LAB WORK REPORT

Name

: Tsabit Albanani

Nim

: 4301413003

Department

: Chemistry

Study program : Chemistry education

Group

:7

Date

: October 08th 2015

Tittle

: Blood

A. Objectives
The purpose of the experiment are:
1. To understand the main component contained in blood,
2. To have skill to make blood plasma and blood serum,
3. To have skill to test the blood plasma, Fe in hemoglobin, albumin and globulin test in
blood serum and non-protein substances in blood serum test.
B. Theory
Blood is a tissue that circulate in in the blood vessels. Blood is composed of the part of
red blood cells, white blood cells, and platelets as well as liquid medium in which the solids
suspended. Liquid medium is called plasma, blood was given an anti-coagulant and left blood
cells settles or centrifuged and separated when bloods allowed to clot without anti-coagulant
and the solid part separated, retrieved the solid called serum. On this serum has noticeably
more fibrinogen, a protein that plays a role in blood clotting. (Hudiyono, 2004)
Blood cell consists of three types of erythrocytes, leukocytes, and platelets. Overall blood
volume is one-twelfth of weight approximately five liters. Approximately 55% is blood
plasma, while the remaining 45% consists of blood cells. The nature of which blood has an
osmotic pressure of 28mmHg, a viscosity of 1.7 at 37 C and a pH of 7.0 to 7.8. (Pearce,
2006)
Blood plasma contains about 90% water. Among the various types of substances that are
dissolved in water inorganic salts. Which is sometimes called blood electrolytes and blood
plasma contained in the form of dissolved ions. The combined concentration of these ions is
important in the maintenance of osmotic equilibrium. Some of these ions assist in supporting
the blood pH. Ions also have a pH of 7.4 in humans. The ability of the brain and nerves to
function normally also depends on the concentration of ions in the interstitial fluid key.
(Campbell, 2006)

The main function of blood in circulation is as a transport medium, regulating body

temperature, maintenance of fluid balance, as well as erythrocytes alkaline balance in his life
remains in the body. Red blood cells capable of transporting effectively without leaving their
functions within the network as well as its presence in the blood just passed away. Blood also
has called blood platelets which play a role in blood clotting process. (Hilman, 2005)
Hemoglobin containing two alpha chains and two beta chains and four heme groups,
each of which binds to the polypeptide chain. Each heme groups can bind one oxygen molecule
in reverse because a large amount of hemoglobin contained in red blood cells of 100 ml of red
blood of mammals. If the full dioxygen can carry oxygen gas 21. The amount of oxygen bound
to hemoglobin depends on four factors:
1. The partial pressure of oxygen
2. The concentration of 2,3 diphosphoglycerate
3. The acidity (pH).
4. The concentration of carbon dioxide also reversible binding of oxygen by hemoglobin
is accompanied by the release of a proton, the reaction :
HHb + O2 HbO2 + H+
Thus the increase in pH will shift the equilibrium pressure and increase hemoglobin binds
oxygen more. Blood also transports carbon dioxide formed as a result of oxidation of the fuel
end of the network to the lungs. Hereinafter carbon dioxide from the lungs was released in the
form of airflow. Blood leaving the heart in carbon dioxide equivalent to about 60 mL of gas
CO2 per 100 mL of blood. While blood in the arteries that leaves the lungs to about 50 mL CO2
per 100 mL (Lehninger, 1982)
Blood clotting events by Dr. Karl Landsteiner in 1901 serve as the basis for classification
of blood by the ABO system. Landsteiner discovered that the erythrocytes of some individuals
will clot when mixed with blood serum from another individual, but this happened to everyone.
Blood clotting mechanism is going to undergo platelet clumping (agglutination) due to tissue
damage or injury. Platelet clots are called thromboplastin. Thrombin binds to Ca ions and in
the presence of thrombin then accelarator plasma globulin serum accelarator from inactive to
active. Prothrombin turns into thrombin and fibrinogen activate. Fibrinogen will turn into fine
threads, called fibrin. (Campbell, 2006)
So much blood flowing through the capillaries so that the cumulative fluid loss could
reach about 4.1 per day. There are also several blood proteins leak though capillary walls less
permeable to large molecules. Fluid and protein are lost will be returned to the blood via the
lymphatic system or lymph system. This fluid entered into the system by way of the lymphatic
capillaries diffuses into the small capillaries that exists between cardiovascular system. Along
the lymph vessels are organs called lymph nodes or lymph nodes that filter the lymph.
(Campell, 2006)
2

C. Equipment and Material

Equipment:

Material:

1. Burner

1. Chicken blood

2. Tripod

2. K4Fe(CN)6

3. Gauze

3. HNO3 concentrated

4. Test tube

4. HCl

5. Beaker glass

5. (NH4)2SO4

6. Pipette

6. Dilute BaCl2

7. Stirrer

7. Dilute AgNO3

8. CH3COOH 2%
9.

KSCN

10. Red chlor phenol

indicator
11. Aquades
12. (NH4)C2O4
13. Glycerol

8. Spatula

14. Na2CO3 solid

9. Porcelain cup

15. Fehling A and B

10. Centrifuge
11. Funnel
D. Work Instruction
1. Fe in hemoglobin test
Start

Add 10 drops
bloods + heated

Add aqua regia into it

Divided by
two
Add potassium
rodanide

Add potassium
ferrocyanide

End
End

2. Non-protein substance test

a. Ca2+ test
Start

1 mL sample

Add 1 mL ammonium
oxalate

End

b. Cl- test
Start

1 mL sample

Add 1 mL AgNO3 +
HNO3

End

c. Glucose test
Start

1 mL sample

Add 2 drops glycerol

Add small amount

of Na2CO3

Add 1 mL
fehling

End

d. SO42- test
Start

1 mL sample

Add 1 mL BaCl2

End

E. Result of Observation
a. Fe in hemoglobin test
Treatment

Observation

Blood (heated)

Become ash (black)

+ HCl + HNO3 (Heated)

Dissolve in the solution (not all)

Add potassium ferrocyanide

Become green solution and precipitate

Add potassium KCNS

Color change to yellowish brown solution

Table 1: Fe in hemoglobin test

b. Non protein substance test

Treatment

Observation

Ca2+ test (filtrate + ammonium oxalate)

Formed precipitate (white crystal)

Cl- test (filtrate + AgNO3 + HNO3)

Formed precipitate (white crystals)

Glucose test (filtrate + glycerol + Formed red brick precipitate, solution

Na2CO3 + fehling)

become green

SO42- test (filtrate + BaCl2)

Colorless

Table 2: Non protein substance test

F. Discussion
1. Fe in hemoglobin test
To prove the presence Fe in blood, that is reacted solid cells blood with ferrocyanide
or rodanide compound. Blood plasma was heated in porcelain cup until all burned and form
ashes. This heating was to oxidize Fe that presence in hemoglobin and to remove water in
blood. After hating, than reacted with mixing of dilute HCl and concentrated HNO3 in 3:1
ratio to separate hemoglobin with precipitate blood component and solution became
browness. The reaction that occured:
Fe(s) + 2HCl(aq) Fe2+(aq) + Cl-(aq) + H2(g)
Fe2+(aq) + HNO3(aq) Fe3+(aq) + NO(g) + 2H2O(g)
The ashes than soluble and filtered. Take the filtrate into 2 test tube. In test tube 1
is reacted with K4Fe(CN)6, the solution became dark green and green precipitate arise.
Fe2+(aq) + [Fe(CN)6 ] (aq) + 4 K+(aq) Fe3+(aq) + [Fe(CN)6 ]4-(aq)
Fe3+(aq) + Fe(CN)6 ]4-(aq) Fe4[Fe(CN)6 ]3

The other tube was added with KCNS, the solution became brownish yellow.
Fe3+(aq) + SCN-(aq) Fe(SCN)3(aq)
From this result can be said that the blood sample contained Fe3+ ions.
2. The test of non-protein substance
The non-protein substance test trials done taking the filtrate from centrifuge
separation chicken blood. The first took 5 ml of the filtrate and add 10 drops of distilled
water and then heated. then added 2% acetic acid to form coagulation. The coagulation
then filtered and the filtrate was divided into 4 tubes.
a. Ca2+ test
The experimental results filtrate was added to a solution of (NH4)2C2O4 obtained white
precipitate CaC2O4 with turbid solution. it shows the test for Ca2+ in the blood test is
positive.
Ca2+(aq) + (NH4)2C2O4(aq) CaC2O4(s) + 2NH4(g)
b. Cl- test
To test for the presence of Cl ions in the blood is used filtrate has been obtained from
the initial experiments were then reacted with a mixture of AgNO3 dilute and dilute
HNO3. The results obtained turbid solution of white precipitate that is AgCl. This
indicates that Cl in the blood test result is positive.
Cl-(aq) +AgNO3(aq) AgCl(s) + NO3-(aq)
c. Glucose test
To test the presence of glucose in the blood was using filtrate has been obtained
previously were then added Fehling A and B. the result is a blue solution with a
brick red precipitate.
C6H12O6 + Cu2+ + 4OH- C6H12O6 + Cu2O(s) + 2H2O
(red-brick ppt)

d. SO42+ test
In this test, blood serum was added with barium chloride 1%. The result indicated
the negative test because the solution still colorless.
SO42-(aq) + BaCl2(aq)
G. Conclusion
Based on this experiment we can conclude:
1. Blood consist of solid cells and blood plasma,
2. There are Fe ions in the blood
3. Blood plasma contains non-protein substances such as Ca2+, Cl-, glucose but not contain
SO42-.

H. Suggestion
In this experiment we should:
1. Understand ways of working or step by step what we must do.
2. Do the experiment quietly and carefully.
3. Use safety equipment when we are doing experiment.
I. Reference
Campbell, N.A., J.B. Reece., M.R. Taylor., and E.J. Simon. 2006. Biology. Concepts and
Connection, Fifth Edition. Pearson Education, Inc. Benjamin Cummins: San Fransisco.
Hilman, R.S., Ault K.A., and Rinder H.M. 2005. Hematology in Clinical Practice 4th ed.
The Washington Manual Hematology: New York.
Hudiyono, sumi. 2004. Biokimia. Depok: UI Press.
Lehninger.1982.Dasar-dasar Biokimia. Jilid 1. Jakarta: Erlangga.
Pearce C.E. 2006. Anatomi dan Fisiologi Untuk Paramedic. PT Gramedia Pustaka Utama:
Jakarta.

Semarang, October 08th 2015

Tsabit Albanani

Answer the question

1. Heating with glacial acetic acid carried out for:
-

Remove the water contained in the blood

Outlines of its binding globulin

2. The function of HCl is as donor Cl- ions in the formation of crystals.

3. The reactions that occur in the test Fe in Hb :
Fe3+ + K4[Fe(CN)6] FeK[Fe(CN)6] + 3K+
Fe3+ + 3KSCN Fe(SCN)3 + 3K+
4. The purpose of heating the blood plasma to burn all to ashes on Fe in hemoglobin test is
fatherly oxidize Fe2+ to Fe3+ and a complete oxidation reaction.
5. The function of HCl and HNO3 mixture (water king) is a solvent other metals contained in
the development of blood so fe easily identified.
6. The protein components in blood serum are albumin and globulin.
7. (NH4)2SO4 saturation and (NH4)2SO4 solid used in the test and to separate proteins in blood
serum protein fractions by salting out method. (NH4)2SO4 saturation to separate albumin
and (NH4)2SO4 solid to separate globulin.
8. Salting out is a method separation of the protein fraction in blood serum using different
concentrations of salt Na and (NH4)2SO4
9. Reactions to the test non-protein substances:
Ca2+ + (NH4)2C2O4 CaC2O4(s) + 2NH4+
HCl + AgNO3 AgCl(s) + HNO3(aq)

10. Fibrinogen
11. The blood serum containing glucose and amino acids because these substances derived
from the metabolism of carbohydrates and protein.