Capillary Electrophoresis The Adenylate Method Shrimp Muscle

Capillary electrophoresis (CE) is one of the most efficient separation techniques today. It has been rapidly developed since the early 1980抯. Capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MECC) have been utilized for the separation of nucleotides. The affecting factors and application of CE analyzing nucleotides are summarized, and the advantages and existing problems in using are discussed. A method for the analysis of adenylates using micellar electrokinetic capillary chromatography (MECC) has been developed. The analysis was accomplished on a bare fused-silica capillary of 48cm(37.Scm effective length) >( 75 1-?m J.D. with on-column UV 254nm detection. The separation of three adenylates was optimized with respect to the concentration of surfactant, pH of running buffer and applied voltage. Optimum conditions were S0mmol/L dibasic sodium phosphate-2Ommol/L hexaclecylerimethylammonium bromide (CTAB)- lmmol/L EDTA buffer (pH 8.0), -15KV applied voltage. Three adenosine phosphates were separated with high resolution within 10 mm. The resolution was higher than 3. The presented method was applied to the determination of adenosine phosphates in the muscles of shrimp. The results demonstrate that MECC provides a simple, sensitive, rapid and reliable way to analyze adenylates in tissues. The method is applied to the research in energy changes during molt cycle. The pattern of changes in the levels of ATP, TAN, AEC and ATP/ADP is S-shape and these parameters are the lowest at ecdysis and the highest at 24 hr after ecdysis (P<0.05). The value of AEC during other times is in a limit rang (0.86- 0.89). The ratio of ATP/ADP is relatively higher on late proecdysis, which may be related to the significantly high value of adenylate kinase equilibrium constant at the same time (p