Ph and Buffer Solution
Title: pH and buffer solutions
Aim
This experiment was carried out to determine the role of buffer solution and the factor which affect the buffer capacity. Besides, this experiment was carried out to investigate the solubility of protein casein over a range of pH concentration. This experiment also was carried out to determine the isoelectric point of the casein and the effect of the isoelectric point toward the casein solution.
Methods
Verification of the Henderson-Hasselbalch equation
Figure 1: Method for Verification of the Henderson-Hasselbalch equation
The tris (hydroxymethyl) aminomethane buffer system
Figure 2: Method The tris (hydroxymethyl) aminomethane buffer system
Effect of temperature and/or dilution on buffering capacity:
Temperature
Figure 3: Method for effect of temperature on buffering capacity
Dilution
Figure 4: Methods for effect of dilution on buffering capacity
Table 1: Table of dilution
Dilution ratios Volume of Phosphate solution (mL) Volume of tris buffer (mL) Volume of distilled water (mL)
1:10 2.0 2.0 18.0
1:50 0.4 0.4 19.6
1:100 0.2 0.2 19.8
Dilution
1:10 dilution
Volume of buffer needed = 1/10 × 20 = 2.0 mL
Volume of distilled water = 20-2.0 =18.0 mL
Concentration of buffer = 0.1/1000 × 2.0 = 2 ×10-4 M
1:50 dilution
Volume of buffer needed = 1/50 × 20 = 0.4 mL
Volume of distilled water = 20-0.4 =19.6 mL
Concentration of buffer = 0.1/1000 × 0.4 = 4 ×10-5 M
1:100 dilution
Volume of buffer needed = 1/100 × 20 = 0.2 mL
Volume of distilled water = 20- 0.2 =19.8 mL
Concentration of buffer = 0.1/1000 × 0.2 = 2 ×10-5 M
Isoelectric precipitation of Casein
Figure 5:Methods for Isoelectric precipitation of Casein
Table 2: The amount of casein solution , distilled water and acetic acid added in finding the isoelectric point of casein. 1 2 3 4 5 6 7
Casein solution (mL) 1.0 1.0 1.0 1.0 1.0 1.0 1.0
Distilled water
Aim
This experiment was carried out to determine the role of buffer solution and the factor which affect the buffer capacity. Besides, this experiment was carried out to investigate the solubility of protein casein over a range of pH concentration. This experiment also was carried out to determine the isoelectric point of the casein and the effect of the isoelectric point toward the casein solution.
Methods
Verification of the Henderson-Hasselbalch equation
Figure 1: Method for Verification of the Henderson-Hasselbalch equation
The tris (hydroxymethyl) aminomethane buffer system
Figure 2: Method The tris (hydroxymethyl) aminomethane buffer system
Effect of temperature and/or dilution on buffering capacity:
Temperature
Figure 3: Method for effect of temperature on buffering capacity
Dilution
Figure 4: Methods for effect of dilution on buffering capacity
Table 1: Table of dilution
Dilution ratios Volume of Phosphate solution (mL) Volume of tris buffer (mL) Volume of distilled water (mL)
1:10 2.0 2.0 18.0
1:50 0.4 0.4 19.6
1:100 0.2 0.2 19.8
Dilution
1:10 dilution
Volume of buffer needed = 1/10 × 20 = 2.0 mL
Volume of distilled water = 20-2.0 =18.0 mL
Concentration of buffer = 0.1/1000 × 2.0 = 2 ×10-4 M
1:50 dilution
Volume of buffer needed = 1/50 × 20 = 0.4 mL
Volume of distilled water = 20-0.4 =19.6 mL
Concentration of buffer = 0.1/1000 × 0.4 = 4 ×10-5 M
1:100 dilution
Volume of buffer needed = 1/100 × 20 = 0.2 mL
Volume of distilled water = 20- 0.2 =19.8 mL
Concentration of buffer = 0.1/1000 × 0.2 = 2 ×10-5 M
Isoelectric precipitation of Casein
Figure 5:Methods for Isoelectric precipitation of Casein
Table 2: The amount of casein solution , distilled water and acetic acid added in finding the isoelectric point of casein. 1 2 3 4 5 6 7
Casein solution (mL) 1.0 1.0 1.0 1.0 1.0 1.0 1.0
Distilled water