Massey Documents by Type

Permanent URI for this communityhttps://mro.massey.ac.nz/handle/10179/294

Browse

Filters

1998 - 199922000 - 20061

Settings

Subject: search.filters.subject.Capillary electrophoresis

Search Results

Now showing 1 - 3 of 3
  • Thumbnail Image
    Item
    The development of a method to determine felinine in body fluids by capillary electrophoresis : a thesis presented in partial fulfilment of the requirements for the degree of Master of Philosophy in Chemistry at Massey University
    (Massey University, 1999) Zhang, Fengying
    Ion-exchange, paper-chromatography and high performance liquid chromatography were used in earlier studies for the determination of felinine in biological fluids. These methods were either inadequate and/or need laborious sample pre-treatments. A new method for the determination of felinine by capillary zone electrophoresis has been developed. Preliminary investigations were carried out to address the conditions required for the separation of felinine. The separation of felinine can be performed on a fused-silica capillary with a 20 mM phosphate buffer (pH 2.0) and detection wavelength 200 nm. The separation principle was based on the different migration times due to the different molecular weights, molecular sizes and charges under an applied potential field. The quantitative determination of felinine levels in cat urine has been achieved. The cat urine analysis was performed directly on the capillary electrophoresis without making any felinine derivative(s). The levels of felinine in different cat genders are reported. The results were compared with the results of an HPLC felinine derivatization method. Felinine levels in entire male cat urine were much higher than those in female and castrated male cat urine. A synthetic felinine was employed as standard felinine. Linear relationships between peak area and concentration of synthetic felinine calibrations are reported. Mean felinine recovery in cat urine was 95.9%. Taurine, urea, creatine and creatinine, which exist in large amounts in cat urine, showed no interference with the analysis of felinine by this method. The new capillary zone electrophoresis method was then applied to the study of felinine stability. Conditions reported to influence the stability of felinine were investigated. These conditions included oxidation, storage temperatures and times, heating, acidic and alkaline solutions. Both synthetic felinine and felinine in cat urine were investigated. Storage temperature (-20°C to 20°C) had no significant influence on the stability of felinine while higher temperatures increased the decomposition of felinine. Felinine degraded at strong acid and base conditions but was relatively stable under mild acid and base conditions. A similar stability of felinine in human urine is also reported. The capillary zone electrophoresis method was also employed to study felinine in plasma and serum. Plasma and serum as well as urine can be analysed directly on the capillary electrophoresis after sufficient dilution. Conditions (eg. protein clean up, changing of injection time, 37°C heating) that might influence of felinine behaviour in plasma and serum are discussed. This study indicated that no traces felinine be found in cat plasma, within the detection limits of this new capillary electrophoresis method.
  • Thumbnail Image
    Item
    Determination of creatinine and creatine by capillary electrophoresis : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Chemistry at Massey University
    (Massey University, 1998) Guo, Hong
    The assessment of creatinine and creatine in biological fluids is important in the evaluation of renal and muscular functions. For routine creatinine determinations in the clinical laboratory, the most frequently used method is the spectrophotometric one based on the Jaffé reaction. However, this reaction is not specific for creatinine. For this reason, several methods have been proposed, but the elimination of interferences in the determination of creatinine has still not been achieved in some of these methods; others solved this problem either with expensive equipment that does not suit routine analysis or necessitates time-waste procedures. In this thesis capillary electrophoresis was the new tool investigated. It was applied in an attempt to achieve both the separation of creatinine from the non-creatinine 'Jaffé- reacting' chromogens and the determination of creatine in serum. Capillary zone electrophoresis was performed with detection at wavelength 480 nm to separate creatinine from the non-creatinine 'Jaffé-reacting' chromogens in urine. The principle was based upon the different migration times due to the different molecule weights, molecular sizes and charges under the applied high voltage. The picric acid was employed as part of the running buffer to allow reaction of creatinine and picrate to take place after the sample injection. This procedure eliminated the negative influence of the reaction time that is controlled manually in the common Jaffé reaction method. Therefore, compared to the Jaffé reaction method, the new method achieved more accuracy and precision in the determination of creatinine. Determination of creatinine in serum and urine were studied at a new wavelength 417 nm, which gave a higher sensitivity of detection than at 480 nm. This wavelength shift made the determination of creatinine in serum possible by capillary zone electrophoresis without the non-creatinine 'Jaffé-reacting' chromogens interfering. In this method, serum only needed a simple filtration before the analysis. Creatine was discovered to have absorption at 417 nm in alkaline medium. Moreover, specific sample stacking was introduced in this method. The sample was dissolved in a mixture of two-volumes acetonitrile and one-volume 3 % ammonium chloride to give a 10-fold enhancement of detection sensitivity.
  • Thumbnail Image
    Item
    Development of methods for capillary isoelectric focusing of dairy proteins : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Chemistry at Massey University
    (Massey University, 2006) Gapper, Leyton William
    Capillary Isoelectric Focusing (CIEF) is a high-resolution technique which can be applied to the separation and characterisation of complex biological mixtures such as dairy proteins. Although dairy proteins are commonly analysed by traditional gel electrophoresis techniques including 2-Dimensional PAGE, CIEF offers the advantages of reduced analysis times, the ability to handle smaller sample volumes and increased sensitivity with improved separation efficiencies. Several methods for capillary isoelectric focusing of dairy proteins have been developed herein. For the analysis of soluble whey proteins methods that can be used with either UV or mass spectrometry (MS) detection have been set up. For MS detection a coaxial sheath flow interface in conjunction with electrospray ionisation has been utilised. For analysis of the inherently insoluble casein proteins with UV detection denaturing and reducing agents have been introduced into the system. Results have shown very close similarities to those obtained by IEF gels.