ACTIVITY NO.

1
pH and Buffers
1.
Introduction
Introduction
◦ pH  
◦ The pH of a solution is the common
logarithm of the reciprocal of the
hydrogen ion concentration expressed as:
◦ 1
◦ pH = log _____
◦ H+

3
Introduction
◦ pH measures the concentration of H+
ions in a solution.
◦ H+ and OH- is = : Neutral
◦ H+ > OH- : acidic
◦ H+ < OH- : Basic

4
Introduction
◦ Buffer – contains weak acid (HA)
and its conjugate base (A-).
◦ Significant in the addition of strong
acid or base without causing large
change in pH

5
2.
Rationale
Rationale
◦ pH of solutions is an important
characteristics in which cells should maintain
constant pH.
◦ Chemical and enzymatic reactions are
typically dependent on a specific pH range.

7
 Rationale
◦ Buffers play important role in stabilizing the
pH of living systems.
◦ Buffer is also important to experimental cell
biology.

8
 Rationale
◦ Cells and organisms should maintain specific
and constant pH in order to keep enzyme in
the optimum state of protonation since
biological processes depends on pH.

9
 Rationale
◦ It is important to determine pH to have an
acid-base balance. When the levels of acid in
your blood are too high, it’s called acidosis.
When your blood is too alkaline, it is called
alkalosis.

10
Objectives
◦ Explain concepts of pH and Buffers
◦ Discuss when a sample is acidic,
neutral or basic

11
DATA
INTERPRETATION
Table A: pH
Test Samples pH Paper Reading pH Meter
Reading
1.Defribinated pH 5 pH 5.27 or 5.29
Blood
pH 6 or pH 7 pH 6.41 or 6.42
2. Fresh Milk

pH 4 or 5 pH 5.64 or 5.65
3. Fresh Urine
13
 Interpretation
◦ Blood has a normal pH range of 7.35 to 7.45
which means that blood is naturally slightly
alkaline or basic. 
◦ Changes in normal blood pH might be a sign
of certain health conditions
◦ Acidosis (below 7.35) and alkalosis (higher
than 7.45)
14
Table A: pH
Test Samples pH Paper Reading pH Meter
Reading
1.Defribinated pH 5 pH 5.27 or 5.29
Blood
pH 6 or pH 7 pH 6.41 or 6.42
2. Fresh Milk

pH 4 or 5 pH 5.64 or 5.65
3. Fresh Urine
15
 Interpretation
◦ Fresh milk has a normal pH of between 6.5-
6.7
◦ If the milk is lower that 6.5 it denotes
presence of bacterial deterioration
◦ Excessive acidity in fermented products will
impart excessive tart to the product and make
it less palatable.
16
Table A: pH
Test Samples pH Paper Reading pH Meter
Reading
1.Defribinated pH 5 pH 5.27 or 5.29
Blood
pH 6 or pH 7 pH 6.41 or 6.42
2. Fresh Milk

pH 4 or 5 pH 5.64 or 5.65
3. Fresh Urine
17
Interpretation
◦ A urine pH of 4 is strongly acidic, 7 is
neutral and 9 is strongly alkaline.
◦ Sometimes urine needs to be either
acidic or alkaline to prevent some types
of kidney stones from forming.  

18
Table A: pH
Test Samples pH Paper Reading pH Meter
Reading
1.Defribinated pH 5 pH 5.27 or 5.29
Blood
pH 6 or pH 7 pH 6.41 or 6.42
2. Fresh Milk

pH 4 or 5 pH 5.64 or 5.65
3. Fresh Urine
19
 Urine pH Value and its Causes
pH Value
Acidic Urine pH (<6-7 pH)
Alkaline Urine pH (>7-8 pH)
Causes
High protein diet
Acidosis
Uncontrolled diabetes
Diarrhea
Starvation and dehydration
Metabolic or respiratory acidosis
Urinary tract obstruction
Ingestion of citrus fruits
Vegetarian diet
Ingestion of milk/dairy products
Pyloric obstruction
Salicylate intoxication
Renal tubular acidosis
Chronic renal failure
Metabolic and respiratory acidosis
20
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
21
Interpretation: Distilled Water
 Original pH is 6.29 which makes it
slightly acidic or somewhere in between
neutral and acidic.
 NaOH + H2O = Na(OH)2 + H2
 The ions float apart leading to extra –OH
ions in the water.

22
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
23
Interpretation: Distilled Water
◦ HCl + H2O H3O + Cl
◦ H+ from the HCl binds to neutral water
molecules to form H3O+ raising the
concentration of H+.

24
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
25
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
26
Interpretation: Sodium Acetate
◦ Sodium acetate react with sodium
hydroxide
◦ CH3COONa + NaOH  CH4 + Na2CO3
◦ NaOH - strong base and CH3COOH
-weak acid= basic

27
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
28
Interpretation: Sodium Acetate
◦ Sodium acetate react with hydrochloric
acid
◦ CH3COONa + HCl = CH3COOH +
NaCl. 

29
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
30
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
31
Interpretation: Phosphate Buffer
◦ Addition of base to a buffer solution
leads to decrease by less than the
amount expected of H+ concentration
◦ The acid and its conjugate base
consume the hydroxide ions.

32
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
33
Interpretation: Phosphate Buffer
◦ HCl added to a buffer solution ; there is
a limit to the quantity of H+ or OH- that
a buffer can absorb without undergoing
a significant change in pH.

34
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
35
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
36
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
37
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
38
ANSWERS TO
QUESTIONS
Question 1:
What is the normal pH
of blood? Milk? Urine?
Answer :
◦
Blood- 7.35 to 7.45
Milk- 6.7 to 6.9
Urine- 4.5 to 8.0

41
Question 2:
What is the
biochemical relevance
of pH?
Answer:
◦ pH is relevant because it gives great effect
on macromolecules specifically on its
structure and activity. For instance,
animals breathe not because of the lack of
oxygen but because CO2 buildup in the
blood increases the blood acidity.

43
Question 3:
Calculate the pH of
the following solutions
Answer:
◦   0.01 M HCl
A.
◦ pH = log or pH = -]
◦ pH = log
◦ pH = 2

45
Answer:
◦   2.2 x 10-3 M NaOH
B.
◦ pH = log or pH = -]
◦ pH = log
◦ pH = 2.66

46
Question 4:
Which sample
exhibited buffer
action? Explain
 Table B
Test Solutions Original pH pH with 0.1 mL 0.1N pH with 0.1 mL
NaOH 0.1N HCl
  pH 6.29 pH 10.47 or 10.48 pH 3.67
1. Distilled Water
 
pH 7.41 pH 9.95 pH 6.29
 
2. 0.1M Sodium Acetate
 
pH 7.01 pH 7.08 pH 6.94
 
3. Phosphate Buffer pH 7.0
 

  pH 7.33 pH 10.18 pH 6.42

4. Albumin Solution
 
48
Question 5:
What are the buffer
systems in whole
blood and plasma?
Answers:
◦ Plasma proteins, phosphate,
bicarbonate and carbonic acid
buffers.

50
Conclusion
◦ The ability of the body to maintain its internal
conditions despite of the changing external
environment is called homeostasis. One of the
body’s example of its ability to maintain its internal
conditions is the presence of buffers. The results of
the experiment are specifically erroneous every time
the pH meter was used. This can be attributed to the
age of the probe, lack of maintenance, lack of pH
meters and their improper use
51
REFERENCES:
◦ pH and Buffers. LibreTexts. Retrieved from
https://bio.libretexts.org/Bookshelves/Biotechnology/Lab_Manual
%3A_Introduction_to_Biotechnology/01%3A_Techniques/1.07%3A_pH_and_Buffe
rs
◦ Buffers, pH, Acids and Bases. Lumen. Retrieved from
https://courses.lumenlearning.com/wm-nmbiology1/chapter/buffers-ph-acids-and-
bases/
◦ pH and Buffers. The chemistry of life. Retrieved from https://dlc.dcccd.edu/biology1-
2/ph-and-buffers
◦ pH , Buffers,Acids and Bases. Lumen. Retrieved from
https://courses.lumenlearning.com/introchem/chapter/ph-buffers-acids-and-
bases/#:~:text=Buffers%20are%20solutions%20that%20contain,
%E2%80%93%20log%5BH%2B%5D.
52
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