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2021 Volume 49 Issue 9
2021, 49(9): 1419-1427
doi: 10.19756/j.issn.0253-3820.211247
Abstract:
Triboelectric nanogenerator (TENG) technology is very important for the rapid development of the intelligent Internet of Things. It can effectively convert mechanical stress into electrical signals or electricity without any additional power supply. Therefore, it has broad prospection in the field of sensors and self-powered detection systems. TENG is characterized by good material compatibility, simple assembly requirements and flexible structure, which effectively promotes their applications in the fields of biomedical, intelligent transportation and chemical sensing. TENG is one of the most promising kind of sensors for commercial application at present. In this paper, the working principle of TENG and its latest research progresses in chemical sensors are introduced. Besides, the challenges for the development of TENG technology in chemical sensors are further discussed.
Triboelectric nanogenerator (TENG) technology is very important for the rapid development of the intelligent Internet of Things. It can effectively convert mechanical stress into electrical signals or electricity without any additional power supply. Therefore, it has broad prospection in the field of sensors and self-powered detection systems. TENG is characterized by good material compatibility, simple assembly requirements and flexible structure, which effectively promotes their applications in the fields of biomedical, intelligent transportation and chemical sensing. TENG is one of the most promising kind of sensors for commercial application at present. In this paper, the working principle of TENG and its latest research progresses in chemical sensors are introduced. Besides, the challenges for the development of TENG technology in chemical sensors are further discussed.
2021, 49(9): 1428-1436
doi: 10.19756/j.issn.0253-3820.211186
Abstract:
The occurrence and development of diseases are often accompanied by abnormal expression of certain microRNAs. MicroRNAs are small and non-coding RNA molecules which play an important role in many basic physiological and pathological processes as post-transcriptional regulators of gene expression. At present, many studies have shown that microRNAs can be used as biomarkers for diagnosis and treatment of diseases. Traditional detection methods of microRNAs often have shortcomings such as low sensitivity and poor selectivity, and cannot achieve real-time detection. Rare earth upconversion nanomaterials have unique anti-stokes shift properties and advantages including good photobleaching resistance, low tissue autofluorescence interference, good biocompatibility and low toxicity, and thus are commonly used for microRNAs detection. Therefore, detecting the changes of expression level of microRNAs using up-conversion nanomaterials in real-time can identify the occurrence and development of diseases, and provide evidence for the diagnosis, treatment and prognosis of diseases. In this papar, the application progress of up-conversion nanomaterials in detection of microRNAs in recent years is reviewed.
The occurrence and development of diseases are often accompanied by abnormal expression of certain microRNAs. MicroRNAs are small and non-coding RNA molecules which play an important role in many basic physiological and pathological processes as post-transcriptional regulators of gene expression. At present, many studies have shown that microRNAs can be used as biomarkers for diagnosis and treatment of diseases. Traditional detection methods of microRNAs often have shortcomings such as low sensitivity and poor selectivity, and cannot achieve real-time detection. Rare earth upconversion nanomaterials have unique anti-stokes shift properties and advantages including good photobleaching resistance, low tissue autofluorescence interference, good biocompatibility and low toxicity, and thus are commonly used for microRNAs detection. Therefore, detecting the changes of expression level of microRNAs using up-conversion nanomaterials in real-time can identify the occurrence and development of diseases, and provide evidence for the diagnosis, treatment and prognosis of diseases. In this papar, the application progress of up-conversion nanomaterials in detection of microRNAs in recent years is reviewed.
2021, 49(9): 1437-1450
doi: 10.19756/j.issn.0253-3820.201568
Abstract:
As the smallest extracellular vesicles, exosomes contain lipids, proteins, mRNA, microRNA and double-stranded DNA, which can be secreted by most cells and circulated in body fluids and are involved in intercellular communication. In recent years, it has been found that exosomes play an extremely important role in tumorigenesis, migration and growth, tissue damage repair, immune antigen presentation and neurodegenerative diseases. Exosomes can be used as biomarker for diagnosis of related diseases. Therefore, it is of great importance to study the separation and detection technology for exosomes. This review summarizes the methods of separation and detection of exosomes based on microfluidic chip.
As the smallest extracellular vesicles, exosomes contain lipids, proteins, mRNA, microRNA and double-stranded DNA, which can be secreted by most cells and circulated in body fluids and are involved in intercellular communication. In recent years, it has been found that exosomes play an extremely important role in tumorigenesis, migration and growth, tissue damage repair, immune antigen presentation and neurodegenerative diseases. Exosomes can be used as biomarker for diagnosis of related diseases. Therefore, it is of great importance to study the separation and detection technology for exosomes. This review summarizes the methods of separation and detection of exosomes based on microfluidic chip.
2021, 49(9): 1451-1460
doi: 10.19756/j.issn.0253-3820.210435
Abstract:
Glycoproteins play an important role in regulating various physiological processes of organisms. Abnormal protein glycosylation is usually associated with the formation and evolution of diseases such as cancer. Many specific glycoproteins can be used as the biomarkers for early cancer diagnosis or prognosis monitoring. Before identifying the glycoproteins with mass spectrometry, effective separation and enrichment of glycoproteins from complex biological samples is a necessary step. Lectins are selective for certain specific glycoproteins, and lectin affinity methods have been widely used in the separation and enrichment of glycoprotein. By taking the four kinds of cancers (breast cancer, lung cancer, colorectal cancer and prostate cancer) with the highest incidence as examples, this article reviews the applications of lectin affinity methods integrated with mass spectrometry in screening and analyzing cancer biomarkers.
Glycoproteins play an important role in regulating various physiological processes of organisms. Abnormal protein glycosylation is usually associated with the formation and evolution of diseases such as cancer. Many specific glycoproteins can be used as the biomarkers for early cancer diagnosis or prognosis monitoring. Before identifying the glycoproteins with mass spectrometry, effective separation and enrichment of glycoproteins from complex biological samples is a necessary step. Lectins are selective for certain specific glycoproteins, and lectin affinity methods have been widely used in the separation and enrichment of glycoprotein. By taking the four kinds of cancers (breast cancer, lung cancer, colorectal cancer and prostate cancer) with the highest incidence as examples, this article reviews the applications of lectin affinity methods integrated with mass spectrometry in screening and analyzing cancer biomarkers.
2021, 49(9): 1461-1469
doi: 10.19756/j.issn.0253-3820.210458
Abstract:
Ion mobility spectrometry (IMS) is a rapid separation technique that has been successfully used for detection of chemical warfare agents, drugs and explosives. The key component of IMS is the drift tube, which plays an important role in effectively separating substances. Traditionally, the drift tube is a fixed-length separation space in which the influence of a uniformly applied weak electric field. In this work, the performance of traveling wave ion mobility spectrometer (TWIMS) was characterized by traveling wave voltage amplitudes, traveling wave speeds, ion gate pulse widths and working pressures. When the traveling wave field voltage amplitude was 50 V, the traveling wave field moving speed was 162.5 m/s, the ion gate pulse width was 13 ms, and the working pressure was 170 Pa, the device demonstrated highly analytical performance and achieved an ion mobility resolution comparable to a commercial traveling wave ion mobility time-of-flight mass spectrometry (TWIMS-MS). Finally, reserpine was used to test the linear response range of the TWIMS. The results showed that the linear response range of the TWIMS was greater than two orders of magnitude and the limit of detection could reach 2.5 ng/mL. The device was expected to be used in conjunction with a time-of-flight mass spectrometer.
Ion mobility spectrometry (IMS) is a rapid separation technique that has been successfully used for detection of chemical warfare agents, drugs and explosives. The key component of IMS is the drift tube, which plays an important role in effectively separating substances. Traditionally, the drift tube is a fixed-length separation space in which the influence of a uniformly applied weak electric field. In this work, the performance of traveling wave ion mobility spectrometer (TWIMS) was characterized by traveling wave voltage amplitudes, traveling wave speeds, ion gate pulse widths and working pressures. When the traveling wave field voltage amplitude was 50 V, the traveling wave field moving speed was 162.5 m/s, the ion gate pulse width was 13 ms, and the working pressure was 170 Pa, the device demonstrated highly analytical performance and achieved an ion mobility resolution comparable to a commercial traveling wave ion mobility time-of-flight mass spectrometry (TWIMS-MS). Finally, reserpine was used to test the linear response range of the TWIMS. The results showed that the linear response range of the TWIMS was greater than two orders of magnitude and the limit of detection could reach 2.5 ng/mL. The device was expected to be used in conjunction with a time-of-flight mass spectrometer.
2021, 49(9): 1470-1477
doi: 10.19756/j.issn.0253-3820.210467
Abstract:
A dispersive detection system based on ultraviolet digital micromirror device (DMD) was used for hydride generation atomic fluorescence spectrometer (HG-AFS) on the basis of the original non-dispersion detection system. The dispersion system was used to suppress the interference when spectral interference from the light source or scattering interference exists. The limit of detection (LOD) of the dispersive system was worse than the non-dispersive one due to the addition of the dispersive components (DMD and grating, etc). In this work, to improve the analytical performance of the dispersive detection method, a cylindrical mirror was installed in front of the photomultiplier tube (PMT), the minimum effective area of the PMT was expanded to enhance the signal intensity, a better fixed location mode of DMD rotation was proposed, and the analysis methods such as peak selection and the instrument parameters affecting the signal intensity were optimized. The results showed that the signal intensity within 180-320 nm was increased more than 2 times, and the spectral resolution was increased more than 1.2 times. Moreover, the emission lines and excitation fluorescence lines of Cd, Te, Sn and Zn were obtained, and the LODs were respectively less than 0.005 ng/mL for Cd, 0.02 ng/mL for Sn, 0.1 ng/mL for As, Bi and Hg, 0.05 ng/mL for Sb, Pb and Te, and 0.2 ng/mL for Se and Zn. The spiked recoveries and the RSDs of As were 93%-101% and 0.6%-3.3% in living and drinking water samples, and 95%-106% and 0.9%-3.4% in food samples, respectively. The LOD and resolution of the dispersive system were effectively improved because of the improvement of the ultraviolet DMD spectrometer, the upgrade of the DMD rotation detection mode and the optimization of the analysis method. As a result, the dispersive system met the detection requirements of toxicological index limits in the Standards for Drinking Water Quality (GB 5749-2006), and also had good accuracy, sensitivity and stability in actual food sample analysis.
A dispersive detection system based on ultraviolet digital micromirror device (DMD) was used for hydride generation atomic fluorescence spectrometer (HG-AFS) on the basis of the original non-dispersion detection system. The dispersion system was used to suppress the interference when spectral interference from the light source or scattering interference exists. The limit of detection (LOD) of the dispersive system was worse than the non-dispersive one due to the addition of the dispersive components (DMD and grating, etc). In this work, to improve the analytical performance of the dispersive detection method, a cylindrical mirror was installed in front of the photomultiplier tube (PMT), the minimum effective area of the PMT was expanded to enhance the signal intensity, a better fixed location mode of DMD rotation was proposed, and the analysis methods such as peak selection and the instrument parameters affecting the signal intensity were optimized. The results showed that the signal intensity within 180-320 nm was increased more than 2 times, and the spectral resolution was increased more than 1.2 times. Moreover, the emission lines and excitation fluorescence lines of Cd, Te, Sn and Zn were obtained, and the LODs were respectively less than 0.005 ng/mL for Cd, 0.02 ng/mL for Sn, 0.1 ng/mL for As, Bi and Hg, 0.05 ng/mL for Sb, Pb and Te, and 0.2 ng/mL for Se and Zn. The spiked recoveries and the RSDs of As were 93%-101% and 0.6%-3.3% in living and drinking water samples, and 95%-106% and 0.9%-3.4% in food samples, respectively. The LOD and resolution of the dispersive system were effectively improved because of the improvement of the ultraviolet DMD spectrometer, the upgrade of the DMD rotation detection mode and the optimization of the analysis method. As a result, the dispersive system met the detection requirements of toxicological index limits in the Standards for Drinking Water Quality (GB 5749-2006), and also had good accuracy, sensitivity and stability in actual food sample analysis.
2021, 49(9): 1478-1487
doi: 10.19756/j.issn.0253-3820.210441
Abstract:
Hepatocellular carcinoma (HCC) is the fourth leading cause of deaths among malignancies, of which 30% is caused by hepatitis B virus (HBV). Most of HCC patients are associated with cirrhosis, while 20% occurs in non-cirrhotic liver. Between HBV-related HCC patients with cirrhosis (C-HBV-HCC) and without cirrhosis (NC-HBV-HCC), the mechanism of HCC is different and not yet clear. This study performed differential proteome analysis of NC-HBV-HCC and C-HBV-HCC from formalin-fixed and paraffin-embedded (FFPE) samples, using LC-MS/MS coupled with label-free quantitation. A total of 4083 proteins were identified, and 283 of them were differentially expressed between these two groups. Six proteins were validated by immunohistochemistry, among which HSD17B13/SOD3/CXCL12 were up-regulated in NC-HBV-HCC group, while the expression level of GPD2/CRTAP/CD276 were higher in C-HBV-HCC group. Overall, 6 proteins were validated as biomarker candidates for guiding mechanism research of HBV-related HCC patients with and without cirrhosis, showing the potential to assist clinical management of these two patient types. Bioinformatic analyses showed that cholesterol homeostasis and adipogenesis pathways were upregulated in NC-HBV-HCC compared with C-HBV-HCC, and the 6 biomarker candidates suggested that PI3K/Akt/MMPs and Wnt/β-catenin pathways might be associated with different cancer progresses in NC-HBV-HCC and C-HBV-HCC, providing data and new ideas for the exploration of the tumorigenesis of HBV-related HCC with and without cirrhosis.
Hepatocellular carcinoma (HCC) is the fourth leading cause of deaths among malignancies, of which 30% is caused by hepatitis B virus (HBV). Most of HCC patients are associated with cirrhosis, while 20% occurs in non-cirrhotic liver. Between HBV-related HCC patients with cirrhosis (C-HBV-HCC) and without cirrhosis (NC-HBV-HCC), the mechanism of HCC is different and not yet clear. This study performed differential proteome analysis of NC-HBV-HCC and C-HBV-HCC from formalin-fixed and paraffin-embedded (FFPE) samples, using LC-MS/MS coupled with label-free quantitation. A total of 4083 proteins were identified, and 283 of them were differentially expressed between these two groups. Six proteins were validated by immunohistochemistry, among which HSD17B13/SOD3/CXCL12 were up-regulated in NC-HBV-HCC group, while the expression level of GPD2/CRTAP/CD276 were higher in C-HBV-HCC group. Overall, 6 proteins were validated as biomarker candidates for guiding mechanism research of HBV-related HCC patients with and without cirrhosis, showing the potential to assist clinical management of these two patient types. Bioinformatic analyses showed that cholesterol homeostasis and adipogenesis pathways were upregulated in NC-HBV-HCC compared with C-HBV-HCC, and the 6 biomarker candidates suggested that PI3K/Akt/MMPs and Wnt/β-catenin pathways might be associated with different cancer progresses in NC-HBV-HCC and C-HBV-HCC, providing data and new ideas for the exploration of the tumorigenesis of HBV-related HCC with and without cirrhosis.
2021, 49(9): 1488-1496
doi: 10.19756/j.issn.0253-3820.191720
Abstract:
A label-free fluorescence detection method was developed for quantitative determination of oxytetracycline (OTC) based on aptamer recognition capability and exonuclease Ⅰ (Exo Ⅰ)-assisted target recycling. In the absence of OTC, SYBR green I (SGI) molecules could insert the double-stranded DNA (dsDNA) of OTC aptamer and its complementary DNA, resulting in strong fluorescence emission of SGI with excitation wavelength of 495 nm. In the presence of OTC, due to the strong affinity of the aptamer to its target, aptamer preferentially bound with OTC and unwound the dsDNA. Thus, SGI molecules released from the hydrogen bonds of dsDNA, accompanied with the obvious decrease of SGI fluorescence. To amplify the fluorescence quenching effect of SGI caused by the addition of OTC, Exo Ⅰ-assisted target recycling was involved in. The results of acrylamide gel electrophoresis confirmed that Exo Ⅰ could digest single stranded DNA selectively and the aptamer bound with OTC. The released OTC participated in the next cycle, continuously destroyed the dsDNA structure, releasled the free SGI, and gradually weakened the fluorescence. Based on this Exo Ⅰ-catalyzed target recycling strategy, the sensitivity of this aptamer-based fluorescent strategy was significantly improved. The linear range of OTC was 0.01-10 μg/mL with a detection limit (3σ) of 6.77 ng/mL. The method realized the detection of OTC in milk and honey sample ssuccessfully. For milk samples, the recoveries were 93.0%-105.1% with relative standard deviations (RSD) of 0.5%-6.2%, and for honey samples, the recoveries were 94.0%-95.8% with RSD of 0.3%-7.7%, respectively. The fluorescence sensor had many advantages such as low cost, high sensitivity and good specificity, showing great potential in the rapid detection of residue of harmful substances in food.
A label-free fluorescence detection method was developed for quantitative determination of oxytetracycline (OTC) based on aptamer recognition capability and exonuclease Ⅰ (Exo Ⅰ)-assisted target recycling. In the absence of OTC, SYBR green I (SGI) molecules could insert the double-stranded DNA (dsDNA) of OTC aptamer and its complementary DNA, resulting in strong fluorescence emission of SGI with excitation wavelength of 495 nm. In the presence of OTC, due to the strong affinity of the aptamer to its target, aptamer preferentially bound with OTC and unwound the dsDNA. Thus, SGI molecules released from the hydrogen bonds of dsDNA, accompanied with the obvious decrease of SGI fluorescence. To amplify the fluorescence quenching effect of SGI caused by the addition of OTC, Exo Ⅰ-assisted target recycling was involved in. The results of acrylamide gel electrophoresis confirmed that Exo Ⅰ could digest single stranded DNA selectively and the aptamer bound with OTC. The released OTC participated in the next cycle, continuously destroyed the dsDNA structure, releasled the free SGI, and gradually weakened the fluorescence. Based on this Exo Ⅰ-catalyzed target recycling strategy, the sensitivity of this aptamer-based fluorescent strategy was significantly improved. The linear range of OTC was 0.01-10 μg/mL with a detection limit (3σ) of 6.77 ng/mL. The method realized the detection of OTC in milk and honey sample ssuccessfully. For milk samples, the recoveries were 93.0%-105.1% with relative standard deviations (RSD) of 0.5%-6.2%, and for honey samples, the recoveries were 94.0%-95.8% with RSD of 0.3%-7.7%, respectively. The fluorescence sensor had many advantages such as low cost, high sensitivity and good specificity, showing great potential in the rapid detection of residue of harmful substances in food.
2021, 49(9): 1497-1505
doi: 10.19756/j.issn.0253-3820.211214
Abstract:
Hg2+ pollutant poses significant risks to human health and the ecological environment. However, the direct and sensitive detection technique for Hg2+ has been lacking at the present time. In this study, Cu/CuO/ZnO wire was fabricated by alkaline oxidation method and hydrothermal method, and PPy was covered on the material surface by electrochemical polymerization method. Based on the principle that the p-n junction barrier drives the electrochemical signal response, the material was used for direct electrochemical detection of Hg2+ and differential pulse voltammetry was performed. The prepared composite was successfully used for detection of Hg2+ in the concentration range from 200 nmol/L to 1600 nmol/L, with a ultra-high sensitivity (1010.82 μA·L/(nmol·cm2)) and a ultra-low detection limit (2.1 pmol/L). The novel sensing mode based on the interface barrier eliminated the interference of other ions and achieved good recoveries (97.3%-105.0%) in tap water, river water and sea water, with RSD of 1.8%-5.6%. This approach could be extended to develop new sensors for other heavy metals by employing p-n junction barrier.
Hg2+ pollutant poses significant risks to human health and the ecological environment. However, the direct and sensitive detection technique for Hg2+ has been lacking at the present time. In this study, Cu/CuO/ZnO wire was fabricated by alkaline oxidation method and hydrothermal method, and PPy was covered on the material surface by electrochemical polymerization method. Based on the principle that the p-n junction barrier drives the electrochemical signal response, the material was used for direct electrochemical detection of Hg2+ and differential pulse voltammetry was performed. The prepared composite was successfully used for detection of Hg2+ in the concentration range from 200 nmol/L to 1600 nmol/L, with a ultra-high sensitivity (1010.82 μA·L/(nmol·cm2)) and a ultra-low detection limit (2.1 pmol/L). The novel sensing mode based on the interface barrier eliminated the interference of other ions and achieved good recoveries (97.3%-105.0%) in tap water, river water and sea water, with RSD of 1.8%-5.6%. This approach could be extended to develop new sensors for other heavy metals by employing p-n junction barrier.
2021, 49(9): 1506-1514
doi: 10.19756/j.issn.0253-3820.211076
Abstract:
Plant esterase-Cu3(PO4)2 hybrid nanoflowers were prepared, which not only achieved the immobilization of plant esterase and maintained its hydrolase activity, but also skillfully used copper phosphate skeleton. The copper phosphate skeleton not only had the supporting role, but also had the peroxidase-like activity. Different from free enzymes, hybrid nanoflowers showed higher stability, and their hydrolase activity and peroxidase-like activity maintained above 80% after storage for 30 days. Meanwhile, the layered nanostructures had larger specific surface area, which facilitated the reaction between enzyme and substrate. Due to the dual enzyme activity, the hybrid nanoflowers were applied to the enzyme inhibition-based colorimetric assay of 1-naphthol-linked bienzymatic reaction for the sensitive detection of organophosphorus pesticides. The limit of detection (LOD) for dichlorvos was as low as 0.26 pmol/L. The recoveries of dichlorvos in vegetables and fruits were 96.8%-107.4%.
Plant esterase-Cu3(PO4)2 hybrid nanoflowers were prepared, which not only achieved the immobilization of plant esterase and maintained its hydrolase activity, but also skillfully used copper phosphate skeleton. The copper phosphate skeleton not only had the supporting role, but also had the peroxidase-like activity. Different from free enzymes, hybrid nanoflowers showed higher stability, and their hydrolase activity and peroxidase-like activity maintained above 80% after storage for 30 days. Meanwhile, the layered nanostructures had larger specific surface area, which facilitated the reaction between enzyme and substrate. Due to the dual enzyme activity, the hybrid nanoflowers were applied to the enzyme inhibition-based colorimetric assay of 1-naphthol-linked bienzymatic reaction for the sensitive detection of organophosphorus pesticides. The limit of detection (LOD) for dichlorvos was as low as 0.26 pmol/L. The recoveries of dichlorvos in vegetables and fruits were 96.8%-107.4%.
2021, 49(9): 1515-1522
doi: 10.19756/j.issn.0253-3820.210480
Abstract:
Cobalt ion (Co2+) plays an important role in human body. But high concentration of Co2+ can cause a series of diseases. Peroxymonosulfate (PMS) is an oxidant with an asymmetric structure, and a certain concentration of Co2+ can catalyze the activation of PMS to produce active free radicals. Studies have shown that there is ligand effect between Co2+ and Ac-. The introduction of electron donor Ac- into Co2+-PMS accelerates electron transfer, so the catalytic activity of Co2+ towards PMS is enhanced and more sulfate radical (SO4·-) and hydroxyl radical (·OH) are produced, which results in corresponding enhancement of the ability of 3,3',5,5'-tetramethylbenzidine (TMB) oxidation. Based on the homogeneous catalytic activity of Co2+ on PMS and the enhanced coordination effect of Ac-, a novel, catalyst free, green and efficient colorimetric detection method for Co2+ was designed in this work. When the concentrations of NaAc, TMB and PMS were 1.0, 0.4 and 0.3 mmol/L, respectively, and the temperature was set at 25℃, the method was specific for the detection of Co2+ and had good anti-interference ability for 20 kinds of interfering ions. The concentration of Co2+ in the concentration range of 0.5-3.0 μmol/L showed a good linear relationship with the absorbance of the system at 652 nm (A652 nm), and the detection limit (S/N=3) was calculated to be 29 nmol/L. Compared with other detection methods for Co2+, this method established a fast, efficient and green platform for Co2+ detection without synthetic materials, large instruments and complex operation procedures.
Cobalt ion (Co2+) plays an important role in human body. But high concentration of Co2+ can cause a series of diseases. Peroxymonosulfate (PMS) is an oxidant with an asymmetric structure, and a certain concentration of Co2+ can catalyze the activation of PMS to produce active free radicals. Studies have shown that there is ligand effect between Co2+ and Ac-. The introduction of electron donor Ac- into Co2+-PMS accelerates electron transfer, so the catalytic activity of Co2+ towards PMS is enhanced and more sulfate radical (SO4·-) and hydroxyl radical (·OH) are produced, which results in corresponding enhancement of the ability of 3,3',5,5'-tetramethylbenzidine (TMB) oxidation. Based on the homogeneous catalytic activity of Co2+ on PMS and the enhanced coordination effect of Ac-, a novel, catalyst free, green and efficient colorimetric detection method for Co2+ was designed in this work. When the concentrations of NaAc, TMB and PMS were 1.0, 0.4 and 0.3 mmol/L, respectively, and the temperature was set at 25℃, the method was specific for the detection of Co2+ and had good anti-interference ability for 20 kinds of interfering ions. The concentration of Co2+ in the concentration range of 0.5-3.0 μmol/L showed a good linear relationship with the absorbance of the system at 652 nm (A652 nm), and the detection limit (S/N=3) was calculated to be 29 nmol/L. Compared with other detection methods for Co2+, this method established a fast, efficient and green platform for Co2+ detection without synthetic materials, large instruments and complex operation procedures.
2021, 49(9): 1523-1530
doi: 10.19756/j.issn.0253-3820.201768
Abstract:
A method based on target proteomics and ultrahigh performance liquid chromatography-high resolution mass spectrometry/tandem mass spectrometry was established for species identification of milks and dairies from cow, buffalo, yak, goat, sheep, horse, donkey and camel. Tryptic hydrolyzed peptides from 6 kinds of milk proteins in 8 species were analyzed and screened through theoretical analysis, instrument examination, investigation of interferences, linearity and digestion speed. A total of 13 specific peptides were finally chosen and applied to identification and detection of multi-species for milk mixture samples and commercial dairies. The results showed that specific peptides maintained good stability and specificity during species analysis. This method provided valuable suggestion in quality control and supervision of multi-species diary industry.
A method based on target proteomics and ultrahigh performance liquid chromatography-high resolution mass spectrometry/tandem mass spectrometry was established for species identification of milks and dairies from cow, buffalo, yak, goat, sheep, horse, donkey and camel. Tryptic hydrolyzed peptides from 6 kinds of milk proteins in 8 species were analyzed and screened through theoretical analysis, instrument examination, investigation of interferences, linearity and digestion speed. A total of 13 specific peptides were finally chosen and applied to identification and detection of multi-species for milk mixture samples and commercial dairies. The results showed that specific peptides maintained good stability and specificity during species analysis. This method provided valuable suggestion in quality control and supervision of multi-species diary industry.
2021, 49(9): 1531-1539
doi: 10.19756/j.issn.0253-3820.211096
Abstract:
Inspired by the fact that phosphorylated histone H2AX (γ-H2AX) has emerged as a useful biomarker for break of double-strand DNA. In this study, an isotope dilution-based liquid chromatography-tandem mass spectrometry screening method for genotoxic impurities (GTI) was established. The detection limits for H2AX and γ-H2AX target peptides were 1 ng/mL and 2 ng/mL, respectively. The accuracy and precision could meet the methodological requirement for biological samples analysis. This study focused on two kinds of important genotoxic impurities, ethylmethylsulfone (EMS) and N-nitrosodimethylamine (NDMA), in two human cell lines (HepG2 and HeLa) with different metabolic capabilities. The results showed that EMS had genotoxic characteristics in the two cell lines, while NDMA had genotoxicity only in HepG2 cells. The sensitivities of the method were 0.06 μg/g for EMS and 0.03 μg/g for NDMA, which satisfied the international detection limit standard (EMS:0.6 μg/g, NDMA:0.3 μg/g). The method was further applied to the screening of GTI in five commercially available tablets. The tablets were dissolved in water, followed by incubating with the cells to detect γ-H2AX level. The results showed that other unconcerned ingredients in these drugs did not interfere with the detection. This method was convenient, accurate and sensitive, and could be used for rapid detection of GTI in drugs.
Inspired by the fact that phosphorylated histone H2AX (γ-H2AX) has emerged as a useful biomarker for break of double-strand DNA. In this study, an isotope dilution-based liquid chromatography-tandem mass spectrometry screening method for genotoxic impurities (GTI) was established. The detection limits for H2AX and γ-H2AX target peptides were 1 ng/mL and 2 ng/mL, respectively. The accuracy and precision could meet the methodological requirement for biological samples analysis. This study focused on two kinds of important genotoxic impurities, ethylmethylsulfone (EMS) and N-nitrosodimethylamine (NDMA), in two human cell lines (HepG2 and HeLa) with different metabolic capabilities. The results showed that EMS had genotoxic characteristics in the two cell lines, while NDMA had genotoxicity only in HepG2 cells. The sensitivities of the method were 0.06 μg/g for EMS and 0.03 μg/g for NDMA, which satisfied the international detection limit standard (EMS:0.6 μg/g, NDMA:0.3 μg/g). The method was further applied to the screening of GTI in five commercially available tablets. The tablets were dissolved in water, followed by incubating with the cells to detect γ-H2AX level. The results showed that other unconcerned ingredients in these drugs did not interfere with the detection. This method was convenient, accurate and sensitive, and could be used for rapid detection of GTI in drugs.
2021, 49(9): 1540-1545
doi: 10.19756/j.issn.0253-3820.211200
Abstract:
Diabetes mellitus, characterized by hyperglycemia, is a worldwide disease which may lead to chronic complications. In the process of diabetes detection and monitoring, blood sampling is essential. In this work, a non-invasive, blood-free method was developed for the quantification of glycated albumin in saliva. The saliva sample first underwent size seizing to obtain proteins with molecular weight of larger than 10 kD, then it was separated using the reversed phase liquid chromatography (RPLC), and 36.0-37.0 min fraction was collected. Glycated salivary albumin was quantified by the glucose phenylosazone spectrophotometric assay, which showed good linearity (R2=0.997) in the range 5.0×10-4-5.0×10-2 mmol/L. With this method, the glycated albumin value (GA value) and reproducibility (RSD) for five parallel experiments were 10.9% and 11.3%, respectively. In addition, the GA values of four healthy volunteers proved the method feasible. In conclusion, this method was sensitive and reliable for the quantification of GA values in human saliva, and was inspiring for the clinical detection of diabetes mellitus.
Diabetes mellitus, characterized by hyperglycemia, is a worldwide disease which may lead to chronic complications. In the process of diabetes detection and monitoring, blood sampling is essential. In this work, a non-invasive, blood-free method was developed for the quantification of glycated albumin in saliva. The saliva sample first underwent size seizing to obtain proteins with molecular weight of larger than 10 kD, then it was separated using the reversed phase liquid chromatography (RPLC), and 36.0-37.0 min fraction was collected. Glycated salivary albumin was quantified by the glucose phenylosazone spectrophotometric assay, which showed good linearity (R2=0.997) in the range 5.0×10-4-5.0×10-2 mmol/L. With this method, the glycated albumin value (GA value) and reproducibility (RSD) for five parallel experiments were 10.9% and 11.3%, respectively. In addition, the GA values of four healthy volunteers proved the method feasible. In conclusion, this method was sensitive and reliable for the quantification of GA values in human saliva, and was inspiring for the clinical detection of diabetes mellitus.
2021, 49(9): 1546-1554
doi: 10.19756/j.issn.0253-3820.210476
Abstract:
All the peptides in the urine is peptidome, which can reflect the pathological changes of the body, thereby providing potential biomarker information. At present, morning urine is most often used in the research of urinary peptidomics. However, whether its composition changes with the sampling time is not clear. In this study, the urine samples from the three healthy people at different time points were separated by the graphene oxide-lanthanum phosphate nanocomposite (LaGM) method and identified by LC-MS/MS. Protein Pilot Software 4.5 was used for database search and analysis. It was found that the peptides and proteins of urine at each time point of the same person were different, which was reflected in the change of number and sequence of peptides in high-abundance degraded proteins and "with" or "without" in low-abundance. The number of peptides detected in urine significantly increased by sampling a plurality of times in a day. On the other hand, the peptidomics was also certain stable. The degradation peptides SGSVEDQSRVLNLGPITR of uromodulin not only existed at every time point but also had higher relative strength than other sequence-related peptides. Collagen was the dominant component in morning urine (urine at 6 o'clock). Further more,the result was verified by single urines, mixed urines of different ages, and literature data. This study showed that the sampling time was the main influencing factor of urinary peptidomics. The 24-h urine samples would be more suitable for sample than the morning urine. Due to the existence of the ladder sequence and the influence of individual differences, the absolute signal intensity of peptide should not be used for direct comparison between different samples and the selection of biomarkers.
All the peptides in the urine is peptidome, which can reflect the pathological changes of the body, thereby providing potential biomarker information. At present, morning urine is most often used in the research of urinary peptidomics. However, whether its composition changes with the sampling time is not clear. In this study, the urine samples from the three healthy people at different time points were separated by the graphene oxide-lanthanum phosphate nanocomposite (LaGM) method and identified by LC-MS/MS. Protein Pilot Software 4.5 was used for database search and analysis. It was found that the peptides and proteins of urine at each time point of the same person were different, which was reflected in the change of number and sequence of peptides in high-abundance degraded proteins and "with" or "without" in low-abundance. The number of peptides detected in urine significantly increased by sampling a plurality of times in a day. On the other hand, the peptidomics was also certain stable. The degradation peptides SGSVEDQSRVLNLGPITR of uromodulin not only existed at every time point but also had higher relative strength than other sequence-related peptides. Collagen was the dominant component in morning urine (urine at 6 o'clock). Further more,the result was verified by single urines, mixed urines of different ages, and literature data. This study showed that the sampling time was the main influencing factor of urinary peptidomics. The 24-h urine samples would be more suitable for sample than the morning urine. Due to the existence of the ladder sequence and the influence of individual differences, the absolute signal intensity of peptide should not be used for direct comparison between different samples and the selection of biomarkers.
2021, 49(9): 1555-1562,1596
doi: 10.19756/j.issn.0253-3820.201724
Abstract:
Isolation and identification of glycoproteins are of great significance for the study of their biological functions, the discovery of more biomarkers and the development of new biological drugs. However, due to the complex composition of biological samples and the low abundance of glycoproteins, it is very challenging to isolate and identify glycoproteins. Therefore, it is very important to develop an efficient method for glycopeptides enrichment. Nitrogen carbamide (C2N), a novel two-dimensional (2D) material, has high specific surface area, good hydrophilicity and abundant nitrogen, which provides the possibility of glycopeptides enrichment. In this study, a novel 2D C2N material was prepared for selective enrichment of glycopeptides. The pore size of the synthesized C2N was 2.5 nm and the specific surface area was 633 m2/g. The enrichment method based on C2N showed many advantages such as high selectivity (glycopeptides could be selectively enriched from the digests of bovine fetuin even mixed with the 200 mass folds of bovine serum albumin), high adsorption capacity (150 mg/g) and high recovery (82.2%±7.0%). The method was used for analysis of glycopeptide in 100 μg of HeLa cell lysates using three technical replicates and a total of 339, 353 and 327 glycosylation sites resulting from 328, 320 and 316 glycopeptides were characterized, respectively. This study provided a new technique for glycoproteomic and a methodological basis for the discovery of potential tumor markers.
Isolation and identification of glycoproteins are of great significance for the study of their biological functions, the discovery of more biomarkers and the development of new biological drugs. However, due to the complex composition of biological samples and the low abundance of glycoproteins, it is very challenging to isolate and identify glycoproteins. Therefore, it is very important to develop an efficient method for glycopeptides enrichment. Nitrogen carbamide (C2N), a novel two-dimensional (2D) material, has high specific surface area, good hydrophilicity and abundant nitrogen, which provides the possibility of glycopeptides enrichment. In this study, a novel 2D C2N material was prepared for selective enrichment of glycopeptides. The pore size of the synthesized C2N was 2.5 nm and the specific surface area was 633 m2/g. The enrichment method based on C2N showed many advantages such as high selectivity (glycopeptides could be selectively enriched from the digests of bovine fetuin even mixed with the 200 mass folds of bovine serum albumin), high adsorption capacity (150 mg/g) and high recovery (82.2%±7.0%). The method was used for analysis of glycopeptide in 100 μg of HeLa cell lysates using three technical replicates and a total of 339, 353 and 327 glycosylation sites resulting from 328, 320 and 316 glycopeptides were characterized, respectively. This study provided a new technique for glycoproteomic and a methodological basis for the discovery of potential tumor markers.
2021, 49(9): 1563-1571
doi: 10.19756/j.issn.0253-3820.210590
Abstract:
The abnormal expression of protein acyltransferase that is responsible for protein palmitoylation in renal clear cell carcinoma suggests that there may be abnormal changes of S-palmitoylation in renal clear cell carcinoma, which is closely related to metabolic disorder or activation of carcinogenic signaling pathway. In the present study, the optimized acyl-biotinyl exchange (ABE) method combined with proteomic techniques was used to analyze S-palmitoylated proteins in clear cell renal cell carcinoma. A total of 92 palmitoylated proteins were identified with high reliability, and the S-palmitoylation of adipose differentiation related protein (ADRP) was verified by Western blotting. This study provides an improved ABE method suitable for analyzing tissue samples. The identification of palmitoylated proteins in clear cell renal cell carcinoma can provide primary data for the molecular targets of early diagnosis, treatment and prognosis of clear cell renal carcinoma, which are of great significance for further study of the pathogenesis of clear cell renal carcinoma.
The abnormal expression of protein acyltransferase that is responsible for protein palmitoylation in renal clear cell carcinoma suggests that there may be abnormal changes of S-palmitoylation in renal clear cell carcinoma, which is closely related to metabolic disorder or activation of carcinogenic signaling pathway. In the present study, the optimized acyl-biotinyl exchange (ABE) method combined with proteomic techniques was used to analyze S-palmitoylated proteins in clear cell renal cell carcinoma. A total of 92 palmitoylated proteins were identified with high reliability, and the S-palmitoylation of adipose differentiation related protein (ADRP) was verified by Western blotting. This study provides an improved ABE method suitable for analyzing tissue samples. The identification of palmitoylated proteins in clear cell renal cell carcinoma can provide primary data for the molecular targets of early diagnosis, treatment and prognosis of clear cell renal carcinoma, which are of great significance for further study of the pathogenesis of clear cell renal carcinoma.
2021, 49(9): 1572-1579
doi: 10.19756/j.issn.0253-3820.210463
Abstract:
A method for determination of silver nanoparticles (AgNPs) in antibacterial composite food packaging by thermal extraction-emulsion injection combining single particle-inductively coupled plasma-mass spectrometry (SP-ICP-MS) was developed. The AgNPs were extracted by breaking polyethylene polymer chain using decalin under the conditions of high temperature, and the size distribution, mass and particle concentration of AgNPs were measured by SP-ICP-MS. The method was assessed to obtain the optimal extraction temperature, efficiency and limit of detection, and eventually applied to the determination of actual sample. The results indicated that, the size distribution and particle concentration would not be affected by thermal extraction process, the optimal extraction temperature of two types of antibacterial composite food packaging (film and bags) was 150℃, and the extraction efficiencies were 94.3% and 90.3%, respectively. The limit of detection for particle size was 23 nm. The detection results of actual sample showed the AgNPs contents in 4 types of antibacterial composite food packaging materials were 0.038-4.884 μg/g, the AgNPs ratio and mean size were 0.69%-3.80% and 35.5-85.1 nm, respectively. The proposed method was accurate and rapid for determination of AgNPs in food packaging materials.
A method for determination of silver nanoparticles (AgNPs) in antibacterial composite food packaging by thermal extraction-emulsion injection combining single particle-inductively coupled plasma-mass spectrometry (SP-ICP-MS) was developed. The AgNPs were extracted by breaking polyethylene polymer chain using decalin under the conditions of high temperature, and the size distribution, mass and particle concentration of AgNPs were measured by SP-ICP-MS. The method was assessed to obtain the optimal extraction temperature, efficiency and limit of detection, and eventually applied to the determination of actual sample. The results indicated that, the size distribution and particle concentration would not be affected by thermal extraction process, the optimal extraction temperature of two types of antibacterial composite food packaging (film and bags) was 150℃, and the extraction efficiencies were 94.3% and 90.3%, respectively. The limit of detection for particle size was 23 nm. The detection results of actual sample showed the AgNPs contents in 4 types of antibacterial composite food packaging materials were 0.038-4.884 μg/g, the AgNPs ratio and mean size were 0.69%-3.80% and 35.5-85.1 nm, respectively. The proposed method was accurate and rapid for determination of AgNPs in food packaging materials.
2021, 49(9): 1580-1586
doi: 10.19756/j.issn.0253-3820.191165
Abstract:
Laboratory spectrometer is a spectrometer that can only obtain spectral signals by spectrometer, and hyperspectral camera acquires image and spectral information by image. Under the condition of using laboratory spectrometer, the calibration transfer of different temperatures, different ordinary spectrometers, different measuring conditions and samples from different regions has been studied. But there are few studies on the calibration transfer between laboratory spectrometer and hyperspectral camera. Taking 164 soil samples from Qingdao as an example, the spectral data of laboratory spectrometer and hyperspectral camera data were obtained in this work. The content models of total nitrogen (TN) and total phosphorus (TP) were established based on the spectral data of laboratory spectrometer. Three methods, namely piecewise direct correction algorithm, model updating and slope/bias correction (PDS-MP-S/B), were used to transfer the hyperspectral data. The transferred hyperspectral data were substituted into the content models of TN and TP to predict their content values and evaluate the prediction effect. Meanwhile, the influence of the number of PDS window and the number of standard set on the prediction results was analyzed. When the number of PDS window was 19 and the number of standard set was 120, the prediction effect after calibration transfer was the best, absolute coefficient of test set (Rt2) was 0.736 and root mean square error of prediction (RMSEP) was 0.274. In the content prediction of TP, when the number of PDS window was 23 and the number of standard set was 80, the prediction effect after calibration transfer was the best, Rt2 was 0.647 and RMSEP was 0.231. The solution of model transfer between laboratory spectrometers and hyperspectral cameras provided a powerful basis for rapid prediction of a large number of image information data collected by hyperspectral cameras, which greatly reduced the workload, and the wide application of hyperspectral cameras in quantitative analysis and rapid measurement technology.
Laboratory spectrometer is a spectrometer that can only obtain spectral signals by spectrometer, and hyperspectral camera acquires image and spectral information by image. Under the condition of using laboratory spectrometer, the calibration transfer of different temperatures, different ordinary spectrometers, different measuring conditions and samples from different regions has been studied. But there are few studies on the calibration transfer between laboratory spectrometer and hyperspectral camera. Taking 164 soil samples from Qingdao as an example, the spectral data of laboratory spectrometer and hyperspectral camera data were obtained in this work. The content models of total nitrogen (TN) and total phosphorus (TP) were established based on the spectral data of laboratory spectrometer. Three methods, namely piecewise direct correction algorithm, model updating and slope/bias correction (PDS-MP-S/B), were used to transfer the hyperspectral data. The transferred hyperspectral data were substituted into the content models of TN and TP to predict their content values and evaluate the prediction effect. Meanwhile, the influence of the number of PDS window and the number of standard set on the prediction results was analyzed. When the number of PDS window was 19 and the number of standard set was 120, the prediction effect after calibration transfer was the best, absolute coefficient of test set (Rt2) was 0.736 and root mean square error of prediction (RMSEP) was 0.274. In the content prediction of TP, when the number of PDS window was 23 and the number of standard set was 80, the prediction effect after calibration transfer was the best, Rt2 was 0.647 and RMSEP was 0.231. The solution of model transfer between laboratory spectrometers and hyperspectral cameras provided a powerful basis for rapid prediction of a large number of image information data collected by hyperspectral cameras, which greatly reduced the workload, and the wide application of hyperspectral cameras in quantitative analysis and rapid measurement technology.
2021, 49(9): 1587-1596
doi: 10.19756/j.issn.0253-3820.191683
Abstract:
Magnetic fullerene (Fe3O4@SiO2@C60) was synthesized as a solid-phase extraction adsorbent for extraction and enrichment of pesticides in Chrysanthemum. Based on this, anultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method was developed for determination of nine kinds of pesticides, including cyprodinil, zoxamide, tolfenpyrad, azoxystrobin, pyraclostrobin, trifloxystrobin, fenpyroximate, pyridaben and phoxim in edible chrysanthemum. The results showed that the Fe3O4@SiO2@C60 had obvious adsorption to cyprodinil, zoxamide, tolfenpyrad, azoxystrobin, pyraclostrobin, trifloxystrobin, fenpyroximate, pyridaben and phoxim. The influence factors of magnetic dispersion solid phase extraction were investigated, and it was found that a best extraction was obtained under the optimal conditions such as 30.0 mg of Fe3O4@SiO2@C60 as adsorbent, extraction for 20 min under shaking, separation of the Fe3O4@SiO2@C60 by an external magnetic field, and elution two times with 1 mL of ethyl acetate. The result showed that Fe3O4@SiO2@C60 had good stability and reusability. All of the tested pesticides showed good linear relationship in the range of concentration 0.005-0.1 mg/L with correlation coefficients of greater than 0.999. The detection limits and the quantitation limits of nine target pesticides were in the range of 0.001-0.003 mg/kg and 0.003-0.01 mg/kg, respectively. When the addition levels of the pesticides were 0.01, 0.05 and 0.10 mg/kg, the recoveries of the method ranged from 71.0% to 94.6% with the relative standard deviations (n=6) ranging from 4.6% to 11.3%. The method was simple, sensitive, fast and with low cost, and suitable for the accurate qualitative and quantitative determination of nine pesticides in chrysanthemum.
Magnetic fullerene (Fe3O4@SiO2@C60) was synthesized as a solid-phase extraction adsorbent for extraction and enrichment of pesticides in Chrysanthemum. Based on this, anultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method was developed for determination of nine kinds of pesticides, including cyprodinil, zoxamide, tolfenpyrad, azoxystrobin, pyraclostrobin, trifloxystrobin, fenpyroximate, pyridaben and phoxim in edible chrysanthemum. The results showed that the Fe3O4@SiO2@C60 had obvious adsorption to cyprodinil, zoxamide, tolfenpyrad, azoxystrobin, pyraclostrobin, trifloxystrobin, fenpyroximate, pyridaben and phoxim. The influence factors of magnetic dispersion solid phase extraction were investigated, and it was found that a best extraction was obtained under the optimal conditions such as 30.0 mg of Fe3O4@SiO2@C60 as adsorbent, extraction for 20 min under shaking, separation of the Fe3O4@SiO2@C60 by an external magnetic field, and elution two times with 1 mL of ethyl acetate. The result showed that Fe3O4@SiO2@C60 had good stability and reusability. All of the tested pesticides showed good linear relationship in the range of concentration 0.005-0.1 mg/L with correlation coefficients of greater than 0.999. The detection limits and the quantitation limits of nine target pesticides were in the range of 0.001-0.003 mg/kg and 0.003-0.01 mg/kg, respectively. When the addition levels of the pesticides were 0.01, 0.05 and 0.10 mg/kg, the recoveries of the method ranged from 71.0% to 94.6% with the relative standard deviations (n=6) ranging from 4.6% to 11.3%. The method was simple, sensitive, fast and with low cost, and suitable for the accurate qualitative and quantitative determination of nine pesticides in chrysanthemum.
