| Because of its high efficiency,fast and little sample volume,capillary electrophoresis has become one of the widely used analysis techniques recently,capillary electrophoresis with contactless conductivity detection method have the advantage of direct quantifcation of nonUV-absorbing compounds without derivatization,simple operation and low cost.The biological samples have complex matrix and usually the sample source is not easy,detection of substances in biological samples has a good guiding role for disease.In this work,we used laboratory-made CE device with capacitively coupled contactless conductivity detector to detect three kinds of biological samples,cells,blood and urine respectively.This method shows an application potential in non-invasively taken samples(urine,saliva,exhaled breath condensate)that may be particularly appealing for fast clinical diagnosis.This thesis is divided into the following chapters:Chapter 1:It was divided into third sections.The first section summarized the capillary electrophoresis system,mainly introduced the development course,principle,separation mode,sample injection technology,detection method and electrophoresis characteristics of capillary electrophoresis.The second section summarized the capillary electrophoresis device with capacitively coupled contactless conductivity detection(CE-C~4D).We introduced the development,principle,characteristics and application of CE-C~4D in pharmaceutical clinical field,food analysis and environmental analysis.The third section summarized the biological samples,mainly introduced the characteristics,separation methods and advantages detected by CE.Chapter 2:In this work,a rapid,sensitive,and practical method was developed for the determination of Na~+and K~+concentration in Hela cell during emodin induced apoptosis by a low-cost capillary electrophoresis device with capacitively coupled contactless conductivity detection(CE-C~4D).Under the optimized condition,both ions were baseline separated in 4 min with 40 mM MES/40 mM His containing 1 mM18-crown-6 as the separation buffer at pH 6.0.The limit of detections(LODs)and limit of quantifications(LOQs)were 0.47-1.15?M and 1.58-3.86?M,respectively.The precision for migration times and peak areas were below 0.56%and 3.74%,respectively.The data proved that the concentration of cations in cells can be accurately quantified.It was found that the K~+concentration decreased from 94.6?M to 39.7?M,and the sodium ion concentration increased from 219.2?M to 572.5?M during the process of apoptosis when the cell density was 1×10~5 cells/mL.The low-cost CE-C~4D provides a convenient way to decipher the interaction of Na~+and K~+in the regulation of cell apoptosis.Chapter 3:Sarcosine,a new marker of prostate cancer discovered by Chinnaiyan et al in 2009,it can not only effectively reflect the aggressive discrimination of prostate cancer cell growth behavior,but also easy to be detected in prostate cancer urine samples.And for L-kynurenine,L-proline and L-cysteine their contents were significantly increased in the process of prostate cancer.In this study,we report a simple,robust,and reproducible CE-C~4D method for the determination of sarcosine and other representative potential biomarkers in pooled urine.In this work,excellent resolution between sarcosine and its isomers(L-alanine)was achieved with no derivatization step.The separation was carried out in normal polarity mode at 20?,18 kV and using hydrodynamic injection(10 cm for 20 s).Under the optimum conditions,a selective separation of the four target analytes from other constituents present in the analyzed biological matrices was achieved in 17 min in a fused silica capillary of 50?m I.D.using an electrolyte of 2.3 M acetic acid.The method presents a good diagnosis tool that can be used for early diagnosis of prostate cancer in situ.Chapter 4:In this paper,a simple and rapid method for the preparation of open capillary coating column has been developed.Based on this method,biological samples can be directly injected for capillary electrophoresis analysis.The biological samples such as blood or serum was injected into fused quartz capillary directly at room temperature,and the proteins and other macromolecular compounds in the biological sample matrix were retained in the inner wall of the capillary,eliminating their adsorption on the analytical capillary during electrophoresis and it ensures stable electroosmotic flow and the migration time of the target analytes.The applicability of this method is demonstrated by the determination of the main inorganic cations in the serum of healthy persons and epileptic patients.Serum samples can be directly injected into capillary after deionized water water dilution.The background electrolyte was 40 mM MES/40 mM L-His containing 1 mM 18-crown-6,The repeatability of capillary tube coated with different biological samples was verified by measuring the same biological sample,and the RSD value of peak area and migration time,for coating column,was in the range of 5.10%and 2.84%,respectively.The sample volume required for each injection is less than 3?L. |
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