| Fluorescence analysis and peroxyoxalate chemiluminescence(PO-CL) analysis all require that the analyte must be fluorescent, but the material molecules having their own fluorescence are not many, which limits its direct determination with fluorescence or chemiluminescence.In order to solve this problem, photochemically induced fluorescence(PIF) and photochemically induced chemiluminescence(PICL) detection technology arise at the historic moment. The research of this aspect in the literature has been many reports, however the main disadvantage is the relatively poor selectivity. Applied in biological samples and complex substances analysis, these methods must be coupled with the efficient separation methods, such as high-performance liquid chromatography, gas chromatography or capillary electrophoresis, or with specific molecular recognition, such as antibodies, ligands, aptamers or molecular imprinting.Peroxyoxalate chemiluminescence system based on bis-(2,4,6-trichlorophenyl) oxalate(TCPO)-H2O2-fluorescent agent, because of its high quantum yield, has become a highly sensitive analytical method for determination of trace hydrogen peroxide or fluorescent materials. Imidazole is often used as enhancer in TCPO chemiluminescence reaction. In our study we found incidentally that when imidazole exists, the structure of photochemically induced fluorescence product changes. In the shorter wavelength position, a new strong fluorescence peak appears, and the distance of two fluorescence peaks wavelength is as long as135nm. More interesting is that in the longer wavelength, fluorescence peak intensity enhances with fluorescent material concentration increases, but in the shorter wavelength, fluorescence peak intensity reduces with fluorescent material concentration increases. This discovery is to bring new possibilities for improving the selectivity of photochemically induced fluorescence method and the accuracy of determination. At the same time, photochemically induced chemilumine-scence analysis reported in the literatureare is almost all using flow injection analysis technology, the analysis efficiency is lower. Considering that the TCPO-H2O2-fluorescent reagent chemiluminescence system is a kind of slow reaction and the luminescence response time is longer, this is to provide the possibility for developing an analysis method which has high throughput, high sensitivity, high selectivity, high detection speed, low detection limit and low cost. Its success will play an important role and have a good application prospect in clinical disease control, environmental monitoring, food analysis field.This paper is composed of two chapters:Chapter1:reviewThis review mainly introduces the principle, method and devices of photochemical reaction, introduces the principle and method of photochemically induced fluorescence and chemiluminescence detection technology, as well as summarizes their development and application in recent years.Chapter2:research reportWe study several kinds of drugs with photochemically induced fluorescence and high-throughput photochemically induced chemiluminescence detection methods. Mainly including the following contents:1. Determination of vitamin Kl by photochemically induced fluorescence analysisVitamin Kl does not have its own fluorescence, can't be directly determined using the fluorescence method. A photochemically induced fluorescence(PIF) method towards vitamin Kl is presented. This method was based on ultraviolet irradiation(254nm/15W) of vitamin K1to produce fluorescent product, which can be determined with fluorescence detection. The influence of imidazole on photochemical product structure and fluorescence spectrum has been studied. The linear range was0.1～10.0μg/mL with a detection limit of0.03μg/mL for vitamin Kl and the relative standard deviation was2.4%(c=1.0μg/mL, n=11). Using this method for determination of vitamin K1injection and human serum, the results are satisfactory. This method has many advantages such as high sensitivity, high selectivity, high accuracy, less reagent consumption and less matrix interference, quick and simple operation. The sensitivity and selectivity of this method were sufficient for clinical and nutritional applications.2. Determination of sulfadiazine by photochemically induced fluorescence analysisSulfadiazine is of a typical sulfa drug and hasn't fluorescence, which can't be determined directly using fluorescence method. A sensitive and selective fluorescence method towards sulfadiazine with photochemically induced fluorescence(PIF) is proposed. This method was based on ultraviolet irradiation of sulfadiazine to produce fluorescent product, which can be determined with fluorescence. Various factors on fluorescent signal including illumination condition, solvent, acidity, sensitizing agent have been optimized. Under the optimum conditions, the analytical performance was obtained. The linear range was0.05-5.Oμg/mL with a detection limit of0.016μg/mL for sulfadiazine and the relative standard deviation was2.1%(c=1.0μg/mL, n=11). Using this method for determination of sulfadiazine tablets and human serum, the results are satisfactory. This method has many advantages such as high sensitivity, high selectivity, less reagent consumption and matrix interference, quick and simple operation, and greatly expand the application range of the fluorescence detection.3. Rapid and high-throughput determination of dipyridamole with chemilum-inescence methodPeroxideoxalate chemiluminescence system(PO-CL), because of its high quantum yield, has become a highly sensitive analysis method for determination of trace hydrogen peroxide or fluorescent materials. An high-throughput, highly sensitive, simple, rapid determination method based on bis-(2,4,6-trichlorophenyl) oxalate(TCPO) chemiluminescence system for determination of dipyridamole is proposed. The analysis conditions including solvent, acidity, catalyst, and the concentration of CL reagents have been investigated. The content of dipyridamole can be determined directly with SynergyTM2SLD chemiluminescence microporous board detecting instrument. Under the optimum conditions, the analytical performance was obtained. The linear range was3.3x10-9～1.7x10-6g/mL with a detection limit of1.1x10-9g/mL for dipyridamole and the relative standard deviation was2.2%(c=3.3xl0-8g/mL, n=l1). The proposed method has been used for successful analysis of dipyridamole in actual samples such as human serum and human urine.4. Rapid and high-throughput determination of vitamin K1with photochemically induced chemiluminescence analysisVitamin K1hasn't own fluorescence and can't be determined directly using chemiluminescence method. A high-throughput determination of vitamin K1by photochemically induced chemiluminescence analysis(PICL) is developed. This method is based on ultraviolet irradiation(254nm/15W) of vitamin K1to produce hydrogen peroxide and a fluorescent product at the same time, which can be determined with peroxideoxalate chemiluminescence system using SynergyTM2SLD microporous board chemiluminescence detecting instrument. The influence of imidazole on photochemical product structure as well as chemiluminescence mechanism has also been studied. Under the optimum conditions, the analytical performance was obtained. The linear range was6.7x10-9～3.3x10-6g/mL with a detection limit of2.2ng/mL for vitamin K1and the relative standard deviation was2.7%(c=1.0x10-1g/mL, n=11). This method has been used for successful analysis of the amount of vitamin K1in actual samples such as human serum and vitamin K1injection. The proposed method has many advantages such as high throughput, high selectivity, high sensitivity, high automaticity, low detection limit and less reagent consumption, thus greatly expand the applications of chemiluminescence detection and has very good development potential. |
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