Thin Layer Chromatography Of Amino Acid

Proteins are the basis of the protoplasm (fluid living content of the cell that contains the cytoplasm and cell nucleus) and are found in all living organisms. Proteins make up the bulk of animals body’s non-skeletal structure. As enzymes, they catalyze biochemical reactions; as antibodies, they prevent the effects of invading organisms; and as hormones, they control metabolic processes (C. Bissonnette, 2011). The Biuret test was used to detect the presence of peptide bonds within proteins, and they were found present in test tube #9 (control for peptide bonds).

Colorimetric assay is a process of determining a concentration of a solution based on absorbance of light. The purpose of this lab is to determine if the Bradford assay is an accurate way to determine an unknown concentration of two samples of protein. The Bradford assay is done by measuring wavelength of light passing through a cuvette filled with Bradford dye and concentrations of PBS and proteins. After the cuvettes are mixed they are placed into a spectrophotometer to measure wavelength. The wavelength given will be used to plot a standard curve based on concentration (x-axis) and wavelength (y-axis). The standard curve is then used to measure an educated guess on the concentrations of unknown protein concentrations. We hypothesized that if we use the Bradford assay and colorimetric spectrophotometry we can determine an accurate concentration of two unknown concentrations of proteins. The results of this lab failed to reject our hypothesis based on accurate measurements of protein concentrations. The standard curves are drawn with a linear increasing slope. The Bradford assay is an accurate way to demine the concentration of an unknown concentration.

2. Four unknown samples were included in the lab kit: flax seed meal, potato starch, egg whites, and dried milk. Using the results of the biochemical testing, determine which number corresponds to the correct unknown. (8 points)

The objective of this lab was to use qualitative and quantitative measurements and observations to properly identify, and then confirm the identity of an unknown compound. After identification, the compound would have to be synthesized, in our case, through an acid-base reaction, followed by confirming its identity with the original unknown compound.

Next, the biuret test for protein turned solutions in test tube 9 and 12 purple, the biuret test was positive, which proves that protein was present in the solutions, whereas test tubes 1,2,3,4,5,6,7,8,10, and 11 had negative results, this is because those solutions do not have peptide bonds and a peptide bond is necessary for the biuret test to be positive. An unknown was also tested in this experiment and we have concluded that the unknown #267 is a protein because it

The objective of this lab was to learn the principle of affinity chromatography by isolating a carbohydrate-binding protein from an extract of jack bean meal.

The role of FXR nuclear receptor in hepatic glutamine has been also investigated. It is believed that this receptor and its ligand are involved in the regulation of glutamine and glutamate metabolism [40, 41]. Some studies have still related the importance of hepatic metabolism of glutamine in hepatic encephalopathy [42] and cancer

It seems these days that the building blocks of proteins, affectionately known as "amino acids", are tiny little gold nuggets that bestow superhuman powers upon anyone lucky enough to stumble upon them in a sports gel, capsule, fizzy drink or cocktail. After all, these little guys are starting to get put by nutrition supplement manufacturers into just about everything, from your engineered pre-workout snack, to your during workout beverage, to your post-workout smoothie mix.

We first added the indicators in five samples containing common organic molecules to learn what a positive and a negative result looked like with each solution. When reacting with Lugol’s reagent, a substance containing starch will turn dark blue or black. If a sample is composed of glucose or other monosaccharides, it will turn an orange or red color when mixed with Benedict’s reagent. A sample containing protein turns purple or dark violet when combined with either Biuret’s reagent or ninhydrin. After identifying the colors indicating positive results, we tested each supplement with the four reagents. We took an equal amount of each sample in a test tube and combined it with a reagent to determine whether the identifying macromolecule was present in the sample. Once the solutions were properly mixed, we watched to see if there was a positive or negative reaction, indicating whether the organic molecule was present in the supplement. We took note of which brands tested positive and then analyzed the

Proteins are an essential part of all living organisms. Amino acids make up proteins which are made of one or more polypeptides. To identify an unknown amino acid or free amino acids, you would use thin layer chromatography and the Biuret test which also can help identify if a substances has peptide or not. The chromatography separates the difference of solubility to help identify the amino acids by showing the polarity. For this lab, I hypothesis that the substance that are being tested will not all have free amino acid. I believe that cysteine and casein will contain free amino acids.

the amino acid is in an acidic or basic salt form). Another point of interest

Amino acids are essential for the human body as they are referred to as the building blocks of proteins (Nutriology 2003). They are compounds that contain a carboxylic acid group (-COOH) and an amino group (-NH2) also joined to them are various side groups which are bonded to the alpha carbon (Thermo-fisher 2015).

B) Proteins Proteins are made up of chains of amino acids. Each protein folds into a characteristic three dimensional shape that is essential to its function. They perform a variety of functions in cells and are probably the most diverse macromolecule in living systems. The Biuret test detects the presence of proteins and short peptides (short chains of amino acids). Biuret reagent contains a strong solution of sodium hydroxide (NaOH) and a very small amount of dilute copper sulfate solution. The reagent changes color in the presence of a protein because the amino groups in the amino acids react with the copper ions producing a violet color. A negative result is blue.

I V B R S I T Applied PhytiCS Laboratory RTDC-TPS-48 1 Laurel M D 207236(199

UPLC-qTOF-MS/MS Waters was used to determine the amino acid sequence of the peptide moiety. The apparatus conditions were: ESI (electrospray ionization), positive mode; capillary temperature, 300oC; capillary voltage, 40 V and carrier gas flow 500 L/h. The lipopeptide and fractions were loaded to qTOF-MS directly without hydrolysis. Tandem-MS was done by collision-induced dissociation-mass spectrometry (CID-MS) using argon as collision gas and collision energy of 40 eV.