfhydryl compounds to a certain extent, and causes the smaller sulfur ions to enter the nucleophilic reaction. Therefore, increasing the steric hindrance of the probe is beneficial to the improvement of the selectivity of the probe. The linear range of probe M1 is 3 脳 10 鈮，
本文編號(hào)：2480763
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