The Application Of Spectral Analysis In The Determination Of Heparin, Coenzyme A And Glucose Of Blood Serum

Heparin, which is produced by some mammal, is polymer of glucide. It exists in the blood excreted by hypertrophy cells of the body. Because of the inhibition of clotting, it was used to prevent thromboses and to cure urgent vein thrombus early in 1935. Now it is still the first—choice to prevent thromboses and to cure urgent vein thrombus because of the absorbability, good antithrombotic function, long half-life and high biological value in use. Furthermore, heparin and its derivants have a variety of biological activities such as anticoagulant, antilipemic, antithrombotic, immunoregulatory, antiphlogistic and antianaphylactic activities, etc. It is a kind of important biochemical medicament in clinic.Coenzyme A, which is synthesized in all cells from pantothenate, cysteine and ATP, is a kind of adenine compounds. It has connection with the metabolism of lipids amino acids and carbohydrates, accelerating the synthesis of acetylcholine, increasing the storage of hepatic glycogen and so on. CoA is often used as a kind of clinical medicine to improve following symptom: anxiety, irritability, inappetence, fatigue, anaesthesia, convulsion and so on. Therefore, it is significant to establish a precise, sensitive and simple method for the determination of CoA in life sciences and clinical research.Horseradish peroxidase is a kind of planting peroxidase. It can be used to determine the concentration of hydrogen peroxide or horseradish peroxidase according to the structure of the substrate. Many biological samples can produce hydrogen peroxide under the catalysis of some enzyme. By coupling with a glucose oxidase-catalyzed reaction, the concentration of glucose in human serum can be determined.In this paper, according to above three objects, using molecule spectroscopy (fluorescence and Uv-Vis spectroscopy), new methods for the spectrofluorimetric determination of heparin and coenzyme A are established. A study on the ArsenazoIII or Zincon as substrate of horseradish peroxidase (HRP)- hydrogen peroxide (H₂O₂) catalytic system is reported. There are three chapters.In the first section, we studied the interaction between heparin and tetracycline analogues-europium ion and the determination of heparin. Tetracycline analogues contain theβ-diketonate configuration and it is the ideal ligand for europium ion. The tetracycline analogues-europium ion binary complex emits the characteristic peak of europium ion at 612 nm. In a certain acidity condition, Hep can remarkably enhance the fluorescence intensity of the tetracycline analogues-europium ion binary complex atλ=612nm and the enhanced fluorescence intensity is in proportion to the concentration of Hep. Optimum conditions for the determination of Hep were investigated. By the Rosenthal graphic method, the binding number (N) and association constant(K) of Hep with the probe are obtained.In the second section, we studied the interaction between coenzyme A and rare metal ion- drug complex and the determination of CoA in the existence of H₅IO₆. CoA can remarkably enhance the fluorescence intensity of the system, and the enhanced fluorescence intensity is proportional to the concentration of CoA. Optimum conditions for the determination of CoA were also investigated. This method is simple, practical and relatively free of interference from coexisting substances, and can be successfully applied to assess of CoA in injection and biological samples.In the third section, A study on the ArsenazoIII or Zincon as substrate of horseradish peroxidase (HRP)- hydrogen peroxide (H₂O₂) catalytic system is reported. A cleavage of covalent bond of -N=N- group was showed. The decreased absorption is in proportion to the concentration of H₂O₂. By coupling with a glucose oxidase-catalyzed reaction, the concentration of glucose can be quantified. The method can be used to quantify the glucose in human serum.