CN108003173A - 一种特异性识别水合肼的荧光探针 - Google Patents

一种特异性识别水合肼的荧光探针 Download PDF

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CN108003173A
CN108003173A CN201711392566.3A CN201711392566A CN108003173A CN 108003173 A CN108003173 A CN 108003173A CN 201711392566 A CN201711392566 A CN 201711392566A CN 108003173 A CN108003173 A CN 108003173A
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宋相志
苏远安
张帆
杨雷
廖立德
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Abstract

本发明公开了一种特异性识别水合肼的荧光探针，其分子结构式如下：该荧光探针与水合肼作用时，在PBS(pH＝7.4，10mM)中荧光由绿色变成蓝色，可以实现比率型检测水合肼。此外，该荧光探针可用于环境或细胞中的水合肼检测，对水合肼的选择性好、灵敏度高、抗干扰能力强，具有很好的应用前景。

Description

一种特异性识别水合肼的荧光探针
技术领域
本发明涉及的是荧光探针领域，涉及一种检测水合肼的荧光探针的制备与应用。
背景技术
水合肼(N2H4)是一种无机二元胺，无色具有氨气味的液体。作为一种具有高反应活性的还原剂、强碱，水合肼及其衍生物在制药、摄影剂、杀虫剂、纺织染料、防腐以及其他精细化工方面有重要的作用。水合肼可以作为植物生长剂、塑料吹塑剂。由于水合肼具有易燃、易爆、燃烧焓高的特性，通常被用于火箭、导弹推进系统。水合肼作为一种重要的工业原料有着广泛的应用，但是它是一种高毒性化学物质，在操作、使用、处理过程中容易造成环境污染以及人体伤害。水合肼作为一种神经毒素、致癌物，可以对肾脏、肺、肝脏、中枢神级系统造成伤害。吸入高浓度的水合肼会造成咽喉、鼻子、眼睛感染，严重的会导致短暂的失明、头晕、恶心、血液病等慢性疾病。在人体内，水合肼虽然不是内生的，但是可以通过一些药物的代谢产生。在饮用水中可以通过氨气与氯气的氯化作用产生。世界健康组织设定了饮用水中容许最高浓度值为10ppb。
用于检测水合肼的常规方法有色谱法、电势法、电化学方法、气质联用法。但是常规方法方法具有一些缺陷如：需要昂贵的仪器设备、复杂的操作过程、样品前处理繁琐等。荧光探针作为一种新型的分析方法，具有高选择性、高灵敏性可以实现在细胞组织内实时检测分析物的优点。但是关于检测水合肼的荧光探针的报道数量有限，比值型检测水合肼的荧光探针数量更是甚少。比值型荧光探针由于可以检测两个发射峰的荧光强度，可以避免由于探针浓度、仪器因素、环境因素造成的误差，从而提高检测的准确性。
发明内容
本发明目的之一是提供一种检测水合肼的荧光探针合成方法；目的之二是提供一种灵敏度高、选择性好、抗干扰能力强能够在生物体外及生物体内检测水合肼的荧光探针。
本发明解决问题采取的技术方案为，一种可以特异性识别水合肼的新型荧光探针，其分子结构式如下：具体合成路线如下：
具体成方法如下：
(1)将400mg化合物2、295mg化合物3溶于10mL甲苯中，回流反应24小时，抽滤，用10mL石油醚洗涤滤饼，烘干得到210mg灰白色粉末为化合物4，产率，54.5％。(2)将51.4mg化合物4溶于10mL无水二氯甲烷中，加入85μL三乙胺，氩气保护冰水浴30分钟，再加入25μL乙酰氯，继续保持冰水浴30分钟。旋干溶剂，柱层析得到40mg黄色粉末为化探针，产率66.9％。
本发明的荧光探针测试方法如下，将探针分子溶解在pH为7.4的PBS(10.0mM)缓冲溶液中，室温下进行测试。可以对水合肼进行定量检测，具体实施方法在实施实例中详细介绍。
本发明的荧光探针的响应机理如下：水合肼与探针分子响应后，水合肼进攻酯基，脱去一个乙酰水合肼分子，释放出染料分子。溶液从黄色变成无色，荧光由绿色变成蓝色。从而实现了荧光比值型检测水合肼过程。探针分子的响应过程如下：
本发明的荧光探针的发射峰在500nm，与水合肼响应后的荧光发射峰在425nm处。探针与水合肼完全响应后425nm处荧光强度与500nm处荧光强度的比值(I425/I500)由0.017变成9.3。
本发明所述的探针分子合成路线简单，操作简单，成本较低，对水合肼的选择性好、抗干扰能力强，该荧光探针在生物诊断、分析化学、环境科学等领域具有实际的应用价值。
附图说明
图1为本发明荧光探针的选择性，荧光探针(1.0×10-5mol/L)在PBS(pH＝7.4，10mM)缓冲溶液中，与不同物质响应后的荧光光谱，横坐标为波长，纵坐标为荧光强度。
图2为本发明荧光探针的抗干扰能力，水合肼与其它干扰物共存时，与荧光探针(1.0×10-5mol/L)在PBS(pH＝7.4，10mM)缓冲溶液中与水合肼响应后425nm处荧光强度与500nm处荧光强度的比值(I425/I500)柱状图。横坐标为水合肼与干扰性混合类别(1,空白；2,NaN3；3,AlF3；4,NaF；5,Na2HPO3；6,Na2S2O3；7,Na2S；8,NaSCN；9,Na2CO3；10,CaCl2；11,KCl；12,MgCl2；13,NaCl；14,NaNO3；15,ZnCl2；16,Na2SO4；17,Na2SO3；18,NaHSO3；19，NaNO2；20,NaACO；21,Hcy；22,Cys；23,GSh；24，NaHCO3)，纵坐标为425nm处荧光强度与500nm处荧光强度的比值(I425/I500)。
图3为本发明的荧光探针(1.0×10-5mol/L)在PBS(pH＝7.4，10mM)缓冲溶液中，与不同浓度水合肼响应后的荧光强度变化，横坐标为波长，纵坐标为荧光强度。
图4为本发明的荧光探针(1.0×10-5mol/L)在PBS(pH＝7.4，10mM)缓冲溶液中，水合肼浓度与的425nm处荧光强度同500nm处荧光强度的比值(I425/I500)的线性关系，横坐标为浓度，纵坐标为425nm处荧光强度与500nm处荧光强度的比值(I425/I500)。
图5为本发明的荧光探针(1.0×10-5mol/L)在在PBS(pH＝7.4，10mM)缓冲溶液中，与水合肼响应过程中425nm处荧光强度与500nm处荧光强度的比值(I425/I500)随时间的变化，横坐标为时间，纵坐标为425nm处荧光强度与500nm处荧光强度的比值(I425/I500)。
图6为本发明的荧光探针(1.0×10-5mol/L)在不同pH值PBS(10.0mM)缓冲溶液中，与水合肼响应前后的425nm处荧光强度与500nm处荧光强度的比值(I425/I500)，横坐标为pH，纵坐标为425nm处荧光强度与500nm处荧光强度的比值(I425/I500)。
图7为本发明的荧光探针(1.0×10-5mol/L)在不同条件下的细胞(Hela细胞)成像：图a探针和水合肼溶液处理的细胞中绿光通成像，b探针和水合肼溶液处理的细胞蓝光通道成像，c探针和水合肼溶液处理的细胞中明场成像，d探针和水合肼溶液处理的细胞复合场成像，图e探针处理的细胞中绿光通成像，f探针溶液处理的细胞蓝光通道成像，g探针处理的细胞中明场成像，h探针处理的细胞复合场成像。
具体实施实例
实施例1：中间产物4的合成
将400mg化合物2、295mg化合物3溶于10mL甲苯中，回流反应24小时，抽滤，用10mL石油醚洗涤滤饼，烘干得到210mg灰白色粉末为化合物4，产率，54.5％。1H NMR(400MHz，DMSO-d6)δ11.75(s，1H)，7.15(s，1H)，5.21(s，1H)，3.23(dd，J＝11.1，5.5Hz，4H)，2.71(dd，J＝8.9，6.1Hz，4H)，1.87(dd，J＝11.9，6.2Hz，4H).13C NMR(100MHz，DMSO-d6)δ167.06，163.29，151.47，146.46，120.35，117.78，105.83，103.57，86.28，49.69，40.43，40.22，40.01，39.80，39.59，27.43，21.49，20.62，＝20.58。
实施例2：探针的合成
将51.4mg化合物4溶于10mL无水二氯甲烷中，加入85μL三乙胺，氩气保护冰水浴30分钟，再加入25μL乙酰氯，继续保持冰水浴30分钟。旋干溶剂，柱层析得到40mg黄色粉末为化探针，产率66.9％。1H NMR(400MHz，CDCl3)δ6.91(s，1H)，5.98(s，1H)，3.27(dd，J＝11.8，6.7Hz，4H)，2.87(d，J＝6.5Hz，2H)，2.76(s，2H)，2.39(s，3H)，2.01–1.91(m，4H).13C NMR(100MHz，CDCl3)δ167.09，163.27，159.93，146.65，119.49，118.40，106.74，98.52，77.39，77.07，76.75，49.97，49.57，27.71，21.39，21.23，20.47。
实施例3：本发明:荧光探针的应用
将探针溶于pH为7.4的PBS(10.0mM)缓冲溶液中,配制成1.0×10-5mol/L的溶液，向溶液中加其它选择项物质(Cys，Hcy，GSH，Na+，k+，Mg2+，Al3+，Zn2+，Ca2+，N3 -，F-，CO3 2-，HPO4 2-，S2O3 2-，S2-，SCN-，HCO3 -，NO3 -，SO3 2-，HSO3 -，NO2 -，ACO-，HSO3 -)，加人水合肼导致溶液500nm处与425nm处荧光强度变化，该荧光探针对水合肼表现出高灵敏性、高选择性的识别。当水合肼与干扰物质NaN3，AlF3，NaF，Na2HPO3，Na2S2O3，Na2S，NaSCN，Na2CO3，CaCl2，KCl，MgCl2，NaCl，NaNO3，ZnCl2，Na2SO4，Na2SO3，NaHSO3，NaNO2，NaACO，Hcy，Cys，GSh，NaHCO3共存时，探针对水合肼响应表现出来很强的抗干扰能力。该探针分子与水合肼响应速度快15分钟内可以观察到溶液500nm处与425nm处荧光强度变化。探针分子在pH为7至9的范围内都可以对水合肼选择性识别，可以在细胞内检测水合肼。

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1.一种特异性识别水合肼的荧光探针，其结构为：
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108689962A (zh) * 2018-06-21 2018-10-23 济南大学 一种检测水合肼的荧光探针及其制备方法和应用
CN108982447A (zh) * 2018-07-19 2018-12-11 曲阜师范大学 一种用于检测肼的比率式荧光探针的制备方法及应用
CN109912612A (zh) * 2019-04-04 2019-06-21 郑州大学 一种基于分子逻辑门识别Cys、GSH和HOCl荧光探针的合成、制备及其应用
CN114605285A (zh) * 2022-03-15 2022-06-10 明士新材料有限公司 一种近红外特异性检测水合肼的荧光探针及其制备方法

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002012867A1 (fr) * 2000-07-28 2002-02-14 Japan Science And Technology Corporation Sonde fluorescente pour analyse d'ions magnesium

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002012867A1 (fr) * 2000-07-28 2002-02-14 Japan Science And Technology Corporation Sonde fluorescente pour analyse d'ions magnesium

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ELISABETE OLIVEIRA，等: "Novel Small Stable Gold Nanoparticles Bearing Fluorescent Cysteine-Coumarin Probes as New Metal-Modulated Chemosensors", 《INORG. CHEM.》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108689962A (zh) * 2018-06-21 2018-10-23 济南大学 一种检测水合肼的荧光探针及其制备方法和应用
CN108982447A (zh) * 2018-07-19 2018-12-11 曲阜师范大学 一种用于检测肼的比率式荧光探针的制备方法及应用
CN109912612A (zh) * 2019-04-04 2019-06-21 郑州大学 一种基于分子逻辑门识别Cys、GSH和HOCl荧光探针的合成、制备及其应用
CN109912612B (zh) * 2019-04-04 2021-05-11 郑州大学 一种基于分子逻辑门识别Cys、GSH和HOCl荧光探针的合成、制备及其应用
CN114605285A (zh) * 2022-03-15 2022-06-10 明士新材料有限公司 一种近红外特异性检测水合肼的荧光探针及其制备方法

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