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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)

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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)

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C. Equipment and Material

Equipment:

1. Burner

2. Tripod

3. Gauze

4. Test tube

5. Beaker glass

6. Pipette

7. Stirrer

8. Spatula

9. Porcelain cup

10. Centrifuge

11. Funnel

Material:

1. Chicken blood

2. K4Fe(CN)6

3. HNO3 concentrated

4. HCl

5. (NH4)2SO4

6. Dilute BaCl2

7. Dilute AgNO3

8. CH3COOH 2%

9. KSCN

10. Red chlor phenol

indicator

11. Aquades

12. (NH4)C2O4

13. Glycerol

14. Na2CO3 solid

15. Fehling A and B

D. Work Instruction

1. Fe in hemoglobin test

Start

Add 10 drops

bloods + heated

Add aqua regia into it

Divided by

two

Add potassium

ferrocyanide

Add potassium

rodanide

End End

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2. Non-protein substance test

a. Ca2+ test

b. Cl- test

Start

1 mL sample

Add 1 mL ammonium

oxalate

End

Start

1 mL sample

Add 1 mL AgNO3 +

HNO3

End

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c. Glucose test

d. SO42- test

Start

1 mL sample

Add 2 drops glycerol

End

Add small amount

of Na2CO3

Add 1 mL

fehling

Start

1 mL sample

Add 1 mL BaCl2

End

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E. Result of Observation

a. Fe in hemoglobin test

Treatment Observation

Blood (heated)

+ HCl + HNO3 (Heated)

Add potassium ferrocyanide

Add potassium KCNS

Become ash (black)

Dissolve in the solution (not all)

Become green solution and precipitate

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 +

Na2CO3 + fehling)

Formed red brick precipitate, solution

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

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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-.

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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

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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.