CN111007049B - 一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法 - Google Patents
一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法 Download PDFInfo
- Publication number
- CN111007049B CN111007049B CN201911365971.5A CN201911365971A CN111007049B CN 111007049 B CN111007049 B CN 111007049B CN 201911365971 A CN201911365971 A CN 201911365971A CN 111007049 B CN111007049 B CN 111007049B
- Authority
- CN
- China
- Prior art keywords
- fmn
- solution
- sample
- phosphate
- detection
- 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
- FVTCRASFADXXNN-UHFFFAOYSA-N flavin mononucleotide Natural products OP(=O)(O)OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-UHFFFAOYSA-N 0.000 title claims abstract description 60
- FVTCRASFADXXNN-SCRDCRAPSA-N flavin mononucleotide Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-SCRDCRAPSA-N 0.000 title claims abstract description 47
- 239000011768 flavin mononucleotide Substances 0.000 title claims abstract description 45
- 235000019231 riboflavin-5'-phosphate Nutrition 0.000 title claims abstract description 45
- 229910019142 PO4 Inorganic materials 0.000 title claims abstract description 40
- 239000010452 phosphate Substances 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 26
- 229940013640 flavin mononucleotide Drugs 0.000 title claims abstract description 16
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 15
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 title claims abstract description 13
- 238000001514 detection method Methods 0.000 claims abstract description 25
- 239000002105 nanoparticle Substances 0.000 claims abstract description 21
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000001179 sorption measurement Methods 0.000 claims abstract description 4
- 239000000523 sample Substances 0.000 claims description 48
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 46
- 239000000243 solution Substances 0.000 claims description 44
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 230000005284 excitation Effects 0.000 claims description 11
- 239000001488 sodium phosphate Substances 0.000 claims description 10
- 229910000162 sodium phosphate Inorganic materials 0.000 claims description 10
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 claims description 10
- 239000012224 working solution Substances 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 7
- 229910021641 deionized water Inorganic materials 0.000 claims description 7
- 239000008351 acetate buffer Substances 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 238000005259 measurement Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 238000011084 recovery Methods 0.000 claims description 4
- 239000012496 blank sample Substances 0.000 claims description 3
- 238000003795 desorption Methods 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 238000010791 quenching Methods 0.000 claims description 2
- 230000000171 quenching effect Effects 0.000 claims description 2
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 238000003891 environmental analysis Methods 0.000 abstract description 2
- 238000004186 food analysis Methods 0.000 abstract description 2
- 239000003814 drug Substances 0.000 abstract 1
- 239000010865 sewage Substances 0.000 description 15
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 8
- 229910052698 phosphorus Inorganic materials 0.000 description 8
- 239000011574 phosphorus Substances 0.000 description 8
- 238000002189 fluorescence spectrum Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 230000007547 defect Effects 0.000 description 3
- 239000008239 natural water Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 238000012851 eutrophication Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 238000004062 sedimentation Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical group OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000000975 co-precipitation Methods 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-M dihydrogenphosphate Chemical compound OP(O)([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-M 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 210000003722 extracellular fluid Anatomy 0.000 description 1
- 229940044631 ferric chloride hexahydrate Drugs 0.000 description 1
- 210000002977 intracellular fluid Anatomy 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- NQXWGWZJXJUMQB-UHFFFAOYSA-K iron trichloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].Cl[Fe+]Cl NQXWGWZJXJUMQB-UHFFFAOYSA-K 0.000 description 1
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 125000005341 metaphosphate group Chemical group 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229940107700 pyruvic acid Drugs 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- 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
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
- C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
- C07F9/65618—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings containing the ring system, e.g. flavins or analogues
-
- 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
- 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
- C09K2211/1074—Heterocyclic compounds characterised by ligands containing more than three nitrogen atoms as heteroatoms
-
- 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"
- G01N2021/6432—Quenching
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Materials Engineering (AREA)
- Optics & Photonics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明公开了一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法,是以黄素单核苷酸FMN为荧光探针,通过磷酸根与FMN在Fe3O4纳米颗粒表面的竞争吸附,实现磷酸根含量的检测。本发明工艺简单、操作方便、选择性好、灵敏度高,在环境分析、食品分析、生命医学等领域,有着广泛的应用前景。
Description
技术领域
本发明涉及分析化学领域,具体涉及一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法。
背景技术
磷是一种自然界中广泛存在的天然非金属元素,在自然水体中和生物体中均发挥着十分重要的作用。磷在人体内主要以磷酸氢根/磷酸二氢根形式存在,在组织液中其浓度达到2.3mM,在细胞内液中其浓度达到29mM,广泛地参与人体内诸多重要生理过程,如:酸碱平衡调节,糖、蛋白质、脂肪的代谢等。在自然水体中,磷主要以正磷酸盐形式存在,其它存在形态的磷(如有机磷、偏磷酸根等)也会在自然水体中逐步氧化水解为正磷酸盐形态。磷是一个衡量水体富营养化的重要指标,当浓度过高时,会造成水体富营养化,使藻类过度生长而快速消耗水中溶解氧,从而导致水生生物死亡、水体毒素上升、水质下降。因此,准确、快速分析磷酸根的含量,在环境分析、食品分析、生物医学等领域都具有极其重要的意义。
目前已有多种磷酸根的分析检测方法,较为成熟的有比色法、离子色谱法、丙酮酸传感器等,但这些方法通常存在样品处理复杂费时、分析检测下限较高、难以实现自动化等不足。
因此,能够实现磷酸根含量的准确高效测定的新方法亟待发展。
发明内容
为避免上述现有技术所存在的不足之处,本发明公开了一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法,以期可以实现水体中磷酸根含量的准确高效测定。
本发明为实现发明目的,采用如下技术方案:
一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法,其特点在于:是以黄素单核苷酸FMN为荧光探针,通过磷酸根与FMN在Fe3O4纳米颗粒表面的竞争吸附,实现磷酸根含量的检测。黄素单核苷酸的分子式如图1所示,其检测原理如图2所示:FMN侧链末端的磷酸根在Fe3O4纳米颗粒表面吸附,造成FMN荧光淬灭;当遇到磷酸根时,磷酸根对Fe3O4表面吸附的FMN进行置换,使FMN从Fe3O4表面解吸附,荧光得以恢复,通过检测荧光强度,实现磷酸根含量的测定。具体包括如下步骤:
步骤1、配置检测用FMN@Fe3O4溶液
将Fe3O4纳米颗粒分散在水中,配成浓度为2mg/mL的Fe3O4纳米颗粒溶液;
配制浓度为5μmol/L的FMN溶液、浓度为4mol/L的氯化钠溶液和浓度为100mmol/LpH=5.8的醋酸盐缓冲溶液;
取0.5mL Fe3O4纳米颗粒溶液、0.2mL FMN溶液、0.125mL氯化钠溶液、0.8mL醋酸盐缓冲溶液和0.175mL去离子水,振荡混合均匀后,再室温静置0.5h,使荧光探针FMN在Fe3O4纳米颗粒表面充分吸附,即获得检测用FMN@Fe3O4溶液;
步骤2、标准工作曲线的绘制
配置浓度范围在2μmol/L-75μmol/L的磷酸钠标准工作溶液,分别取0.2mL加入到1.8mL所述检测用FMN@Fe3O4溶液中,室温静置1h,使FMN充分解吸附,然后在激发波长为450nm条件下测定521nm处的荧光强度,记为I标;
将0.2mL去离子水加入到1.8mL所述检测用FMN@Fe3O4溶液中,室温静置1h,作为空白样品,在激发波长为450nm条件下测定521nm处的荧光强度,记为I0;
计算ΔI=I标-I0,以ΔI对磷酸根的浓度关系,做标准工作曲线;
步骤3、待测样品检测
取0.2mL待测样品加入到1.8mL所述检测用FMN@Fe3O4溶液中,室温静置1h,使FMN充分解吸附,然后在激发波长为450nm条件下测定521nm处的荧光强度,记为I样品;
计算ΔI样品=I样品-I0,再根据步骤2的标准工作曲线,获得所述待测样品的磷酸根含量。
图3为黄素单核苷酸的三维荧光光谱图,从图中可以看出当激发波长为450nm时,521nm的荧光发射最强。因此,本发明在激发波长为450nm条件下测定521nm处的荧光发射强度,作为磷酸根含量检测的依据。
与已有技术相比,本发明的有益效果体现在:
本发明以FMN为荧光探针,利用FMN侧链末端的磷酸官能团与磷酸根在Fe3O4纳米颗粒表面竞争吸附,通过荧光强度的变化实现磷酸根含量的测定,避免了传统方法存在的样品处理费时、检测灵敏度差等不足,具有操作简单快速、检测下限低(2μmol/L,62μg/L)、灵敏度高的优势。且对于不同离子,具有良好的抗干扰能力,选择性好。
附图说明
图1为黄素单核苷酸的分子式;
图2为本发明以黄素单核苷酸为荧光探针测定磷酸根含量的原理示意图;
图3为黄素单核苷酸的三维荧光光谱图;
图4为本发明实施例中各磷酸钠标准工作溶液的荧光光谱图;
图5为本发明实施例中所得标准工作曲线图(2-75μmol/L);
图6为图5中浓度2~10μmol/L之间曲线的线性拟合工作曲线(c=0.11272ΔI+5.739);
图7为本发明实施例中城市污水厂各水样的荧光光谱图。
具体实施方式
下面对本发明的实施例作详细说明,本实施例在以本发明技术方案为前提下进行实施,给出了详细的实施方式和具体的操作过程,但本发明的保护范围不限于下述的实施例。
下述实施例所用Fe3O4纳米颗粒通过水热共沉淀法制得,具体步骤为:称取13.9g七水合硫酸亚铁和27g六水合氯化铁,溶解于除去溶解氧的50mL去离子水中,氮气保护下60℃水浴加热搅拌10min,再缓慢加入8mL质量浓度为25~28%的浓氨水,继续恒温搅拌20min,再80℃水浴静置30min。所得反应液置于高温反应釜中于145℃对流烘箱加热6h,然后离心得到Fe3O4纳米颗粒粗品。将粗品依次水洗两次、酸洗两次、水洗两次、无水乙醇洗涤一次,最后置于60℃对流烘箱干燥24h,即获得Fe3O4纳米颗粒。
下述实施例所用黄素单核苷酸FMN购买自SIGMA
实施例1
本实施例的待测水样取自城市污水处理厂,具体检测方法如下:
步骤1、配置检测用FMN@Fe3O4溶液
将Fe3O4纳米颗粒分散在水中,配成浓度为2mg/mL的Fe3O4纳米颗粒溶液;
配制浓度为5μmol/L的FMN溶液、浓度为4mol/L的氯化钠溶液和浓度为100mmol/LpH=5.8的醋酸盐缓冲溶液;
取0.5mL Fe3O4纳米颗粒溶液、0.2mL FMN溶液、0.125mL氯化钠溶液、0.8mL醋酸盐缓冲溶液和0.175mL去离子水,振荡混合均匀后,再室温静置0.5h,使荧光探针FMN在Fe3O4纳米颗粒表面充分吸附,即获得检测用FMN@Fe3O4溶液;
步骤2、标准工作曲线的绘制
配置浓度范围在2μmol/L-75μmol/L的磷酸钠标准工作溶液(2、3、4、5、7.5、10、20、30、50、75μmol/L),分别取0.2mL加入到1.8mL检测用FMN@Fe3O4溶液中,室温静置1h,使FMN充分解吸附,然后在激发波长为450nm条件下测定521nm处的荧光强度,记为I标;
将0.2mL去离子水加入到1.8mL检测用FMN@Fe3O4溶液中,室温静置1h,作为空白样品,在激发波长为450nm条件下测定521nm处的荧光强度,记为I0;
计算ΔI=I标-I0,以ΔI对磷酸根的浓度关系,做标准工作曲线。
图4为各磷酸钠标准工作溶液的荧光光谱图,图5为标准工作曲线,图6为图5中浓度2~10μmol/L之间曲线的线性拟合工作曲线(c=0.11272ΔI+5.739)。
步骤3、待测样品检测
取0.2mL待测样品加入到1.8mL检测用FMN@Fe3O4溶液中,室温静置1h,使FMN充分解吸附,然后在激发波长为450nm条件下测定521nm处的荧光强度,记为I样品;
计算ΔI样品=I样品-I0,再根据步骤2的标准工作曲线,获得所述待测样品的磷酸根含量。
为验证本发明方法的灵敏度,对如下待测水样进行检测:
1、模拟水样
配置浓度为0.5μmol/L、1μmol/L、5μmol/L的磷酸钠溶液,按上述的方法进行检测,测得浓度分别为0.495μmol/L、0.964μmol/L、5.18μmol/L,确认本发明的方法准确、可靠。
2、城市污水厂水样检测
在城市污水厂的不同工段进行取样:污水厂进水口水样(样品1)、污水处理好氧段水样(样品2)、污水处理厌氧段水样(样品3)、污水处理沉淀池水样(样品4)、污水处理出水口水样(样品5)。
按上述方法进行检测,测得磷酸根的摩尔浓度,并计算出其质量浓度(以磷计),结果如表1所示。
表1.城市污水处理厂水样磷酸根分析检测
样品序号 | 磷酸根浓度(mg/L) | 水样来源 |
1 | 2.235 | 污水厂进水口水样 |
2 | 0.479 | 污水处理好氧段水样 |
3 | 0.813 | 污水处理厌氧段水样 |
4 | 0.398 | 污水处理沉淀池水样 |
5 | 0.355 | 污水处理出水口水样 |
以上述的样品5为基质,加入浓度为0.5μmol/L磷酸钠标准溶液,进行加标回收实验。三个平行样品,得回收率分别为96.7%,98.2%,97.8%,进一步确认本发明的分析方法准确、可靠。
3、湖水、河水样品
取巢湖水样和南淝河水样,按上述方法进行检测,测得磷酸根的摩尔浓度,并计算出其质量浓度(以磷计),结果如表2所示。
表2
水样来源 | 磷酸根浓度(mg/L) |
巢湖-水样 | 0.185 |
南淝河水样 | 0.254 |
以上所述仅为本发明的示例性实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等,均应包含在本发明的保护范围之内。
Claims (3)
1.一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法,其特征在于:是以黄素单核苷酸FMN为荧光探针,通过磷酸根与FMN在Fe3O4纳米颗粒表面的竞争吸附,实现磷酸根含量的检测。
2.根据权利要求1所述的方法,其特征在于:FMN侧链末端的磷酸根在Fe3O4纳米颗粒表面吸附,造成FMN荧光淬灭;当遇到磷酸根时,磷酸根对Fe3O4表面吸附的FMN进行置换,使FMN从Fe3O4表面解吸附,荧光得以恢复,通过检测荧光强度,实现磷酸根含量的测定。
3.根据权利要求1或2所述的方法,其特征在于,包括如下步骤:
步骤1、配置检测用FMN@Fe3O4溶液
将Fe3O4纳米颗粒分散在水中,配成浓度为2mg/mL的Fe3O4纳米颗粒溶液;
配制浓度为5μmol/L的FMN溶液、浓度为4mol/L的氯化钠溶液和浓度为100mmol/L pH=5.8的醋酸盐缓冲溶液;
取0.5mL Fe3O4纳米颗粒溶液、0.2mL FMN溶液、0.125mL氯化钠溶液、0.8mL醋酸盐缓冲溶液和0.175mL去离子水,振荡混合均匀后,再室温静置0.5h,使荧光探针FMN在Fe3O4纳米颗粒表面充分吸附,即获得检测用FMN@Fe3O4溶液;
步骤2、标准工作曲线的绘制
配置浓度范围在2μmol/L-75μmol/L的磷酸钠标准工作溶液,分别取0.2mL加入到1.8mL所述检测用FMN@Fe3O4溶液中,室温静置1h,使FMN充分解吸附,然后在激发波长为450nm条件下测定521nm处的荧光强度,记为I标;
将0.2mL去离子水加入到1.8mL所述检测用FMN@Fe3O4溶液中,室温静置1h,作为空白样品,在激发波长为450nm条件下测定521nm处的荧光强度,记为I0;
计算ΔI=I标-I0,以ΔI对磷酸根的浓度关系,做标准工作曲线;
步骤3、待测样品检测
取0.2mL待测样品加入到1.8mL所述检测用FMN@Fe3O4溶液中,室温静置1h,使FMN充分解吸附,然后在激发波长为450nm条件下测定521nm处的荧光强度,记为I样品;
计算ΔI样品=I样品-I0,再根据步骤2的标准工作曲线,获得所述待测样品的磷酸根含量。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201911365971.5A CN111007049B (zh) | 2019-12-26 | 2019-12-26 | 一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201911365971.5A CN111007049B (zh) | 2019-12-26 | 2019-12-26 | 一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN111007049A CN111007049A (zh) | 2020-04-14 |
CN111007049B true CN111007049B (zh) | 2022-06-14 |
Family
ID=70117999
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201911365971.5A Active CN111007049B (zh) | 2019-12-26 | 2019-12-26 | 一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN111007049B (zh) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS63105697A (ja) * | 1986-10-21 | 1988-05-10 | Toyobo Co Ltd | 化学発光法によるnadh又はnadphの定量法 |
US4806415A (en) * | 1983-12-21 | 1989-02-21 | Miles Inc. | Method and system for determining the presence of adenosine triphosphate or flavin mononucleotide |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20060172320A1 (en) * | 2004-07-13 | 2006-08-03 | Stojanovic Milan N | Modular aptametric sensors without covalently attached fluorophores |
-
2019
- 2019-12-26 CN CN201911365971.5A patent/CN111007049B/zh active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4806415A (en) * | 1983-12-21 | 1989-02-21 | Miles Inc. | Method and system for determining the presence of adenosine triphosphate or flavin mononucleotide |
JPS63105697A (ja) * | 1986-10-21 | 1988-05-10 | Toyobo Co Ltd | 化学発光法によるnadh又はnadphの定量法 |
Non-Patent Citations (2)
Title |
---|
光动力活性氧的研究进展;蒋昕鹏等;《科学通报》;20180630(第18期);全文 * |
测定外周血核黄素及其衍生物含量的HPLC方法研究;韦京豫等;《营养学报》;20060225(第01期);全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN111007049A (zh) | 2020-04-14 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN111269715B (zh) | 一种比率荧光探针及其在可视化检测谷胱甘肽中的应用 | |
CN110987843B (zh) | 基于双金属mof纳米类氧化酶的磷酸根比色检测法 | |
CN113916858A (zh) | 一种利用氮掺杂碳量子点荧光探针检测Cr6+的方法 | |
CN108120831A (zh) | 一种基于链置换及纳米金检测汞离子的比色生物传感器 | |
CN110484242B (zh) | 一种检测水中三价铁离子的荧光探针及其制备和检测方法 | |
CN110907589B (zh) | 一种基于GQDs光催化可视化检测Cu2+的方法 | |
Moll et al. | Complexation of curium (III) by adenosine 5′-triphosphate (ATP): A time-resolved laser-induced fluorescence spectroscopy (TRLFS) study | |
CN110596061A (zh) | 基于BPEI-CuNCs荧光探针快速检测铜离子的方法 | |
CN111007049B (zh) | 一种以黄素单核苷酸为荧光探针测定磷酸根含量的方法 | |
CN113138185A (zh) | 基于mof的sers技术检测牛奶中硫氰酸钠的方法 | |
CN105928912B (zh) | 一种肝素的检测方法 | |
CN110514625A (zh) | 一种人体血清叶酸的测定方法 | |
CN110715915A (zh) | 一种基于鸡蛋蛋清制备的碳量子点检测三价铁离子的方法 | |
CN110655919B (zh) | 一种铜离子荧光探针及其制备方法与应用 | |
CN115980010A (zh) | 一种以氮掺杂碳点为荧光探针检测环境水体中Fe3+的方法 | |
CN113024595B (zh) | 1,3,5,7-四甲基环四硅氧烷基环丙沙星荧光探针及其在铁离子检测中的应用 | |
CN113758908B (zh) | 一种快速荧光检测铬含量的方法 | |
Lei et al. | Nano-fluorescent probes based on DNA-templated copper nanoclusters for fast sensing of thiocyanate | |
CN114813698A (zh) | 一种基于表面增强拉曼光谱检测养殖水中磷酸盐的方法 | |
CN103207160A (zh) | 以纳米金为显色探针的硫氰酸盐快速测定方法 | |
CN108752272B (zh) | 8-氨基喹啉酰胺衍生物、制备方法、应用及其荧光分析的方法 | |
CN103185737B (zh) | 一种检测水样中铅离子的方法 | |
CN112697780A (zh) | 基于锇纳米粒子氧化酶活性的汞离子比色检测方法 | |
CN110698390A (zh) | 一种识别亚硫酸氢根的荧光探针及其制备方法和检测方法 | |
CN109254067A (zh) | 一种基于罗丹明b/还原氧化石墨烯修饰的玻碳电极及其制备和应用 |
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 |