The Application And Study Of Spectrometry In The Hydrolysates Of Nucleic Acid Analysis

Nucleic acid is a vital substance, it is the basic of the taker and genic expression of the descendiblity information. It dominates the composition of protein, the growth, manifold, aberrance and heredity of all flesh. Along with the development of modern molecular biology and molecular biology technology, the investigation of nucleic acid and the hydrolysates of nucleic acid, their interaction with other molecules become an important field.Fluorimetry is developing in the analysis of biologic molecular, due to its highly sensitivity,good selectivity and simple procedure. In recent years, fluorimetry developed rapidly and used more and more abroad in analysis. Obviously, fluorimetry have a favorable application future in the analysis field.The fluorescence intensity of the hydrolysates of nucleic acid is very feeble, so their character, configuration and determination cannot be investigated by fluorimetry directly. Therefore, to improve the sensitivity, in this paper, we used fluorescence quenching method to determine adenine, uracil, thymine and hypoxanthine, we also studied the fluorescence quenching mechanism. In addition, a method was described for the determination of adenine by fluorimetry with 1,3-cyclohexanediono- formaldehyde as fluorescent derivative reagent.The first chapter reviews the character and application of the hydrolysates of nucleic acid, the application and prospect of fluorimetry.In the second chapter to the fifth chapter, we found fluorescence quenching method to determine adenine, uracil, thymine and hypoxanthine, studied the quenching mechanism between adenine, uracil, thymine, hypoxanthine and Evans blue by the Stern-Volmer plot and the fluorescence lifetimes of Evans blue respectively. The resultant constants of Evans blue with adenine, uracil, thymine and hypoxanthine were obtained based on the connections of log [(F0-F)/F] and log [Q]. Effects of pH, foreign ions, stability time and the presence of hydrolysates of nucleic acid on the determination have been examined. This method was simple, rapid and the detection limit were low, it can be used in the determination of adenine, uracil, thymine and hypoxanthine in samples, the results obtained by the fluorescence quenching method agreed with those by the HPLC method.In the sixth chapter, we studied the determination of adenine by fluorimetry with 1,3-cyclohexanediono-formaldehyde as fluorescent derivative reagent. The measurement was carried out at 453 nm with excitation at 394 nm . A good linear relationship was obtained between the fluorescence intensity and the concentration of adenine, so adenine can be determined by this method. Effects of pH, foreign ions, stability of the production and the presence of hydrolysates of nucleic acid on the determination have been examined. This method was simple, rapid and the detection limit were low, it can be used in the determination of adenine in samples...