ICPs POLYMER

TECHNICAL FIELD

The invention relates to a gold electrode-based triglyceride biosensor prepared by modifying self-conducting ICPs polymer, which enables the determination of triglyceride in a practical, fast and wide measurement range.

PRIOR ART

The main form of all kinds of fat in food is triglycerides. All of the saturated and unsaturated fats, animal and vegetable fats, are first decomposed into triglycerides and transported to the liver and undergo different processes in the liver. If you take in too many calories from either carbohydrate, fat or protein sources, your body converts the excess energy sources into triglycerides and stores them in your fat cells. Thus, fat cells swell, grow, and cause weight gain and invite obesity. Excess triglyceride that is not stored in fat cells circulates in your veins. LDL; increases cholesterol (harmful cholesterol), HDL; reduces cholesterol (useful cholesterol). Excess energy consumption and increased alcohol use lead to rapid and unexpected increases in triglyceride levels.

The increase in triglyceride level causes coronary heart disease, atherosclerosis, stroke, memory loss, and terminal vascular diseases to be seen more frequently.

The blood triglyceride level is very variable. Therefore, it is important to have an accurate and safe triglyceride measurement. For this, in addition to the personal precautions to be taken before the measurement, the kits used for the measurement are important. In the absence of accurate test results, it is difficult for physicians to diagnose or provide effective treatment. The total testing process in clinical laboratories is a complex and multi-step process, starting and ending with the patient, from the medical decision for testing to the reporting of results. Total testing process; test selection and test request, sample collection, identification, transport of the sample to the laboratory, sample preparation, analysis, reporting of test results and interpretation of test results. LDL cannot be reported if the triglyceride level cannot be measured in this process, which provides the greatest assistance to the physician in making the right decision and applying the treatment. This situation causes the clinician not to evaluate a parameter that is very important in the follow-up and treatment of cardiovascular diseases.

DETAILED DESCRIPTION OF THE INVENTION

The invention is a gold electrode-based triglyceride biosensor prepared by modifying the self-conducting ICPs polymer, which enables the determination of triglyceride quantity in a practical, fast and wide measurement range.

The preparation steps of the biosensor subject to the invention are as follows; After the synthesis of the new generation conductive polythiophene polymer, which stands out with its electron donor structure, from the thiophene monomer, its synthesis characterization is controlled by 1 H-NMR, 13CNMR. Then, the lipase enzyme is immobilized to the gold (Au: 99.9999%) electrode, where Thiophene is modified by electropolymerization, by cross-linking method. Optimum working conditions and performance factors of the prepared enzyme electrode are determined.

For coating, the electropolymerization method of the Thiophene Monomers on the surface is used with the Cyclic Voltammetry method in the CHI 660C Potentiostat device on the electrode surfaces. Potential value ranges of 0.3-1.5 volts are taken as a basis for this. During this process, the Scanrate value is 10 mv/s and 80 conversions are applied. Solution used to polymerize thiophene monomers; Prepared to contain 0.1 M distilled Thiophene, Pure Acetonitrile, Tetra Butyl Ammonium Hexa Fluoro Phosphate. The ready electrode was left to dry for 1 day. Pure Lipase Enzyme was dropped on the dried electrode surface and left to dry for 1 day. After applying Glutaraldehyde as a crosslinker to the dried electrode surface, it was dried for 1 day. Thus, the completed electrode was made ready as a Biosensor. The inventive biosensor used in the experiments was tested in comparison with Platinum, Silver/Silver Chloride electrodes.

A working solution was prepared to test the biosensor subject to the invention and the biosensors to be used in the comparison. This solution; It contains 0.5 grams of triglyceride and 0.4 grams of Triton X100, 10 ml of Ultrapure water. The solution was stirred at 60 °C for 1 hour to ensure homogenization of the mixture. The volume was completed to 50 ml by adding distilled water to the solution again. Again, Phosphate Buffer Saline (PBS) solution was prepared for the solution. This solution; It was made by weighing 8 gr Sodium Chloride, 200mg Potassium Chloride, 1.44 grams Disodium Hydrogen Phosphate and 245mg Potassium Dihydrogen Phosphate and completing it with distilled water to 1000 ml volume. Potassium Ferric Cyanide was added to this solution at a concentration of 5 millimolar in order to facilitate electron exchange. The solution was adjusted to pH=7.4 with Sodium Hydroxide (0.1 N) and Citric Acid (0.1 M).

Trials; The inventive biosensor and counter-electrodes were immersed in the PBS, in which the working solution was added at the concentrations determined for the study. After the CHI 660 device connections were made to the electrodes, experiments were carried out with the Cyclic Voltammetry method. As a result of this process, the calibration curve was drawn and controlled depending on the triglyceride concentration. Experiments have shown the operability of the method.

The test results showed that the linear measurement ranges of the prepared triglyceride lipase biosensor are sufficient for routine analysis, its reusability is high, its operational stability is high, and it is economical and useful.