Guinea Pig Ileum Experiment

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AN EXPERIMENT TO DETERMINE THE PREVALENT CONCENTRATIONS OF ACETYLCHOLINE, HISTAMINE AND 5-HT IN URTICA INCISA STRAIN OF NETTLE PLANT SPECIES AND THE EXTENT TO WHICH THEY SHOW BIOLOGICAL ACITIVITY IN A GUINEA PIG ILEUM ABSTRACT 1. INTRODUCTION The nettle plan is an annually growing shrub which has been used for hundreds of years for medicinal and nutritional purpose by humans. Recently, published experiments have identified three active compounds present in this species of plant, chiefly: Histamine, Serotonin (5-HT) and Acetylcholine, all of which are active neurotransmitters in the mammalian central nervous system. Generally, Histamine is important for inflammatory mediation as part of the human immune response; 5-HT is crucial for appetite, sleep and mood regulation; and Acetylcholine is the only neurotransmitter present in the somatic nervous system, which is key for human motor function. The aim of this experiment is determine the presence of these neurotransmitters in Urtica incisa nettle plant extract, and the concentrations which they prevalent. Then their biological activity will be measured based on their contractility effects of a guinea pig ileum sample. 2. MATERIALS AND METHODS 2.1 NETTLE EXTRACT The Urtica incisa nettle sample was finely cut and ground in 15 ml of chilled water. This is a noted change from the initial recommended 25 ml of chilled water, because it would give a more concentrated sample. The homogenate was then placed in the centrifuge at 3000 rpm for 5 minutes. Following this, the Silica plates were marked with 3 parallel lines; for the Ach, Histamine and 5-HT samples. Similarly, 1 mg/ml of Ach, 0.1mg/ml Histamine and 0.1mg/ml of 5-HT were blotted evenly for comparison. The silica plate was then placed in the solvent for 1 hour to allow chromatography separation. Solvent line was then marked and Rf values calculated

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