CHAPTER I

INTRODUCTION
A. Background
Proteins are composed of peptides to form a polymer called polypeptides.
Each monomer is composed of amino acids. The role of proteins such as
catalysts, support, backup, immune system, etc.
Almost all amino acids except glycine have chiral carbon atoms. Chiral
amino acids have two forms isomeri. Have similar physical and chemical
properties, except the ability to distinguish the polarization direction of the play
field.
Proteins are composed of amino acid chains will have a wide variety of
typical structure of each protein. The protein structure includes primary
structure, secondary structure, tertiary structure and quaternary structure. The
primary structure is a structure composed of amino acid sequence does not
occur linearly and chain branching. Secondary structure is a combination of the
primary structure of the linear and has segments in polypeptide entwined. The
tertiary structure of a protein is an overlapping layer above the secondary
structure pattern consisting of irregular distortion of the bond between the side
chain (R group) of various amino acids are quaternary structure of proteins to
form complex molecules, some protein chains joined together to form a ball
(Carey , 2006).
B. Formulation of the problem
How the peptide bonds that form proteins?
C. Purpose
Students can provd there are amino acid

CHAPTER II
LITERATURE

Amino acids derived from the hydrolysis of amino acid protein is also
called acid or -aminokarboksilat. Amino acid that occurs naturally as the building
blocks of protein has an amino group (NH2) and the carboxylic group (COOH) are
bound to the same atom, namely the carbon atom alpha.
Amino acid is a carboxylic acid having an amino group. Amino acids
contained as protein components have -NH2 the carbon atom of position -COOH
group.
The general formula for amino acids is: R - CH - COOH
NH2
Of the general formula can be seen that the carbon atom is asymmetric
carbon atoms, except when R is atomic H. The difference between an amino acid
with another amino acid that is caused by differences in the R group called side
chains. There are 20 amino acids that act as the builder of a protein molecule,
namely glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine,
threonine, cysteine, methionine, proline, phenylalanine, tyrosine, tryptophan,
aspartic acid, glutamic acid, asparagine, glutamine, lysine, arginine and histidine.
In general, the amino acid is soluble in water and insoluble in non-polar
organic solvents such as ether, acetone and chloroform. Amino acids have a higher
melting point than the carboxylic acid or amine. Both of these physical properties
indicate that the amino acids tend to have a structure that is charged and has a high
polarity and not just a compound having -COOH and -NH2 groups. It also appears
on the nature of the amino acid as the electrolyte.
Amino acid containing an amino group which is alkaline and acidic karoksil
group in the same molecule. Amino acids undergo an acid-base reaction that
generates an internal dipolar ions, which are also called zwitterionic or amphoteric
ion.
If the water-soluble amino acids, carboxylic group will release H + ions,
while the amine group will accept H + ions, as written below:
-COOH -COO- + H +
-NH2 + H + -NH3 +

At the amino acid test there is general test and the test is specifically based
on the type of amino acid. Ninhydrin test of a general nature which reacted
positively to produce a violet color of all amino acids with primary amino groups.
And xantroproteat test positive reaction for amino acids containing a benzene
nucleus.
If ninhydrin heated with amino acids, it will form a color complex. For one
amino acid can be determined quantitatively by observing the intensity of the color
formed is proportional to the concentration of the amino acids. In this case NH3
and CO2 removed so that the possibility can be measured quantitatively. reaction:
RCH (NH2) COOH RCHO + NH3 + CO2 (purple)
Such as alanine, valine, leucine, isoleucine, phenylalanine and methionine
produce different colored complex with other amino acids. Color complex formed
contains 2 ninhydrin molecules that react with ammonia after amino acids are
oxidized.
Ninhydrin compound is a hydrate of triketon cyclic, and when it reacts with
amino acids, produces a purple color. Only the nitrogen atom of purple dye derived
from amino acids, amino acids and the rest is converted into aldehyde carbon
dioxide. Thus, the same purple dye produced from all amino acids with primary
amino groups and the intensity of the color is directly proportional to the
concentration of the amino acid. Only proline, which has a secondary amino
group, react differently and resulted in a yellow dye, but it also can be used for
analysis.

CHAPTER III
RESEARCH METHODS

A. Tools and Materials

Tools :

Reaction tube
pipette
test tube rack
clamp test tube
measuring cup
Bunsen

Materials

:
Protein solution
Solution of ninhidrin 0,1 %

B. Procedures
1.

Add 5 drops of 0.1% ninhydrin solution in 1 ml of protein

solution in a test tube

2.

Heat until boiling, then let it cool and observe the color
change.

CHAPTER IV
RESULT AND DISCUSSION

A. Results

No
1.

2.

3.

4.

5.

6.

Activity
Arginin + ninhidrin
solution,heated

Result of Observed
Before
After
Arginin :
Arginin + ninhidrin
colorless
solution = colorless
Ninhidrin :
blue

Arginin + ninhidrin
solution,heated = soft
purple (+)

Albumin :
colorless

Albumin + ninhidrin
solution = colorless

Ninhidrin :
blue

Albumin + ninhidrin
solution,heated =purple
(++)

Tofu extract :
white

Tofu extract + ninhidrin

solution = colorless

Ninhidrin :
Blue

Tofu extract + ninhidrin

solution,heated = white

mung bean extract +

5 drops of 0.01%
ninhidrin
solution,Then boiled
it

Mungbean extract
color:
White

Mungbean extract +
ninhidrin :
white

Ninhidrin:
light turqoise

Chicken heart
extract +
ninhidrine,heated

Chicken heart
extract color:
maroon

after heated :
dark purple with
brownie color.
Chicken heart extract +
ninhidrine + heated =
purple

Albumin + ninhidrin
solution,heated

Tofu extract +
ninhidrin
solution,heated

peanut extract +
ninhidrine,heated

Ninhidrine color:
ligth turqoise
Color extract =
white muddy
solution
Ninhidrin =
colorless

7.

Extract tempe+
ninhidrin solution
,heated

extract tempe
color:
white muddy
Ninhidrin solution
color:

Extract tempe+
ninhidrin solution +
heated:
purple muddy
Many vapor (+++)
Extract tempe+
ninhidrin solution +
heated:
grey purple

8.

9.

Fish meat extract +

ninhidrin solution
,heated

chicken meat extract

+ ninhidrin
solution ,heated

clear
Fish meat extract
color: cream
muddy
Ninhidrine color:
colorless
Chicken heart
extract color:
muddy,cream
pinkish
Ninhidrine color:
colorless

B.

Fish meat extract +

ninhidrin solution +
heated:
purplish blue
chicken meat extract +
ninhidrin solution
,heated:
blue purplelish
there is precipitation (+
+)
water vapor (+)

Analysis
Based on the above observations obtained results that are arginine
indicator changes color to purple after undergoing a process of combustion or
dipanaskan.kemudian for albumin indicator is also changing to purple color but
the color purple in albumin solution is darker than the laruran arginine. While in
ekstract ekstract we tested using ninhydrin test produces visible changes, such as a
change in color and there is water vapor.
In ekstract maroon colored chicken livers, if added to a solution of
ninhydrin after it is heated will produce a brown color which has not shown the
presence of amino acids. In peanut extract white when added to a solution of
ninhydrin after it is heated will produce muddy purple color and many vapor
which indicates amino acids on these materials. In the fish meat extract white
colored muddy when added to a solution of ninhydrin after it is heated will
produce purplish blue color which indicates the presence of amino acids.
In tempeh extract cream colored when added to a solution of ninhydrin
after it is heated will produce a purple color which also showed the presence of
amino acids in the soybean extract and the extract of chicken meat soft pink color
produces a color change to purple after adding 0.1% ninhydrin solution and
heated, the color change indicates the presence of amino acids. In extracts of
white tofu did not produce a change in color to white after adding 0.1% ninhydrin
solution and heated, the color change has not shown the presence of amino acids

in the tofu extracts. And on the white colored mungbean extract produces a color
change into dark purple after adding 0.1% ninhydrin solution and heated, the color
perubabahan indicate amino acids.
C. Discussion of the result
Can we know that ekstract that changes color to purple to prove that the
protein solution containing amino acids. The darker the purple color of the peptide
bond in the solution more and kuat.akan but if the color is not dark purple then
show the peptide bond in the solution is not much and not as strong as the solution
changed color to a darker purple.
However, in our experiments that use ekstract know to get the color
changes to white after adding 5 drops of 0.1% ninhydrin solution. And on chicken
liver extract obtain Warne change into brown. The possibility that happened was
during the making of the state ekstract know know at the time is not good or is
already happening fermentasi.sehingga process causes the loss of peptide bond
know that amino acids in the know is missing too. And what happened to the
chicken liver extract, at the time of making the chicken liver extract, not stirred
first but took on the bottom / top of the chicken liver extracts, which could result
in the loss of chicken liver extract amino acids.

D. Question
1. Why ninhydrin reagent can be used to determine the presence of amino
acids? Explain!
Because ninhydrin an oxidizing cause oxidative decarboxylation
and amino acid by removing the CO2, NH3, and aldehydes, so that the
reduced ninhydrin reacts with NH3-free form of Compounds blue
complex. And ninhydrin a very strong oxidizing agent that can cause

oxidative decarboxylation of -amino acids. Ninhydrin reduced, then

reacts with amino loose form complexes purple blue. The intensity of the
resulting blue color purple in a raw state is the basis for quantitative ter
very useful for amino acids and amines are not -amino acids.

CHAPTER V
CLOSING
A. Conclusion
Based on the observation that has been done got the result that most of the
extracts proved the presence of an amino acid, the extract of chicken meat, fish,
tempeh, beans, and mung bean, because at the moment with the addition of 0.1%
ninhydrin solution and heated produces purple because Ninhydrin compound is a

hydrate of triketon cyclic, and when it reacts with amino acids, produces a purple
color. Tus, the same purple dye produced from all amino acids with primary
amino groups and the intensity of the color is proportional to the concentration
Directly of the amino acid. But the tofu and chicken liver extract did not produce
a purple color which indicates the absence of amino asm, due to errors and lack of
prudence in the lab.
B. Suggestion
Next to the lab work was recommended to one who performs lab work to be
more careful in conducting a particular experiment, either in pouring reagent
solution, and in terms of reacting solutions of further maintaining the
cleanliness examiners.And of lab work tools so as not Contaminated by
another chemical solution. And As for suggestions to improve the quality of
research:
a. It should be more careful in mixing the ingredients so that the results
agree with the theory that has been taught.
b. Tools and materials need to be added Because The test process must
use a considerable reaction tube, so do not spend a lot of time.
c. Maintain hygiene before and after use laboratory.
d. The existence of material preparation before entering the study

Bibliography
Aksara Winarno, F. G. 2004. Kimia Pangan dan Gizi. Gramedia : Jakarta
Brown, Wiliam H. 1994. Biokimia jilid II. EGC : Jakarta
Dawn,B. 2000. Biokimia Kedokteran Dasar. EGC : Jakarta
Hawab, H. M. 2003. Pengantar Biokimia. Bayumedia. Publishing : Malang
Ramsden.1994. Biokimia dalam kehidupan. Gramedia : Jakarta

Riawan, S. 1990. Kimia Organik. Binarupa: Jakarta

Tim Biokimia Jurusan Biologi FMIPA UNESA. 2014. Petunjuk Praktikum
Biokimia. Surabaya. UNIPRESS.