CHAPTER 3
Methods and Procedures This chapter presents the methodology and procedures used to determine the renal protective property of crude extract in stingless bee propolis.
Research Methodology Experimental methods and techniques will be used for the determination of the renal protective property. Systematic procedures will be observed in the determination of the renal protective property of crude extract in stingless bee propolis. Chemical tests will be used to perform to determine presence of phenolic compounds that are 50% present in the propolis extract. Instrumental analysis was done to identify the components of the crude extract. Biological and histopathological testing will be conducted to determine its renal protective property in creatinine and BUN by Gentamicin induced in rats. The preparation, extraction, physical, chemical tests and biological test of the crude extract from propolis will be
Blood samples will be collected via Axillary bleed, allowed to clot. The serum will be separated by centrifuge at 2500 rpm for 15 minutes and will be analyzed for various chemical parameters. After the extraction of blood the rats will be euthanized by CO2 Inhalation. The kidneys will be isolated to view it under the microscope for examination. 4.5 Histopathological Examination One representative per group will be used in histopathological examination. The kidneys will be isolated, weighed and sectioned longitudinally and kept in 10% neutral formalin solution. The kidneys obtained will be embedded in paraffin wax and will be stained with hematoxylin and eosin. The kidneys will be observed under a microscope and photographs will be taken, The histopathological study and blood extraction will be performed by the same IACUC representative and veterinary pathologist who will be assisting the researchers throughout the biological study.
5. Statistical