| Before the 1980s,chemiluminescence analysis method was widely used in the analysis and testing of some inorganic substances due to its advantages such as high sensitivity,simple operation and fast analysis speed.Since the early 1980s,the application of chemiluminescence analysis in the field of life analysis has gradually attracted researchers’ attention.Compared to radioisotope labeling,chemiluminescence is safer and its reagents are more stable.Compared to fluorescence,because the chemiluminescence process does not require an exogenous excitation light,it has high signal-to-noise ratio and high sensitivity.Compared to bioluminescence,chemiluminescence has low cost,flexible application,and a wide range of detection objects.The above advantages make chemiluminescence analysis a powerful technical support for modern life science and medical research.Designing and synthesizing new chemiluminescence probes,establishing a new chemiluminescence system,and significantly improving analytical properties and application fields are the current hot focus in chemiluminescence research.This work aims to design and synthesize a new chemiluminescence probe PMPHB-I and study its chemiluminescence properties and in vivo imaging applications.The content of this paper consists of review and research report.The review section briefly introduces the mechanism and development history of chemiluminescence,and summarizes the applications of chemiluminescence in the field of life analysis in the past five years.This research report is divided into the following two parts:1.Synthesis and Chemiluminescence Properties of Probe PMPHB-I and Its Application in In Vivo ImagingSynthesis and Chemiluminescence Properties of Probe PMPHB-I.Based on the experience of synthesizing hydrazide chemiluminescence reagents in the laboratory,a new probe PMPHB-I was designed and synthesized,and its structure was characterized by nuclear magnetic resonance and mass spectrometry.The chemiluminescence properties of PMPHB-I were study and the results are as follows:(1)The emission wavelength of PMPHB-I is 530 nm,and it emits a bright yellow-green chemiluminescence when oxidized.(2)pH has an effect on PMPHB-I chemiluminescence.The chemiluminescence intensity is the strongest when the solution pH is 8.0.(3)Among several reactive oxygen species,PMPHB-I has high selectivity to singlet oxygen(102)under physiological conditions(pH=7.4,in PBS buffer).(4)The prepared PMPHB-I solution has good chemiluminescence stability,and and can be used as a stock solution.Probe PMPHB-I is used in In Vivo Imaging.(1)Firstly,the linearity of PMPHB-I detection for 1O2 under physiological conditions was studied.Within the range of 1.0×10-7-5.0×10-6 mol·L-1,PMPHB-I can quantitatively respond to 102 and the detection limit is 4.2×10-8 mol·L-1.(2)Based on the high selectivity and sensitivity of PMHPB-I to 1O2 under physiological conditions,PMPHB-I was used for chemiluminescence in vivo imaging of 1O2-enriched peritonitis inflammation sites.Achieced the highly sensitive detection of the nude mice peritonitis.These results illustrate that PMPHB-I can provide a new method support for in vivo inflammation detection.2.Rapid Detection of Sudan Red in FoodThe thin-layer chromatography(TLC)technology and matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS)technology are matched to meet the needs of on-site rapid inspection and laboratory qualitative analysis.First,a non-toxic,green eluent capable of completely separating Sudan Red Ⅰ,Ⅱ,Ⅲ,Ⅳ and capsanthin was selected,and the proportion of the eluent was optimized.N-hexane:ethyl acetate=7:0.5.By comparing the Rf value of the suspected spots with the Sudan red standard sample spots,realize the rapid on-site detection of illegal addition of Sudan red in the sample.Subsequently,the suspected spots of Sudan Red in the thin-layer plate were separated,and the pigment in silica gel was dissolved with ethyl acetate.After filtering,the molecular ion peaks were detected by matrix-assisted laser desorption ionization time-of-flight mass spectrometry,and the quasi-determinism of the illegal addition of Sudan Red was achieved.This work develops a simple,green,non-professional,and suitable for field application of Sudan Red detection technology.A laboratory-qualified matrix-assisted laser desorption ionization time-of-flight mass spectrometry method is also provided.This research progress has also been applied to undergraduate experimental teaching,teaching students to establish fast detection methods for illegal additives in food,strengthening the concept of green environmental protection,and realizing the learning and function of matrix-assisted laser desorption ionization time-of-flight mass spectrometry. |
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