CN114113057B - 氧化铜纳米棒-血红素功能化石墨烯及其制备方法和应用 - Google Patents
氧化铜纳米棒-血红素功能化石墨烯及其制备方法和应用 Download PDFInfo
- Publication number
- CN114113057B CN114113057B CN202111338935.7A CN202111338935A CN114113057B CN 114113057 B CN114113057 B CN 114113057B CN 202111338935 A CN202111338935 A CN 202111338935A CN 114113057 B CN114113057 B CN 114113057B
- Authority
- CN
- China
- Prior art keywords
- cuo
- bpf
- heme
- peroxidase
- concentration
- 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
- QPLDLSVMHZLSFG-UHFFFAOYSA-N Copper oxide Chemical compound [Cu]=O QPLDLSVMHZLSFG-UHFFFAOYSA-N 0.000 title claims abstract description 73
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 11
- 229910021389 graphene Inorganic materials 0.000 title claims abstract description 11
- 239000005751 Copper oxide Substances 0.000 title claims abstract description 8
- 229910000431 copper oxide Inorganic materials 0.000 title claims abstract description 8
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- PXKLMJQFEQBVLD-UHFFFAOYSA-N bisphenol F Chemical compound C1=CC(O)=CC=C1CC1=CC=C(O)C=C1 PXKLMJQFEQBVLD-UHFFFAOYSA-N 0.000 claims abstract description 74
- 238000000034 method Methods 0.000 claims abstract description 16
- 102000003992 Peroxidases Human genes 0.000 claims abstract description 8
- 108040007629 peroxidase activity proteins Proteins 0.000 claims abstract description 8
- 150000003278 haem Chemical class 0.000 claims abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 13
- 238000001514 detection method Methods 0.000 claims description 13
- 239000000047 product Substances 0.000 claims description 11
- 239000000725 suspension Substances 0.000 claims description 10
- 238000002835 absorbance Methods 0.000 claims description 9
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 239000010949 copper Substances 0.000 claims description 9
- BTIJJDXEELBZFS-QDUVMHSLSA-K hemin Chemical compound CC1=C(CCC(O)=O)C(C=C2C(CCC(O)=O)=C(C)\C(N2[Fe](Cl)N23)=C\4)=N\C1=C/C2=C(C)C(C=C)=C3\C=C/1C(C)=C(C=C)C/4=N\1 BTIJJDXEELBZFS-QDUVMHSLSA-K 0.000 claims description 9
- 229940025294 hemin Drugs 0.000 claims description 9
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims description 9
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims description 9
- 239000001267 polyvinylpyrrolidone Substances 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 8
- 239000007853 buffer solution Substances 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 6
- 239000002244 precipitate Substances 0.000 claims description 5
- 238000009210 therapy by ultrasound Methods 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 230000035484 reaction time Effects 0.000 claims description 4
- 238000010521 absorption reaction Methods 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- NWZSZGALRFJKBT-KNIFDHDWSA-N (2s)-2,6-diaminohexanoic acid;(2s)-2-hydroxybutanedioic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O.NCCCC[C@H](N)C(O)=O NWZSZGALRFJKBT-KNIFDHDWSA-N 0.000 claims description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 2
- SXTLQDJHRPXDSB-UHFFFAOYSA-N copper;dinitrate;trihydrate Chemical compound O.O.O.[Cu+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O SXTLQDJHRPXDSB-UHFFFAOYSA-N 0.000 claims description 2
- IKDUDTNKRLTJSI-UHFFFAOYSA-N hydrazine monohydrate Substances O.NN IKDUDTNKRLTJSI-UHFFFAOYSA-N 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 5
- 238000011534 incubation Methods 0.000 claims 2
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 11
- 230000003197 catalytic effect Effects 0.000 abstract description 8
- 239000000463 material Substances 0.000 abstract description 8
- 238000004737 colorimetric analysis Methods 0.000 abstract description 4
- 239000002114 nanocomposite Substances 0.000 abstract description 4
- 239000002086 nanomaterial Substances 0.000 abstract description 4
- 239000000758 substrate Substances 0.000 abstract description 4
- 230000003647 oxidation Effects 0.000 abstract description 3
- 238000007254 oxidation reaction Methods 0.000 abstract description 3
- 238000001027 hydrothermal synthesis Methods 0.000 abstract description 2
- 239000002073 nanorod Substances 0.000 abstract description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 abstract 1
- 230000002708 enhancing effect Effects 0.000 abstract 1
- 239000002135 nanosheet Substances 0.000 abstract 1
- 238000002474 experimental method Methods 0.000 description 5
- NIQCNGHVCWTJSM-UHFFFAOYSA-N Dimethyl phthalate Chemical compound COC(=O)C1=CC=CC=C1C(=O)OC NIQCNGHVCWTJSM-UHFFFAOYSA-N 0.000 description 4
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical group NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 3
- 238000006555 catalytic reaction Methods 0.000 description 3
- 230000002195 synergetic effect Effects 0.000 description 3
- 238000000026 X-ray photoelectron spectrum Methods 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- FBSAITBEAPNWJG-UHFFFAOYSA-N dimethyl phthalate Natural products CC(=O)OC1=CC=CC=C1OC(C)=O FBSAITBEAPNWJG-UHFFFAOYSA-N 0.000 description 2
- 229960001826 dimethylphthalate Drugs 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- HQFLTUZKIRYQSP-UHFFFAOYSA-N 3-ethyl-2h-1,3-benzothiazole-6-sulfonic acid Chemical compound OS(=O)(=O)C1=CC=C2N(CC)CSC2=C1 HQFLTUZKIRYQSP-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010033128 Ovarian cancer Diseases 0.000 description 1
- 206010061535 Ovarian neoplasm Diseases 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 208000024313 Testicular Neoplasms Diseases 0.000 description 1
- 206010057644 Testis cancer Diseases 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- IISBACLAFKSPIT-UHFFFAOYSA-N bisphenol A Chemical compound C=1C=C(O)C=CC=1C(C)(C)C1=CC=C(O)C=C1 IISBACLAFKSPIT-UHFFFAOYSA-N 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000000598 endocrine disruptor Substances 0.000 description 1
- 231100000049 endocrine disruptor Toxicity 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 125000005842 heteroatom Chemical group 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 239000002064 nanoplatelet Substances 0.000 description 1
- 229910000069 nitrogen hydride Inorganic materials 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 201000003120 testicular cancer Diseases 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
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
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems 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
- G01N21/78—Systems 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
-
- 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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
-
- 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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N2021/0106—General arrangement of respective parts
- G01N2021/0112—Apparatus in one mechanical, optical or electronic block
Landscapes
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Engineering & Computer Science (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
本发明涉及一种氧化铜纳米棒‑血红素功能化石墨烯(CuO/H‑Gr)及其制备方法和应用。通过对氧化铜纳米材料(CuO)简单的掺杂，可提高CuO的过氧化物酶活性，进而增强与过氧化氢(H2O2)之间的亲和力。本发明通过一步水热法合成了血红素功能化石墨烯纳米片(H‑Gr)，CuO纳米棒附着在H‑Gr上，形成CuO/H‑Gr。该纳米复合材料的类过氧化物酶活性明显高于单一的CuO和H‑Gr。在H2O2存在下，CuO/H‑Gr可催化氧化底物3,3',5,5'‑四甲基联苯胺(TMB)生成蓝色产物(oxTMB)。但当双酚F(BPF)存在时，该材料的类过氧化物酶催化活性显著降低，从而可成功构建一种比色分析方法，实现对BPF方便、快速的检测。
Description
技术领域
本发明涉及纳米复合材料，具体属于一种氧化铜纳米棒-血红素功能化石墨烯(CuO/H-Gr)的制备方法和应用。
背景技术
双酚F(BPF)作为双酚A(BPA)的替代品，广泛存在于饮料包装、塑料、食品罐等消费品中。研究表明，它是一种具有毒性的内分泌干扰物，会对人体健康造成不可逆转的影响，比如会导致生育能力下降、卵巢癌和睾丸癌等多种疾病。因此，简单、快速地检测BPF对人类的生存和健康有重要意义。比色分析方法因其响应快速、成本低廉和操作模式简单而受到广泛关注。
纳米酶克服了天然酶的诸多缺陷，将其应用于许多领域。氧化铜纳米材料(CuO)在弱酸性条件下可以模拟过氧化物酶，但单一CuO催化性能不高，因此现在市场上引入了杂原子、空位或缺陷位调控其催化活性。然而，当CuO转移到溶液中时，它们变得不稳定、不易分散，导致聚集并且催化效率降低。因此，提供一种既能提高CuO的分散性又能改善其过氧化物酶活性的技术方案，具有实用意义。
发明内容
针对现有技术存在的问题，本发明的目的在于提供一种氧化铜纳米棒-血红素功能化石墨烯(CuO/H-Gr)的制备方法，以及CuO/H-Gr在检测双酚F(BPF)中的应用。
本发明的目的是通过以下技术方案实现的：
一种类过氧化物酶CuO/H-Gr的制备方法，包括如下步骤：
(1)将血红素(hemin)、石墨烯氧化物(GO)均匀的分散在二次水中搅拌10～20min；然后依次加入氨水(NH3·H2O)和水合肼(N2H4·H2O)；在50～70℃下水浴3.0～5.0h，静置冷却至室温；在10000～12000rpm下离心10～20min，得到的产物离心洗涤两次；最后将获得的沉淀分散在10mL的二次水中超声处理，得到血红素功能化石墨烯(H-Gr)悬浮液；
所述的NH3·H2O和N2H4·H2O的质量分数分别为25％和80％，hemin、GO、NH3·H2O、N2H4·H2O和H2O的质量比为1.2-1.6:1.0:8.0-10:13-16:1800-2000；
(2)将三水合硝酸铜(Cu(NO3)2·3H2O)、聚乙烯吡咯烷酮(PVP)加入到步骤(1)制得的H-Gr悬浮液中，搅拌10～20min；该混合溶液加热至90～110℃后，将新鲜配置的NaOH溶液快速加入，持续搅拌5～10min，静置冷却至室温；在3000～4000rpm下离心5～10min，得到的产物离心洗涤两次；最后将获得的沉淀分散在2.0mL的二次水中超声处理，制成CuO/H-Gr；
所述的Cu(NO3)2·3H2O、PVP、H-Gr、NaOH和H2O的质量比为1.0:0.16-0.24:1.0:20:1800-2000。
进一步优选：
所述步骤(1)中hemin、GO、NH3·H2O、N2H4·H2O和H2O的质量比为1.4:1.0:9.1:13:2000。
所述步骤(1)中水浴温度为60℃，反应时间为3.5h。
所述步骤(1)中在12000rpm下离心15min。
所述步骤(2)中Cu(NO3)2·3H2O、PVP、H-Gr、NaOH和H2O的质量比为1.0:0.20:1.0:20:2000。
所述步骤(2)中反应温度为100℃，反应时间为5min。
所述步骤(2)中在4000rpm下离心10min。
所述CuO/H-Gr在酸性条件下作为具有过氧化物酶活性的催化剂，比单一H-Gr和CuO具有更好的催化性能。
所述CuO/H-Gr在酸性条件下作为具有过氧化物酶活性的催化剂，用于催化氧化不同底物，显示不同的颜色；当BPF存在时，该材料的类过氧化物酶催化活性显著降低，显色反应减弱；因此该材料可用于对BPF的定量检测。
与现有技术相比，本发明的有益效果：本发明通过简单的水热法制得CuO/H-Gr纳米复合材料。该复合材料将GO、hemin和CuO三者的优良性能结合，不仅具有较大的比表面积，而且改善了材料分散性，还起到协同催化效应，增强了纳米材料的类过氧化物酶活性。与单一H-Gr和CuO相比，CuO/H-Gr展现出更高的催化性能。在H2O2存在下，CuO/H-Gr可催化氧化TMB生成oxTMB。当BPF存在时，CuO/H-Gr的类过氧化物酶活性显著降低。因此，成功地构建了一种比色分析方法，实现了对BPF方便、快速的检测。
附图说明
图1是本发明制备的CuO/H-Gr透射电子显微镜图和X-射线光电子能谱图。
图2是本发明制备的CuO/H-Gr以及其他对比材料在TMB和H2O2存在下的吸光度图。
图3是本发明制备的CuO/H-Gr在不同底物和H2O2存在下显色反应图。
图4是本发明制备的CuO/H-Gr检测BPF的紫外吸收光谱图。
图5是本发明制备的CuO/H-Gr检测BPF的特异性比较图。
具体实施方式
下面结合实施例和附图对本发明技术解决方案作进一步详细说明，这些实施例不能理解为是对本发明技术解决方案的限制。
实施例1制备CuO/H-Gr，具体步骤：
(1)按照Hummers方法合成GO，将制备好的5.0mg GO超声分散于10mL二次水中，得到0.50mg·mL-1的GO悬浮液；
(2)将7.0mg hemin溶于步骤(1)制备的GO悬浮液中，搅拌10min，依次加入200μLNH3·H2O和80μL N2H4·H2O(NH3·H2O和N2H4·H2O的质量分数分别为25％和80％)；在60℃下水浴3.5h，静置冷却到室温；在12000rpm下离心15min；得到的产物用二次水洗涤两次，之后加入二次水制成0.50mg·mL-1的H-Gr悬浮液；
(3)将200μL 0.020M Cu(NO3)2·3H2O和100μL 2.0mg·mL-1PVP添加到2.0mL步骤(2)制备的H-Gr悬浮液中，磁力搅拌15min；在剧烈搅拌下，将混合物加热至100℃，然后快速加入1.0mL 0.50M NaOH溶液，持续加热5min，静置冷却至室温；在4000rpm下离心10min，得到的产物离心洗涤两次；最后将获得的沉淀分散在2.0mL的二次水中超声处理，得到CuO/H-Gr悬浮液。
制备的CuO/H-Gr的透射电子显微镜图和X-射线光电子能谱图，见图1。从图1A中可以看出轻薄且带有褶皱结构的H-Gr成功合成；从图1B中可以看出CuO纳米棒均匀分布在H-Gr纳米片表面；图1C为CuO/H-Gr全谱，证明CuO/H-Gr成功合成；图1D中934.6eV和953.9eV处的特征峰分别代表CuO中Cu 2p3/2和Cu 2p1/2的自旋轨道能级，在943.7eV和962.2eV处的峰进一步证实了存在Cu2+。
实施例2 CuO/H-Gr具有类过氧化物酶活性的验证，具体为：
在1590μL 0.20M HAc-NaAc缓冲溶液(pH 4.0)中加入10μL 0.50mg·mL-1上述实施例1获得的CuO/H-Gr；然后加入200μL 10mM TMB和200μL 100mM H2O2，产生蓝色产物oxTMB；用紫外-可见吸收分光光度计测量λ＝652nm处的吸光度。
对照实验1：将实施例2反应体系中的CuO/H-Gr替换为H-Gr，其余实验体系同实施例2。
对照实验2：将实施例2反应体系中的CuO/H-Gr替换为CuO，其余实验体系同实施例2。
如图2所示，实施例2的吸光度高于对照实验1和对照实验2，说明CuO/H-Gr具有协同催化作用。
实施例3 CuO/H-Gr作为类过氧化物酶催化H2O2氧化底物的应用实验，具体步骤如下：
在1590μL 0.20M HAc-NaAc缓冲溶液(pH 4.0)中加入10μL 0.50mg·mL-1上述实施例1获得的CuO/H-Gr；然后加入200μL 12mM TMB和200μL 120mM H2O2，产生蓝色产物oxTMB；观察反应颜色变化。
对照实验1：将实施例3反应体系中的TMB替换为邻苯二胺(OPD)，其余实验体系同实施例3。
对照实验2：将实施例3反应体系中的TMB替换为2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)(ABTS)，其余实验体系同实施例3。
如图3所示，在H2O2存在下，CuO/H-Gr可将TMB、OPD和ABTS分别催化氧化为蓝色、黄色和绿色产物。
实施例4 CuO/H-Gr作为类过氧化物酶检测BPF含量，具体步骤如下：
在1390μL 0.20M HAc-NaAc缓冲溶液(pH 4.0)中加入10μL 0.50mg·mL-1上述实施例1获得的CuO/H-Gr和200μL不同浓度的BPF(0.10、0.32、1.0、3.2、10、32ng·mL-1),孵育5min。再依次加入200μL 12mM TMB和200μL 120mM H2O2，产生蓝色产物oxTMB；利用紫外分光光度计测其400-800nm内的吸光度。
如图4为本发明制备的CuO/H-Gr在加入不同浓度BPF的λ-A光谱图，以及lg浓度BPF-A曲线。从图中可以看出，在0.10-32ng·mL-1范围内，λ＝652nm处的吸光度与lg浓度BPF呈现良好的线性关系，方程为：A＝-0.1867lg浓度BPF+0.4501(R2＝0.9983)，检出限为0.10ng·mL-1。说明该方法线性范围宽，灵敏度高，可以利用该方法检测BPF。表1为本发明制备的CuO/H-Gr在含有1.0％实际样品的HAc-NaAc缓冲溶液中，加标回收法测试结果。结果显示，样品回收率在97.5％-99.6％之间，RSD在1.20％–6.88％之间，满足实际样品检测的要求。说明CuO/H-Gr可用于实际样品中BPF的检测。
表1实际水样中的检测结果
实施例5 CuO/H-Gr作为类过氧化物酶检测BPF的特异性比较，具体步骤如下：
在1390μL 0.20M HAc-NaAc缓冲溶液(pH 4.0)中加入10μL 0.50mg·mL-1上述实施例1获得的CuO/H-Gr和200μL 1.0ng·mL-1BPF，孵育5min。再依次加入200μL 12mM TMB和200μL 120mM H2O2，产生蓝色产物oxTMB；用紫外-可见吸收分光光度计测量λ＝652nm处的吸光度。
对照试验1：将实施例5中的1.0ng·mL-1BPF替换为100ng·mL-1Hg2+,其余同实施例5。
对照试验2：将实施例5中的1.0ng·mL-1BPF替换为100ng·mL-1Ni2+,其余同实施例5。
对照试验3：将实施例5中的1.0ng·mL-1BPF替换为100ng·mL-1Na+,其余同实施例5。
对照试验4：将实施例5中的1.0ng·mL-1BPF替换为100ng·mL-1PO4 3-,其余同实施例5。
对照试验5：将实施例5中的1.0ng·mL-1BPF替换为100ng·mL-1邻苯二甲酸二甲酯(DMP),其余同实施例5。
图5为CuO/H-Gr作为类过氧化物酶检测BPF的特异性比较图，可以看出干扰物的加入几乎不影响BPF的检测。
本发明方法将CuO与H-Gr结合，制备了CuO/H-Gr纳米复合材料。该材料结合了GO、hemin和CuO三者的优良性质，不仅具有较大的比表面积，而且改善了材料分散性，还起到协同催化效应，增强了纳米材料的类过氧化物酶活性。本发明构建的比色分析方法操作简单、可行性强，可简单快速的检测BPF。
Claims (10)
1.一种氧化铜纳米棒-血红素功能化石墨烯(CuO/H-Gr)的制备方法，其特征在于，包括如下步骤：
(1)将血红素(hemin)、石墨烯氧化物(GO)均匀的分散在二次水中搅拌10～20min；然后依次加入氨水(NH3·H2O)和水合肼(N2H4·H2O)；在50～70℃下水浴3.0～5.0h，静置冷却至室温；在10000～12000rpm下离心10～20min，得到的产物离心洗涤两次；最后将获得的沉淀分散在10mL的二次水中超声处理，得到血红素功能化石墨烯(H-Gr)悬浮液；
所述的NH3·H2O和N2H4·H2O的质量分数分别为25％和80％，hemin、GO、NH3·H2O、N2H4·H2O和H2O的质量比为1.2-1.6:1.0:8.0-10:13-16:1800-2000；
(2)将三水合硝酸铜(Cu(NO3)2·3H2O)、聚乙烯吡咯烷酮(PVP)加入到步骤(1)制得的H-Gr悬浮液中，搅拌10～20min；该混合溶液加热至90～110℃后，将新鲜配置的NaOH溶液快速加入，持续搅拌5～10min，静置冷却至室温；在3000～4000rpm下离心5～10min，得到的产物离心洗涤两次；最后将获得的沉淀分散在2.0mL的二次水中超声处理，制成CuO/H-Gr；
所述的Cu(NO3)2·3H2O、PVP、H-Gr、NaOH和H2O的质量比为1.0:0.16-0.24:1.0:20:1800-2000。
2.如权利要求1所述的CuO/H-Gr的制备方法，其特征在于，所述步骤(1)中hemin、GO、NH3·H2O、N2H4·H2O和H2O的质量比为1.4:1.0:9.1:13:2000。
3.如权利要求1所述的CuO/H-Gr的制备方法，其特征在于，所述步骤(1)中水浴温度为60℃，反应时间为3.5h。
4.如权利要求1所述的CuO/H-Gr的制备方法，其特征在于，所述步骤(1)中在12000rpm下离心15min。
5.如权利要求1所述的CuO/H-Gr的制备方法，其特征在于，所述步骤(2)中Cu(NO3)2·3H2O、PVP、H-Gr、NaOH和H2O的质量比为1.0:0.20:1.0:20:2000。
6.如权利要求1所述的CuO/H-Gr的制备方法，其特征在于，所述步骤(2)中反应温度为100℃，反应时间为5min。
7.如权利要求1所述的CuO/H-Gr的制备方法，其特征在于，所述步骤(2)中在4000rpm下离心10min。
8.如权利要求1-7任一所述方法制得的CuO/H-Gr。
9.如权利要求8所述的CuO/H-Gr作为类过氧化物酶在检测双酚F(BPF)中的应用。
10.如权利要求9所述的CuO/H-Gr作为类过氧化物酶在检测BPF中的应用，其特征在于，操作步骤如下：
(1)将CuO/H-Gr悬浮液和不同浓度的BPF加入醋酸-醋酸钠(HAc-NaAc)缓冲溶液，孵育；然后加入固定浓度的TMB及H2O2，产生oxTMB；
(2)用紫外-可见吸收分光光度计测定步骤(1)得到的oxTMB的吸光度，得到不同浓度BPF所对应的吸光度；
所述步骤(1)的CuO/H-Gr浓度保持在2.5μg·mL-1；HAc-NaAc缓冲溶液浓度保持在0.20M，pH为4.0；孵育温度为室温，孵育时间为5min；TMB浓度保持在1.2mM，H2O2浓度保持在12mM；
所述步骤(2)吸光度是在λ＝652nm处测量。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111338935.7A CN114113057B (zh) | 2021-11-12 | 2021-11-12 | 氧化铜纳米棒-血红素功能化石墨烯及其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111338935.7A CN114113057B (zh) | 2021-11-12 | 2021-11-12 | 氧化铜纳米棒-血红素功能化石墨烯及其制备方法和应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114113057A CN114113057A (zh) | 2022-03-01 |
| CN114113057B true CN114113057B (zh) | 2024-05-24 |
Family
ID=80379016
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111338935.7A Active CN114113057B (zh) | 2021-11-12 | 2021-11-12 | 氧化铜纳米棒-血红素功能化石墨烯及其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114113057B (zh) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108273554A (zh) * | 2018-01-09 | 2018-07-13 | 西安交通大学 | 一种g-C3N4@Hemin复合物模拟酶的制备与应用 |
| CN108872204A (zh) * | 2018-04-27 | 2018-11-23 | 中国科学院兰州化学物理研究所 | 一种二维多孔石墨烯/氧化亚铜复合材料的制备和应用 |
| AU2020100704A4 (en) * | 2020-05-05 | 2020-06-11 | Chen, Shumeng Miss | A method of synthesis of ultrathin palladium nanosheet with peroxidase mimetic activity for the colorimetric detection of H2O2 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4252629A3 (en) * | 2016-12-07 | 2023-12-27 | Biora Therapeutics, Inc. | Gastrointestinal tract detection methods, devices and systems |
-
2021
- 2021-11-12 CN CN202111338935.7A patent/CN114113057B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108273554A (zh) * | 2018-01-09 | 2018-07-13 | 西安交通大学 | 一种g-C3N4@Hemin复合物模拟酶的制备与应用 |
| CN108872204A (zh) * | 2018-04-27 | 2018-11-23 | 中国科学院兰州化学物理研究所 | 一种二维多孔石墨烯/氧化亚铜复合材料的制备和应用 |
| AU2020100704A4 (en) * | 2020-05-05 | 2020-06-11 | Chen, Shumeng Miss | A method of synthesis of ultrathin palladium nanosheet with peroxidase mimetic activity for the colorimetric detection of H2O2 |
Non-Patent Citations (2)
| Title |
|---|
| 基于氧化石墨烯模拟酶比色法检测汞离子;林茜;陆一松;杨胜园;刘露群;李菲菲;何顺珍;;光谱学与光谱分析;20181015(10);全文 * |
| 纳米酶在污染物去除和生物分子快速检测中的应用;冉繁鹏;《中国优秀硕士论文全文数据库(工程科技Ⅰ辑)》;20210915;全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114113057A (zh) | 2022-03-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Tang et al. | Fe3O4 and metal–organic framework MIL-101 (Fe) composites catalyze luminol chemiluminescence for sensitively sensing hydrogen peroxide and glucose | |
| CN107890873B (zh) | 一种空心状铂铜钴三元合金纳米颗粒模拟酶及其制备和应用 | |
| Guan et al. | Synthesis of copper sulfide nanorods as peroxidase mimics for the colorimetric detection of hydrogen peroxide | |
| Dong et al. | Graphene oxide–Fe 3 O 4 magnetic nanocomposites with peroxidase-like activity for colorimetric detection of glucose | |
| Chen et al. | N, N′-di-caboxy methyl perylene diimide (PDI) functionalized CuO nanocomposites with enhanced peroxidase-like activity and their application in visual biosensing of H 2 O 2 and glucose | |
| Wang et al. | Cobalt oxyhydroxide nanoflakes with intrinsic peroxidase catalytic activity and their application to serum glucose detection | |
| CN108445142B (zh) | 一种铑片纳米酶在模拟生物酶催化中的应用 | |
| Munyemana et al. | Deep eutectic solvent-assisted facile synthesis of copper hydroxide nitrate nanosheets as recyclable enzyme-mimicking colorimetric sensor of biothiols | |
| Wang et al. | A double carbon dot system composed of N, Cl-doped carbon dots and N, Cu-doped carbon dots as peroxidase mimics and as fluorescent probes for the determination of hydroquinone by fluorescence | |
| Yu et al. | A nano-sized Cu-MOF with high peroxidase-like activity and its potential application in colorimetric detection of H 2 O 2 and glucose | |
| Basiri et al. | Green synthesis of reduced graphene oxide-Ag nanoparticles as a dual-responsive colorimetric platform for detection of dopamine and Cu2+ | |
| Zhao et al. | A copper (II)/cobalt (II) organic gel with enhanced peroxidase-like activity for fluorometric determination of hydrogen peroxide and glucose | |
| CN111203221B (zh) | 铁酸钴纳米团簇模拟酶及制备方法以及用其检测亚硫酸根的方法 | |
| Ju et al. | Modification and application of Fe3O4 nanozymes in analytical chemistry: A review | |
| Chen et al. | Porous surface MnO 2 microspheres as oxidase mimetics for colorimetric detection of sulfite | |
| Lin et al. | Luminol and gold nanoparticle-co-precipitated reduced graphene oxide hybrids with long-persistent chemiluminescence for cholesterol detection | |
| Chen et al. | Palygorskite@ Co3O4 nanocomposites as efficient peroxidase mimics for colorimetric detection of H2O2 and ascorbic acid | |
| Han et al. | Bioinspired Nanozymes with pH‐Independent and Metal Ions‐Controllable Activity: Field‐Programmable Logic Conversion of Sole Logic Gate System | |
| Luo et al. | Bimetallic Fe-Co nanoalloy confined in porous carbon skeleton with enhanced peroxidase mimetic activity for multiple biomarkers monitoring | |
| CN108872204B (zh) | 一种二维多孔石墨烯/氧化亚铜复合材料的制备和应用 | |
| Hajiagha et al. | Synthesis of cobalt-modified MSN as a model enzyme: Evaluation of the peroxidatic performance | |
| Cai et al. | MoS2‐Pt3Au1 Nanocomposites with enhanced peroxidase‐like activities for selective colorimetric detection of phenol | |
| CN113105646B (zh) | 双金属-有机无限配位聚合物纳米微球的制备方法和应用 | |
| Li et al. | In situ synthesis of fluorescent polydopamine polymer dots based on Fenton reaction for a multi-sensing platform | |
| CN114113057B (zh) | 氧化铜纳米棒-血红素功能化石墨烯及其制备方法和应用 |
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 |