pH and pKa

By- Pankaj Singh

pH
Outline of presentation
We will discuss:
Introduction of pH Importance of pH The pH meter Calibration of pH meter Some Useful Trips while measuring pH of the Solution Effect of Temperature on pH Effect of pH on Solubility

pKa
Outline of presentation
We will discuss:
Relation between pH and pKa Importance of pKa Determination of pKa by Analytical methods

Introduction of pH
Arrhenius 1887 was the first person to give a definition of an acid and a base, namely that an Acid gives rise to excess of H+ in aqueous solution whereas a Base gives rise to excess of OH- in solution.
The theory was refined by Bronsted-Lowry in 1923 such that a proton donor was defined as an acid and a proton acceptor as a base. They also introduced the familiar concept of the conjugate acid - case pair. The final refinement to acid base theory was completed by Lewis in 1923, who extended the concept to be an acid is an e -pair acceptor and a base an e -pair donor. Hydrogen Ion Concentration Pure water or an aqueous solution contains hydrogen and hydroxyl ions. An aqueous solution is a mixture of an acid and its Salt. In aqueous solution, hydrogen ion concentration can cover a wide range, from about 1 M or more to 10-14 or less. To avoid the use of negative exponents, Sorensen introduced the pH scale. pH is defined as the negative logarithm of the hydrogen ion concentration in solution. i.e., pH= - log10 C H+ or pH= log10 1/C H+

Importance of pH
Many of natures processes are highly dependent on pH Chemical Reactions which take place in industry or in Laboratory The higher (more alkaline) the pH of a substance or solution, the more electrical resistance that substance or solution holds. Therefore, electricity travels slower with higher pH. Ideally, the pH of the blood should be maintained at 7.4. If the pH drops below 6.8 or rises above 7.8, death may occur.

Introduction of pH
Many drugs have one (or more) chemical parts of their structure that are capable of donating or receiving a proton
In situation A, the drug (referred to below as A) has a hydrogen ion attached which it is willing to share (a

chemical philanthropist!) with water, thus producing two slightly different compounds than those at the start.

A-H + H2O H3O+ + A situation B, the drug (referred to below as B) is willing to accept a hydrogen ion (a chemical non-profit
organization) from water, thus producing two slightly different compounds than those at the start. B + H2O B-H + + OH

The pH meter
Principle of a pH meter:
A pH meter is essentially a voltmeter with high input impedance which measures the voltage of an electrode sensitive to the hydrogen ion concentration, relative to another electrode which exhibits a constant voltage. The key feature of the pH-sensitive electrode is a thin glass membrane whose outside surface contacts the solution to be tested. The inside surface of the glass membrane is exposed to a constant concentration of hydrogen ions (0.1 M HCl).
Inside the glass electrode assembly, a silver wire, coated with silver chloride and immersed in the HCl solution, is called an Ag/AgCl electrode. This electrode carries current through the half-cell reaction. The potential between the electrode and the solution depends on the chloride ion concentration, but, since this is constant (0.1 M), the electrode potential is also constant.

A reference electrode is needed to complete the electrical circuit. A common choice is to use another Ag/AgCl electrode as the reference. The Ag/AgCl electrode is immersed in an 0.1 M KCl solution which makes contact with the test solution through a porous fiber which allows a small flow of ions back and forth to conduct the current. The potential created at this junction between the KCl solution and the test solution is nearly zero and nearly unaffected by anything in the solution, including hydrogen ions.

The pH meter
Glass electrode Reference electrode Combined electrode

Calibration of pH meter
pH meters must be regularly calibrated for accuracy using standard pH buffer solutions that are made to an internationally agreed recipe.
It is essential to calibrate using at least two pH buffers that differ in pH by at least three pH units over the range being used. Use the two buffers that bracket the expected range of measurements. For example, if your samples are expected to be acidic, use the pH 7 and pH 4 buffers to calibrate. If they are expected to be basic use the pH 7 and pH 10 buffers. Ensure to calibrate first with pH buffer 7.0 followed by pH buffer 4.0

Some Useful Trips

A pH meter may require a warm up time of several minutes. When a pH meter is routinely used in the laboratory, it is better to leave it ON with the function switch at standby. Set the temperature control knob to the temperature of your buffer solution. Before you begin make sure the electrode is well rinsed with deionized water and wiped off with a clean absorbent paper. Always rinse and wipe the electrode when switching from one solution to another. Calibrate your pH meter by using at least two standard buffer solutions.

Do not allow the electrode to touch the sides or bottom of your container.
When using a magnetic bar to stir the solution make sure the electrode tip is high enough to prevent any damage. Do not stir the solution while taking the reading. Inspect your electrode periodically. The liquid level should be maintained as per the specification provided with the instrument .

Some Useful Trips

pH Electrode
Storage:
When not in use, the pH electrode should always be stored in a storage solution of 1 gram of KCl per 200ml of either the Buffer 4 or 7.

Cleaning :
1. soak electrode in 0.1 M HCl for 15-20 minutes.
2. Rinse with DI H2O 3. Recondition by allowing to soak in either a 4 or 7 buffer for 30 minutes

Effect of Temperature on pH
Generally when we consider the use of buffers we make the following two assumptions.
(a) The activity coefficients of the buffer ions is approximately equal to 1 over the useful range of buffer concentrations. (b) The value of Ka is constant over the working range of temperature. However, in real practice one observes that pH changes slightly with change in temperature. This might be very critical in biological systems where a precise hydrogen ion concentration is required for reaction systems to operate with maximum efficiency.

Effect of pH on Solubility
Solubility Equilibrium: Effect of pH on Solubility A brief discussion of the effect of pH on solubility is of significant importance when dissolution of compounds into solvents is under consideration. Changes in pH can affect the solubility of partially soluble ionic compounds. Example: Mg(OH)2 Mg2+ + 2OH Here K = [Mg2+] [OH ]2 / [Mg(OH)2 ]

As a result of the common ion effect, the solubility of Mg(OH)2 can be increased or decreased. When a base is added, the concentration of OH increases and shifts the solubility equilibrium to the left causing a diminution in the solubility of Mg(OH)2 . When an acid is added to the solution, it neutralizes the OH and shifts the solubility equilibrium to the right. This results in increased dissolution of Mg(OH)2 .

Relation B/W pH and pKa

The famous Henderson-Hasselbach equation defines the relationship between pH, and pKa. It allows us to calculate the relative concentrations of an acid and its salt.

where [A-] is the molar concentration of the salt (dissociated species) and [HA] is the concentration of the un dissociated acid. When the concentrations of the salt and acid are equal, the pH of the system equals the pKa of the acid.

Importance of pKa
In the pharmaceutical industry pKa is an important factor in drug design and development. Most APIs have acidic and/or basic functionalities, their ionization state is controlled by both solution pH and acidic dissociation constants (i.e. K a values) The chromatographic retention and electrophoretic behavior of ionizable compounds strongly depend on the pKa of the compound and the mobile phase pH Moreover, the acidbase property of a drug molecule is the key parameter for drug development because it governs solubility, absorption, distribution, metabolism and elimination. Particularly for developing new APIs, the pKa has become of great importance because the transport of drugs into cells and across other membranes is a function of physicochemical properties, and of the pKa of the drugs .

Determination of pKa
There are several methods for the determination of dissociation constants. Traditionally, potentiometry and UVVIS absorption spectrometry have been the most useful techniques for the determination of equilibrium constants, due to their accuracy and reproducibility. In the past decade, some alternative techniques for dissociation-constant determination, based on separation methods (e.g., LC and CE) have been developed. 1. Potentiometric titration: In the past, potentiometric titration was the standard method for pKa measurement. In a potentiometric titration, a sample is titrated with acid or base using a pH electrode to monitor the course of titration. The pKa value is calculated from the change in shape of the titration curve compared with that of blank titration without a sample present. Potentiometric titration is a high-precision technique for determining the pKa values of substances. Limitations: Solutions of at least 10-4 M are required in order to detect a significant change in shape of the titration curve. To avoid errors, especially for measurements at neutral-to-high pH, carbonate-free solutions must be prepared laboriously.

Determination of pKa
2. Spectro photometric methods (Importance):
An alternative to potentiometric titration is UVVIS spectro photometry because it can handle compounds with lower solubility and lower sample concentrations.

The main advantage is higher sensitivity (>10-6 M) to compounds with favorable molar absorption coefficients.

Determination of pKa
2. Spectro photometric methods (Principle):
Spectral data are recorded continuously during the course of titration by a diode-array spectrometer. The absorption spectra of the sample changes during the course of the titration to reflect the concentration of neutral and ionized species present. The largest change in absorbance occurs at the pH corresponding to a pKa value. These changes are usually identified from the first derivative of the absorbance against time plot or from overlay plots of the different spectra.

Determination of pKa
Spectrophotometric method Limitations:
compound must contain a UV-active chromophore close enough to the site of the acidbase function in the molecule. Spectrophotometric methods offer excellent precision, as in potentiometry, but they require different spectra for different species and reagents must be pure.

To use this method, the absorption spectra of individual species must be characterized beforehand and the molar absorptivities of protonated and de protonated species are thus required.
These measurements are non-trivial if acid-base equilibria comprise more than two ionization steps or if reacting components are not stable within two pH units of the pKa value, so a multi-wavelength Spectrophotometric approach has been developed to determine acid dissociation.

Determination of pKa
3. Liquid Chromatography (LC):
LC is used as a powerful technique for the determination of dissociation constants, as it requires only a small quantity of compounds, studied samples do not need to be pure and poor water solubility is not a serious drawback. The determination of pKa values by means of LC is based on the relationships between capacity factors and the pH of the mobile phase. For each compound and for every mobile phase composition and pH considered, retention time values, TR, are determined.. For mono protic acid, the observed capacity factors at different pH values can be described as a function of the capacity factors of neutral and anionic species (KHA and KA-) and their corresponding molar fractions (XHA and XA-) K = KHA10-pH + KA-10-pKa / 10-pH + 10-pKa

This equation is related to the chromatographic retention of a mono protic substance and the pH of the mobile phase, and can be used to determine pKa values and also to predict the chromatographic behavior of substances.

Determination of pKa
3. Liquid Chromatography (LC) (Limitations):
One of the most important disadvantages of the LC method is that the pH of the mobile phase and, therefore, the range of pKa values that can be determined are limited by the stability of the column package. Moreover, due to the large retention times observed, it is not easy to determine pKa values in water and aqueousorganic mixtures with low contents of organic solvent..

Test for Knowledge

1. 10 mL of 0.2M acetic acid and 4 mL of 0.2M NaOH are mixed. What is the pH of the Solution ? (pKa of Acetic acid is 4.75)

2. The solubility product of CaF2 is 1.7 10-10 . Calculate its Solubility.

3. 50 mL of 0.0010 M NaCl is mixed with 50 mL of 0.00020 M AgNO3. Will a precipitate form? The Solubility product of AgCl is 1.6 x 10-10 4. A Solution contains 0.1M Cl- and 0.01M CrO2-4. AgNO3 is now added to the above solution. Which will be precipitate first, AgCl or Ag2Cro4? Solubility products of AgCl and Ag2Cro4 are respectively 2.8 10-10 and 1.9 10-12.

THANKS