CHEM 132.

2 – BIOCHEMISTRY (LABORATORY)
LABORATORY REPORT

Name _John Allan T. Pasana_______ Date Submitted: _01/26/21______

Lab Schedule: MTh at 10:00-12:00_____ Date Performed: _01/26/21______

Exercise No. 5
Amino Acids
I. OBJECTIVES:
 To identify the R group in amino acids.
 To determine the pH of various amino acids in water.
 To use chromatography to separate amino acids.
 To calculate Rf value for amino acids.
 Use Rf values to identify amino acids.

II. RESULTS AND DISCUSSION:

A. STRUCTURE OF AMINO ACIDS (Choose 1 amino acid)

Name:___Alanine_______________ Abbreviation: ___Ala, A_______________

Draw the structure of the dipeptide in your combined model

Note: I chose Alanine and Leucine for this one.
Name:__ l-alanyl-l-leucine_________________ Abbreviation: _____Ala-Leu____________

B. pH OF AMINO ACIDS

Color of Blue Litmus Color of Red Litmus Acidic, Neutral or

Amino acid Paper Paper Basic
Glycine Blue Red Neutral
Glutamic acid Red Red Acidic
Tyrosine Blue Red Neutral
Valine Blue Red Neutral
Aspartic acid Red Red Acidic
Lysine Blue Blue Basic
Alanine Blue Red Neutral

Discussion:
This experiment was conducted to determine the pH of these given amino acids. To know
whether they are basic, acidic, or netural, a litmus paper was utilized. Litmus paper is paper that
has been treated with a specific indicator - a mixture of 10 to 15 natural dyes obtained from
lichens that turns to either blue or red in specific conditions. After these amino acids were tested,
it was found out that 4 out of 7 of them were neutral by nature, and these are glycine, tyrosine,
valine, and alanine. They were neutral because the colors of the blue and red litmus papers
respectively remained as they are and did not change. On the other hand, 2 of these amino acids
were shown to be of acidic nature and these are glutamic and aspartic acids. They were acidic
since the color of the blue litmus paper turned into red while the red litmus paper had the same
color it possesses. Lysine, on the contrary, was the only one which is basic by nature. It was
categorized as basic since the blue litmus paper remained blue while the color of the red litmus
paper turned blue.

C. PAPER CHROMATOGRAPHY
Attached or sketch the results of the paper chromatogram.

Distance Distance
Solution/Mixtur
Amino acid Travelled by Travelled by Rf
e
Solute Solvent
A Leucine 2.7cm 7.5cm 0.36cm
B Valine 4.5cm 7.5cm 0.6cm
Leucine 2.9cm 7.5cm 0.39cm
Unknown Valine 4.6cm 7.5cm 0.613cm or

0.61cm

Discussion:

Paper chromatography is a technique for separating dissolved chemical substances by

taking advantage of their different rates of migration across sheets of paper. There were three
solvents utilized in this experiment which contain different amino acids. For solution A, the
amino acid leucine was present. For solution B, the amino acid valine was present. And for the
unknown solution, it is comprised by mixing small volumes of solutions A (which contains
leucine) and B (which contains valine). The solvent penetrates the paper by capillary
action and, in passing over the sample spot, carries along with it the various components of the
sample. The components move with the flowing solvent at velocities that are dependent on
their solubilities in the stationary and flowing solvents. Separation of the components is
brought about if there are differences in their relative solubilities in different solvents. Before
the flowing solvent reaches the farther edge of the paper, both solvents are evaporated, and
the location of the separated components is identified. The retention factor (R f) was also
gotten through dividing the distance travelled by the solute by the distance travelled by the
solvent. That is exactly why we got the results which are unveiled above.

D. THIN-LAYER CHROMATOGRAPHY
Sketch the results of the thin-layer chromatogram:

Discussion:
 Thin layer chromatography, or TLC, is a method for analyzing mixtures by separating the
compounds in the mixture. TLC can be used to help determine the number of components in a
mixture, the identity of compounds, and the purity of a compound. Thin layer chromatography
(TLC) is similar to paper chromatography but instead of paper, the stationary phase is a thin
layer of an inert substance (eg silica) supported on a flat, unreactive surface (eg a glass plate).
TLC tends to produce more useful chromatograms than paper chromatography, which show
greater separation of the components in the mixture - and are therefore easier to analyze. The
distance a sample travels can depend on the size or the polarity of the molecules involved.
Larger molecules take longer to move up the chromatography paper or TLC plate, whereas
smaller molecules are more mobile. Likewise, the polarity of the molecules can affect how far
the spots travel, depending on the type of solvent used. Polar molecules will be more strongly
attracted to polar solvents, and so would move further if a polar solvent was used as opposed
to a non-polar solvent. The distance that spots move can be compared to the overall distance
the solvent has moved and comparisons and measurements made. The behavior of an
individual compound in TLC is characterized by a quantity Known as Rƒ and is expressed as a
decimal fraction. The Rƒ is calculated by dividing the distance the compound traveled from the
original position by the distance the solvent travelled from the original position (the solvent
front). Overall, the advantages of this type of chromatography are the facts that it’s time-
saving, provides rigid support and so much more.

III. CONCLUSIONS:

Amino acids are a group of 20 organic compounds that share specific formation
traits. They are known as the ‘building blocks’ of proteins in both plants and animals.
Because they play such a foundational role, they are involved in many chemical
reactions throughout your body to help maintain your body’s normal functions. Each
amino acid has the same fundamental structure , which consists of a central carbon atom, also
known as the alpha (α) carbon, bonded to an amino group (NH 2), a carboxyl group (COOH),
and to a hydrogen atom. In the aqueous environment of the cell, the both the amino group and
the carboxyl group are ionized under physiological conditions, and so have the structures
-NH3+ and -COO–, respectively. Every amino acid also has another atom or group of atoms
bonded to the central atom known as the R group. This R group, or side chain, gives each
amino acid proteins specific characteristics, including size, polarity, and pH. Through the
different types of experiment performed with regards to this topic, I have become well-aware
on how the process of paper chromatography, thin-layer chromatography and the way to get
the retention factor (Rf) values of different solutions/mixtures. Amino acids is a necessity to
have in life which means taking them and including them in the diet is a must because their
deficiency results in decrease formation of protein that ultimately leads to a disease or health
conditions.