Cell Membrane Permeability in Beetroot
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Cell Membrane Permeability in BeetrootByNadir al-Kabra BIO 1045 HypothesisI hypothesize that as the temperature increase, there will be greater damage to the cell membrane causing increased pigmentation in the solution, and therefore an increasing absorbance rate reading on the spectrophotometer as the temperature goes up.Method and Materials Materialsfresh beetrootsharp bladecork borer11 test tubes and a boiling rack (with spare tubes)measuring cylinder (25ml)beakers (x1 500cm3 and x1 250cm3)timerspectophotometer cuvettesdeionized waterwater baths (45, 50, 55, 50, 65, & 70 Celcius)liquid nitrogen access20cm3 chloroform saturated solution20cm3 70% IBM solutionMethod(s)Effect of Temperature on Beet Membrane PermeabilityBeetroot cylinders were cut into 2x 1cm strips, then rinsed for 10 minutes under running water to remove pigmentation from damaged cells7 boiling tubes are prepared by placing 15 cm3 of distilled water in each tubeOne beetroot cylinder is transferred into a beaker in a 70 Celsius waterbath for one minute, and then transferred again into a tube of 15cm3 distilled water.Step 3 repeated at 45, 50, 55, 60, and 65 Celcius. Control tube is place in distilled water at 21 Celcius.Each cylinder is left for 45 minutes.After 45minutes, tubes are each agitated and the solution transferred into a cuvette.Each cuvette, in turn, is placed in the spectrophotometer set at 540nm.Effect of Organic Solvents on Beet Membrane PermeabilityTwo boiling tubes are prepared by placed 15cm3 of chloroform-saturated water in one, and 15cm3 of 70% alcohol IBM in the other.A prepared beetroot cylinder is placed in each, and these are left to sit for 45 minutes with occasional agitation.Spectrophotometer readings are taken as above, at 540nm, calibrated by the control solution, for each tube.Results are then recorded and runt through DataAnalysisEffect of freezing on membrane permeabilityBeetroot cylinders are frozen in liquid nitrogenThe beetroot cylinders are then placed in 15cm3 of distilled water at room temperature.The absorbance is then compared with the control solution.Results are then recorded and runt through DataAnalysisData Table 1: The effect of temperature on beetroot cell membranes.
TubeNo.Temperature at which beetroot cylinder was immersed for one minuteTime beetroot cylinder was placed in 15 cm3 of distilled waterTime beetroot cylinder was removed from 15 cm3 of distilled waterTime in minutes beetroot cylinder was in 15 cm3 of distilled waterAbsorbance value (at 540 nm) of 15 cm3 of distilled water after approx 45 mins immersion12111:0011:45450.03024511:0311:48450.04435011:0511:49440.13445511:0711:52450.19556011:1011:55450.11566511:1211:57450.23877011:1311:58450.500Table 2: The effect of organic solvents on beetroot cell membranes.TubeNoSolvent in which beetroot cylinder was immersedTime beet cylinder placed in boiling tubeTime beet cylinder retrieved from boiling tubeTime in minutes beetroot cylinder was immersed in solventAbsorbance value (at 540 nm)8Chloroform11:2312:08450.0509IBM (70% Alcohol)11:2012:05451.38510Table 3: The effect of rapid freezing (by liquid nitrogen) on beet cell membranesTubeNo.Treatment of beetroot cylinderTime beet cylinder placed in 15 cm3 of distilled waterTime beet cylinder removed from 15 cm3 of distilled waterTime (in minutes) beet cylinder was in 15 cm3 of distilled H2O after freezingAbsorbanceValue(at 540 nm)11Rapid freezing11:3912:24451.947GRAPH 1