CN107699233B - 一种双激发型荧光探针的制备及在检测水中肼的应用 - Google Patents
一种双激发型荧光探针的制备及在检测水中肼的应用 Download PDFInfo
- Publication number
- CN107699233B CN107699233B CN201710979613.8A CN201710979613A CN107699233B CN 107699233 B CN107699233 B CN 107699233B CN 201710979613 A CN201710979613 A CN 201710979613A CN 107699233 B CN107699233 B CN 107699233B
- Authority
- CN
- China
- Prior art keywords
- fluorescein
- bis
- dtpa
- fluorescence probe
- bidifly
- 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
Links
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 title claims abstract description 114
- 239000000523 sample Substances 0.000 title claims abstract description 78
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 22
- 238000001514 detection method Methods 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 claims abstract description 70
- 230000005284 excitation Effects 0.000 claims abstract description 38
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims abstract description 30
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract description 24
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims abstract description 18
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 11
- 238000010438 heat treatment Methods 0.000 claims abstract description 9
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000007853 buffer solution Substances 0.000 claims abstract description 8
- RAZLJUXJEOEYAM-UHFFFAOYSA-N 2-[bis[2-(2,6-dioxomorpholin-4-yl)ethyl]azaniumyl]acetate Chemical compound C1C(=O)OC(=O)CN1CCN(CC(=O)O)CCN1CC(=O)OC(=O)C1 RAZLJUXJEOEYAM-UHFFFAOYSA-N 0.000 claims abstract description 7
- 238000001914 filtration Methods 0.000 claims abstract description 6
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims abstract description 6
- RPNUMPOLZDHAAY-UHFFFAOYSA-N Diethylenetriamine Chemical compound NCCNCCN RPNUMPOLZDHAAY-UHFFFAOYSA-N 0.000 claims abstract description 5
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 claims abstract description 5
- 229960003330 pentetic acid Drugs 0.000 claims abstract description 5
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims abstract description 4
- 238000002156 mixing Methods 0.000 claims abstract description 4
- 238000003756 stirring Methods 0.000 claims abstract description 4
- 238000002189 fluorescence spectrum Methods 0.000 claims description 15
- 239000000243 solution Substances 0.000 claims description 14
- 239000000872 buffer Substances 0.000 claims description 6
- 238000001291 vacuum drying Methods 0.000 claims description 6
- 125000004177 diethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- 238000004090 dissolution Methods 0.000 claims description 2
- 239000000706 filtrate Substances 0.000 claims description 2
- WTBIAPVQQBCLFP-UHFFFAOYSA-N N.N.N.CC(O)=O.CC(O)=O.CC(O)=O.CC(O)=O.CC(O)=O Chemical compound N.N.N.CC(O)=O.CC(O)=O.CC(O)=O.CC(O)=O.CC(O)=O WTBIAPVQQBCLFP-UHFFFAOYSA-N 0.000 claims 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims 1
- GTDPSWPPOUPBNX-UHFFFAOYSA-N ac1mqpva Chemical compound CC12C(=O)OC(=O)C1(C)C1(C)C2(C)C(=O)OC1=O GTDPSWPPOUPBNX-UHFFFAOYSA-N 0.000 claims 1
- 238000005070 sampling Methods 0.000 claims 1
- OALHHIHQOFIMEF-UHFFFAOYSA-N 3',6'-dihydroxy-2',4',5',7'-tetraiodo-3h-spiro[2-benzofuran-1,9'-xanthene]-3-one Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 OALHHIHQOFIMEF-UHFFFAOYSA-N 0.000 abstract description 2
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 abstract description 2
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 abstract description 2
- 229940116269 uric acid Drugs 0.000 abstract description 2
- 238000001816 cooling Methods 0.000 abstract 2
- 238000005406 washing Methods 0.000 abstract 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 10
- 238000010521 absorption reaction Methods 0.000 description 10
- 239000011550 stock solution Substances 0.000 description 9
- 239000000126 substance Substances 0.000 description 6
- 239000000463 material Substances 0.000 description 5
- 238000001228 spectrum Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 230000002708 enhancing effect Effects 0.000 description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 238000000862 absorption spectrum Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 210000003739 neck Anatomy 0.000 description 3
- 239000012488 sample solution Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- RPTUSVTUFVMDQK-UHFFFAOYSA-N Hidralazin Chemical compound C1=CC=C2C(NN)=NN=CC2=C1 RPTUSVTUFVMDQK-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 229910052771 Terbium Inorganic materials 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- -1 hydralazine Chemical class 0.000 description 2
- 238000010248 power generation Methods 0.000 description 2
- 238000004451 qualitative analysis Methods 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- GZCRRIHWUXGPOV-UHFFFAOYSA-N terbium atom Chemical compound [Tb] GZCRRIHWUXGPOV-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical class N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 206010067125 Liver injury Diseases 0.000 description 1
- 101710138657 Neurotoxin Proteins 0.000 description 1
- 231100000766 Possible carcinogen Toxicity 0.000 description 1
- 206010061481 Renal injury Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000000908 ammonium hydroxide Substances 0.000 description 1
- 150000008064 anhydrides Chemical class 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 230000003178 anti-diabetic effect Effects 0.000 description 1
- 230000002365 anti-tubercular Effects 0.000 description 1
- 239000003472 antidiabetic agent Substances 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 238000005260 corrosion Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 231100000784 hepatotoxin Toxicity 0.000 description 1
- 230000002363 herbicidal effect Effects 0.000 description 1
- 239000004009 herbicide Substances 0.000 description 1
- 229960002474 hydralazine Drugs 0.000 description 1
- 150000002429 hydrazines Chemical class 0.000 description 1
- 238000006698 hydrazinolysis reaction Methods 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000001459 mortal effect Effects 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 239000003471 mutagenic agent Substances 0.000 description 1
- 231100000707 mutagenic chemical Toxicity 0.000 description 1
- 239000002581 neurotoxin Substances 0.000 description 1
- 231100000618 neurotoxin Toxicity 0.000 description 1
- 230000000269 nucleophilic effect Effects 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 239000005648 plant growth regulator Substances 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000011343 solid material Substances 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/18—Metal complexes
- C09K2211/182—Metal complexes of the rare earth metals, i.e. Sc, Y or lanthanide
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
本发明公开了一种双激发型荧光探针及其制备方法和在检测肼中的应用。其制备方法如下:取二乙三胺五乙酸,乙酸酐,吡啶,在65℃下搅拌加热24h,冷却,过滤,洗涤,干燥,得到的二乙三胺五乙酸二酐与三乙胺,DMF,荧光素,混合均匀,恒温100℃加热搅拌24h,冷却,过滤,干燥,得到的二乙三胺五乙酸‑双(荧光素)溶解于pH=7.5的Tris‑HCl/DMSO(10mmol·L‑1,v/v=9:1)缓冲溶液中,与Tb(NO3)3·6H2O混合,加热或放置两小时以上,得目标产物。将Tb3+‑dtpa‑bis(fluorescein)作为双激发探针结合荧光方法检测尿酸。本发明方法简单新颖,效率高,成本低,且可应用在实际水样当中。
Description
技术领域
本发明属于分析化学领域,尤其涉及新型荧光探针的合成及其对肼的检测。
背景技术
肼是重要的工业原料,被广泛应该用在农业、药物、化学领域。由于肼的易燃易爆性质,它还是一种熟悉的高能燃料,应用于火箭的推进剂。从化学结构上来看,肼具有两个官能团,并且都能表现出亲核性质,这使得肼可以合成很多的衍生物产品,如抗糖尿病药物、抗结核药物、植物生长调节剂、除草剂、缓蚀剂和染料等。最近,还发现了一种常见的联氨衍生物,如肼酞嗪,它用于治疗心脏衰竭和高血压。尽管有广泛的应用,但我们应该高度注意的是,肼通常被认为是一种神经毒素、致癌物质、诱变剂、肝毒素,对人类生命非常有害。肼是剧毒的,可以很容易地吸收通过口腔、皮肤或吸入途径。此外,如果人长期暴露在肼的环境中,它会对肝脏、肾脏和中枢神经系统造成致命伤害。事实上,根据美国环境保护署(EPA)的报道,肼被列为可能的致癌物质,其暴露上限为10ppb(3.2×10-7mol/L)。环境中肼的危害主要存在于,含肼废水的排放和运输中肼的泄漏,国内目前对肼中毒案例的报道呈现增多的趋势。因此对肼的检测是至关重要的。
荧光探针就是以荧光物质作为指示剂,并在一定波长光的激发下使指示剂产生荧光,通过检测所产生的荧光实现对被检测物质的定性或者定量分析。荧光探针具有灵敏度高,选择性好,使用方便,成本低,不需预处理,不受外界干扰等优点。特别是在分子生物学、生物化学、医学等领域中有较广泛的应用。
发明内容
本发明的目的在于设计合成一种可用于有效检测水样品中肼的新型双激发荧光探针Tb3+-dtpa-bis(fluorescein)。本发明所涉及化合物属于新型荧光探针,将其应用于检测肼操作简单,成本低,无污染,且选择性好。
本发明采用的技术方案是:一种双激发型荧光探针,所述的新型双激发荧光探针是Tb3+-dtpa-bis(fluorescein)。
上述的双激发型荧光探针的制备方法,方法如下:
1)二乙三胺五乙酸(dtpa)、乙酸酐和吡啶,混合均匀于三颈烧瓶,在60-70℃下,缓慢搅拌加热22-25h,冷却至室温,过滤,用乙酸酐和无水乙醚洗涤,抽滤,干燥,得二乙三胺五乙酸二酐(dtpaa);
2)二乙三胺五乙酸二酐(dtpaa)、三乙胺、无水DMF和荧光素,混合均匀于三颈烧瓶,95-105℃下,搅拌加热22-25h,静置,冷却到室温,过滤,真空干燥,得淡黄色固体二乙三胺五乙酸-双荧光素(dtpa-bis(fluorescein));
3)将二乙三胺五乙酸-双荧光素用pH=7.5的Tris-HCl/DMSO(10mmol·L-1,v/v=9:1)缓冲溶液溶解,得dtpa-bis(fluorescein)溶液,与Tb(NO3)3·6H2O混合,过滤,滤液于70-80℃下加热20-30min或于室温下放置1-2天,得Tb3+-dtpa-bis(fluorescein)。
优选的,上述的双激发型荧光探针的制备方法,步骤1),用于反应过程的乙酸酐的加入量为,按摩尔比,二乙三胺五乙酸:乙酸酐:吡啶=1:4:6。
优选的,上述的双激发型荧光探针的制备方法,按摩尔比,二乙三胺五乙酸二酐:三乙胺:荧光素=1:3:2。
优选的,上述的双激发型荧光探针的制备方法,按质量比,二乙三胺五乙酸-双(荧光素):Tb(NO3)3·6H2O=1:0.45-0.5。
本发明的双激发型荧光探针可应用于检测水样中的肼。定性分析方法如下:取含肼水样,加入上述的新型双激发荧光探针Tb3+-dtpa-bis(fluorescein)的pH=7.5的Tris-HCl/DMSO(10mmol·L-1,v/v=9:1)缓冲溶液,搅拌均匀,在247nm和500nm的激发波长下观察荧光光谱的变化。
定量分析方法如下:取含肼水样0.5mL于50mL容量瓶中,加入5mL浓度为5.0×10- 4mol/L的含上述新型荧光探针Tb3+-dtpa-bis(fluorescein)的pH=7.5的Tris-HCl/DMSO(10mmol·L-1,v/v=9:1)缓冲溶液,用缓冲溶液定容,在247nm和500nm的激发波长下观察荧光光谱的变化。
本发明的有益效果是:
1.本发明,对dtpa进行了修饰,在dtpa两端接上荧光素,引入酯键用来识别肼,通过酯键的肼解作用,形成两个发光基团,dtpa配合物和荧光素。使用247nm和500nm分别激发,得到荧光增强的效果,从而达到检测肼的目的。
2.本发明,针对肼的化学特点,设计了一种新型的双激发荧光探针。通过本发明的方法,该探针可以对肼进行特异性检测并且应用在实际水样中。与其它检测肼的荧光探针相比,具有简单,快速,成本低等特点。
附图说明
图1是双激发型荧光探针Tb3+-dtpa-bis(fluorescein)的合成反应的流程图。
图2a是dtpa的核磁氢谱(1H-NMR)图。
图2b是fluorescein的核磁氢谱(1H-NMR)图。
图2c是dtpa-bis(fluorescein)的核磁氢谱(1H-NMR)图。
图3a是dtpa的傅里叶变换红外光谱(FT-IR)图。
图3b是dtpaa的傅里叶变换红外光谱(FT-IR)图。
图3c是荧光素的傅里叶变换红外光谱(FT-IR)图。
图3d是dtpa-bis(fluorescein)的傅里叶变换红外光谱(FT-IR)图。
图4a是Tb3+-dtpa-bis(fluorescein),Tb3+-dtpa-bis(fluorescein)-N2H4,fluorescein的紫外吸收光谱图。
图4b是Tb3+-dtpa,Tb3+-dtpa-DHz在247nm激发下的荧光光谱图。
图5a是荧光探针在247nm的激发波长下对肼检测的荧光光谱图。
图5b是荧光探针在500nm的激发波长下对肼检测的荧光光谱图。
图6a是荧光探针在247nm的激发波长下对肼分别与不同物质混合的干扰荧光光谱图。
图6b是荧光探针在500nm的激发波长下对肼分别与不同物质混合的干扰荧光光谱图。
图7a是荧光探针在247nm的激发波长下对真实水样中肼检测的荧光光谱图。
图7b是荧光探针在247nm的激发波长下对真实水样中肼检测的荧光光谱图。
具体实施方式
实施例1新型双激发荧光探针TbIII-dtpa-bis(fluorescein)
(一)制备方法
1、二乙三胺五乙酸二酐(dtpaa)的制备
称取dtpa 7.8100g(0.02mmol),乙酸酐16.0mL(0.08mmol),吡啶10.0mL(0.12mmol)置于三颈圆底烧瓶中,混合均匀,在65℃下,搅拌加热24h。冷却至室温,将反应混合物过滤,并用少量乙酸酐和无水乙醚洗涤两次,用真空泵抽滤,所得物于真空干燥箱中80℃真空干燥,即得dtpaa。
2、二乙三胺五乙酸双荧光素(dtpa-bis(fluorescein))的制备
取1.9610g的dtpaa(5.5mmol),三乙胺8.0mL(16.5mmol),无水DMF(50mL),荧光素1.6515g(11mmol),于三颈圆底烧瓶,混合均匀。恒温100℃加热,快速搅拌24h。反应完全后静置,冷却到室温后,得淡黄色固体物质,过滤,50℃真空干燥,即得dtpa-bis(fluorescein)。
3、双激发荧光探针Tb3+-dtpa-bis(fluorescein)的制备
称取0.1236g的dtpa-bis(fluorescein)于200.0mL的pH=7.4的Tris-HCl缓冲溶液中溶解。称取0.0566g的Tb(NO3)3·6H2O置于烧杯中。然后用上面配制的dtpa-bis(fluorescein)溶溶解,待溶解后移入250mL容量瓶中,pH=7.5的Tris-HCl/DMSO(10mmol·L-1,v/v=9:1)缓冲溶液定容。把容量瓶中的溶液加热或长时间放置,形成Tb3+-dtpa-bis(fluorescein),此时浓度为5.0×10-4mol/L,作为双激发荧光探针储备液。合成过程如图1所示。
(二)检测
1.Dtpa,fluorescein,dtpa-bis(fluorescein)的FT-IR图如图2a,图2b,图2c所示。对比发现,在图2a出现的dtpa的特征峰出现在δ=2~3ppm,在图2b中fluorescein的特征峰出现在δ=6~8ppm,这些特征峰在图2c中均可以发现。这得注意的是,明显的位移出现紧挨在酯键旁边的氢质子上面,在图2c中出现了一个明显的多重峰信号在δ=3.25–2.85ppm处。这证实了酯键的存在,表明dtpa-bis(fluorescein)已经被合成。
2.Dtpa,dtpaa,fluorescein,dtpa-bis(fluorescein)的FT-IR图如图3a,图3b,图3c和图3d所示。对比发现,在图3a和图3b中,1821cm-1和1774cm-1的峰归于C=O,1118cm-1的峰吸收峰来自于C-O,2979cm-1的峰来自于-CH2-CH2-,通过这个比较可以确定dtpaa已经合成。图3d展示了dtpa-bis(fluorescein)的红外光谱图,通过与图3a和图3c比较可以观察到,图3d没有出现1774cm-1-1821cm-1的酸酐的吸收峰而新的吸收峰出现在1616cm-1、1250cm-1和1108cm-1处。1616cm-1是源于酯键上C=O的伸缩振动,1250cm-1和1108cm-1是源于酯分子中的C-O-C的反对称伸缩振动和对称伸缩振动。图3d中两波数之差为142cm-1,满足通常两吸收带波数之差为130~170cm-1的条件。此外,在847cm-1处出现了苯环上C-H的弯曲振动吸收峰,图3c和图3d相比较,847cm-1处C-H的峰没有发生位移,由此可以确定荧光素已经和dtpa连接起来,再次证明dtpa-bis(fluorescein)已经合成。值得注意的是,图3d中出现了3412cm-1的酚羟基O-H的伸缩振动吸收峰,这表明dtpa-bis(fluorescein)的合成仅仅占用了fluorescein的一个酚羟基-OH,还有一个酚羟基-OH被保留下来,这为以后Tb3+的配位奠定了理论基础。
3.Tb3+-dtpa-bis(fluorescein),Tb3+-dtpa-bis(fluorescein)-N2H4,fluorescein的紫外吸收光谱图如图4a所示。由紫外吸收光谱可以看出,Tb3+-dtpa-bis(fluorescein)的溶液在400nm-500nm处只有很低的吸收峰,而Tb3+-dtpa-bis(fluorescein)-N2H4在500nm处有非常强的吸收峰,这个峰与Tb3+-dtpa-bis(fluorescein)相比较,强烈增强。并且Tb3+-dtpa-bis(fluorescein)-N2H4溶液的特征吸收峰和fluorescein溶液的特征吸收峰几乎相同,说明Tb3+-dtpa-bis(fluorescein)在加入肼之后可以形成荧光素,才使得其和fluorescein溶液有相同的吸收峰。这表明Tb3+-dtpa-bis(fluorescein)具有在500nm处激发下作为荧光探针检测肼的潜在能力。
4.Tb3+-dtpa,Tb3+-dtpa-DHz在247nm波长激发下的荧光光谱图如图4b所示。由荧光光谱可以看出,Tb3+-dtpa的溶液在550nm处几乎没有荧光特征峰,而Tb3+-dtpa-DHz有一个显著的铽配合物的荧光峰在550nm处。这个峰与Tb3+-dtpa相比较,明显增强。说明Tb3+-dtpa-bis(fluorescein)加入肼之后形成的Tb3+-dtpa-DHz可以形成新的配位场使Tb3+电子结构发生变化,在247nm波长激发下发出铽配合物的荧光。这表明Tb3+-dtpa-bis(fluorescein)具有在247nm波长激发下作为荧光探针检测肼的潜在能力。
实施例2双激发荧光探针Tb3+-dtpa-bis(fluorescein)在检测肼中的应用
1.双激发荧光探针肼检测的荧光光谱
实验条件:取一定量的肼(N2H4)用蒸馏水配制成浓度为1.0×10-3mol/L的溶液,作为肼储备液。
分别取5.0mL肼储备液于50mL容量瓶中,再分别加入实施例1制备的5.0mL的双激发型荧光探针储备液,用缓冲液定容。此时探针浓度为5.0×10-5mol/L,各检测物质浓度为1.0×10-4mol/L。在247nm和500nm的激发波长下观察荧光光谱的变化。
结果如图5a、图5b所示。如图5a所示,在247nm激发下,荧光探针在520nm处有较弱发射峰。而Tb3+配合物在550nm处也有很弱的荧光。当把肼加入到探针溶液时,荧光强度在520nm处被明显增强。并且这个荧光与荧光素以及荧光素和Tb3+-dtpa-DHz两者混合的溶液荧光几乎相同。如图5b所示,在500nm激发下,荧光探针在520nm处仍然可以保持较弱发射峰。此时,Tb3+-dtpa-DHz配合物的荧光不能被激发。当把肼加入到探针溶液时,荧光强度在520nm处表现出了更加明显的增强,并且这个荧光与荧光素以及荧光素和Tb3+-dtpa-DHz两者混合的溶液荧光几乎相同。在247nm和550nm激发下,荧光探针均可达到检测肼的效果,使用这种方法可以排除更多的干扰因素,检测更稳定。
2.肼与不同检测物混合对双激发荧光探针Tb3+-dtpa-bis(fluorescein)检测的影响
实验条件:分别取浓度为1.0×10-3mol/L的葡萄糖,双氧水,氨水,金属离子(K+,Na+,Zn2+,Mg2+,Ba2+,Ni2+,Mn2+,Ca2+,Cr3+,Al3+and Fe3+)和阴离子(Cl-,ClO-and SO4 2-)储备液5.0mL于50mL容量瓶中,分别加入5.0mL的肼储备液,再分别加入5.0mL的荧光探针储备液,用缓冲液定容。此时探针浓度为5.0×10-5mol/L,尿酸和各检测物质浓度都为1.0×10- 4mol/L的溶液。在247nm和500nm的激发波长下观察荧光光谱的变化。
结果如图6a、图6b所示。如图6a,在247nm激发下,不同检测物的混合液对荧光探针的检测几乎没有影响,都达到了使荧光强度得明显增强。其他混合物质对探针检测几乎没有影响。如图6b,在500nm激发下,只有Mn2+的混合液对荧光探针的检测有微弱影响,但是和荧光探针的荧光强度相比,Mn2+的混合液显示出了明显的荧光增强效果,并且这个影响可以更好的排除通过使用另一个激发波长的检测。这在检测中不会造成影响。由此可见,双激发型荧光探针在有其他物质干扰的情况下,对肼依然具有特异性。
表1
3.荧光探针Tb3+-dtpa-bis(fluorescein)在真实水样中对肼的检测
实验条件:真实水样取自(sample1:自来水;sample2:饮用水;sample3:参窝水库出口水;sample4:参窝水库库中水;sample5:纯水)。首先准备第一组浓度:50mL的容量瓶中分别加入肼,使肼浓度为1.0×10-3mol/L,之后分别使用真实水样定容,得到还有肼浓度为1.0×10-3mol/L浓度的真实样品溶液。依次准备肼浓度为5.0×10-4mol/L,2.0×10-4mol/L的真实样品溶液。分别取真实样品溶液5mL和5mL荧光探针贮备液于15支50mL的容量瓶中,用缓冲液定容。再取一支容量瓶,加入5mL荧光探针贮备液,用缓冲液定容。在247nm和500nm的激发波长下观察荧光光谱的变化。
结果如图7a、图7b所示。如图7a,在247nm激发下,在真实样品中,荧光探针只有微弱的荧光。加入不同浓度的肼,荧光强度都有增强,并且这个增强值与在纯水中检测肼得结果相同。如图7b,在500nm激发下,荧光的增强在各个浓度之间保持了更好的一致性。由此可以判断,该探针可以在真实水样中对肼进行检测,并且不受到水中其他物质干扰。
表2
Claims (7)
1.一种双激发型荧光探针的制备方法,其特征在于,所述的荧光探针是Tb3+-dtpa-bis(fluorescein),其制备过程如下:
1)二乙三胺五乙酸、乙酸酐和吡啶,混合均匀于三颈烧瓶,在60-70 ℃下,缓慢搅拌加热22-25 h,冷却至室温,过滤,用乙酸酐和无水乙醚洗涤,抽滤,50 ℃真空干燥,得白色固体二乙三胺五乙酸二酐dtpaa;
2)二乙三胺五乙酸二酐、三乙胺、无水DMF和荧光素,混合均匀三颈烧瓶,于95-105 ℃下,搅拌加热22-25 h,静置,冷却到室温,过滤,50 ℃真空干燥,得淡黄色固体二乙三胺五乙酸-双荧光素dtpa-bis(2,6-fluorescein);
3)将二乙三胺五乙酸-双荧光素用10 mmol∙L-1, v/v = 9:1,pH=7.5的Tris-HCl/DMSO缓冲溶液溶解,得dtpa-bis(fluorescein)溶液,与Tb(NO3)3•6H2O混合,过滤,滤液于70-80℃下加热搅拌20-30 min或于室温下放置1 - 2天,得Tb3+-dtpa-bis(fluorescein)。
2.如权利要求1所述的双激发型荧光探针的制备方法,其特征在于:按摩尔比,二乙三胺五乙酸:乙酸酐:吡啶 = 1 : 4 : 6。
3.如权利要求1所述的双激发型荧光探针的制备方法,其特征在于:按摩尔比,二乙三胺五乙酸二酐:三乙胺: 荧光素= 1 : 3 :2。
4.如权利要求1所述的双激发型荧光探针的制备方法,其特征在于:按质量比,二乙三胺五乙酸-双(荧光素): Tb(NO3)3•6H2O = 1 : 0.45-0.5。
5.权利要求1所述的双激发型荧光探针Tb3+-dtpa-bis(fluorescein)在检测水样肼中的应用。
6.如权利要求5所述的应用,其特征在于,方法如下:取水样,加入权利要求1所述的双激发型荧光探针TbШ-dtpa-bis(fluorescein),缓冲溶液浓度为10 mmol∙L-1, v/v = 9:1,pH=7.5的Tris-HCl/DMSO,搅拌均匀,在247 nm和500 nm的激发波长下观察荧光光谱的变化。
7.如权利要求5所述的应用,其特征在于,方法如下:取含肼水样0.5 mL于50 mL容量瓶中,加入5 mL 浓度为5.0 × 10-4 mol/L的权利要求1所述的双激发型荧光探针TbШ-dtpa-bis(fluorescein),缓冲溶液浓度为10 mmol∙L-1, v/v = 9:1,pH=7.4的Tris-HCl/DMSO,用缓冲溶液定容,在247 nm和500 nm的激发波长下观察荧光光谱的变化。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710979613.8A CN107699233B (zh) | 2017-10-19 | 2017-10-19 | 一种双激发型荧光探针的制备及在检测水中肼的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710979613.8A CN107699233B (zh) | 2017-10-19 | 2017-10-19 | 一种双激发型荧光探针的制备及在检测水中肼的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN107699233A CN107699233A (zh) | 2018-02-16 |
CN107699233B true CN107699233B (zh) | 2019-08-09 |
Family
ID=61181879
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201710979613.8A Active CN107699233B (zh) | 2017-10-19 | 2017-10-19 | 一种双激发型荧光探针的制备及在检测水中肼的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN107699233B (zh) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108982447B (zh) * | 2018-07-19 | 2020-12-01 | 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 |
CN110437245B (zh) * | 2019-08-26 | 2022-04-05 | 辽宁大学 | 双激发双发射荧光探针CDs-COO-F及其在检测水和细胞中肼的应用 |
CN110437175A (zh) * | 2019-08-26 | 2019-11-12 | 辽宁大学 | 双激发荧光探针EuШ-dtpa-bis(HBT)及其在检测肼中的应用 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107163068A (zh) * | 2017-05-08 | 2017-09-15 | 辽宁大学 | 一种新型荧光探针及其制备方法和在检测尿酸中的应用 |
-
2017
- 2017-10-19 CN CN201710979613.8A patent/CN107699233B/zh active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107163068A (zh) * | 2017-05-08 | 2017-09-15 | 辽宁大学 | 一种新型荧光探针及其制备方法和在检测尿酸中的应用 |
Non-Patent Citations (1)
Title |
---|
Fast and ratiometric "naked eye"" detection of hydrazine for both solid and vapour phase sensing;Goswami Shyamaprosad等;《NEW JOURNAL OF CHEMISTRY》;20150116;第39卷(第3期);全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN107699233A (zh) | 2018-02-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107699233B (zh) | 一种双激发型荧光探针的制备及在检测水中肼的应用 | |
US8962339B2 (en) | Fluorescent probe compounds, preparation method and application thereof | |
Kessler et al. | ET (33), a solvatochromic polarity and micellar probe for neutral aqueous solutions | |
Zhang et al. | A ruthenium (II) complex–cyanine energy transfer scaffold based luminescence probe for ratiometric detection and imaging of mitochondrial peroxynitrite | |
Prodi et al. | Dansylated polyamines as fluorescent sensors for metal ions: photophysical properties and stability of copper (II) complexes in solution | |
Hou et al. | Detection of hydrazine via a highly selective fluorescent probe: A case study on the reactivity of cyano-substituted CC bond | |
CN108752377B (zh) | 一种检测过氧亚硝基阴离子的荧光探针、合成方法和应用 | |
Freiherr von Richthofen et al. | Probing the radialene-character in triplesalophen ligands by spectroscopic and structural analysis | |
CN110229165A (zh) | 上转换荧光探针罗丹明衍生物及其应用 | |
Pope et al. | Metal-to-ligand charge-transfer sensitisation of near-infrared emitting lanthanides in trimetallic arrays M 2 Ln (M= Ru, Re or Os; Ln= Nd, Er or Yb) | |
Hu et al. | A ratiometric fluorescence sensor for Fe3+ based on FRET and PET processes | |
Shen et al. | A mitochondria-targeting ratiometric fluorescent probe for the detection of hypochlorite based on the FRET strategy | |
CN106497547B (zh) | 一种稀土发光材料及其制备方法、应用 | |
Zhou et al. | A fluorescent probe based on modulation of ESIPT signaling for the highly selective detection of N2H4 and cell-imaging | |
CN107698557B (zh) | 基于吡啶联吡唑酰腙衍生物的荧光探针及其制备方法和应用 | |
Fang et al. | Highly sensitive and selective recognition behaviour for fluoride based on a homoditopic curcumin-difluoroboron receptor | |
Chen et al. | Improving anion sensing ability of the indolocarbazole-based fluorescence turn-on sensor by increasing salicylaldehyde response unit | |
Li et al. | Design and synthesis of dual-excitation fluorescent probe, Tb3+-dtpa-bis (fluorescein), and application in detection of hydrazine in environmental water samples and live cells | |
CN110128328A (zh) | 一种识别钼酸根离子的荧光探针及其制备方法和识别方法 | |
Yang et al. | A novel fluorescent probe (dtpa-bis (cytosine)) for detection of Eu (III) in rare earth metal ions | |
CN105152971B (zh) | 一种双萘西弗碱氰根离子传感器及其合成和应用 | |
CN103242328A (zh) | 对N-乙酰氨基苯基罗丹明6GpH荧光分子探针及其制备方法与用途 | |
CN107722008A (zh) | 一种识别HepG2细胞中Ag+的2‑芳基咪唑邻菲啰啉探针及其制备方法 | |
CN112142753B (zh) | 异硫氰酸荧光素衍生物的制备方法 | |
CN107286171A (zh) | 一种探针及应用该探针同时检测微量Al3+和/或I‑的方法 |
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 |