Objective To determine the pKa of methyl red by measuring the absorbance spectra as a function of pH Summary

Introduction An acid dissociation constant is equilibrium constant for the dissociation of an acid. It is denoted by Ka. Methly red provides a particular good system which study acid-base equilibrium since both [HMR] and [MR-] have strong absorption peaks in the visible of the spectrum HMR +H2O Ka is defined, subject to certain conditions, as Ka = {[H3O+][MR-]}/[HMR] MR- + H30+

where [HMR], [MR-] and [H3O+] are equilibrium concentrations of the reactants. The term acid dissociation constant is also used for pKa, which is equal to -log10 Ka (Lim You, 2008).The term pKb is used in relation to bases, though pKb has faded from modern use due to the easy relationship available between the strength of an acid and the strength of its conjugate base. The value of pKa indicates the strength of an acid, the larger the value the weaker the acid. In aqueous solution, simple acids are appreciably dissociated in the pH range pKa 2. The actual extent of the dissociation can be calculated if the acid concentration and pH are known. Knowledge of pKa values is essential for the understanding of the behavior of acids and bases in solution. For example, many compounds used for medication are weak acids or bases, so a knowledge of the pKa and log p values is essential for an understanding of how the compound

enters (or does not enter) the blood stream. Other applications include buffer solutions, acid-base homeostasis and certain kinds of enzyme kinetics, such as Michaelis-Menten kinetics, which involve a pre-equilibrium step. Also, knowledge of pKa values is a prerequisite for a quantitative understanding of the interaction between acids or bases and metal ions to form complexes in solution. In spectroscopy, an isosbestic point is a specific wavelength at which two (or more) chemical species have the same molar absorptivity ().When an isosbestic plot is constructed by the superposition of the absorption spectra of two species (whether by using molar absorptivity for the representation, or by using absorbance and keeping the same molar concentration for both species), the isosbestic point corresponds to a wavelength at which these spectra cross each other.

Figure1.1: Graph to find Isosbestic Point Figure 1.1 above show that the absorption spectrum of HMR and MR-. There are 3 graphs show the changes in the spectrum for the system with the change in the pH. Sometime at the certain wavelength the molar extinction coefficient (), for the species HMR and MR- is the same because the absorption of both species cannot be differentiated. therefore at this wavelength the absorption for overall system will be constant because the concentration of both HA and A- is constant. At this point the absorption of the system is always constant, this point is known as the isobectic point.

Equipments: Ultraviolet- Visible Spectrophotometer, Silica cell 10 mm, Pipette 1, 5, 10 ml, Beaker 50, 100 ml, Dropper, Ph-meter Volumetric Flask (12 x 50mL)

Materials: Methyl red, Ethanol (95%), HCl (0.1M), Sodium hydroxide (1.0M), Sodium acetate (0.2M) Acetic acid (0.2M).

Procedure Part A: Preparation Standard Solution of Methyl Red

1. 5.0 mL of methyl red (MR) stock solution is pipette into a 100mL volumetric flask

2. 50 mL of Ethanol is added and distilled water is used to dilute to obtain a 100 mL solution

Part B: Preparation Acid and Basic Solution of Methyl Red

I.

Acid solution of MR (pH 1)

1. 6.0 mL MR standard solution is pipette into 50 mL volumetric flask

2. 10.0 mL of HCL (0.1M) 3. The solution is diluted to 50 mL with distilled water 4. The pH of the solution is check with a pH meter and the colour of the solution is recorded 5. The absorption spectrum for the solution is recorded in the wavelength range of 350-600 nm.

II.

Basic Solution of MR (pH 13) 1. 5.0 mL of MR standard solution is pipette into a 50 mL volumetric flask 2. 10.0 mL of NaOH (1.0 M) is added 3. The solution is diluted to 50 mL volumetric flask 4. The pH value is checked and the colour of the solution is recorded
5. The absorption spectrum of the solution is recorded as above

Part C: Absorption Spectrum of MR at various pH

1. The absorption spectrum at eight different pH values besides the two pHs in the Part B is

recorded
2. The volume of mix solution from table below can be used as a guideline to obtain

specific pH value
3. The quantity of substance as in table below is pipette into a volumetric flask and is

diluted with distilled water up to the 50 mL mark to prepare a solution with a specific pH.

4. The flask is shacked well to homogenize the solution 5. The absorption spectrum is recorded for each solution on a different paper. The recording should be carried out immediately after preparing the solution 6. The remaining of the solution is kept to determine the pH 7. The true pH value for all solution is determined using the pH meter. Solution Volume of MR/ml 6.0 6.0 6.0 6.0 6.0 6.0 6.0 6.0 Volume of Acetic acid 0.2M 12.0 10.0 8.0 6.0 4.0 3.0 2.0 1.0 Volume of Sodium Acetate 0.2M/ml 8.0 10.0 12.0 14.0 16.0 17.0 18.0 19.0 pH Abs

a b c d e f g h

4.63 4.74 4.94 5.10 5.34 5.50 5.58 5.88

Results Table 1 Solution pH 1 A B C D E F G H pH 13 pH 1.58 4.42 4.63 4.83 5.05 5.33 5.52 5.71 6.14 13.32 Absorption Max 1.873 1.425 1.276 1.076 0.843 0.611 0.627 0.671 0.717 0.602

Table 2 Wavelength Absorption

(nm) 600 550 500 450 400

pH 1 0.045 1.457 1.648 0.422 0.049

A 0.075 1.324 1.171 0.466 0.248

B 0.067 1.184 1.059 0.493 0.296

C 0.049 0.997 0.910 0.518 0.348

D 0.037 0.775 0.740 0.551 0.414

E 0.030 0.554 0.576 0.607 0.502

F 0.023 0.419 0.476 0.627 0.542

G 0.020 0.309 0.400 0.663 0.594

H 0.01 4 0.14 5 0.28 3 0.69 3 0.64 8

pH 13 0.013 0.020 0.161 0.571 0.547

Table 3 Solution pH Absorptio n Max A B C D E F G H 4.42 4.63 4.83 5.05 5.33 5.52 5.71 6.14 1.425 1.276 1.076 0.843 0.611 0.627 0.671 0.717 0.448 0.597 0.797 1.030 1.262 1.246 1.202 1.156 0.823 0.674 0.474 0.241 9x10-3 0.025 0.069 0.115 MRHMR (MR-)/ (HMR) 0.544 0.886 1.681 4.274 140.222 49.840 17.420 10.052 Log [(MR-)/ (HMR)] -0.264 -0.053 0.226 0.631 2.147 1.694 1.241 1.002 4.684 4.683 4.604 4.419 3.183 3.826 4.469 5.138 pKa

Graph 1

Graph 2: Absorption Vs Wavelength

Absorpti on

Wavelengt h

Graph 3