Analysis of Amino Acids by Paper Chromatography
Introduction-
Proteins may be thought of natural polymers of amino acids, as the
composition of proteins is of amino acids.
The technique known as paper chromatography is used to separate amino
acids for analysis. In this technique small spots of amino acids are
introduced to a piece of porous filter paper. The bottom of the paper
is then placed in a small bath of an appropriate solvent. The solvent
is allowed to rise up the paper. The various amino acids are attracted
to the paper to different degrees due to their differences in
polarity.
Various amino acids travel at different rates.
The location of the amino acids can be determined by spraying the
paper with a 2% solution of Ninhydrin in ethanol. The Ninhydrin forms
a purple complex with an amino acid which is readily identified.
Materials-
Solutions of amino acids used are -
Amino acid mixture 'X'
Aspartic Acid
Leucine
Proline
Asparagine
Phenylalanine
Solvent (ammonia and propan-2-ol in the ratio 2:1)
2 % solution of Ninhydrin in ethanol
Method-
Setting up the apparatus
1. This section of the experiment is done in a fume cupboard as the
solvent is dangerous. Safety glasses should also be worn. The
solvent is poured into a chromatography jar to a depth that is
enough for the chromatography paper end to be dipped into. A lid
is placed over the jar and left in the fume cupboard. This gives
time for the air in the jar to be saturated with the solvent of
the vapour.
2. Before handling the chromatography paper, hands are washed. The
chromatography paper mustn't be touched all over the place
constantly because amino acids that is smeared on the hand may get
Catecholase is an enzyme formed by catechol and oxygen used to interlock oxygen at relative settings, and it is present in plants and crustaceans (Sanyal et. al, 2014). For example, in most fruits and vegetables, the bruised or exposed area of the pant becomes brown due to the reaction of catechol becoming oxidized and oxygen becoming reduced by gaining hydrogen to form water, which then creates a chain that is is the structural backbone of dark melanoid pigments (Helms et al., 1998). However, not all fruits and plants darken at the same rate. This leads to question the enzymatic strength of catecholase and how nearby surroundings affect its activity. The catecholase enzyme has an optimal temperature of approximately 40°C (Helms et al., 1998). Anything above that level would denature the tertiary or primary structure of the protein and cause it to be inoperable. At low temperatures, enzymes have a slower catalyzing rate. Enzymes also function under optimal pH level or else they will also denature, so an average quantity of ions, not too high or low, present within a solution could determine the efficiency of an enzyme (Helms et al., 1998). Also, if more enzymes were added to the concentration, the solution would have a more active sites available for substrates and allow the reaction rate to increase if excess substrate is present (Helms et al., 1998). However, if more
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J.C. Biro, B. Benyó, C. Sansom, Á. Szlávecz, G. Fördös, T. Micsik, and Z. Benyó; A common periodic table of codons and amino acids. Biochemical and Biophysical Research Communications 306 (2003) 408–415.
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Defying Robinson Appleby 's agreement to never teach a Negro how to read, Mamed, the overseer with a raised cane, proposes Aminata an offer which she cannot decline. Being the overseer in Appleby 's plantation, Mamed 's duty is to ensure that the plantation is properly functioning while Master Appleby is absent. In fear of being disciplined for divulging the prohibited prayer, Aminata recognizes Mamed 's humanity as he murmurs, “Allahu Akbar”. With Mamed 's offer to Aminata to tutor her how to read and write, perhaps if “Georgia was teaching [Aminata] how to survive in the land of the buckra, but maybe Mamed could teach [her] how to get out” (Hill, 216). Mamed 's teachings start with the pronunciation of Aminata 's name to eventually forming a sentence with basic, common words to the rules and procedures established in Appleby 's hacienda. As Aminata 's fear of Mamed eventually subsides, Aminata apprehends the idea of Mamed being a different man, “like a man who was willing to teach” (Hill, 218).