CN110452687A - 一种连二亚硫酸钠比色荧光探针及其制备方法和应用 - Google Patents
一种连二亚硫酸钠比色荧光探针及其制备方法和应用 Download PDFInfo
- Publication number
- CN110452687A CN110452687A CN201910530662.2A CN201910530662A CN110452687A CN 110452687 A CN110452687 A CN 110452687A CN 201910530662 A CN201910530662 A CN 201910530662A CN 110452687 A CN110452687 A CN 110452687A
- Authority
- CN
- China
- Prior art keywords
- probe
- sodium dithionite
- colorimetric
- added
- molecular probe
- 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
Links
- 239000000523 sample Substances 0.000 title claims abstract description 53
- JVBXVOWTABLYPX-UHFFFAOYSA-L sodium dithionite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])=O JVBXVOWTABLYPX-UHFFFAOYSA-L 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 239000003068 molecular probe Substances 0.000 claims abstract description 25
- 238000001514 detection method Methods 0.000 claims abstract description 16
- 235000013305 food Nutrition 0.000 claims abstract description 14
- 239000000654 additive Substances 0.000 claims abstract description 13
- 230000000996 additive effect Effects 0.000 claims abstract description 12
- 239000011734 sodium Substances 0.000 claims description 26
- JLTDJTHDQAWBAV-UHFFFAOYSA-N N,N-dimethylaniline Chemical compound CN(C)C1=CC=CC=C1 JLTDJTHDQAWBAV-UHFFFAOYSA-N 0.000 claims description 6
- 229930014669 anthocyanidin Natural products 0.000 claims description 6
- 150000001452 anthocyanidin derivatives Chemical class 0.000 claims description 6
- 235000008758 anthocyanidins Nutrition 0.000 claims description 6
- 230000008859 change Effects 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 239000007850 fluorescent dye Substances 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 3
- 238000004090 dissolution Methods 0.000 claims description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- LNOPIUAQISRISI-UHFFFAOYSA-N n'-hydroxy-2-propan-2-ylsulfonylethanimidamide Chemical compound CC(C)S(=O)(=O)CC(N)=NO LNOPIUAQISRISI-UHFFFAOYSA-N 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- 229940125904 compound 1 Drugs 0.000 claims 2
- 238000000034 method Methods 0.000 abstract description 8
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 229910004878 Na2S2O4 Inorganic materials 0.000 description 32
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 19
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 230000004044 response Effects 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 9
- 235000019441 ethanol Nutrition 0.000 description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 8
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 8
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 8
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 239000012043 crude product Substances 0.000 description 4
- 235000013312 flour Nutrition 0.000 description 4
- 235000013373 food additive Nutrition 0.000 description 4
- 239000002778 food additive Substances 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 238000005457 optimization Methods 0.000 description 4
- 239000012074 organic phase Substances 0.000 description 4
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 238000005292 vacuum distillation Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000002189 fluorescence spectrum Methods 0.000 description 3
- 238000010898 silica gel chromatography Methods 0.000 description 3
- 229910052938 sodium sulfate Inorganic materials 0.000 description 3
- 238000011895 specific detection Methods 0.000 description 3
- XFVZSRRZZNLWBW-UHFFFAOYSA-N 4-(Diethylamino)salicylaldehyde Chemical compound CCN(CC)C1=CC=C(C=O)C(O)=C1 XFVZSRRZZNLWBW-UHFFFAOYSA-N 0.000 description 2
- 239000007832 Na2SO4 Substances 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000012044 organic layer Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- WROMPOXWARCANT-UHFFFAOYSA-N tfa trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.OC(=O)C(F)(F)F WROMPOXWARCANT-UHFFFAOYSA-N 0.000 description 2
- 238000000954 titration curve Methods 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- ZENKESXKWBIZCV-UHFFFAOYSA-N 2,2,4,4-tetrafluoro-1,3-benzodioxin-6-amine Chemical group O1C(F)(F)OC(F)(F)C2=CC(N)=CC=C21 ZENKESXKWBIZCV-UHFFFAOYSA-N 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- PSDYQSWHANEKRV-UHFFFAOYSA-N [S]N Chemical compound [S]N PSDYQSWHANEKRV-UHFFFAOYSA-N 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000007844 bleaching agent Substances 0.000 description 1
- 206010006451 bronchitis Diseases 0.000 description 1
- 239000005321 cobalt glass Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000012875 competitive assay Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 238000000695 excitation spectrum Methods 0.000 description 1
- 230000004438 eyesight Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000012886 linear function Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- YIXJRHPUWRPCBB-UHFFFAOYSA-N magnesium nitrate Inorganic materials [Mg+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O YIXJRHPUWRPCBB-UHFFFAOYSA-N 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000004043 responsiveness Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 229920002379 silicone rubber Polymers 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/78—Ring systems having three or more relevant rings
-
- 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
-
- 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
-
- 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
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1007—Non-condensed systems
-
- 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
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1088—Heterocyclic compounds characterised by ligands containing oxygen as the only heteroatom
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Materials Engineering (AREA)
- Engineering & Computer Science (AREA)
- Optics & Photonics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
本发明提供了一个比色荧光双功能分子探针,及其制备方法与在食品样品中非法添加剂连二亚硫酸钠检测上的应用,属于分析化学技术领域。该探针分子式为C29H31N4O+,其结构式如式I所示:
Description
技术领域
本发明属于分析检测领域,具体涉及比色荧光双功能有机小分子探针的构建及利用其检测分析食品中非法添加剂连二亚硫酸钠的新方法。
背景技术
食品添加剂是指用于防止食物变质延长食物存放期、方便食品加工、保持提高食物营养价值和改善食物感官性状的一类天然或者化学合成物质。国家对添加剂的使用是有严格的规定的,不得使用非法添加剂。连二亚硫酸钠(Na2S2O4•2H2O)俗称保险粉,是一种白色砂状结晶或淡黄色粉末化学用品,是一种工业用还原性漂白剂,对人体非常有害的化学物质,并不是可食用物质,国家明文规定禁止将其用作食品添加剂,但仍有企业非法使用。消费者若长期食用会破坏人体的肝脏、肠胃以及视力,易引起头疼、腹泻和支气管炎等多种疾病,使得人体的免疫功能急剧下降,甚至造成癌变,近年来,社会上出现的食品安全问题,引起了消费者对非法添加剂的关注,因此,设计一种特异性检测Na2S2O4的比色荧光分子探针来直接检测非法添加剂Na2S2O4的含量,对于防止消费者误食有害食品具有十分重要的指导意义。
然而,目前现有的大部分检测方法均是针对Na2S2O4用作工业还原剂时使用的,而作为食品添加剂,Na2S2O4含量检测方法研究的比较少,并且已有的检测方法存在以下有待改进的地方,不能实现对Na2S2O4的直接快速检测,易受到其他物质的干扰,操作不便,耗时较长等。从而缺少研究一种具有高选择性并且能够快速直接检测Na2S2O4的方法。
因此,设计一种特异性检测Na2S2O4的比色荧光双功能分子探针来应用于食品检测分析方面具有很好的发展前景,也能达到快速检测、分析准确的目的。
发明内容
本发明的目的是提供一种具有特异性的比色荧光双功能分子探针直接、准确、快速检测非法添加剂连二亚硫酸钠的方法。
本发明的技术方案是,一种检测连二亚硫酸钠的比色荧光分子探针,其结构式如下:
一种检测连二亚硫酸钠的比色荧光分子探针的制备方法,步骤如下:
(1)类花青素荧光染料的合成:将1份N,N-二乙基-4-氨基-2-羟基-苯甲醛与1份6-氨基-3,4-二氢-1(2H)-萘酮依次加入到装有浓硫酸(5 mL)的反应容器中,在氮气保护的条件下加热至90 ℃,反应5小时。反应完成并冷却后,在冰浴的条件下加入高氯酸(70%),使用萃取剂 (CH2Cl2/MeOH, 20:1)对生成物进行萃取。每次使用30 mL溶液进行萃取,收集有机相,重复萃取四次,待萃取完成后,往所收集的有机相中加入过量的无水硫酸镁干燥1小时,抽滤除去硫酸镁,减压蒸馏得到粗产物。用硅胶柱色谱提纯,得到类花青素荧光染料。
(2)检测非法添加剂连二亚硫酸钠比色荧光分子探针的制备:将类花青素荧光染料用含有1%三氟乙酸(TFA)的MeCN /CH2Cl2(1:4)溶液溶解,将反应进行氮气保护,并在0℃下进行搅拌,向其中加入NaNO2,在0℃下继续搅拌10分钟。然后加入氨基磺酸并继续搅拌5分钟。接着,加入MeCN(1mL)中的N,N-二甲基苯胺,再将所得混合物在0℃下搅拌反应1小时。待反应完成并冷却后,先用H2O稀释产物,然后对产物进行萃取。使用CH2Cl2(30 mL)进行萃取,收集有机层,重复萃取三次,待萃取完成后,加入过量的Na2SO4干燥1小时,抽滤除去Na2SO4,转入烧瓶中,减压蒸馏得到粗产物。用硅胶柱色谱提纯,得到检测非法添加剂连二亚硫酸钠的比色荧光双功能分子探针。在本说明书的实施例中更详细地说明了该探针的合成和检测方法。
本发明具有如下的技术效果,探针合成步骤简单,目标产物产率高,容易分离提纯;其激发和发射光谱在可见光区,化学稳定性好,具有较好的水溶性,可以使用于中性及偏碱性条件下的食品体系中Na2S2O4的检测;以发射波长处荧光强度的比值I620/I620为信号参数,显著提高了检测的特异性与灵敏性,在0- 3mM对Na2S2O4有很好的线性关系;此外,该探针对Na2S2O4具有高选择性,与其它含硫离子作用时溶液颜色和荧光强度比值I620/I620都基本不变。因此该发明方法具有原始创新性、良好的社会价值和应用前景。
附图说明
图1 连二亚硫酸钠比色荧光分子探针设计原理(A)与合成路线图(B)。
图2 (A)2µM探针(a)与其响应10mM Na2S2O4(b)荧光光谱图;(B)2µM探针与其响应10mM Na2S2O4吸收光谱图。
图3 2µM探针在不同溶剂条件下响应10mM Na2S2O4最大荧光发射强度增强对比图。
图4 2µM探针在不同EtOH/H2O条件下响应10mM Na2S2O4最大荧光发射强度增强对比图。
图5 2µM探针在不同pH条件下响应10mM Na2S2O4最大荧光发射强度增强对比图。
图6 2µM探针与10mM Na2S2O4反应响应动力学曲线图。
图7 连二亚硫酸钠比色荧光探针选择性考察。
图8 (A)2µM探针在70%EtOH体系中对不同浓度Na2S2O4 (0-3 mM)响应吸收光谱变化;(B)2µM探针对不同Na2S2O4浓度响应颜色可视化分析图。
图9 (A)2µM探针在70%EtOH体系中对不同浓度Na2S2O4 (0-3 mM)响应荧光光谱变化;(B)2µM探针对不同Na2S2O4浓度响应信背比(S/B)线性关系图。
具体实施方式
下面结合附图和具体实施例对本发明进行详细说明,但不限于此。
实施例1:连二亚硫酸钠比色荧光分子探针的合成,合成路线如图1(B)所示,基本操作过程如下。
(1)类花青素荧光染料的合成:将N,N-二乙基-4-氨基-2-羟基-苯甲醛(200mg,1.03mmol)与6-氨基-3,4-二氢-1(2H)-萘酮(185mg,1.13mmol)依次加入到装有浓硫酸(5mL)的烧瓶(50mL)中,在氮气保护的条件下加热至90 ℃,反应5小时。反应完成并冷却后,在冰浴的条件下加入高氯酸(70%,7 mL),使用萃取剂 (CH2Cl2/MeOH, 20:1)对生成物进行萃取。每次使用30 mL溶液进行萃取,收集有机相,重复萃取四次,待萃取完成后,往所收集的有机相中加入过量的无水硫酸镁干燥1小时,抽滤除去硫酸镁,减压蒸馏得到粗产物。用硅胶柱色谱提纯,得到得到最终产物, 红色粉末状固体215 mg,产率为56.3%。
(2)连二亚硫酸根荧光探针的制备:将类花青素荧光染料(23.0mg,56µmol)用含有1%三氟乙酸(TFA)的MeCN /CH2Cl2(1:4,5.0mL)溶液溶解,将反应进行氮气保护,并在0℃下进行搅拌,向其中加入NaNO2(7.5mg,109µmol),在0℃下继续搅拌10分钟。然后加入氨基磺酸(10.1mg,104µmol)并继续搅拌5分钟。接着,加入MeCN(1mL)中的N,N-二甲基苯胺(46µL,361µmol),再将所得混合物在0℃下搅拌反应1小时。待反应完成并冷却后,先用H2O稀释产物,然后对产物进行萃取。使用CH2Cl2(30 mL)进行萃取,收集有机层,重复萃取三次,待萃取完成后,加入过量的Na2SO4干燥1小时,抽滤除去Na2SO4,转入烧瓶中,减压蒸馏得到粗产物。用硅胶柱色谱提纯,最终得到深蓝色固体即特异性检测连二亚硫酸钠比色荧光分子探针21.9mg,产率为72%。
1HNMR(400MHz,DMSO) δ(ppm)8.71(s,1H), 8.34(d,1H), 7.98(d,1H), 7.90(s,1H), 7.86(s,1H),7.84(s,1H),7.80(s,1H),7.52(d,1H),7.37(s,1H),6.86(d,2H),3.73(m,4H),3.18(m,2H),3.11(s,6H),3.08(s,2H),1.26(m,6H).13CNMR(100MHz,DMSO):δ161.65,158.81,156.11,155.78,153.65,148.62,143.33,132.46,127.17,126.70,126.07,122.17,121.63,119.66,118.94,112.10,96.14,46.08,31.62,30.30,26.73,24.92。结果表明,所得产物结构正确。
实施例2:连二亚硫酸钠比色荧光分子探针的可行性验证
为了验证得到的目标探针理是否具有预期理想的功能,进行了初步试验。如图2(A)所示,连二亚硫酸钠比色荧光分子探针在620nm处有强吸收峰,在加入Na2S2O4(10mM)之后,吸收峰明显蓝移,而吸收池中探针与Na2S2O4响应前后,颜色由蓝色明显变成了红色。如图(B)所示,在570nm激发下,连二亚硫酸钠比色荧光分子探针表现出极小的荧光发射峰,但加入Na2S2O4(10mM)之后,探针的荧光发射强度有很明显的上升,表明该探针对于Na2S2O4的响应具有高灵敏度。而且同时随着Na2S2O4的加入,可以看到荧光池中溶液由微弱的荧光到明显的红色荧光颜色的改变。这些现象证明了所设计的目标探针能够与Na2S2O4响应,Na2S2O4使得目标探针的偶氮键断裂,生成了具有强荧光的新物质,吸收峰发生明显的蓝移。
实施例3:连二亚硫酸钠比色荧光分子探针响应条件的优化
为优化出最佳检测体系,进行了常用溶剂:乙腈(CH3CN), N,N-二甲基甲酰胺(DMF),四氢呋喃(THF),二甲基亚砜(DMSO), 乙醇(EtOH), 甲醇(MeOH)的筛选实验。得到了在不同溶剂中于620nm处2µM探针以及加入10mM Na2S2O4后的荧光响应强度比,由图3可知,EtOH为溶剂时,探针与Na2S2O4响应情况最佳。
无机盐Na2S2O4易溶于水不易溶于EtOH,因此实验接下来探究了乙醇与水比例对探针响应的影响。得到了在EtOH/H2O不同时于620nm处2µM探针以及加入10mM Na2S2O4后的荧光响应强度比。由图4可知,在70% EtOH的条件下,探针和待测物溶解性都比较好,两者之间充分作用,探针与Na2S2O4响应情况最佳。
Na2S2O4的还原性太强,导致其水溶液极其不稳定。为尽量避免因此带来的分析误差,本实验体系所需待测Na2S2O4采用现配现测的方法。另据文献报道,S2O4 2-在碱性条件下稳定性会有所增强。所以考察了探针在不同pH值条件下荧光探针与待测物连Na2S2O4的响应情况。得到了在不同pH条件下于620nm处2µM探针以及加入10mM Na2S2O4后的荧光响应强度比。由图5可知,在pH=11时,荧光信号强度比例F/F0达到最大值,符合文献中的结论,碱性条件下Na2S2O4的稳定性较好。
实施例4:连二亚硫酸钠比色荧光分子探针响应动力学考察
对探针反应动力学进行考察,进行时间扫描实验,得到2µM探针与各浓度Na2S2O4反应时荧光信号与时间对应的关系图。由图6可知,加入 Na2S2O4后荧光信号随着时间不断地增大且在大约5s的时候达到最大值,如此快速的响应十分适合进行对于食品添加剂的检测。
实施例5:连二亚硫酸钠比色荧光分子探针的选择性考察
为了评价该探针的选择特性,本实验测定了连二亚硫酸钠比色荧光分子探针对食品中可能含有的具有代表性的各种无机盐(NaCl, KCl, Na2S, Na2S2O3, NaHCO3, Mg(NO3)2 ,NaNO3, Na2SO3, Na2CO3, Na2S2O4(图中数字序号依次对应上述物质顺序)) 的荧光光谱。如图7所示,相对于Na2S2O4而言,其他的竞争底物对探针的响应程度很小,其中在可见光和荧光下的插图均说明,只有加入连二亚硫酸钠才会发生明显的颜色变化,表明连二亚硫酸钠比色荧光分子探针能够很灵敏地特异性响应食品中添加剂中的Na2S2O4。此外,竞争实验也表明该探针能够在有其他竞争底物存在的情况下与Na2S2O4特异性反应。
实施例6:连二亚硫酸钠比色荧光分子探针的吸收曲线
在优化得到的条件下进行吸收曲线的测定。图8(A)表明,加入不同浓度Na2S2O4后,探针在620nm处的吸收峰不断降低,而在450nm处的吸收峰不断增强。图8(B)表明,在0-3mM范围内随着Na2S2O4的浓度不断加大,溶液的颜色由蓝色逐渐变为黄色最后变为红色。该探针能够很好的通过颜色的改变,大致判断其含量。
实施例7:连二亚硫酸钠比色荧光分子探针的滴定曲线
在优化得到的条件下进行滴定曲线的测定。图9(A)表明,经过不同浓度Na2S2O4(0-3mM)后,探针的荧光强度随Na2S2O4浓度的升高而明显增加。图9(B)表明,荧光探针在0-3 mM的范围内与待测物连二亚硫酸钠的信号-背景比(S / B)呈现较好的线性关系,线性函数式为y = 18.535x +8.029,R² = 0.9935。该探针可以达到对食品中添加剂成分Na2S2O4的直接快速准确的检测效果。
实施例8:连二亚硫酸钠比色荧光分子探针的实际应用
将所制的荧光探针用于面粉(市面上购买的)中Na2S2O4回收率的测定。将所制备的面粉样品经离心取其上层清液后用所得到的探针测定,得知面粉样品中不含Na2S2O4,然后加入不同浓度的Na2S2O4样品溶液。结果列于表1。从表中可以看出,探针对所有面粉样中Na2S2O4回收率的测定结果令人满意,看来所制探针可有效的应用于实际样品中Na2S2O4的分析测定。
表1
a三个样品的平均值,b标准偏差。
Claims (3)
1.一种检测连二亚硫酸钠的比色荧光分子探针,其结构式如式I所示:
(式I)。
2.根据权利要求1所述的一种检测连二亚硫酸钠比色荧光分子探针的制备方法,其特征在于,它如下步骤:将类花青素荧光染料(化合物1)溶解后,向其中加入NaNO2,搅拌10分钟,然后加入氨基磺酸并继续搅拌5分钟,接着加入N,N-二甲基苯胺进行反应,待反应完成并冷却后,分离提纯得到目标探针,
(化合物1)。
3.一种权利要求1所述的比色荧光分子探针的应用,其特征在于,所述探针用于食品中非法添加剂连二亚硫酸钠含量的检测,能够产生肉眼可见的显著颜色变化和荧光信号增强,并具有直接简单快速检测的特点。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910530662.2A CN110452687B (zh) | 2019-06-19 | 2019-06-19 | 一种连二亚硫酸钠比色荧光探针及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910530662.2A CN110452687B (zh) | 2019-06-19 | 2019-06-19 | 一种连二亚硫酸钠比色荧光探针及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN110452687A true CN110452687A (zh) | 2019-11-15 |
CN110452687B CN110452687B (zh) | 2022-12-27 |
Family
ID=68480768
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910530662.2A Expired - Fee Related CN110452687B (zh) | 2019-06-19 | 2019-06-19 | 一种连二亚硫酸钠比色荧光探针及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN110452687B (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113354583A (zh) * | 2021-06-15 | 2021-09-07 | 上海大学 | 用于检测乏氧水平的荧光探针、其制备方法及其应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107121417A (zh) * | 2017-04-19 | 2017-09-01 | 中国科学院理化技术研究所 | 一种检测连二亚硫酸钠的荧光化学传感器及其制备方法和应用 |
CN107418556A (zh) * | 2017-05-31 | 2017-12-01 | 济南大学 | 一种检测硫化氢的荧光探针及其制备方法和应用 |
CN108164494A (zh) * | 2018-02-11 | 2018-06-15 | 山东师范大学 | 一种检测过氧亚硝酸根的双光子荧光探针及其制备方法和应用 |
CN108949155A (zh) * | 2018-08-29 | 2018-12-07 | 济南大学 | 一种检测细胞内连二亚硫酸钠的荧光探针及其应用 |
-
2019
- 2019-06-19 CN CN201910530662.2A patent/CN110452687B/zh not_active Expired - Fee Related
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107121417A (zh) * | 2017-04-19 | 2017-09-01 | 中国科学院理化技术研究所 | 一种检测连二亚硫酸钠的荧光化学传感器及其制备方法和应用 |
CN107418556A (zh) * | 2017-05-31 | 2017-12-01 | 济南大学 | 一种检测硫化氢的荧光探针及其制备方法和应用 |
CN108164494A (zh) * | 2018-02-11 | 2018-06-15 | 山东师范大学 | 一种检测过氧亚硝酸根的双光子荧光探针及其制备方法和应用 |
CN108949155A (zh) * | 2018-08-29 | 2018-12-07 | 济南大学 | 一种检测细胞内连二亚硫酸钠的荧光探针及其应用 |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113354583A (zh) * | 2021-06-15 | 2021-09-07 | 上海大学 | 用于检测乏氧水平的荧光探针、其制备方法及其应用 |
Also Published As
Publication number | Publication date |
---|---|
CN110452687B (zh) | 2022-12-27 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US11505530B2 (en) | 1,8-naphthalimide derivative, preparation method therefor and use thereof | |
Li et al. | Highly selective colorimetric/fluorometric dual-channel sensor for cyanide based on ICT off in aqueous solution | |
Zhang et al. | An effective ICT-based and ratiometric fluorescent probe for sensing sulfite | |
Huang et al. | Highly selective and sensitive fluorescent probe for mercury ions based on a novel rhodol-coumarin hybrid dye | |
Qin et al. | A thiocoumarin-based colorimetric and ratiometric fluorescent probe for Hg 2+ in aqueous solution and its application in live-cell imaging | |
CN109678763B (zh) | 一种近红外生物硫醇荧光探针及其制备方法和应用 | |
CN103804369B (zh) | 一种裸眼和荧光比率检测含氰离子荧光分子探针的合成及应用 | |
CN109535147B (zh) | 一种快速响应的甲醛荧光探针及其制备方法和应用 | |
CN109867611B (zh) | 一种用于红酒和活体内硫化氢检测的水溶性双光子硫化氢荧光探针及其制备方法和应用 | |
Elmas et al. | Selective and sensitive fluorescent and colorimetric chemosensor for detection of CO32-anions in aqueous solution and living cells | |
Wang et al. | A dual-mode turn-on fluorescent BODIPY-based probe for visualization of mercury ions in living cells | |
CN108658838B (zh) | 一种基于七甲川吲哚菁的甲醛荧光探针及其制备方法和使用方法 | |
CN110092773B (zh) | 一种氧杂蒽类衍生物及其制备方法和应用 | |
CN109761978A (zh) | 一种检测苯硫酚的近红外荧光探针及其合成方法与应用 | |
CN109988560A (zh) | 一种新型香豆素衍生物的肼荧光探针 | |
CN105985769B (zh) | 一种苯硫酚荧光探针的制备及应用 | |
Ning et al. | A novel colorimetric and fluorescence turn-on pH sensor with a notably large Stokes shift for its application | |
CN110452687A (zh) | 一种连二亚硫酸钠比色荧光探针及其制备方法和应用 | |
CN114516836A (zh) | 一种荧光探针材料及其制备方法和检测硫化物的方法 | |
CN108558839B (zh) | 一种香豆素-吡啶化合物,制备方法及其应用 | |
CN111138431A (zh) | 一种检测苯硫酚的反应型荧光探针及其合成方法与应用 | |
CN107831165B (zh) | 一种双通道铜离子检测试纸及其制备方法 | |
CN107843578B (zh) | 一种基于香豆素铜离子配合物的荧光探针,制备方法及其在选择性识别焦磷酸盐中的应用 | |
CN109761969A (zh) | 一种水溶性萘酰亚胺类化合物的合成及其用途 | |
CN112479930B (zh) | α-苯基肉桂腈衍生物及制备方法及应用 |
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: 20221227 |
|
CF01 | Termination of patent right due to non-payment of annual fee |