CN107698600A - 一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料及其制备方法和应用 - Google Patents

一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料及其制备方法和应用 Download PDF

Info

Publication number
CN107698600A
CN107698600A CN201710699740.2A CN201710699740A CN107698600A CN 107698600 A CN107698600 A CN 107698600A CN 201710699740 A CN201710699740 A CN 201710699740A CN 107698600 A CN107698600 A CN 107698600A
Authority
CN
China
Prior art keywords
rhodamine
fluorescent sensing
cyanobiphenyl
preparation
response
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201710699740.2A
Other languages
English (en)
Other versions
CN107698600B (zh
Inventor
唐旭
王赟
韩娟
李春梅
刘仁杰
李程
王蕾
倪良
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jiangsu University
Original Assignee
Jiangsu University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Jiangsu University filed Critical Jiangsu University
Priority to CN201710699740.2A priority Critical patent/CN107698600B/zh
Publication of CN107698600A publication Critical patent/CN107698600A/zh
Application granted granted Critical
Publication of CN107698600B publication Critical patent/CN107698600B/zh
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07DHETEROCYCLIC COMPOUNDS
    • C07D491/00Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10Spiro-condensed systems
    • C07D491/107Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00Luminescent, e.g. electroluminescent, chemiluminescent materials
    • C09K11/06Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N21/643Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • G01N21/80Indicating pH value
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/10Non-macromolecular compounds
    • C09K2211/1003Carbocyclic compounds
    • C09K2211/1007Non-condensed systems
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/10Non-macromolecular compounds
    • C09K2211/1018Heterocyclic compounds
    • C09K2211/1025Heterocyclic compounds characterised by ligands
    • C09K2211/1029Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/10Non-macromolecular compounds
    • C09K2211/1018Heterocyclic compounds
    • C09K2211/1025Heterocyclic compounds characterised by ligands
    • C09K2211/1088Heterocyclic compounds characterised by ligands containing oxygen as the only heteroatom
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6439Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pathology (AREA)
  • Optics & Photonics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Engineering & Computer Science (AREA)
  • Plasma & Fusion (AREA)
  • Molecular Biology (AREA)
  • Materials Engineering (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

本发明涉及一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料及其制备方法和应用,属于生物化学荧光传感材料技术领域;本发明以罗丹明B和氰基联苯酚两种对pH敏感的光学基团作为基础原料,首先将罗丹明B与水合肼进行反应制得罗丹明B酰肼,之后通过席夫碱反应将其与醛基修饰过的氰基联苯酚相结合制得荧光传感材料;本发明的方法制备条件温和,原料成本低,制得的材料自身化学稳定性良好,该材料对不同的pH具有独特的响应性质,而且这种响应能够通过肉眼可见的荧光和外观颜色呈现出来。本发明制得的pH响应型的荧光传感材料可以用于水体及生物细胞中pH的测定。

Description

一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料及 其制备方法和应用
技术领域
本发明涉及一种荧光化学传感材料的制备方法和用途,特别涉及一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料及其制备方法和应用,属于生物化学荧光传感材料技术领域。
背景技术
通常一些化学反应的进行和完成以及生物体中的各种生理过程都与pH值密切相关。其中细胞内pH值的大小在重要的生物过程如离子的传输,细胞的吞噬作用,细胞增值或凋亡及体内平衡中都扮演着重要的角色。生物细胞内不同组织的pH环境也有所不同,溶酶体和胞内体内的pH值呈酸性,在4.5–6.8范围内;细胞质和活性线粒体内的pH则呈偏碱性,在6.8–8范围内。当细胞内某一部分的pH 出现异常时,就会引起不适的生理反应,因此常常将细胞内pH的异常作为一些疾病的重要指标,如癌症,肿瘤,阿尔茨海默病等等。例如细胞在缺血的情况下其pH 会降低到6左右。甚至0.01-0.02的pH偏差都会引起心肺及神经系统出现问题,更严重的可能会危及生命。由此可见,细胞内pH值出现异常或者些许的波动都会影响健康。pH值规律性的波动能够帮助更好的理解pH值在生理中的角色和作用,并且也能很好的帮助研究相应的病理环境。所以,有效的监控细胞内的pH显得尤为重要。
通常会采用玻璃电极来测定pH值,但在测定过程中会存在一些电化学干扰,不规范的操作极易导致电极污染影响测量结果的准确度,有一定的适用寿命,而且玻璃电极对于极端的pH并不能精确测量。对于活体pH的监测而言,玻璃电极会造成机械损伤,所以并不适用于活体细胞内pH的监控和检测。众所周知,一些有机荧光基团的荧光或吸光性质会随pH的变化而不同,这种在不同酸碱条件下呈现出的不同的光学信号可以用来作为测定pH的有效手段,利用光学信号而构建的荧光测定pH的方法,可以利用荧光参数如荧光发射波长,荧光强度的变化来测定pH值,灵敏度更高,不仅便于荧光显微学研究,而且可实时检测细胞内pH的动态分布和区域变化。目前可用于检测pH的荧光材料已有大量报道,但其中大多数都只能对或酸或碱小范围pH值进行测定,很少有材料能够实现由酸性到中性再到碱性的宽范围pH的测定。因此,设计和制备宽范围的pH响应型荧光传感材料仍然是目前研究的重要方向之一。
发明内容
本发明考虑到玻璃电极测定pH的不足,目的在于提供一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料及其制备方法和应用,能够很好的对水体及生物细胞内的pH进行测定。具有成本低,合成简单,响应灵敏,测定范围宽,抗干扰性强等特点。
本发明采用的技术方案是:
本发明首先提供一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料,所述材料是一个有机分子化合物,呈浅褐色粉末状,将其溶解在HEPES缓冲溶液(0.05M, pH=7.0)/甲醇体系中,溶液呈现无色,在365nm的紫外灯照射溶液体系无荧光发射。
本发明还提供一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料的制备方法,其特征在于,包括如下步骤:
S1.罗丹明B酰肼的制备:
参考文献Tang X, Han J, Wang Y, et al. A multifunctional Schiff base as afluorescence sensor for Fe3+ and Zn2+ ions, and a colorimetric sensor for Cu2+and applications[J]. Spectrochimica Acta Part A: Molecular and BiomolecularSpectroscopy, 2017, 173: 721-726进行制备。
将罗丹明B完全溶解在甲醇中,边搅拌边滴加85%的水合肼,滴加完毕后置于油浴锅中加热搅拌回流,待反应完成后,冷却至室温,减压移除溶剂得粗产品,在甲醇中重结晶进行纯化。
上述合成方法中:罗丹明B,85%的水合肼,甲醇的用量分别为1~2g(2~4mmol),2.5~5mL,20~40mL,所述的回流反应温度为60~70℃,反应时间为8~12h。
S2. 3-甲酰基-4-羟基联苯氰的制备:
参考文献Chen S, Li H, Hou P. A novel cyanobiphenyl benzothiazole-basedfluorescent probe for detection of biothiols with a large Stokes shift andits application in cell imaging[J]. Tetrahedron, 2017, 73(5): 589-593进行制备。
将对氰基联苯酚和乌洛托品用三氟乙酸完全溶解并置于圆底烧瓶中,在油浴锅中加热搅拌回流。待反应完成后将体系冷却至室温,向体系中加入1.0 M 的稀HCl进行酸化,然后用二氯甲烷对产品进行萃取,收集二氯甲烷相并用蒸馏水洗涤三次,并用无水硫酸镁进行干燥,过滤后减压移除溶剂得3-甲酰基-4-羟基联苯氰。
上述合成方法中,对氰基联苯酚,乌洛托品和三氟乙酸的用量分别为0.39~1.17 g(2~6 mmol), 1.41g ~4.23g (10~30 mmol), 30~70 mL, 回流反应温度为100~120 ℃,反应时间为4~8 h。
S3. 制备基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料:将罗丹明B酰肼和3-甲酰基-4-羟基联苯氰用无水乙醇将其溶解,置于圆底烧瓶中,滴加2滴冰乙酸作为催化剂,油浴锅中加热搅拌回流,待反应结束后冷却至室温,减压旋蒸移除溶剂得粗产品,在乙醇中重结晶纯化得浅褐色固体。
上述合成方法中,罗丹明B酰肼、3-甲酰基-4-羟基联苯氰和无水乙醇的用量分别为0.468~1.404g (1~3mmol),0.227~0.681g ( 1~3mmol),20~50mL, 回流反应温度为65~75℃,反应时间为8~12h。
本发明还提供一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料用于水环境中pH检测的用途。
本发明还提供一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料用于生物体细胞中pH的实时检测分析。
本发明的技术效果为:
(1)本发明提供了一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料、其制备方法及应用。其中罗丹明B作为一种具有良好光学性质的荧光染料自身结构具有独特的内酰胺环,在酸性条件下,内酰胺环处于开环状态,溶液体系会因内酰胺环的打开而发射荧光,外观颜色也由无色变为粉红色。对氰基联苯酚结构中的酚羟基在碱性条件下会脱氢而且极易发生异构化,同时其芳香环共轭体系又会诱发荧光的发射。采用水合肼作为桥梁将两者结合形成一个稳定的席夫碱化合物,长时间放置自身不会发生分解,结构中的罗丹明B和对氰基联苯酚基团在酸碱条件下所呈现出的不同光学性质为测定pH提供了前提。
(2)本发明中的荧光传感材料的合成制备简单,条件温和易于控制,制备成本低廉,制备过程中的加料量是基于大量实验研究而确定的最佳的反应摩尔比。反应温度和时间是基于反应速率和产率来确定的。在最佳的参数范围内,合成产率高达70%以上。
(3)本发明制备的荧光传感材料对pH 具有宽范围的双响应性,在水环境处于中性条件下,加入荧光传感材料溶液,体系呈现无色无荧光;在酸性条件下(pH<6), 溶液体系颜色变为粉红色并伴随着橙红色荧光发射,颜色或荧光会随着pH值的减小而加深和增强;在碱性条件下(pH>8),溶液体系颜色会变为黄色并伴随着黄绿色荧光发射,同样的颜色或荧光会随着pH值的增大而加深和增强,呈现出的光学信号肉眼可见,利用荧光风光光度计和紫外分光光度计对不同波长下的荧光强度和紫外吸收进行测定,基于pH和光学参数之间的线性关系即可实现对pH 的准确测定。相比传统的玻璃电极等测定pH方法而言,灵敏度更高,不存在电化学干扰,更具有应用价值。
附图说明
图1为实施例3所制备的基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料的合成过程示意图;
图2为实施例3所制备的pH响应型荧光传感材料的1H NMR,其中溶剂为DMSO-D6;
图3为实施例3所制备的pH响应型荧光传感材料的13C NMR图,其中溶剂为DMSO-D6;
图4为实施例3所制备pH响应型荧光传感材料的MS图;
图5为实施例3所制备pH响应型荧光传感材料在不同的pH条件水体环境中的荧光光谱图,激发波长为500nm,狭缝宽度均为5nm;
图6为实施例3所制备pH响应型荧光传感材料在不同的pH条件下水体环境中的紫外可见吸收光谱;
图7为实施例3所制备pH响应型荧光传感材料在对应发射波长下的荧光强度与水体pH之间的线性关系图;
图8为实施例3所制备荧光传感材料用于生物细胞中pH的荧光成像图。
具体实施方式
为使本领域技术人员更好的理解本发明的技术方案,下面将结合附图说明对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例是本发明一部分实施例,而不是全部的实施例,基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动的前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1:
S1.罗丹明B酰肼的合成:将1g (2mmol)罗丹明B完全溶解在20mL甲醇中,边搅拌边滴加2 mL 85%的水合肼,滴加完毕后置于60℃油浴锅中加热搅拌回流8h,待反应完成后,冷却至室温,减压移除溶剂得粗产品,在甲醇中重结晶进行纯化。
S2. 3-甲酰基-4-羟基联苯氰的制备:将0.39g(2mmol)对氰基联苯酚和1.41g(10mmol)乌洛托品用30 mL三氟乙酸完全溶解并置于圆底烧瓶中,100℃油浴锅中加热搅拌回流4h。待反应完成后将体系冷却至室温,向体系中加入100mL 1.0 M 的稀HCl进行酸化,然后用二氯甲烷对产品进行萃取,收集二氯甲烷相并用蒸馏水洗涤三次,并用无水硫酸镁进行干燥,过滤后减压移除溶剂得3-甲酰基-4-羟基联苯氰。
S3. 制备基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料:将0.468g(1mmol) 罗丹明B酰肼和0.227g (1mmol) 3-甲酰基-4-羟基联苯氰用20mL无水乙醇将其溶解,置于圆底烧瓶中,滴加2滴冰乙酸作为催化剂,在65℃油浴锅中搅拌回流8h,待反应结束后冷却至室温,减压旋蒸移除溶剂得粗产品,在乙醇中重结晶纯化得浅褐色固体。
实施例2:
S1. 罗丹明B酰肼的合成:将1.5g (3mmol) 罗丹明B完全溶解在30mL甲醇中,边搅拌边滴加3.7 mL 85%的水合肼,滴加完毕后置于65℃油浴锅中加热搅拌回流10h,待反应完成后,冷却至室温,减压移除溶剂得粗产品,在甲醇中重结晶进行纯化。
S2. 3-甲酰基-4-羟基联苯氰的制备:将0.78g(4mmol)对氰基联苯酚和2.82g(20mmol)乌洛托品用50 mL三氟乙酸完全溶解并置于圆底烧瓶中,110℃油浴锅中加热搅拌回流6h。待反应完成后将体系冷却至室温,向体系中加入100mL 1.0 M 的稀HCl进行酸化,然后用二氯甲烷对产品进行萃取,收集二氯甲烷相并用蒸馏水洗涤三次,并用无水硫酸镁进行干燥,过滤后减压移除溶剂得3-甲酰基-4-羟基联苯氰。
S3. 制备基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料:将0.936g(2mmol)罗丹明B酰肼和0.454g (2mmol) 3-甲酰基-4-羟基联苯氰用35mL无水乙醇将其溶解,置于圆底烧瓶中,滴加2滴冰乙酸作为催化剂,70℃油浴锅中加热搅拌回流10h,待反应结束后冷却至室温,减压旋蒸移除溶剂得粗产品,在乙醇中重结晶纯化得浅褐色固体。
实施例3:
S1. 罗丹明B酰肼的合成:将2g (4mmol) 罗丹明B完全溶解在40mL甲醇中,边搅拌边滴加5 mL 85%的水合肼,滴加完毕后置于70℃油浴锅中加热搅拌回流12h,待反应完成后,冷却至室温,减压移除溶剂得粗产品,在甲醇中重结晶进行纯化。
S2. 3-甲酰基-4-羟基联苯氰的制备:将1.17 g(6mmol)对氰基联苯酚和4.23g(30mmol)乌洛托品用70 mL三氟乙酸完全溶解并置于圆底烧瓶中,120℃油浴锅中加热搅拌回流8h。待反应完成后将体系冷却至室温,向体系中加入100mL 1.0 M 的稀HCl进行酸化,然后用二氯甲烷对产品进行萃取,收集二氯甲烷相并用蒸馏水洗涤三次,并用无水硫酸镁进行干燥,过滤后减压移除溶剂得3-甲酰基-4-羟基联苯氰。
S3. 制备基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料:将1.404g(3mmol)罗丹明B酰肼和0.681g (3mmol) 3-甲酰基-4-羟基联苯氰用50mL无水乙醇将其溶解,置于圆底烧瓶中,滴加2滴冰乙酸作为催化剂,70℃油浴锅中加热搅拌回流10h,待反应结束后冷却至室温,减压旋蒸移除溶剂得粗产品,在乙醇中重结晶纯化得浅褐色固体。
如图1所示是基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料的合成过程示意图。
如图2所示为基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料的1H NMR 图,其中溶剂为CDCl3。光谱解析:1H NMR (400 MHz, CDCl3) δ 8.06 – 7.98 (m, 1H), 7.73(d, J = 5.5 Hz, 1H), 7.68 (d, J = 8.2 Hz, 2H), 7.58 (d, J = 8.5 Hz, 2H), 7.55(s, 1H),7.53 (dd, J = 7.9, 6.2 Hz, 1H), 7.44(m, 1H), 7.34 (t, J = 7.0 Hz,1H), 7.20 (dd, J = 15.4, 7.3 Hz, 1H), 6.98 (d, J = 8.6 Hz, 1H), 6.50 (m, 4H),6.28 (m, 2H), 3.35 (dd, J = 13.7, 6.8 Hz, 8H), 1.40 – 0.95 (m, 12H)。如图3所示为基于罗丹明B衍生物和氰基联苯酚的多重响应型荧光传感材料的13C NMR图,其中溶剂为CDCl3, 光谱解析:13C NMR (101 MHz, CDCl3) δ 164.38, 159.27, 153.65, 153.46,152.21, 150.67, 149.18, 149.05, 144.75, 132.58, 132.41, 129.92, 129.79,128.68, 128.10, 127.90, 126.94, 124.25, 123.41, 119.17,117.92, 108.24,108.09, 105.37, 98.18, 98.06, 66.65, 44.36, 12.59。
通过核磁谱图解析可以确定制备合成的pH响应型荧光传感材料分子结构与图1中预期的结构是一致的。
如图4所示为荧光传感材料(C42H39N5O3, Mn=661.782)的质谱图,其中,662.61为[M+1]对应的荧光传感材料的分子量。
实施例4:本发明制备的荧光传感材料对不同pH的水体的酸碱度测定验证
将实施例3制备的荧光传感材料用甲醇配制成10mM的储备液待用。用蒸馏水分别配制不同pH (pH=1,2,3,4,5,6,7,8,9,10,11,12,13,14)的水溶液,其中采用 0.5M的氢氧化钠和0.5M的盐酸来进行调节。移取0.05mL的荧光传感材料储备液置于5mL的上述配好的不同pH 水溶液体系中,混合均匀,采用荧光分光光度计和紫外可见分光光度计对溶液体系的荧光发射光谱和吸收光谱进行测定。
图5为不同pH水溶液体系在加入荧光传感材料溶液后的荧光光谱图,由图可以看出,当溶液体系pH<7时,其荧光发射光谱在590nm处出现了强的荧光发射峰并伴随着橙红色荧光发射,对应的荧光强度随着pH值的减小而增强;当溶液体系pH>7时,在558nm处出现一强的发射峰并伴随着黄绿色荧光,荧光强度随着pH值的增大而增强;当水溶液体系pH=7处于中性时,其发射光谱并没有出现发射峰。
图6为不同pH水溶液体系在加入荧光传感材料溶液后的紫外吸收光谱,不同的pH体系不仅会引起荧光传感材料发射不同的荧光信号,溶液的外观颜色也随pH 不同而不同。当pH<7时,溶液颜色呈粉红色,在557nm处出现了一个吸收峰,pH值越小,吸收峰越强;当pH>7时,溶液颜色呈现黄色,557nm处的吸收峰消失,但却在358nm处出现了一个新吸收,随着pH值增大吸收峰逐渐增强。
实施例5:本发明制备的荧光传感材料呈现出的荧光发射强度与水体中pH的线性验证
根据实施4中测得的荧光传感材料对不同pH 水溶液体系的荧光发射光谱,将相应的荧光发射强度与对应的pH值进行作图,分析两者之间的关系。
图7a为水体pH<7时,加入荧光传感材料后体系荧光发射光谱中对应发射波长为590nm处的荧光强度与pH值之间的线性关系,得到的线性方程为:Y=862.75378-119.75622X,R2=0.991;图7b为水体pH>7时,加入荧光传感材料后体系荧光发射光谱中对应发射波长为559nm处的荧光强度与pH值之间的线性关系,得到的线性方程为:Y=122.04812X-954.12892,R2=0.996;可以看出荧光强度与pH之间存在良好的线性关系,这说明本发明制备的荧光传感材料可以通过光学参数荧光强度来对水体pH进行测定。
实施例6:本发明制备的荧光传感材料对细胞中的pH成像分析
将人B淋巴瘤细胞(购买于Sigma-aldrich西格玛奥德里奇(上海)贸易有限公司)在RPMI-1640培养液中在37℃的培养箱中进行培养24h,然后加入50μM的实例3中制备的荧光传感材料继续在37℃条件下培养50min,之后用PBS缓冲溶液洗涤三次移除未进入细胞内部残余的荧光传感材料,然后该细胞分别用pH=5,pH=6,pH=7,pH=8,pH=9的PBS缓冲溶液培养30min, 然后细胞荧光成像被获得通过荧光倒置显微镜,其中激发光分别采用470–545 nm(绿)和545–650 nm(红)两个波段。
不同pH条件下,荧光传感材料对细胞的成像如图8所示,不难看出,当细胞内pH<7时,细胞成像以红色荧光为主,采用470–545 nm(绿)激发时,几乎看不到荧光成像;当细胞内pH>7时,成像以绿色荧光为主,此时采用545–650 nm(红)激发时,同样看不到细胞荧光成像;细胞pH处于中性时,两个波段都很难看到荧光成像;结果与水体中pH检测的现象是一致的,这说明本发明制备的荧光传感材料能够用于生物细胞中pH的实时监控。

Claims (9)

1.一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料,其特征在于,所述材料为浅褐色粉末状固体,溶解在pH=7.0 (0.5M, HEPES缓冲溶液)的甲醇/水混合溶液时,溶液体系无色无荧光发射。
2.一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料的制备方法,其特征在于,包括如下步骤:
S1. 罗丹明B酰肼的合成;
S2. 3-甲酰基-4-羟基-4-联苯基氰的制备;
S3.制备基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料:
将罗丹明B酰肼和3-甲酰基-4-羟基联苯氰用无水乙醇将其溶解,置于圆底烧瓶中,滴加冰乙酸作为催化剂,油浴锅中加热搅拌回流,待反应结束后冷却至室温,减压旋蒸移除溶剂得粗产品,在乙醇中重结晶纯化得浅褐色固体。
3.根据权利要求2所述的一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料的制备方法,其特征在于,步骤S3中所述罗丹明B酰肼和3-甲酰基-4-羟基联苯氰的质量比为0.468~1.404g:0.227~0.681g。
4.根据权利要求2所述的一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料的制备方法,其特征在于,步骤S3中所述的无水乙醇用量为:20~50mL。
5.根据权利要求2所述的一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料的制备方法,其特征在于,步骤S3中所述回流反应温度为65~75℃。
6.根据权利要求2所述的一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料的制备方法,其特征在于,步骤S3中所述回流反应时间为8~12 h。
7.一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料对pH检测的应用。
8.根据权利要求7所述的应用,其特征在于,所述应用为对水体中pH的灵敏检测。
9.根据权利要求7所述的应用,其特征在于,所述应用为一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料对生物细胞中pH的实时监测分析。
CN201710699740.2A 2017-08-16 2017-08-16 一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料及其制备方法和应用 Expired - Fee Related CN107698600B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710699740.2A CN107698600B (zh) 2017-08-16 2017-08-16 一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料及其制备方法和应用

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710699740.2A CN107698600B (zh) 2017-08-16 2017-08-16 一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料及其制备方法和应用

Publications (2)

Publication Number Publication Date
CN107698600A true CN107698600A (zh) 2018-02-16
CN107698600B CN107698600B (zh) 2020-02-21

Family

ID=61171037

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710699740.2A Expired - Fee Related CN107698600B (zh) 2017-08-16 2017-08-16 一种基于罗丹明B和氰基联苯酚的pH响应型荧光传感材料及其制备方法和应用

Country Status (1)

Country Link
CN (1) CN107698600B (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108892672A (zh) * 2018-08-28 2018-11-27 安徽工业大学 一种用于pH荧光传感的罗丹明6G席夫碱衍生物的制备方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104341347A (zh) * 2014-10-21 2015-02-11 苏州福来兹检测科技有限公司 一种用于制备广泛pH荧光探针的有机化合物及其应用
CN106496067A (zh) * 2015-09-07 2017-03-15 香港科技大学深圳研究院 基于间位链接的联肼基吖嗪结构化合物及其制备方法和应用
CN106588966A (zh) * 2016-11-14 2017-04-26 济南大学 一种开型过氧化氢荧光探针化合物的制备与应用
CN106966924A (zh) * 2017-04-17 2017-07-21 济南大学 一种基于席夫碱的锌离子荧光探针的制备与应用

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104341347A (zh) * 2014-10-21 2015-02-11 苏州福来兹检测科技有限公司 一种用于制备广泛pH荧光探针的有机化合物及其应用
CN106496067A (zh) * 2015-09-07 2017-03-15 香港科技大学深圳研究院 基于间位链接的联肼基吖嗪结构化合物及其制备方法和应用
CN106588966A (zh) * 2016-11-14 2017-04-26 济南大学 一种开型过氧化氢荧光探针化合物的制备与应用
CN106966924A (zh) * 2017-04-17 2017-07-21 济南大学 一种基于席夫碱的锌离子荧光探针的制备与应用

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
SONG CHEN ET AL.: "A novel cyanobiphenyl benzothiazole-based fluorescent probe for detection of biothiols with a large Stokes shift and its application in cell imaging", 《TETRAHEDRON》 *
谭甲莲: "新型罗丹明类pH荧光探针的设计及其生物应用", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108892672A (zh) * 2018-08-28 2018-11-27 安徽工业大学 一种用于pH荧光传感的罗丹明6G席夫碱衍生物的制备方法
CN108892672B (zh) * 2018-08-28 2021-05-28 安徽工业大学 一种用于pH荧光传感的罗丹明6G席夫碱衍生物的制备方法

Also Published As

Publication number Publication date
CN107698600B (zh) 2020-02-21

Similar Documents

Publication Publication Date Title
Li et al. A ratiometric fluorescence probe for lysosomal polarity
Zhang et al. A ratiometric lysosomal pH probe based on the coumarin–rhodamine FRET system
Zhang et al. A ratiometric lysosomal pH probe based on the naphthalimide–rhodamine system
Fang et al. Fluorescent probes for sensitive and selective detection of pH changes in live cells in visible and near-infrared channels
US11505530B2 (en) 1,8-naphthalimide derivative, preparation method therefor and use thereof
Liu et al. The ESIPT fluorescent probes for N2H4 based on benzothiazol and their applications for gas sensing and bioimaging
Vegesna et al. pH-activatable near-infrared fluorescent probes for detection of lysosomal pH inside living cells
Dong et al. A series of highly sensitive and selective fluorescent and colorimetric “off-on” chemosensors for Cu (II) based on rhodamine derivatives
Shen et al. A rhodamine B-based probe for the detection of HOCl in lysosomes
Yang et al. A rhodamine-based fluorescent probe selective for bisulfite anion in aqueous ethanol media
Lv et al. Highly selective and sensitive pH-responsive fluorescent probe in living Hela and HUVEC cells
Liu et al. A super-sensitive ratiometric fluorescent probe for monitoring intracellular subtle pH fluctuation
Mukherjee et al. A novel pyrrole fused coumarin based highly sensitive and selective fluorescence chemosensor for detection of Cu2+ ions and applications towards live cell imaging
Li et al. A near-infrared fluorescent probe for Cu2+ in living cells based on coordination effect
Hua et al. A novel turn off fluorescent sensor for Fe (III) and pH environment based on coumarin derivatives: the fluorescence characteristics and theoretical study
Wang et al. 5-Hydroxymethylfurfural modified rhodamine B dual-function derivative: Highly sensitive and selective optical detection of pH and Cu2+
Sen et al. A water soluble FRET-based ratiometric chemosensor for Hg (ii) and S 2− applicable in living cell staining
Fan et al. Novel colorimetric and fluorescent off–on enantiomers with high selectivity for Fe3+ imaging in living cells
Leng et al. New rhodamine B-based chromo-fluorogenic probes for highly selective detection of aluminium (III) ions and their application in living cell imaging
Jin et al. Novel near-infrared pH-sensitive cyanine-based fluorescent probes for intracellular pH monitoring
Wu et al. A red-to-near-infrared fluorescent probe for the detection of thiophenol based on a novel hydroxylflavone-quinoline-amino molecular system with large Stokes shift
CN103555317A (zh) 一种pH敏感近红外荧光分子探针及其制备方法与用途
CN103320120A (zh) 一种含半胱氨酸乙酯结构的罗丹明B类靶向溶酶体pH荧光探针及应用
KR101472318B1 (ko) 넓은 범위의 pH 측정이 가능한 비율계량적 pH 프로브
Jiao et al. A fluorescein derivative-based fluorescent sensor for selective recognition of copper (II) ions

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20200221

Termination date: 20200816

CF01 Termination of patent right due to non-payment of annual fee