In order to isolate -carotene from spinach, we started out with a completely dehydrated solution of spinach extract. All of the water in the solution needed to be removed because the high boiling point of water would have made it difficult to evaporate in the RotoVap. An anhydrous sodium sulfate salt was added to the liquid spinach extract, which absobed all of the water. The solution was filtered with gravity filtration removing all of the salt. The remaining spinach solution was then RotoVaped to almost completely dry off the solvent, but a small amount of water, ethyl acetate, or both was left in the RBF so that the mostly-dry solution could be used in thin layer chromatography (TLC).
One of the best methods of separation of components
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Components of a solution with polarity similar to that of the solvent used in chromatography will move further across the adsorbant used, while components with different polarity will move less because they are not as dissolved by the developing liquid. With thin layer chromatography, a TLC plate is prepared coated with an adsorbant. Samples are dotted at the bottom of the plate on a line called the origin, and then are set in a consciously chosen devloping liquid in a jar so that the adsorbent picks up the liquid and moves up the plate. As the liquid moves and the plate develops, different components are dissolved or not dissolved in each sample dot, and the ones that are more dissolved move up the plate. The location where the solvent stops when the plate is removed from the developing jar is called the solvent front. Some of the components that move are not necessarily visible in normal light, so the adsorbent on the plates are is …show more content…
The column was wet with hexane as a solvent. The spinach extract was added to the top of the column so that its components could separate and run down it, and were developed by constantly adding hexane with a drop of ethyl acetate on top. The drop of ethyl acetate was added to increase the polairty of the solution and increase the speed at which the column developed. When all of the components started to separate down the column into bands, the eluents were intentionally colleced in separate flasks so that each compoment would be separated. -carotene was colleced seperately as it ran off as the first band. This isolated -carotene solution was then RotoVaped so that only a very small amount of hexane ethyl-acetate remained to keep the -carotene barely wet enough to use in thin layer
Iodine is used to distinguish starch from mono/di/polysaccharides. In the test only two of the six solutions showed any signs of starch being present. The potato juice turned a brownish color with a precipitate indicating a slight presence of starch. The stach solution turned a dark blue/black color indicating a very high presence of starch in the solution. The presence of starch in the potato juice indicates that it does not have as high an amount of carbohydrates as onion juice, but a higher concentration of starch.
To uncover organic compounds like carbohydrates, lipids, proteins and nucleic acid, by using tests like Benedict, Lugol, Biuret and Beta Carotene. Each test was used to determine the presents of different organic molecules in substances. The substances that were tested for in each unknown sample were sugars, starches, fats, and oils. Moreover, carbohydrates are divided into two categories, simple and complex sugars. Additionally, for nonreducing sugars, according to Stanley R. Benedict, the bond is broken only by high heat to make make the molecules have a free aldehydes (Benedict). As for Lipids, there are two categories saturated and unsaturated fats. One of the difference is that saturated fats are mostly solids and have no double bond (Campbell Biology 73). The Beta Carotene test works by dissolving in a lipid, thus giving it color to make it visible. Moreover, proteins are made out of amino acids that are linked by a polypeptide bond (Campbell Biology 75). The purpose of this experiment was to determine whether an unknown class sample or food sample had any carbohydrates, lipids, or proteins in it. The expected result of the lab was that some substances would be present while other would be absent.
The objective of this experiment was to perform extraction. This is a separation and purification technique, based on different solubility of compounds in immiscible solvent mixtures. Extraction is conducted by shaking the solution with the solvent, until two layers are formed. One layer can then be separated from the other. If the separation does not happen in one try, multiple attempts may be needed.
The procedure for this laboratory assignment is on the following pages. 3) Separation of Carboxylic Acid, Phenol and Neutral Substance. The purpose of this acid/base extraction is to separate a mixture of equal parts of benzoic acid(strong acid) and 2-naphthanol(weak base) and 1,4- dimethoxybenzene(neutral) by extracting from tert-butylmethyl ether(very volatile).The goal of this experiment was to identify the three components in the mixture and to determine the percent recovery of each from the mixture. 4) Separation of a Neutral and Basic Substance. A mixture of equal parts of a neutral substance containing either naphthalene or benzoin and a basic substance containing either 4-chloroaniline or ethyl 4-aminobenzoate were to be separated by extraction from an ether solution. Once the separation took place, and crystallization was carried out, it became possible to determine what components were in the unknown mixture, by means of a melting point determination.
This experiment requires four tubes with an enzyme solution, chelating agent and deionized water. Also a fifth tube that is the calibration tube for the spectrophotometer, which only has 5ml of dH2O. The calibration tube is used to level out the spectrophotometer to zero before each trial. The spectrophotometer was set at 540 nm, “since green is not a color seen with the conversion of catechol to benzoquinone.” The enzyme solution was made by using potato that was peeled so that the golden color of the skin wouldn’t react or interfere with the red color needed in the spectrophotometer. After it was peeled, it was cut into chunks to minimize excess heat created while it was blended. It was put in a chilled blender and 500ml of deionized water was added. Chilled, deionized water was used because it created a hypotonic environment that caused the cells from the potato to burst and release the catecholase. It was chilled
The experiment was conducted using carbon dioxide to see how it affected the rate of photosynthesis in spinach leaves. Carbon dioxide should increase the rate of photosynthesis because there will be more carbon dioxide, a reactant in the photosynthesis formula.
Experiment #3: The purpose of this experiment to test the chromatography of plant pigments the alcohol test strip test will be used.
To allow the solution to go into the leaf discs completely, add one drop of liquid soap in each solution. Use the photometer to measure the light intensity near the beakers. If the light intensities are not similar, relocate the beaker where the light intensity is similar. Cut at least 10 spinach leaf disks for each beaker.
Once the mixture had been completely dissolved, the solution was transferred to a separatory funnel. The solution was then extracted twice using 5.0 mL of 1 M
Saturated sodium chloride solution, also known as brine solution, is used to wash the distillate mixture. The distillate mixture is the phosphoric acid the co-distilled with the product. The brine solution also removes most of the water from the 4-methylcyclohexane layer. When six drops of 4-methylcyclohexene were added with two
In our Biology Lab we did a laboratory experiment on fermentation, alcohol fermentation to be exact. Alcohol fermentation is a type of fermentation that produces the alcohol ethanol and CO2. In the experiment we estimated the rate of alcohol fermentation by measuring the rate of CO2 production. Both glycolysis and fermentation consist of a series of chemical reactions, each of which is catalyzed by a specific enzyme. Two of the tables substituted some of the solution glucose for two different types of solutions. They are as followed, Table #5 substituted glucose for sucrose and Table #6 substituted the glucose for pH4. The equation for alcohol fermentation consists of 6 Carbons 12 Hydrogens 6 Oxygen to produce 2 pyruvates plus 2 ATP then finally the final reaction will be 2 CO2 plus Ethanol. In the class our controlled numbers were at Table #1; their table had 15 mL Glucose, 10 mL RO water, and 10 mL of yeast which then they placed in an incubator at 37 degrees Celsius. We each then measured our own table’s fermentation flasks every 15 mins for an hour to compare to Table #1’s controlled numbers. At
HPLC technology works on the principle of conventional chromatography where in there is a stationary phase and a mobile phase. The sample containing the mixture of components is introduced in a column packed
· The beetroot piece is then placed into a tube of 5 cm of distilled
Researchers then hypothesized that the results would indicate the greatest amount of potato enzyme activity level will take place at room temperature. In this experiment, researchers used potato extract and different temperature levels to test the hypothesis. Moreover, researchers wanted to test the color intensity scale and how specific catechol oxidase is for catechol. In this experiment, researchers used dH2O, catechol solution, hydroquinone, and potato extract. Lastly, researchers tested the substrate concentration and how it has an effect on enzyme activity.
Three different sweet potato and carrot samples were finely diced up separately, using a scalpel. Upon cutting them into smaller pieces, they were measured into 50 gram quantities using a scale. Then, 50 g of sweet potato was added to the blender along with 50mL of distilled water and a small pinch of ice. Once blended until it looked smooth, we prepared the container it would go in. There was a funnel extending into a graduated cylinder with cheesecloth over the top opening of the funnel. The sweet potato mixture was then poured over the cheesecloth so that chunks of potato would be separated from the liquid. Liquid was poured into the graduated cylinder until a volume of 50mL was reached. For a few samples, squeezing of the cheesecloth was necessary