ACTIVITY 5 BUFFERING ACTION OF THE BLOOD SERUM Buffer action is the ability of the buffer solution to resist the changes in pH value on the addition of small amount of an acid or a base is known as buffer action. Human blood contains a buffer of carbonic acid (H2CO3) and bicarbonate anion (HCO3-) in order to maintain blood pH between 7.35 and 7.45, as a value higher than 7.8 or lower than 6.8 can lead to death. In this buffer, hydronium and bicarbonate anion are in equilibrium with carbonic acid. Furthermore, the carbonic acid in the first equilibrium can decompose into CO2 gas and water, resulting in a second equilibrium system between carbonic acid and water. Because CO2 is an important component of the blood buffer, its regulation in the body, as well as that of O2, is extremely important. The effect of this can be important when the human body is subjected to strenuous conditions.
Buffer solutions are extremely important in biology and medicine because most biological reactions and enzymes need very specific pH ranges in order to work properly.
Objectives: 1. To observe the buffering action of blood serum 2. To determine the pH of blood serum
Materials/Reagents: 20ml distilled water 1 graduated cylinder centrifuge machine 1ml 0.1M NaOH 10 test tubes medicine dropper 1ml 0.1M HCl 1 test tube rack alcohol 1ml phenolphthalein 1 syringe 1 tourniquet 1ml methyl orange 5 cotton
Procedure A. 1. Withdraw 5ml venous blood (make sure to follow aseptic technique) and place it into a test tube. Centrifuge the sample. 2. Extract at least 1mL of serum from the blood and use it to procedure B. B. 1. Prepare four test tubes as follows: Test Tubes 1 2 3 4 Water 5mL 5mL 4.5mL 4.5mL 0.1 M NaOH 1 Drop - - - 0.1 M HCl - 1 Drop - - phenolphthalein 1 Drop - 1 Drop - Methyl orange - 1 Drop - 1 Drop Serum - - 0.5mL 0.5 mL
2. To test tube 3, add 0.1M NaOH dropwise until it matches the color of test tube 1. 3. To test tube 4, add 0.1M HCl dropwise until it matches the color of test tube 2. 4. Record the total number of drops of 0.1M NaOH and 0.1M HCl in each case.
2. Give at least two factors that would account for the buffering action of blood serum.
_________________________________________________________________________ _________________________________________________________________________ _________________________________________________________________________ 3. Give the physiologic importance of buffers. _________________________________________________________________________ _________________________________________________________________________ _________________________________________________________________________ 4. What are the symptoms if the blood pH becomes lower or becomes higher than its normal value? _________________________________________________________________________ _________________________________________________________________________ _________________________________________________________________________ REFERENCES: ____________________________________________________________________________ ____________________________________________________________________________ ____________________________________________________________________________
