CN106188074B - 一类水溶性铁离子荧光探针及其制备方法 - Google Patents

一类水溶性铁离子荧光探针及其制备方法 Download PDF

Info

Publication number
CN106188074B
CN106188074B CN201610528177.8A CN201610528177A CN106188074B CN 106188074 B CN106188074 B CN 106188074B CN 201610528177 A CN201610528177 A CN 201610528177A CN 106188074 B CN106188074 B CN 106188074B
Authority
CN
China
Prior art keywords
water
iron ion
fluorescence probe
soluble iron
precipitate
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.)
Active
Application number
CN201610528177.8A
Other languages
English (en)
Other versions
CN106188074A (zh
Inventor
祁秀秀
陈文华
陈柏
刘巧云
李耀中
刘文斌
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.)
Changzhou Vocational Institute of Engineering
Original Assignee
Changzhou Vocational Institute of Engineering
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 Changzhou Vocational Institute of Engineering filed Critical Changzhou Vocational Institute of Engineering
Priority to CN201610528177.8A priority Critical patent/CN106188074B/zh
Publication of CN106188074A publication Critical patent/CN106188074A/zh
Application granted granted Critical
Publication of CN106188074B publication Critical patent/CN106188074B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic 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
    • C07D491/02—Heterocyclic 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
    • C07D491/10—Spiro-condensed systems
    • C07D491/107—Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/34—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from hydroxy compounds or their metallic derivatives
    • C08G65/48—Polymers modified by chemical after-treatment
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells

Landscapes

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

Abstract

本发明涉及一类水溶性铁离子荧光探针，该荧光探针分子具有如下图所示结构。本发明还公开了所述水溶性铁离子荧光探针的制备方法。

Description

一类水溶性铁离子荧光探针及其制备方法
技术领域
本发明属于离子探针新材料领域。具体涉及一类OFF-ON型的三价铁离子荧光探针及其制备方法，该探针可用于全水体系中的三价铁离子的测定。
背景技术
铁离子是生命体必需的离子之一，在机体中起到运输和贮存氧的作用，有极其重要的生理意义。另一方面，铁虽然是机体必需的元素，但是在人体内铁过量，则会产生过量的氧自由基，从而对机体造成损害。因此，开发性能优异的Fe3+荧光探针是很有必要的，以便于实时、在线动态的检测生物体或环境中Fe3+的含量和分布。目前大多数的铁离子荧光探针水溶性较差，需要加入一定比例的有机溶剂，很大程度上限制其应用，尤其限制了在含有活生物体的水性环境中的使用，因为生物体内缺乏这样的有机溶剂。若另加有机溶剂，因为有机溶剂往往具有毒性，会对活生物体造成损害。
因此，急需开发一类能在全水环境下使用的水溶性铁离子荧光探针。
发明内容
本发明的目的是提供一类新的OFF-ON型的罗丹明衍生物作为水溶性铁离子荧光探针的合成方法。这类方法得到可溶性的罗丹明衍生物，在全水环境中检测铁离子方面具有很好的应用前景。
本发明第一方面涉及一类水溶性铁离子荧光探针，该铁离子荧光探针分子具有如下结构：
本发明第二方面涉及所述水溶性铁离子荧光探针的制备方法，包括以下步骤：
(1)以乙醇作为溶剂，加入罗丹明6G至浓度为0.02mol/L～0.2mol/L，加入1,3-丙二胺与之进行反应，其中1,3-丙二胺与罗丹明6G的摩尔比＝0.5:1～10:1，，反应温度控制在40～80℃，反应4～9小时。冷却析出沉淀，用乙腈重结晶得到白色晶体A；
(2)以二氯甲烷为溶剂，加入聚乙二醇单甲醚至浓度为0.025mol/L～0.25mol/L、按与聚乙二醇单甲醚摩尔比为10:1～1:1的量加入三乙胺，按与聚乙二醇单甲醚摩尔比为10:1～1:1的量滴加氯乙酰氯，在黑暗中搅拌过夜，在乙醚中沉淀出产物，将产物溶于水，调节pH值至6，用20ml二氯甲烷萃取3次，加乙醚洗出沉淀B；
.(3)以二氯甲烷为溶剂，按摩尔比为0.2:1～1:0.2的量加入前述白色晶体A和沉淀B，并按照与白色晶体A的摩尔比为0.1:1～1:0.1的量加入KI，并加入碳酸钾0.002mol/L～0.2mol/L，反应2～48h，过滤，滤液加到乙醚中析出沉淀C，该沉淀C即为所述水溶性铁离子荧光探针。
上述制备步骤涉及的反应图示如下：
本发明的有益效果：
1、本发明的水溶性铁离子荧光探针具有良好的水溶性，使用时不需要加入有机溶剂，这使得其可以直接用于对含有活生命体的水性环境中的铁离子进行测定。
2、本发明的荧光离子探针对铁离子具有优异的选择性，对Fe3+、Fe2+、Cu2+、Zn2+、Ni2 +、Ca2+、Cd2+、Mg2+、Pb2+、Hg2+、K+、Na+等常见金属离子具有很好的抗干扰能力。
3、本发明的荧光离子探针对铁离子的检测可以通过肉眼观察，不需要复杂的仪器。
4、本发明的荧光离子探针能够穿透细胞膜，与细胞内的Fe3+结合，可用于监测活细胞内部环境中的Fe3+。
附图说明
图1为在本发明的水溶性铁离子荧光探针的水溶液中加入不同金属离子后溶液的荧光发射光谱(激发波长为500nm)。
图2为不同光线下(左图日光下，右图紫外灯下)两个样品瓶的照片(两瓶中均为本发明的水溶性铁离子荧光探针的水溶液，左瓶中未加入Fe3+，右瓶中加入了Fe3+)。
图3为用含不同样品的培养基培养5小时后HeLa细胞的共聚焦荧光照片。其中，第一行照片为仅用本发明的水溶性铁离子荧光探针培养过的细胞，第二行照片为先后用Fe3+和本发明的水溶性铁离子荧光探针培养过的细胞。第一列照片为荧光照片，第二列为明场照片，第三列为前两列叠加后的照片。
具体实施方式
实施例1铁离子荧光探针化合物C的制备方法步骤如下：
罗丹明6G衍生物(A)的合成：罗丹明6G 2mmol溶于20ml热乙醇中，逐滴加入1,3-丙二胺，加热回流6小时。溶液冷却至室温，析出固体，用乙腈进行重结晶，得到白色晶体A。(1H-NMR(CDCl3):δ7.95(d,1H),7.47(t,2H),7.05(d,1H),6.34(s,2H),6.23(s,2H),3.50(t,2H),3.24(t,4H),2.39(t,2H),1.90(s,6H),1.36(t,6H),1.19(m,2H))
甲氧基氯乙酸聚乙二醇酯(B)的合成：聚乙二醇单甲醚(例如MPEG2000，其分子量为2000左右，当然，也可以采用其它分子量的MPEG)10g，5mmol，溶于无水二氯甲烷20ml，三乙胺2.09ml，15mmol，在N2氛下逐滴加入氯乙酰氯7.95ml，100mmol，在黑暗中搅拌过夜，在乙醚中沉淀出产物，将产物溶于水，调节pH值至6，用20ml二氯甲烷萃取3次，加乙醚析出沉淀B。(1H-NMR(CDCl3)(4.20(COOCH2),4.15(CH2Cl),3.3–3.7(OCH2CH2O))
荧光探针(C)的合成：罗丹明6G衍生物10mg，0.02mmol，KI 4.4mg，0.026mmol，碳酸钾15mg，0.11mmol，PEG-10mg，0.005mmol，二氯甲烷5ml，N2下反应10h，过滤，滤液加到乙醚中析出沉淀C。(1H-NMR(CDCl3)(δ7.95(COCCH),7.47(CHCHCH),7.05(CHCHC),6.34(CCHCC),6.23(CCHCO),4.20(COOCH2).4.15(CH2Cl),3.3–3.7(OCH2CH2O,NCH2,OCH3),3.24(CH2CH3),2.39(NHCH2),1.90(CCH3),1.36(CH2CH3),1.19(NCH2CH2))
实施例2铁离子荧光探针化合物性能实验
1)铁离子探针的选择性
在平行的几组pH值为7.0的水中，分别加入上述沉淀C，依次在各组中加入Fe3+、Fe2 +、Cu2+、Zn2+、Ni2+、Ca2+、Cd2+、Mg2+、Pb2+、Hg2+、K+、Na+，各5μM,最后一组不加作为对照组。测各组的荧光发射光谱(激发波长为500nm)。发现与不加离子的对照组相比，加入Fe3+之后，荧光强度显著增强；而加入其他离子之后溶液荧光强度基本不变(如图1所示)。结果表明该沉淀C对Fe3+具有优异的选择性。
2)铁离子荧光探针对铁离子的检测
如图2左图所示，日光下左边的样品瓶中加入沉淀C的水样，右边样品瓶中的水样加入沉淀C和Fe3+，用紫外灯照射两个样品瓶就得到右图的照片。由此可见，沉淀C在水中为无色澄清透明状，证明其水溶性比一般的基于罗丹明结构的荧光探针水溶性好。另一方面，沉淀C在水中碰到Fe3+即由无色变为肉粉色，颜色变化非常明显，肉眼可见。在紫外灯照射下发出明亮的黄色荧光。由此可见，用沉淀C检测Fe3+可以不用借助仪器来判断，肉眼即可。
3)铁离子探针的细胞实验
在37℃恒温培养箱中用培养基DMEM培养HeLa细胞5小时，一组在培养基中加Fe3+(10μM)，一组不加Fe3+作为对照组。然后分别用pH为7.4的磷酸缓冲溶液洗去未进入细胞的Fe3+。将两组细胞分别转移到加入C(10μM)的DMEM中继续培养。30分钟后细胞用磷酸缓冲溶液润洗，然后分别用共聚焦荧光显微镜观察。两组的荧光成像如图3所示。发现对照组(图3第一行照片)即未事先用Fe3+培养的细胞几乎没有观察到荧光，而用含Fe3+培养基培养过的细胞(见图3第二行照片)有荧光。结果显示本发明的铁离子荧光探针能够穿透HeLa细胞膜与细胞内的Fe3+结合。这种对细胞膜的穿透能力以及由于与Fe3+结合而导致荧光增强的性能对于监测细胞内Fe3+的浓度研究活细胞中的毒性或生物活性将起到很重要的作用。

Claims (2)

1.一类水溶性铁离子荧光探针，其特征在于具有如下结构：
其中n为正整数。
2.根据权利要求1所述的水溶性铁离子荧光探针的制备方法，其特征在于包括以下步骤：
(1)以乙醇作为溶剂，加入罗丹明6G至浓度为0.02mol/L～0.2mol/L，加入1,3-丙二胺与之进行反应，其中1,3-丙二胺与罗丹明6G的摩尔比＝0.5:1～10:1，反应温度控制在40～80℃，反应4～9小时，冷却析出沉淀，用乙腈重结晶得到白色晶体A；
(2)以二氯甲烷为溶剂，加入聚乙二醇单甲醚至浓度为0.025mol/L～0.25mol/L、按与聚乙二醇单甲醚摩尔比为10:1～1:1的量加入三乙胺，按与聚乙二醇单甲醚摩尔比为10:1～1:1的量滴加氯乙酰氯，在黑暗中搅拌过夜，在乙醚中沉淀出产物，将产物溶于水，调节pH值至6，用20ml二氯甲烷萃取3次，加乙醚洗出沉淀B；
(3)以二氯甲烷为溶剂，按摩尔比为0.2:1～1:0.2的量加入前述白色晶体A和沉淀B，并按照与白色晶体A的摩尔比为0.1:1～1:0.1的量加入KI，并加入碳酸钾0.002mol/L～0.2mol/L，反应2～48h，过滤，滤液加到乙醚中析出沉淀C，该沉淀C即为所述水溶性铁离子荧光探针。
CN201610528177.8A 2016-07-06 2016-07-06 一类水溶性铁离子荧光探针及其制备方法 Active CN106188074B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610528177.8A CN106188074B (zh) 2016-07-06 2016-07-06 一类水溶性铁离子荧光探针及其制备方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610528177.8A CN106188074B (zh) 2016-07-06 2016-07-06 一类水溶性铁离子荧光探针及其制备方法

Publications (2)

Publication Number Publication Date
CN106188074A CN106188074A (zh) 2016-12-07
CN106188074B true CN106188074B (zh) 2018-01-16

Family

ID=57465574

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610528177.8A Active CN106188074B (zh) 2016-07-06 2016-07-06 一类水溶性铁离子荧光探针及其制备方法

Country Status (1)

Country Link
CN (1) CN106188074B (zh)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107727620B (zh) * 2017-09-26 2020-04-07 盐城师范学院 一种检测亚硝酸根离子的试剂盒及其应用
CN108440547B (zh) * 2018-03-31 2021-07-27 浙江工业大学 一种罗丹明6g席夫碱类荧光探针及其制备与应用
CN108285454A (zh) * 2018-04-26 2018-07-17 福建师范大学泉港石化研究院 一种含氨基酸的Fe3+荧光探针及其合成方法
CN108676307B (zh) * 2018-05-17 2020-07-31 江苏大学 基于两亲性温敏嵌段共聚物混合化学传感器的制备方法及其应用
CN108997363B (zh) * 2018-07-20 2020-12-25 山西大学 一种基于罗丹明b的衍生物及其制备方法和应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101135644A (zh) * 2007-08-16 2008-03-05 南京大学 一种汞离子荧光显色剂及检测方法、测定试纸及其用途
CN103320119A (zh) * 2013-06-05 2013-09-25 上海电力学院 一种具有荧光识别功能的碳钢缓蚀剂及其应用

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101135644A (zh) * 2007-08-16 2008-03-05 南京大学 一种汞离子荧光显色剂及检测方法、测定试纸及其用途
CN103320119A (zh) * 2013-06-05 2013-09-25 上海电力学院 一种具有荧光识别功能的碳钢缓蚀剂及其应用

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Selective Detection of Iron(III) by Rhodamine-Modified Fe3O4 Nanoparticles;Baodui Wang,et al.,;《Angew. Chem. Int. Ed.》;20100520;第49卷;第4576-4579页 *
基于罗丹明衍生物的Fe(Ⅲ) 荧光探针的合成及其性能研究;成春文,等，;《化学试剂》;20151231;第37卷(第3期);第255-258页 *

Also Published As

Publication number Publication date
CN106188074A (zh) 2016-12-07

Similar Documents

Publication Publication Date Title
CN106188074B (zh) 一类水溶性铁离子荧光探针及其制备方法
Cheng et al. “Reactive” probe for hydrogen sulfite: good ratiometric response and bioimaging application
Tian et al. A series of naphthalimide derivatives as intra and extracellular pH sensors
Du et al. A low cytotoxic and ratiometric fluorescent nanosensor based on carbon-dots for intracellular pH sensing and mapping
Zhao et al. Fluorescent/phosphorescent dual-emissive conjugated polymer dots for hypoxia bioimaging
US10351707B2 (en) Water-soluble iron ion fluorescent probe and preparation method thereof
Zhang et al. A ratiometric lysosomal pH probe based on the coumarin–rhodamine FRET system
Fan et al. An ICT-based ratiometric probe for hydrazine and its application in live cells
Hu et al. A FRET approach for luminescence sensing Cr3+ in aqueous solution and living cells through functionalizing glutathione and glucose moieties
Luo et al. Highly sensitive and selective turn-on fluorescent chemosensor for Hg2+ in pure water based on a rhodamine containing water-soluble copolymer
Li et al. A “turn-on” fluorescent chemosensor for zinc ion with facile synthesis and application in live cell imaging
CN106833625B (zh) 一种双光子溶酶体pH荧光探针及其制备方法和应用
Wang et al. Synthesis of an amphiphilic copolymer bearing rhodamine moieties and its self-assembly into micelles as chemosensors for Fe3+ in aqueous solution
Guo et al. An “OFF–ON” fluorescent chemosensor for highly selective and sensitive detection of Al (III) in aqueous solution
CN107245334B (zh) 一种检测汞离子的水溶性高分子荧光素类荧光探针及其制备方法
El Kaoutit et al. Chromogenic and fluorogenic detection of cations in aqueous media by means of an acrylic polymer chemosensor with pendant Rhodamine-based dyes
Li et al. A reversible fluorescent chemosensor for selective and sequential detection of copper ion and sulfide
Zhang et al. A tissue-permeable fluorescent probe for Al (III), Cu (II) imaging in vivo
Zhou et al. Luminescence modulation of two individual fluorophores over a wide pH range and intracellular studies
Haldar et al. A fluorimetric water-soluble polymeric pH chemosensor for extremely acidic conditions: Live-cell and bacterial imaging application
CN104629754A (zh) 咔唑类比率型pH荧光探针及其制备方法和应用
CN108350479A (zh) 细胞内递送媒介物
Kim et al. A renovation of non-aqueous Al3+ sensor to aqueous media sensor by simple recyclable immobilize electrospun nano-fibers and its uses for live sample analysis
Niu et al. A novel pH fluorescent probe based on indocyanine for imaging of living cells
CN110229165A (zh) 上转换荧光探针罗丹明衍生物及其应用

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant