Spectrometry Analysis of Biological Membranes

Measurements of absorbance and color concentrations were taken to observe the affect of three stress factors on biological membranes. The detergents, pH, and concentration gradients in water were experimented on beet membranes. The results observed proved the validity that the factors, if not at the right conditions, cause cell death by disruption of the cellular membrane.
Introduction
The physical properties of membranes are important to chemical and biological studies. Membranes are formed from phospholipids, made up of water-fearing tails and water-loving heads, which are found throughout living cells. These phospholipids allow smaller neutral molecules like water to move through the membrane to the inside of the cell.1 Three stress factors that affect membranes are osmosis, detergents, and pH levels.2 Osmosis is termed the water mobility from high concentration to low concentration. There are three forms of water movement hypertonic, hypotonic, and isotonic. Hypertonic is when cell shrinkage results from the water inside the cell moving outside the cell where there is a lower water concentration. When the water concentration gradient is reversed, the end result is the swelling of a cell that can lead to bursting. Isotonic is when the flow of a water is at equilibrium.3 Detergents can be harmful or helpful when it comes to the solubility of lipids. For example, if a ketchup stain was on a shirt and the shirt was put into the washing machine, the detergent is used to lift that stain from the shirt using this chemical reaction. Because phospholipids have a phosphate in its hydrophilic head making it dissolvable in water, detergents cause lipid membranes to become soluble in water that results in the membranes failure to function.42 The pH crucially affects the dwelling place of all that is living. Level of pH can be measured using a scale where 7 is neutral, 0 is extremely acidic, and 14 extremely basic. These levels can also be color coded for red/yellow is acidic, purple is neutral, and green/blue is basic.5 The relationship between the concentration and absorbance of various factors were examined to determine the stress it had on a beet’s biological membrane.
Experimental Procedure
Throughout the experiment laboratory goggles and gloves were worn. 10 mL of tap water were added to six wells on a well plate while 10 mL of 15% salt solution were obtained. Six salt solutions were prepared using either graduated or beral pipets to add the water amounts to 4 or 5 wells with the concentration measured by percentage respectively, 0%, 3%, 6%, 9%, 12%, and 15%. Another pipet was used to add 15% salt solution to the 5 wells. While the solutions were being prepared, someone chopped a piece of beet into 6 squares each 0.5 cm. The beet needed to have all skin removed, smooth, same size, and remain moist. A little of water was used to wash the pieces of beet so pigment is washed off. A timer was set for 15 minutes once all the beets were added to the wells and stirred every 2-3 minutes with a stir rod. While waiting for timer, another partner setup and calibrated the Spectrometer using water in a cuvette as a blank. After the calibration process, the beets were removed and rinsed with water for the next preparation. The maximum wavelength collected was at 540 nm. All 0% solution was added to cuvette and data was collected and recorded (Table 1). The solution was then discarded, dried, and filled with the 3% solution by beral pipet. The data was collected and kept once the absorbance value leveled. These steps were repeated until all values were recorded. Once finished, the solutions were discarded in the waste.
Results
The data in Table 1 showed the fluctuation in absorbance as the salt