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Chromatography of amino acids lab report

Owusu Asante United Care Roland Clinic, Gambia Posters & Accepted Abstracts: J Chromatogr Sep Tech Abstract : The technique assists in the analysis, detection, purification and quantitative analysis of unknown separating mixtures. The mobile phase is either liquid or gas that moves the solvent through a stationary phase during the process. A
stationary phase is a liquid or solid component that is fixed in place for the procedure. Paper chromatography in considerable work works on capillary attractions. Capillary gravity, which depends on glue and cohesive forces, allows the mobile phase to move up through the stationary phase due to the created interaction of surface tension from forces. The
main types are paper chromatography, thin layer, gas chromatography, column chromatography, high-emission liquid chromatography, paper chromatography and thin layer chromatography. There are several applications of paper chromatography and other basic types of chromatography methods. This technique is used in the pharmaceutical industry,
hospitals, forensic science, environmental science and manufacturing plants. This report describes an experiment conducted using paper chromatography to detect an unknown mixture. This will be done by comparing four known amino acids with two unknown mixtures to identify unknown mixtures. The experiment will also help to master the technique and
analyze the movements made by both unknown mixtures and known amino acids. It is necessary to determine the materials of gloves, glasses, laboratory coat, filter paper, toothpicks, nihydrin solution, mixtures. Laboratory procedures entail various steps that ultimately lead to the detection of unknown mixtures. This procedure is largely divided into
stationary phase training, preparation of mobile phases and the development of a chromatograph. For a stationary phase of preparation on paper, the necessary markings are made to identify and create a baseline. Baseline marks are 1.7 cm from the shorter left edge and 1.0 cm from the lower part of the longer edge. Known amino acid mark symbols on
paper. The spotting of the known four amino acids and two unknown mixtures is then carried out with the help of separate toothpicks, which will help prevent contamination. Preparation of the mobile phase was made by pouring 10 ml of soluble mixture into a glass of Berzelius 400 ml, until the development of chromatography was done after the filter paper
has already been dried. Biography : Email: [email protected] Share this: Facebook Twitter Reddit LinkedIn Chromatography WhatsApp can be described as a wide class of biochemical methods in which a mixture of substances can be separated by different properties that include charge, size, hydrophobicity, nonvalent bonding affinity or some other
properties, allowing the mixture to divide between phase and stationary phase. (Alberts et al., 2008). Get Help Your essay If you need help writing essays, our professional essay writing service is here to help! To learn more paper chromatography is a technique that includes two phases; phase and mobile phase. As the name suggests, the stationary phase
does not move. In this experiment, the stationary phase was filter paper, and the mobile phase was solvent. In ascending chromatography, the solvent moves upwards, and in downward chromatography the solvent moves down. This experiment uses ascending chromatography. The solvent (mobile phase) rises up with capillary paper. Other components in
the mixture rise up with paper at a speed proportional to the partition in the solvent. The process when the mobile phase moves through a stationary phase is called development. Since different components travel at different rates, the end result of the chromatograph will be dyed by components at different distances. Using this information, it was possible to
calculate the Rf values of components. Formula used: Since neither of the two components has the same RF values, components can be identified. (Clark, 2007). Chromatography of size exclusion is a chromatographic technique in which molecules are separated according to their size, or rather their hydrodynamic volume. In this experiment, sugar was a
stationary phase, while oil ether with pigments was a mobile phase. When the mobile phase moves down, components of different sizes begin to separate as they move at different speeds. The advantage of chromatography of size exclusion is that the components are separated, retaining their biological properties. (McNight & Wilkinson, 2006).
Experiment 1A: Identification of unknown amino acid by paper chromatography method Method of apparatus Chromatography Paper paper tape materials Ninhydrin Butanol / Glacial acetate / water (12/3/5) solvent Various amino acids Procedure Pencil line was first drawn through a piece of filter paper. This line was about 2 cm from the edge. Toothpick was
used to detect 5 drops of amino acid solution. Then these drops were dried and then the next drop with another amino acid was applied. The above procedure was repeated for each amino acid solution, as well as for the unknown. Then the chromatographic tank was filled with a mixture of butanol solvent / vineic acid glacier / water (12/3/5) to a depth of
about 0.5 cm. Spotted paper has been placed in a stained tank at the base. The solvent was allowed to rise upwards with paper within 2cm of the top of the paper. The paper was removed, and the front of the solvent was marked with a pencil. The paper was allowed to dry. Then it was sprayed with a solution of ningydrin. The paper was allowed to dry again.
The distance was measured from the point of application anterior solvent and to the center of any spots detected by a solution of ningydrin. These values were recorded, and the overall color of the spots observed was also observed. According to the obtained results, the values of the Russian Federation of amino acid standards and unknown ones were
calculated. Consequently, it was possible to identify the composition of the unknown. Precautions The line on the filter paper has been drawn with a pencil. This was done because if ink was used, it would be dissolved in solvent. The spots did not allow to touch the solvent in the chromatography of the tank. The filter paper was processed from the edges to
prevent staining of the results. Filter paper was stored vertically in the chromatography of the tank. Ninhydrin was sprayed in the hood because it is a known carcinogen. The container containing the solvent was hermetically sealed. Errors Solvent in front does not move up in a perfectly straight line. Solvent leaks from the container. Results Of Equation used
to calculate RF values: Amino acid distance displaced (cm) Rf Phenylalanine 5.3 0.589 Tyrosine 3.8 0.422 Cauldrin 4.8 0.533 Threonine 2.6 0.289 Tryptofan 4.3 0.478 Unknown A 5.5 0.611 Unknown B 2.4 0.267 Total distance, Overcame solvent = 9.00cm Unknown A of RF 0.611 is Phenylalanine. Unknown B with rf 0.267 is Threonine. Debatable amino
acids are compounds that are monomers of proteins. There are about 20 different types of amino acids. Each amino acid is built around the same nucleus structure through which it is easy to make a connection in a standard way with other amino acids. The main structure of the amino acid can be seen below: Figure 1. The main structure of the amino acid
All amino acids have a carboxyl group and an amino acid, which are both associated with a carbon atom called α carbon. Each protein or polypeptide is essentially a chain of amino acids strung together. However, the protein then consists of a three-dimensional structure that is unique to each type of protein. The covalent bond between the two adjacent
amino acids in the protein chain forms amide. (Alberts et al., 2008). In order to discuss the amino acid that will be used in reaction with Ninghydrin is alanine. Alanine is a simple amino acid with the formula C3H7NO2. Figure 2:10 Alanine Figure 3. Ninhydrin Ninghydrin was the dye used in this experiment. Since amino acids are not colored dye was necessary
so that you could see the amino acids and calculate the value of the Russian Federation. Figure 4:10 The mechanism of the reaction of ningydriin with amino acid The value of the RF chemical or component is very unproductive and, as can be observed from the results, although the same chemical was used on the same sheet, the results were not quite the
same. In this case, the most similar meaning of the Russian Federation was taken as unknown. From the results obtained, we can conclude that Unknown A was the most Phenylalanine while Unknown B was Threonine. Experiment 1B: Size Exclusion Chromatography Method 5.1 The device dividing the funnel glass column plunger glass sparks glass
Spatula Scalpel 5.2 Materials Methanol / Oil ether (2/1) Leaves Fine powdered sugar 5.3 Procedure Green leaves were originally cut into very small pieces with a sharp knife. Up to 5 grams of material was added 150 ml of solvent mixture, consisting of two parts of absolute methanol and one part of oil ether. This was left for 5-15 minutes to ensure that all or
most of the pigments were removed. Then the extract was filtered through a cloth in one liter separating the funnel. Then 500-600 ml of filtering 10% sodium chloride solution was added. All pigments were transferred to the top layer, an oil ether that separated from methanol. The contents were mixed, gently rotating the funnel. Then a dark green layer of oil
ether and a chromatograph were drawn on a column of adsorbent powder on adsorbent paper. A small cotton bug was put in the glass tube and pressed flat with a plunger. About 2-3cm of powdered sugar have been placed in a tube and shaking by pressing on the side of the tube while using the plunger to firmly pack down the tube. The tube was filled with
sugar until the column was 2 cm on top. A glass rod was inserted into the tube until the end of the rod touched sugar. A small sample of pigment was poured out of a glass rod. This was done to the top 0.5 cm to 1.5 cm of dark green sugar column. Most of the solvent was allowed to move into sugar. Added a small amount of 0.5 percent solution of n-propilot
alcohol in oil elyse. The developing solution caused the pigments to separate from each other. The speed of movement of the solvent through the sugar column depended on pigments, solvent, tube packing and suction power. Different pigments were adsorbed to sugar particles with different tightness and this was carried around the column at different
speeds. 5.4 Precautions The sugar column has been compressed into the column as much as possible. Sugar refrained from drying, constantly adding a solution containing pigments. 5.5 Errors There were some holes in the package due to the presence of a wide glass column. Layers did not move evenly, causing bands to be wrapped. This could be due to
the packaging not being close enough. The Results of Discussion Photosynthesis is the process by which carbon dioxide is converted into organic compounds for plants using energy from sunlight. Photosynthetic pigments are needed to absorb this energy from sunlight. Since the pigment can absorb only a limited range of organisms, the photosynthesis of
which usually has a number of different compounds, so that they make the best use of all the random sunlight. The three main Pigments found in photosynthetic plants are carotenoids, chlorophylls and phycobilins, the latter found in some unicellular photosynthetic organisms. Below is a diagram showing the different absorption rate of these different
pigments: Figure 5. Absorption of frequencies of various photosynthetic pigments As can be observed from the graph, the main pigments found in plants do not absorb most of the green-yellow frequencies, and this is reflected, therefore providing a typical green color of plants. From the results it can be seen that carotenoids are the smallest, as they moved
the fastest down the glass column. Chlorophyll b is the largest, and chlorophyll A is of medium size. Conclusion Experiments were successfully carried out. Both chromatography methods used are important because they are used in different applications. Experiment A: It was found that the unknown compounds were Threonine and Phenylalanine.
Experiment B: Three groups were released in a glass column, and carotenoids were found in the lower lane, and Chlorophyll B was found in the higher group. The middle layer was recognized as Chlorophyll A. Share this: Facebook Twitter Reddit LinkedIn WhatsApp

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