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2021 Volume 49 Issue 12
2021, 49(12): 1945-1954
doi: 10.19756/j.issn.0253-3820.210465
Abstract:
Infectious diseases caused by pathogens seriously threaten human life and health. Rapid and accurate detection of pathogens is an important prerequisite for effective prevention and control of infectious diseases. However, traditional methods for detection of pathogen nucleic acid often require professional personnel and specialized instruments and equipments, which challenges the realization of field detection of pathogen. Using paper as matrix to construct biosensor has the advantages of light and portable, low cost and intuitive results, and is suitable for rapid detection of pathogens in the field by combining with nucleic acid amplification method. Currently, the paper-based pathogen nucleic acid detection technique mainly includes two types:lateral flow strips and paper-based microfluidic devices. Here, antibody-dependent and antibody-independent lateral flow strips, two- and three-dimensional paper-based microfluidic devices and fabrication techniques are reviewed, and the strategies of developing paper-based integrated devices for nucleic acid extraction, amplification and detection of pathogens are introduced, which can provide a reference for the development of new techniques for the field detection of pathogens.
Infectious diseases caused by pathogens seriously threaten human life and health. Rapid and accurate detection of pathogens is an important prerequisite for effective prevention and control of infectious diseases. However, traditional methods for detection of pathogen nucleic acid often require professional personnel and specialized instruments and equipments, which challenges the realization of field detection of pathogen. Using paper as matrix to construct biosensor has the advantages of light and portable, low cost and intuitive results, and is suitable for rapid detection of pathogens in the field by combining with nucleic acid amplification method. Currently, the paper-based pathogen nucleic acid detection technique mainly includes two types:lateral flow strips and paper-based microfluidic devices. Here, antibody-dependent and antibody-independent lateral flow strips, two- and three-dimensional paper-based microfluidic devices and fabrication techniques are reviewed, and the strategies of developing paper-based integrated devices for nucleic acid extraction, amplification and detection of pathogens are introduced, which can provide a reference for the development of new techniques for the field detection of pathogens.
2021, 49(12): 1955-1969
doi: 10.19756/j.issn.0253-3820.210487
Abstract:
Upconversion nanoparticles (UCNPs) can convert low-energy excitation into high-energy emission via multiphoton absorption processes. UCNPs have some unique photophysical properties including large anti-Stokes shift, strong penetration into biological tissues, resistance to photobleaching, low background fluorescence, great chemical stability and low toxicity, therefore, they draw great attention and provide a variety of possibilities for sensing detection, bioimaging, and bioanalysis, and promoting the development of fluorescently labeled probes. In this review, the synthesis methods and surface functionalization strategies of UCNPs were summarized, also the recent progress of UCNPs on food safety detection was reviewed. Finally, the challenges faced by UCNPs and the opportunities for future development were discussed.
Upconversion nanoparticles (UCNPs) can convert low-energy excitation into high-energy emission via multiphoton absorption processes. UCNPs have some unique photophysical properties including large anti-Stokes shift, strong penetration into biological tissues, resistance to photobleaching, low background fluorescence, great chemical stability and low toxicity, therefore, they draw great attention and provide a variety of possibilities for sensing detection, bioimaging, and bioanalysis, and promoting the development of fluorescently labeled probes. In this review, the synthesis methods and surface functionalization strategies of UCNPs were summarized, also the recent progress of UCNPs on food safety detection was reviewed. Finally, the challenges faced by UCNPs and the opportunities for future development were discussed.
2021, 49(12): 1970-1976
doi: 10.19756/j.issn.0253-3820.210474
Abstract:
The development and improvement of instrument for diagnosis of high energy ion generated by the method such as vaccum discharge is very important for improving the research level of high energy plasma. In this work, a time-of-flight mass spectrometer (TOF-MS) for diagnosis of high-energy ion was developed, and the qualitative analysis of high energy ions (H+, H2+, C2+, Al3+, C+, Al2+, O+ and Al+) between 10 and 150 keV was carried out on the basis of aluminum vacuum discharge ion source. The influence of pulse width and voltage of ion gate on performance of TOF-MS was optimized, and it was found that, when the pulse width was 100 ns and the accelerating voltages were 20 kV and 50 kV, respectively, the best voltages of ion gate were ±500 V and ±2500 V. Besides, it was found that the mass resolution was 22 FWHM (Full width at half maximum) when the accelerating voltage was 50 kV. Temporal variation of intensity of different ions was further studied by changing the delay time of pulse of ion gate.
The development and improvement of instrument for diagnosis of high energy ion generated by the method such as vaccum discharge is very important for improving the research level of high energy plasma. In this work, a time-of-flight mass spectrometer (TOF-MS) for diagnosis of high-energy ion was developed, and the qualitative analysis of high energy ions (H+, H2+, C2+, Al3+, C+, Al2+, O+ and Al+) between 10 and 150 keV was carried out on the basis of aluminum vacuum discharge ion source. The influence of pulse width and voltage of ion gate on performance of TOF-MS was optimized, and it was found that, when the pulse width was 100 ns and the accelerating voltages were 20 kV and 50 kV, respectively, the best voltages of ion gate were ±500 V and ±2500 V. Besides, it was found that the mass resolution was 22 FWHM (Full width at half maximum) when the accelerating voltage was 50 kV. Temporal variation of intensity of different ions was further studied by changing the delay time of pulse of ion gate.
2021, 49(12): 1977-1985
doi: 10.19756/j.issn.0253-3820.201379
Abstract:
Deep-sea in-situ chemical analyzer can obtain data with high frequency, which is very useful for scientific research and necessary for deep-sea transportation equipment. In this work, an in-situ analyzer was developed for automated determination of dissolved Fe, Fe, Mn and sulfide simultaneously in deep sea based on continuous flow analysis and spectrophotometric detection. The integrated system consisted of a pressure-resistant housing an oil-filled pressure-compensated vessel, and a self-developed hardware to control two multi-channel peristaltic pumps, four solenoid valves and a photoelectric acquisition module. Experimental conditions related to the analysis, including flow path module, spectrometric regent compositions and pH of buffer solution were evaluated and optimized. This deep-sea in-situ chemical analyzer was designed to work at the depth of 7000 m with a measuring frequency of 1 Hz, and showed a limit of detection as low as 0.013, 0.024, 0.014, and 0.012 μmol/L for dissolved Fe, Fe, Mn and sulfide, respectively. The calibration curves prepared in standard solutions were consistent over the linear range of 0.1-60, 0.2-100, 0.1-40 and 0.1-40 μmol/L for dissolved Fe, Fe, Mn and sulfide, respectively. After the test of shallow sea, hydrostatic pressure and tank test, this analyzer was implemented on tool shed of Jiaolong deep manned submersible, and performed sea trial. Spectra and signal intensity changes of the maximum absorption wavelength were obtained in-situ at the depth of 3196 m. This compact automatic analyzer was suitable for deep sea in-situ determination of the chemical environment of hydrothermal vent habitats.
Deep-sea in-situ chemical analyzer can obtain data with high frequency, which is very useful for scientific research and necessary for deep-sea transportation equipment. In this work, an in-situ analyzer was developed for automated determination of dissolved Fe, Fe, Mn and sulfide simultaneously in deep sea based on continuous flow analysis and spectrophotometric detection. The integrated system consisted of a pressure-resistant housing an oil-filled pressure-compensated vessel, and a self-developed hardware to control two multi-channel peristaltic pumps, four solenoid valves and a photoelectric acquisition module. Experimental conditions related to the analysis, including flow path module, spectrometric regent compositions and pH of buffer solution were evaluated and optimized. This deep-sea in-situ chemical analyzer was designed to work at the depth of 7000 m with a measuring frequency of 1 Hz, and showed a limit of detection as low as 0.013, 0.024, 0.014, and 0.012 μmol/L for dissolved Fe, Fe, Mn and sulfide, respectively. The calibration curves prepared in standard solutions were consistent over the linear range of 0.1-60, 0.2-100, 0.1-40 and 0.1-40 μmol/L for dissolved Fe, Fe, Mn and sulfide, respectively. After the test of shallow sea, hydrostatic pressure and tank test, this analyzer was implemented on tool shed of Jiaolong deep manned submersible, and performed sea trial. Spectra and signal intensity changes of the maximum absorption wavelength were obtained in-situ at the depth of 3196 m. This compact automatic analyzer was suitable for deep sea in-situ determination of the chemical environment of hydrothermal vent habitats.
2021, 49(12): 1986-1994
doi: 10.19756/j.issn.0253-3820.210677
Abstract:
A multifunctional hemostatic sponge (CIP-HNTs/SAs) with rapid hemostasis and high efficiency antibacterial property was built by freeze-drying based on ciprofloxacin (CIP), halloysite (HNTs) and sodium alginate (SA). Fourier transform infrared (FTIR) spectroscopy, scanning electron microscope (SEM), density method and weighing method were used to detect the composition, porous structure, porosity and water absorption of the sodium alginate sponge (HNTs/SAs). Blood clotting time (BCT) and blood clotting index (BCI) were applied to test the hemostatic of HNTs/SAs and the optimal halloysite loading ratio was selected by these tests. Furthermore, the activated partial thrombin time (APTT) and prothrombin time (PT) were used to explore the mechanism of sponge hemostasis. The halloysite hollow tube structure could load with antibacterial drugs. The components of halloysite loaded with antibacterial drugs were characterized by energy dispersive spectroscopy (EDS) and FTIR spectroscopy, and the loaded antibacterial drugs were quantitatively detected by ultraviolet (UV) absorption spectrum. The long-term antibacterial activity of CIP-HNTs/SAs was tested by the agar plate counting methodand inhibition zone, and the biocompatibility of CIP-HNTs/SAs was tested by the cell proliferation test kit (CCK-8) and the hemolysis rate of red blood cells. The results showed that the porous and hydrophilia of sodium alginate sponge could quickly absorb water in the blood to promote blood cell aggregation. Moreover, calcium ions (Ca2+) and halloysite could enhance the body's own coagulation cascade activity. Comparing with the coagulation time in vitro of the commercial gelatin hemostatic sponge, the sodium alginate sponge (HNTs/SA) containing halloysite could achieve rapid hemostasis in only 18 s. Meanwhile, CIP-HNTs/SAs had great antibacterial property and 7-day long-lasting antibacterial performance.
A multifunctional hemostatic sponge (CIP-HNTs/SAs) with rapid hemostasis and high efficiency antibacterial property was built by freeze-drying based on ciprofloxacin (CIP), halloysite (HNTs) and sodium alginate (SA). Fourier transform infrared (FTIR) spectroscopy, scanning electron microscope (SEM), density method and weighing method were used to detect the composition, porous structure, porosity and water absorption of the sodium alginate sponge (HNTs/SAs). Blood clotting time (BCT) and blood clotting index (BCI) were applied to test the hemostatic of HNTs/SAs and the optimal halloysite loading ratio was selected by these tests. Furthermore, the activated partial thrombin time (APTT) and prothrombin time (PT) were used to explore the mechanism of sponge hemostasis. The halloysite hollow tube structure could load with antibacterial drugs. The components of halloysite loaded with antibacterial drugs were characterized by energy dispersive spectroscopy (EDS) and FTIR spectroscopy, and the loaded antibacterial drugs were quantitatively detected by ultraviolet (UV) absorption spectrum. The long-term antibacterial activity of CIP-HNTs/SAs was tested by the agar plate counting methodand inhibition zone, and the biocompatibility of CIP-HNTs/SAs was tested by the cell proliferation test kit (CCK-8) and the hemolysis rate of red blood cells. The results showed that the porous and hydrophilia of sodium alginate sponge could quickly absorb water in the blood to promote blood cell aggregation. Moreover, calcium ions (Ca2+) and halloysite could enhance the body's own coagulation cascade activity. Comparing with the coagulation time in vitro of the commercial gelatin hemostatic sponge, the sodium alginate sponge (HNTs/SA) containing halloysite could achieve rapid hemostasis in only 18 s. Meanwhile, CIP-HNTs/SAs had great antibacterial property and 7-day long-lasting antibacterial performance.
2021, 49(12): 1995-2004
doi: 10.19756/j.issn.0253-3820.210527
Abstract:
A nanozyme-motor (Cu/PDA) with peroxidase-like and catalase-like characteristics was constructed by in situ chelation between Cu2+ and polydopamine (PDA). Cu/PDA can efficiently catalyze the decomposition of hydrogen peroxide to produce oxygen for the movement of nano-motors and to generate highly toxic reactive oxygen species (ROS) for killing bacteria. Promoted by the photothermal effect of PDA, the diffusion range of nanozyme-motor was significantly expanded, which enhanced the damage range of ROS effectively. Antimicrobial experiments results showed that Cu/PDA could inhibit bacterial growth potently. Moreover, with the functional surface modification, Cu/PDA antibacterial system exhibited more distinct antimicrobial activity against the resistant Gram-positive bacteria. The damage range of ROS generated by antibacterial nanozyme was optimized through the combination with nanomotor, which addressed the issue on limited bactericidal efficiency of traditional nanozymes, and showed great potential in the development of new generation of antibacterial nanoagents.
A nanozyme-motor (Cu/PDA) with peroxidase-like and catalase-like characteristics was constructed by in situ chelation between Cu2+ and polydopamine (PDA). Cu/PDA can efficiently catalyze the decomposition of hydrogen peroxide to produce oxygen for the movement of nano-motors and to generate highly toxic reactive oxygen species (ROS) for killing bacteria. Promoted by the photothermal effect of PDA, the diffusion range of nanozyme-motor was significantly expanded, which enhanced the damage range of ROS effectively. Antimicrobial experiments results showed that Cu/PDA could inhibit bacterial growth potently. Moreover, with the functional surface modification, Cu/PDA antibacterial system exhibited more distinct antimicrobial activity against the resistant Gram-positive bacteria. The damage range of ROS generated by antibacterial nanozyme was optimized through the combination with nanomotor, which addressed the issue on limited bactericidal efficiency of traditional nanozymes, and showed great potential in the development of new generation of antibacterial nanoagents.
2021, 49(12): 2005-2014
doi: 10.19756/j.issn.0253-3820.210432
Abstract:
A novel photoelectrochemical (PEC) sensor was constructed based on carbon quantum dots decorated titanium dioxide nanotube arrays (TiO2 NTAs/CQDs). Highly ordered self-supporting titanium dioxide nanotube arrays (TiO2 NTAs) were prepared on the surface of a titanium foil by electrochemical anodization. The diameter and the length of TiO2 nanotube were around 80 nm and 8 μm, respectively. CQDs were prepared by electrochemical anodization of a graphite electrode, with a diameter of 3-9 nm. Uniformly distributed CQDs were decorated on the inner walls of TiO2 NTAs by hydrothermal treatment, examined by TEM observation. CQDs could effectively reduce the band gap of TiO2 NTAs and enhanced its response under visible light. CQDs were electron acceptors that could promote the separation of photo-generated electron-hole pairs. The conductance of TiO2 NTAs was also enhanced by CQDs decoration. Under visible light, TiO2 NTAs/CQDs exhibited excellent PEC response toward 5-hydroxytryptamine (5-HT), with a linear range of 20-300 μmol/L and a detection limit of 11.5 μmol/L (S/N=3). Moreover, the PEC sensor showed good selectivity and stability for determination of 5-HT and was successfully applied to determination of 5-HT in human serum.
A novel photoelectrochemical (PEC) sensor was constructed based on carbon quantum dots decorated titanium dioxide nanotube arrays (TiO2 NTAs/CQDs). Highly ordered self-supporting titanium dioxide nanotube arrays (TiO2 NTAs) were prepared on the surface of a titanium foil by electrochemical anodization. The diameter and the length of TiO2 nanotube were around 80 nm and 8 μm, respectively. CQDs were prepared by electrochemical anodization of a graphite electrode, with a diameter of 3-9 nm. Uniformly distributed CQDs were decorated on the inner walls of TiO2 NTAs by hydrothermal treatment, examined by TEM observation. CQDs could effectively reduce the band gap of TiO2 NTAs and enhanced its response under visible light. CQDs were electron acceptors that could promote the separation of photo-generated electron-hole pairs. The conductance of TiO2 NTAs was also enhanced by CQDs decoration. Under visible light, TiO2 NTAs/CQDs exhibited excellent PEC response toward 5-hydroxytryptamine (5-HT), with a linear range of 20-300 μmol/L and a detection limit of 11.5 μmol/L (S/N=3). Moreover, the PEC sensor showed good selectivity and stability for determination of 5-HT and was successfully applied to determination of 5-HT in human serum.
2021, 49(12): 2015-2022
doi: 10.19756/j.issn.0253-3820.210626
Abstract:
It is an effective attempt to increase the level of reactive oxygen species (ROS) to abrogate the redox balance and successively result in sever cells oxidative damage and death in cancer treatment. Herein, biodegradable, safe and tumor-specific hollow manganese carbonate (MnCO3) nanocarriers (HMC NPs) were synthesized by hydrothermal method. HMC NPs loaded with sonosensitizer protoporphyrin (PpIX) constituted HMC-PpIX nanoparticles (HMC-P NPs), which could be activated in the acidic tumor microenvironment (TME). By degrading and releasing Mn2+ and sonosensitizer, the overloaded Mn2+ in the tumor triggered the Fenton reaction under the physiological buffer environment of HCO3-/CO2, which converted the over-expressed endogenous hydrogen peroxide (H2O2) to highly toxic hydroxyl radicals (·OH). Moreover, under the irradiation of ultrasound, PpIX could convert oxygen in cells to singlet oxygen (1O2), forming continuous and accumulated oxygen stress. The synergistic treatment of chemodynamic therapy (CDT) and sonodynamic therapy (SDT) could produce a large amount of reactive oxygen species in tumor cells, which broke the self-regulation ability of malignant tumor cells, and caused oxidative damage to multiple intracellular organelles, and ultimately led to the death of cancer cells. In addition, under the adjuvant treatment of anticancer drugs doxorubicin (DOX) and ultrasound irradiation, the killing efficiency of cancer cell was about 90%. And the experimental group also achieved good tumor inhibition effects in vivo.
It is an effective attempt to increase the level of reactive oxygen species (ROS) to abrogate the redox balance and successively result in sever cells oxidative damage and death in cancer treatment. Herein, biodegradable, safe and tumor-specific hollow manganese carbonate (MnCO3) nanocarriers (HMC NPs) were synthesized by hydrothermal method. HMC NPs loaded with sonosensitizer protoporphyrin (PpIX) constituted HMC-PpIX nanoparticles (HMC-P NPs), which could be activated in the acidic tumor microenvironment (TME). By degrading and releasing Mn2+ and sonosensitizer, the overloaded Mn2+ in the tumor triggered the Fenton reaction under the physiological buffer environment of HCO3-/CO2, which converted the over-expressed endogenous hydrogen peroxide (H2O2) to highly toxic hydroxyl radicals (·OH). Moreover, under the irradiation of ultrasound, PpIX could convert oxygen in cells to singlet oxygen (1O2), forming continuous and accumulated oxygen stress. The synergistic treatment of chemodynamic therapy (CDT) and sonodynamic therapy (SDT) could produce a large amount of reactive oxygen species in tumor cells, which broke the self-regulation ability of malignant tumor cells, and caused oxidative damage to multiple intracellular organelles, and ultimately led to the death of cancer cells. In addition, under the adjuvant treatment of anticancer drugs doxorubicin (DOX) and ultrasound irradiation, the killing efficiency of cancer cell was about 90%. And the experimental group also achieved good tumor inhibition effects in vivo.
2021, 49(12): 2023-2031
doi: 10.19756/j.issn.0253-3820.210692
Abstract:
This work demonstrated an oxidase mimic, e.g. a composite consisting of poly(vinyl alcohol) aerogel (PAA) and CeO2 nanorods in-situ modified on the aerogel surface through a hydrothermal reaction. The introduction of PAA fixed the CeO2 nanorods, making its easy to separate with reaction solution in results of an improved recycling rate. The oxidase-like activity of the composite (CeO2-PAAC) could be elevated by fluorid ion (F-), which was utilized to construct a colorimetric sensor for F-, exhibiting a detection range of 80-4000 μmol/L and a detection limit (3σ) of 63.7 μmol/L. Such a value was below the Chinese criterion for the Class V surface water (80 μmol/L). Compared to other detection methods, this method had the merits of wider detection range and proportionate detection limit. Additionally, this method possessed eligible anti-interference ability and selectivity, and was adequate in detection of F- in lake water with recovery rate of 97.1%-120.4%, validating the potential application in the real scene.
This work demonstrated an oxidase mimic, e.g. a composite consisting of poly(vinyl alcohol) aerogel (PAA) and CeO2 nanorods in-situ modified on the aerogel surface through a hydrothermal reaction. The introduction of PAA fixed the CeO2 nanorods, making its easy to separate with reaction solution in results of an improved recycling rate. The oxidase-like activity of the composite (CeO2-PAAC) could be elevated by fluorid ion (F-), which was utilized to construct a colorimetric sensor for F-, exhibiting a detection range of 80-4000 μmol/L and a detection limit (3σ) of 63.7 μmol/L. Such a value was below the Chinese criterion for the Class V surface water (80 μmol/L). Compared to other detection methods, this method had the merits of wider detection range and proportionate detection limit. Additionally, this method possessed eligible anti-interference ability and selectivity, and was adequate in detection of F- in lake water with recovery rate of 97.1%-120.4%, validating the potential application in the real scene.
2021, 49(12): 2032-2038
doi: 10.19756/j.issn.0253-3820.210679
Abstract:
Hydrogen production by water electrolysis has the advantages such as high efficiency and green development, and higher hydrogen purity. The development of efficient electrocatalysts for hydrogen evolution reaction (HER) must be based on the minimal overpotential to trigger proton reduction and fast kinetics. Palladium-based catalysts are used as one of the electrocatalysts for HER. However, they typically exhibit low activity in electrocatalytic hydrogen evolution reaction as too strong Pd-H bonding makes the electronic desorption of H adatoms (Had) hardly occur. Based on this, a highly efficient catalyst with uniformly dispersed palladium phosphide nanoparticles (Pd3P NP) embedded in a nitrogen-phosphorus co-doped carbon material (Pd3P/NPC) was designed and synthesized in this work. The Pd3P/NPC presented a small overpotential of 11 mV to delivering 10 mA/cm2, and a robust stability, which was much better than Pd/C, other reported Pd-based catalysts, and even Pt/C. Moreover, the Pd3P/NPC electrocatalyst exhibited high turnover frequencies at 20 mV (5.95 H2/s). It was higher than that of commercial Pt/C and Pd/C catalysts. Experiments evidence revealed that the introduction of P atoms into the Pd nanocrystals formed a Pd-P bond, which restructured the electronic structure of Pd and changed the content of Pd(0) and Pd(+2), promoting the desorption of H atoms. Meanwhile, the introduction of P increased the active site and intrinsic activity.
Hydrogen production by water electrolysis has the advantages such as high efficiency and green development, and higher hydrogen purity. The development of efficient electrocatalysts for hydrogen evolution reaction (HER) must be based on the minimal overpotential to trigger proton reduction and fast kinetics. Palladium-based catalysts are used as one of the electrocatalysts for HER. However, they typically exhibit low activity in electrocatalytic hydrogen evolution reaction as too strong Pd-H bonding makes the electronic desorption of H adatoms (Had) hardly occur. Based on this, a highly efficient catalyst with uniformly dispersed palladium phosphide nanoparticles (Pd3P NP) embedded in a nitrogen-phosphorus co-doped carbon material (Pd3P/NPC) was designed and synthesized in this work. The Pd3P/NPC presented a small overpotential of 11 mV to delivering 10 mA/cm2, and a robust stability, which was much better than Pd/C, other reported Pd-based catalysts, and even Pt/C. Moreover, the Pd3P/NPC electrocatalyst exhibited high turnover frequencies at 20 mV (5.95 H2/s). It was higher than that of commercial Pt/C and Pd/C catalysts. Experiments evidence revealed that the introduction of P atoms into the Pd nanocrystals formed a Pd-P bond, which restructured the electronic structure of Pd and changed the content of Pd(0) and Pd(+2), promoting the desorption of H atoms. Meanwhile, the introduction of P increased the active site and intrinsic activity.
2021, 49(12): 2039-2047
doi: 10.19756/j.issn.0253-3820.210737
Abstract:
Taking 7 genotoxic compounds (GCs) and 7 non-DNA damaging compounds (Non-DCs) as examples, mass spectrometry (MS) and high-content analysis (HCA) were used for detecting phosphorylated histone γ-H2AX in two human cell lines (HepG2 and HeLa) to assess the genotoxicity of compound. The results showed that the both techniques could complete the experimental process within 2 days and have high-throughput characteristics. For the selected 5 GCs, the minimum effective concentration (MEC) value of compound that give rise significant phosphorylation of H2AX determined by MS and HCA was same; for the remaining 2 GCs, the MEC value determined by MS was lower than that by HCA. MS could detect the significant changes in γ-H2AX levels at as low as 10 μmol/L chlorambucil and 0.1 μmol/L mitomycin C, while HCA detected significant changes of γ-H2AX levels at the concentration of 100 μmol/L chlorambucil and 1 μmol/L mitomycin C, indicating that MS had higher sensitivity. We also found that MS could specifically detect the changes of γ-H2AX levels induced by GCs. However, HCA could not distinguish γ-H2AX focus between GCs and Non-DCs. That is, HCA was prone to false positive results for Non-DCs. Taken together, these results suggested that MS analysis of γ-H2AX could not only detect compound genotoxicity more sensitively and specifically, but also could dynamically monitor the detail profiles of DNA damage and repair processes, which showed feasibility and superiority in the genotoxicity testing of the compounds.
Taking 7 genotoxic compounds (GCs) and 7 non-DNA damaging compounds (Non-DCs) as examples, mass spectrometry (MS) and high-content analysis (HCA) were used for detecting phosphorylated histone γ-H2AX in two human cell lines (HepG2 and HeLa) to assess the genotoxicity of compound. The results showed that the both techniques could complete the experimental process within 2 days and have high-throughput characteristics. For the selected 5 GCs, the minimum effective concentration (MEC) value of compound that give rise significant phosphorylation of H2AX determined by MS and HCA was same; for the remaining 2 GCs, the MEC value determined by MS was lower than that by HCA. MS could detect the significant changes in γ-H2AX levels at as low as 10 μmol/L chlorambucil and 0.1 μmol/L mitomycin C, while HCA detected significant changes of γ-H2AX levels at the concentration of 100 μmol/L chlorambucil and 1 μmol/L mitomycin C, indicating that MS had higher sensitivity. We also found that MS could specifically detect the changes of γ-H2AX levels induced by GCs. However, HCA could not distinguish γ-H2AX focus between GCs and Non-DCs. That is, HCA was prone to false positive results for Non-DCs. Taken together, these results suggested that MS analysis of γ-H2AX could not only detect compound genotoxicity more sensitively and specifically, but also could dynamically monitor the detail profiles of DNA damage and repair processes, which showed feasibility and superiority in the genotoxicity testing of the compounds.
2021, 49(12): 2048-2054
doi: 10.19756/j.issn.0253-3820.210602
Abstract:
The polarized micro-Raman spectra (PMRS) technique can provide information about the orientations and ordering of molecules in samples without any special sample preparation or staining. In this work, PMRS was used to collect Raman spectra of normal and cancerous breast tissues. By spectral analysis and difference analysis, it was found that PMRS could significantly amplify the differences of main characteristic bands other than amide Ⅰ band, even the bands of proline (921 cm-1) and phenylalanine (1032 cm-1) which had unobvious difference in conventional micro-Raman spectroscopy. The results indicated that PMRS was much efficient to identify breast cancer tissue. The bimodal intensity ratio of the amide Ⅲ band (I1247/I1269) was lower than 1 in cancerous tissues, no matter excited with parallel or perpendicular polarized light, but it was lower and greater than 1 in normal tissue when excited with parallel and perpendicular polarized light, respectively, indicating that collagen fibers lost their original orientation in cancerous tissues. Meanwhile, it was proved that this phenomenon was related to the increase in the content of hydroxylated amino acids of collagen fibers after canceration. The information obtained in this study helped to elucidate the molecular mechanism of induced intravasation during breast cancer invasion more clearly. It suggested that PMRS had more potential to be developed as an effective tool for clinical diagnosis of breast cancer, to assist in the clinical diagnosis and treatment of invasive breast cancer.
The polarized micro-Raman spectra (PMRS) technique can provide information about the orientations and ordering of molecules in samples without any special sample preparation or staining. In this work, PMRS was used to collect Raman spectra of normal and cancerous breast tissues. By spectral analysis and difference analysis, it was found that PMRS could significantly amplify the differences of main characteristic bands other than amide Ⅰ band, even the bands of proline (921 cm-1) and phenylalanine (1032 cm-1) which had unobvious difference in conventional micro-Raman spectroscopy. The results indicated that PMRS was much efficient to identify breast cancer tissue. The bimodal intensity ratio of the amide Ⅲ band (I1247/I1269) was lower than 1 in cancerous tissues, no matter excited with parallel or perpendicular polarized light, but it was lower and greater than 1 in normal tissue when excited with parallel and perpendicular polarized light, respectively, indicating that collagen fibers lost their original orientation in cancerous tissues. Meanwhile, it was proved that this phenomenon was related to the increase in the content of hydroxylated amino acids of collagen fibers after canceration. The information obtained in this study helped to elucidate the molecular mechanism of induced intravasation during breast cancer invasion more clearly. It suggested that PMRS had more potential to be developed as an effective tool for clinical diagnosis of breast cancer, to assist in the clinical diagnosis and treatment of invasive breast cancer.
2021, 49(12): 2055-2066
doi: 10.19756/j.issn.0253-3820.210604
Abstract:
The nickel nanoparticles/graphene (Ni/Gr) composite synthesized here was served as a novel matrix and adsorbent for highly efficient matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy (MALDI-TOF MS) analysis of various pharmaceutical molecules and amino acids. In comparison with 2,5-dihydroxybenzoic acid (DHB) and the control substances (Gr, Ni-1/Gr, Ni-5/Gr), the Ni-2/Gr matrix exhibited excellent performances in detection of resveratrol, 6-gingerol and rutin under positive-ion mode, including low background noise, high ion intensity, high signal to noise ratio and wide linear range (0.01-50 μmol/L, 0.2-50 μmol/L and 0.05-60 μmol/L). Meanwhile, the low limit of detection (LODs) of resveratrol, 6-gingerol and rutin were 0.0038 μmol/L, 0.09 μmol/L and 0.02 μmol/L, respectively. Benefiting from high surface area, abundant mesoporous structures, lots of sp2 structure, strong UV absorption and high saturation magnetization (Ms) value, Ni-2/Gr composite also acted as a favorable adsorbent for effectively analysis of L-phenylalanine, L-tryptophan and L-tyrosine with magnetic enrichment. Further, the low concentration of L-tryptophan spiked in mice serum bio-samples was directly enriched.
The nickel nanoparticles/graphene (Ni/Gr) composite synthesized here was served as a novel matrix and adsorbent for highly efficient matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy (MALDI-TOF MS) analysis of various pharmaceutical molecules and amino acids. In comparison with 2,5-dihydroxybenzoic acid (DHB) and the control substances (Gr, Ni-1/Gr, Ni-5/Gr), the Ni-2/Gr matrix exhibited excellent performances in detection of resveratrol, 6-gingerol and rutin under positive-ion mode, including low background noise, high ion intensity, high signal to noise ratio and wide linear range (0.01-50 μmol/L, 0.2-50 μmol/L and 0.05-60 μmol/L). Meanwhile, the low limit of detection (LODs) of resveratrol, 6-gingerol and rutin were 0.0038 μmol/L, 0.09 μmol/L and 0.02 μmol/L, respectively. Benefiting from high surface area, abundant mesoporous structures, lots of sp2 structure, strong UV absorption and high saturation magnetization (Ms) value, Ni-2/Gr composite also acted as a favorable adsorbent for effectively analysis of L-phenylalanine, L-tryptophan and L-tyrosine with magnetic enrichment. Further, the low concentration of L-tryptophan spiked in mice serum bio-samples was directly enriched.
2021, 49(12): 2067-2074
doi: 10.19756/j.issn.0253-3820.210684
Abstract:
Ion mobility spectrometry (IMS) has been widely used in the field of detecting explosives, chemical agents and environmental pollutants due to the advantages such as high sensitivity, fast response ability and portability. How to improve the sensitivity and resolution of IMS has always been a research hot spot. The sensitivity and resolution are closely related to the characteristics of the initial ion current that is controlled by the voltage waveform applied to the ion gate. In this work, the voltage waveforms of different Bradbury-Nielsen (BN) ion gates were simulated by SIMION software. It was found that the performances of IMS including the sensitivity and resolution could be improved by adjusting the voltage waveform. The homemade research platform of IMS was built, and the effects of switching pulse waveform time (Δt1), pulse voltage width (GPW) and pulse voltage difference (GVD) on the signal intensity and resolution of IMS were systematically investigated. The results showed that when GPW=80 μs, the acetone ion RIP intensity was increased by 3 times and the resolution was increased by 13%. When detecting dimethyl methylphosphate (DMMP) sample (5.8 μg/L), the ion intensities of (Ac)2H+, (Ac)(DMMP) H+ and (DMMP)2H+ were increased by 81%, 156% and 260%, respectively. Meanwhile, the signal-to-noise ratio of (Ac)(DMMP) H+ was increased by 2.5 times and that of (DMMP)2H+ was increased by 3 times. The results were helpful to develop high resolution ion mobility spectrometer.
Ion mobility spectrometry (IMS) has been widely used in the field of detecting explosives, chemical agents and environmental pollutants due to the advantages such as high sensitivity, fast response ability and portability. How to improve the sensitivity and resolution of IMS has always been a research hot spot. The sensitivity and resolution are closely related to the characteristics of the initial ion current that is controlled by the voltage waveform applied to the ion gate. In this work, the voltage waveforms of different Bradbury-Nielsen (BN) ion gates were simulated by SIMION software. It was found that the performances of IMS including the sensitivity and resolution could be improved by adjusting the voltage waveform. The homemade research platform of IMS was built, and the effects of switching pulse waveform time (Δt1), pulse voltage width (GPW) and pulse voltage difference (GVD) on the signal intensity and resolution of IMS were systematically investigated. The results showed that when GPW=80 μs, the acetone ion RIP intensity was increased by 3 times and the resolution was increased by 13%. When detecting dimethyl methylphosphate (DMMP) sample (5.8 μg/L), the ion intensities of (Ac)2H+, (Ac)(DMMP) H+ and (DMMP)2H+ were increased by 81%, 156% and 260%, respectively. Meanwhile, the signal-to-noise ratio of (Ac)(DMMP) H+ was increased by 2.5 times and that of (DMMP)2H+ was increased by 3 times. The results were helpful to develop high resolution ion mobility spectrometer.
2021, 49(12): 2075-2085
doi: 10.19756/j.issn.0253-3820.210543
Abstract:
Human chorionic gonadotropin (hCG) is a marker of early pregnancy and diseases such as ovarian cancer. Therefore, the rapid, accurate and real-time determination of hCG is of great significance. In this work, an electrochemical immunosensor for detection of hCG based on three-dimensional Au nanoparticles anchored on cobalt boride (AuNPs/CoB) nanosheet array electrode was developed. AuNPs/CoB nanosheet arrays were synthesized by an electrodeposition method, which was first explored for immunoassay of hCG. AuNPs/CoB nanosheeet arrays could enhance the performance of electrocatalytic reduction of thionine to amplify signal. Thionine (Thi) was electro-polymerized on the AuNPs/CoB nanosheet arrays, thus forming Thi/AuNPs/CoB nanosheet array-electrode. HCG antibody (anti-hCG) could be successfully attached to the Thi/AuNPs/Co-B nanosheet array-electrode by Au-N bond of AuNPs or the amino group of thionine, which caused an obvious decrease in the catalytic current due to the insulated anti-hCG. The current signal decreased linearly with the increase of hCG concentration due to the formation of antibody-antigen immunocomplex. The immunosensor possessed a good linearity with logarithm of hCG concentration in the range of 10-5.0×104 pg/mL, and the detection limit was 2.80 pg/mL (S/N=3). The immunosensor based on Thi/AuNPs/Co-B nanosheet arrays could exactly detect hCG in human serum samples, providing a simple and reliable evaluation method for the clinical diagnosis of early pregnancy and gynecological diseases.
Human chorionic gonadotropin (hCG) is a marker of early pregnancy and diseases such as ovarian cancer. Therefore, the rapid, accurate and real-time determination of hCG is of great significance. In this work, an electrochemical immunosensor for detection of hCG based on three-dimensional Au nanoparticles anchored on cobalt boride (AuNPs/CoB) nanosheet array electrode was developed. AuNPs/CoB nanosheet arrays were synthesized by an electrodeposition method, which was first explored for immunoassay of hCG. AuNPs/CoB nanosheeet arrays could enhance the performance of electrocatalytic reduction of thionine to amplify signal. Thionine (Thi) was electro-polymerized on the AuNPs/CoB nanosheet arrays, thus forming Thi/AuNPs/CoB nanosheet array-electrode. HCG antibody (anti-hCG) could be successfully attached to the Thi/AuNPs/Co-B nanosheet array-electrode by Au-N bond of AuNPs or the amino group of thionine, which caused an obvious decrease in the catalytic current due to the insulated anti-hCG. The current signal decreased linearly with the increase of hCG concentration due to the formation of antibody-antigen immunocomplex. The immunosensor possessed a good linearity with logarithm of hCG concentration in the range of 10-5.0×104 pg/mL, and the detection limit was 2.80 pg/mL (S/N=3). The immunosensor based on Thi/AuNPs/Co-B nanosheet arrays could exactly detect hCG in human serum samples, providing a simple and reliable evaluation method for the clinical diagnosis of early pregnancy and gynecological diseases.
2021, 49(12): 2086-2095
doi: 10.19756/j.issn.0253-3820.201398
Abstract:
Peroxynitrite anion (ONOO-) is one of the most important molecules in biological systems and is usually maintained at a very low concentration level under normal physiological condition. However, under oxidative stress, the body will produce high concentration of ONOO- abnormally, which may damage many biomolecules seriously and affect the physiological functions of cells. Therefore, it is very important to construct a sensor that can detect ONOO- released by cells sensitively and rapidly. In this work, a novel ONOO- electrochemical sensor was constructed based on carbon nanospheres (CNS) composite materials modified with platinum nanoparticles (PtNPs). Electron microscopes and electrochemical technology were used for characterization of the prepared CTS/PtNPs/CNS. The sensor showed excellent analytical performance for ONOO- detection, with a wide linear range from 0.615 nmol/L to 0.139 mmol/L, and an extremely low detection limit of 0.205 nmol/L (S/N=3). In addition, it also showed good selectivity, stability and reproducibility, and was applied to detect ONOO- released from living cells. The released ONOO- of cells induced by Cd2+ was investigated, and it was found that more ONOO- released from cells at higher Cd2+ concentration. Finally, the protective effects of the α-lipoic acid (ALA) and glutathione (GSH) on cells were investigated by electrochemical method, and it was found that the antioxidants mixture (ALA+GSH) had better protect cells due to their synergistic effect. The experimental results showed that the developed sensing platform and efficient antioxidation strategy provided the possibility for the future application in the biomedical field and cancer diseases.
Peroxynitrite anion (ONOO-) is one of the most important molecules in biological systems and is usually maintained at a very low concentration level under normal physiological condition. However, under oxidative stress, the body will produce high concentration of ONOO- abnormally, which may damage many biomolecules seriously and affect the physiological functions of cells. Therefore, it is very important to construct a sensor that can detect ONOO- released by cells sensitively and rapidly. In this work, a novel ONOO- electrochemical sensor was constructed based on carbon nanospheres (CNS) composite materials modified with platinum nanoparticles (PtNPs). Electron microscopes and electrochemical technology were used for characterization of the prepared CTS/PtNPs/CNS. The sensor showed excellent analytical performance for ONOO- detection, with a wide linear range from 0.615 nmol/L to 0.139 mmol/L, and an extremely low detection limit of 0.205 nmol/L (S/N=3). In addition, it also showed good selectivity, stability and reproducibility, and was applied to detect ONOO- released from living cells. The released ONOO- of cells induced by Cd2+ was investigated, and it was found that more ONOO- released from cells at higher Cd2+ concentration. Finally, the protective effects of the α-lipoic acid (ALA) and glutathione (GSH) on cells were investigated by electrochemical method, and it was found that the antioxidants mixture (ALA+GSH) had better protect cells due to their synergistic effect. The experimental results showed that the developed sensing platform and efficient antioxidation strategy provided the possibility for the future application in the biomedical field and cancer diseases.
2021, 49(12): 2096-2105
doi: 10.19756/j.issn.0253-3820.201728
Abstract:
The chitosan-sodium alginate composite nanofibers were prepared by freeze-drying. The nanofibers had a uniform filamentous structure with a width of about 0.4 μm. The aptamer of ochratoxin A was immobilized on the surface of the fiber by chemical bonding to obtain the aptamer-modified chitosan-sodium alginate composite nanofiber, and the bonding amount of the aptamer could reach 2.3 μg/mg. It could be used as an adsorbent for dispersive solid phase extraction technology. The adsorbent showed good extraction ability and high selectivity for ochratoxin A, the extraction capacity was about 3.1 ng/mg, and the extraction amount was 2.44-12.8 times of the structural analogue ochratoxin B and the five reference molecules. Compared with the composite nanofiber modified by the scrambled oligonucleotide and composite nanofiber, the extraction amount of ochratoxin A was increased by 4.88 and 13.0 times, respectively. Under the optimal extraction conditions, an analytical method based on aptamer modified composite nanofiber dispersion solid phase extraction-high performance liquid chromatography for determination of ochratoxin A was established. The linear range was 0.05-3.0 μg/L, the detection limit was 13 ng/L (S/N=3), and the recoveries of standard addition were 86.7%-101.0%. This method had good selectivity and high sensitivity, and could be applied to the detection of trace ochratoxin A in peanut, corn and wheat samples.
The chitosan-sodium alginate composite nanofibers were prepared by freeze-drying. The nanofibers had a uniform filamentous structure with a width of about 0.4 μm. The aptamer of ochratoxin A was immobilized on the surface of the fiber by chemical bonding to obtain the aptamer-modified chitosan-sodium alginate composite nanofiber, and the bonding amount of the aptamer could reach 2.3 μg/mg. It could be used as an adsorbent for dispersive solid phase extraction technology. The adsorbent showed good extraction ability and high selectivity for ochratoxin A, the extraction capacity was about 3.1 ng/mg, and the extraction amount was 2.44-12.8 times of the structural analogue ochratoxin B and the five reference molecules. Compared with the composite nanofiber modified by the scrambled oligonucleotide and composite nanofiber, the extraction amount of ochratoxin A was increased by 4.88 and 13.0 times, respectively. Under the optimal extraction conditions, an analytical method based on aptamer modified composite nanofiber dispersion solid phase extraction-high performance liquid chromatography for determination of ochratoxin A was established. The linear range was 0.05-3.0 μg/L, the detection limit was 13 ng/L (S/N=3), and the recoveries of standard addition were 86.7%-101.0%. This method had good selectivity and high sensitivity, and could be applied to the detection of trace ochratoxin A in peanut, corn and wheat samples.
2021, 49(12): 2106-2116
doi: 10.19756/j.issn.0253-3820.210623
Abstract:
O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) is a single carbohydrate moiety post-translational modification that occurs on serine and threonine side chains of intracellular protein and plays an important role in nutrient metabolism. However, the great research challenges are the inherently low stoichiometry, poor ionization efficiency of O-GlcNAc peptides and no specific animo acid sequence. In this study, a strategy for quantitative detection of O-GlcNAc sites was developed by combining pseudo-isobaric dimethyl strategy, which was applied for the O-GlcNAcylation sites profiling towards the mice liver. In total, 783 O-GlcNAc sites were unambiguously quantified from high-fat fed mice liver. Among which, 122 O-GlcNAc sites were differentially expressed, corresponding to 85 O-GlcNAc proteins. Finally, the biological function of the differentially expressed O-GlcNAc proteins was analyzed.
O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) is a single carbohydrate moiety post-translational modification that occurs on serine and threonine side chains of intracellular protein and plays an important role in nutrient metabolism. However, the great research challenges are the inherently low stoichiometry, poor ionization efficiency of O-GlcNAc peptides and no specific animo acid sequence. In this study, a strategy for quantitative detection of O-GlcNAc sites was developed by combining pseudo-isobaric dimethyl strategy, which was applied for the O-GlcNAcylation sites profiling towards the mice liver. In total, 783 O-GlcNAc sites were unambiguously quantified from high-fat fed mice liver. Among which, 122 O-GlcNAc sites were differentially expressed, corresponding to 85 O-GlcNAc proteins. Finally, the biological function of the differentially expressed O-GlcNAc proteins was analyzed.
2021, 49(12): 2117-2125
doi: 10.19756/j.issn.0253-3820.210725
Abstract:
Ethanol reflux extract of Murraya exotica L was separated by macroporous resin, and 80% methanol elution component was determined to be the flavonoids-rich component as shown by the HCl-Mg reaction assay. N-Hexane-ethylacetate-methanol-water (4:6:4:6, V/V) was selected as the two-phase solvent system for high-speed countercurrent chromatography (HSCCC) based on the partition coefficients (K) and separation factors (α) values that calculated from main compounds dissolved in n-hexane, ethyl acetate, methanol and water. Purity of the isolated product was analyzed by high performance liquid chromatography. Five polymethoxylated flavonoids were isolated and determined by nuclear magnetic resonance and mass spectrometry methods. Among them, 5,7,3',4'-tetramethoxyflavone, 5,7,8,3',4',5'-hexamethoxyflavone and 5,7,3',4',5'-pentamethoxyflavone were obtained from Murraya exotica L for the first time, three compounds (5,7,3',4'-tetramethoxyflavone, 5,7,3',4',5'-pentamethoxyflavone and 5-hydroxy-6,7,3',4'-tetramethoxyflavone) potentiated cystic fibrosis transmembrane conductance regulator (CFTR) activities in dose-dependent ways. The present study established a HSCCC method for separation and analysis of low content of polymethoxylated flavonoids from Murraya exotica L., which provided foundation for the in-depth and systematic analysis of the effective substances and active chemical constituents of Murraya exotica L.
Ethanol reflux extract of Murraya exotica L was separated by macroporous resin, and 80% methanol elution component was determined to be the flavonoids-rich component as shown by the HCl-Mg reaction assay. N-Hexane-ethylacetate-methanol-water (4:6:4:6, V/V) was selected as the two-phase solvent system for high-speed countercurrent chromatography (HSCCC) based on the partition coefficients (K) and separation factors (α) values that calculated from main compounds dissolved in n-hexane, ethyl acetate, methanol and water. Purity of the isolated product was analyzed by high performance liquid chromatography. Five polymethoxylated flavonoids were isolated and determined by nuclear magnetic resonance and mass spectrometry methods. Among them, 5,7,3',4'-tetramethoxyflavone, 5,7,8,3',4',5'-hexamethoxyflavone and 5,7,3',4',5'-pentamethoxyflavone were obtained from Murraya exotica L for the first time, three compounds (5,7,3',4'-tetramethoxyflavone, 5,7,3',4',5'-pentamethoxyflavone and 5-hydroxy-6,7,3',4'-tetramethoxyflavone) potentiated cystic fibrosis transmembrane conductance regulator (CFTR) activities in dose-dependent ways. The present study established a HSCCC method for separation and analysis of low content of polymethoxylated flavonoids from Murraya exotica L., which provided foundation for the in-depth and systematic analysis of the effective substances and active chemical constituents of Murraya exotica L.
2021, 49(12): 2126-2133
doi: 10.19756/j.issn.0253-3820.210541
Abstract:
A new type of amine functionalized polymer adsorption material was synthesized by using polyacrylonitrile as the raw material and triethylenetetramine (TETA) as the modified group. The morphology and structure of polymer materials were characterized by scanning electron microscope (SEM) and Fourier transform infrared spectrometer (FT-IR). A fast on-line analysis method integrating separation, enrichment and determination was established by combining with a home-made solid phase extraction column and inductively coupled plasma mass spectrometry technology. The effects of pH, flow rate, volume and coexisting interfering ions on the separation and preconcentration of copper and lead were investigated. Under the conditions of pH 7, flow rate of 5 mL/min and sample volume of 50 mL, the adsorption efficiency of the metal ions could reach 95% within 10 min, and the interfering ions in real environmental water could be ignored. The dynamic adsorption saturation capacities of copper and lead were 15.1 and 22.5 mg/g, respectively. At a flow rate of 1 mL/min, 1% HNO3 solution was adopted to quantitatively elute the trace elements, and high enrichment factors were obtained. The limits of detection (LODs) of the method for copper and lead were 1.7 and 2.5 μg/L, respectively, and the relative standard deviation (RSD%) was less than 3.5%. Finally, the environmental Certified Reference Material GSB07-3186-2014 (CRM) was selected to verify the feasibility and accuracy of the on-line method. The prepared amine functionalized polyacrylonitrile had many advantages such as low cost, environmental friendliness, high adsorption efficiency and reusability. The online integrated method had the merits of easy to operate, fast, stable, and was suitable for the analysis of trace copper and lead in real environmental water samples.
A new type of amine functionalized polymer adsorption material was synthesized by using polyacrylonitrile as the raw material and triethylenetetramine (TETA) as the modified group. The morphology and structure of polymer materials were characterized by scanning electron microscope (SEM) and Fourier transform infrared spectrometer (FT-IR). A fast on-line analysis method integrating separation, enrichment and determination was established by combining with a home-made solid phase extraction column and inductively coupled plasma mass spectrometry technology. The effects of pH, flow rate, volume and coexisting interfering ions on the separation and preconcentration of copper and lead were investigated. Under the conditions of pH 7, flow rate of 5 mL/min and sample volume of 50 mL, the adsorption efficiency of the metal ions could reach 95% within 10 min, and the interfering ions in real environmental water could be ignored. The dynamic adsorption saturation capacities of copper and lead were 15.1 and 22.5 mg/g, respectively. At a flow rate of 1 mL/min, 1% HNO3 solution was adopted to quantitatively elute the trace elements, and high enrichment factors were obtained. The limits of detection (LODs) of the method for copper and lead were 1.7 and 2.5 μg/L, respectively, and the relative standard deviation (RSD%) was less than 3.5%. Finally, the environmental Certified Reference Material GSB07-3186-2014 (CRM) was selected to verify the feasibility and accuracy of the on-line method. The prepared amine functionalized polyacrylonitrile had many advantages such as low cost, environmental friendliness, high adsorption efficiency and reusability. The online integrated method had the merits of easy to operate, fast, stable, and was suitable for the analysis of trace copper and lead in real environmental water samples.
