1. Pharmaceutical Analysis 501722 Dr. Wael Abu Dayyih Group (3) 20111

2. Physical and Chemical Properties of Drug Molecules 2

3. Introduction The physical properties of organic molecules, such as pKa and partition coefficient, are dealt with extensively in pharmacy courses but do not feature greatly in analytical chemistry courses. It is often surprising that analytical chemists cannot distinguish between, for instance, basic, weakly basic, acidic, weakly acidic and natural nitrogen functions. The physical properties of drug molecules, along with simple chemical dramatization and degradation reactions, play an important part in the design of analytical methods. Drug molecules can be complex, containing multiple functional groups that in combination produce the overall properties of the molecules. The chapter will serve as a starting point for understanding the chemical and physio-chemical behavior of drug molecules, which influence the development of analytical methods. The latter part of the chapter focuses on some typical drugs that are representative of a class of drug molecules and lists their physical properties and the properties of their functional groups in so far as they are known. 3

4. Calculation of pH value of aqueous solutions of strong and weak acids and bases Dissociation of water The pH of a solution is defined as –log [H + ], where [H + ] is the concentration of hydrogen ions in solution. In pure water the concentration of hydrogen ions is governed by the equilibrium Ka is the dissociation constant for the equilibrium, is known as Kw in the case of the dissociation of water and is determined by the following expression: 4

5. Dissociation of water Since the concentration of water does not change appreciably as a result of ionization, its concentration can be regarded as not having an effect on the equilibrium and it can be omitted from the equation, and this means that in pure water: [H + ] = [HO - ] = 10 -7 The pH of water is thus given by –log 10 -7 = 7.00 5

6. Strong Acids and Bases If an acid is introduced into an aqueous solution the [H + ] increases. If the pH of an aqueous solution is known, the [H + ] is given by the expression 10 -pH, e.g. [H + ] in pH 4 solution = 10 -4 M = 0.0001 M. Since [H + ] [OH - ] = 10 -14 for water, the concentration of [OH - ] in this solution is 10 -6 M. A strong acid is completely ionized in water and [H + ] is equal to its molarity, e.g. 0.1M HCl contains 0.1 M H + (10 -1 H + ) and has a pH of – log 0.1 = 1. For a solution of a strong base such as 0.1 M NaOH, [OH - ] = 0.1M and [0.1] [H + ] = 10 -14, therefore, [H + ] = 10 -13 M and the pH of the solution = 13. Although the pH range is regarded as being between 0 and 14, it does extend above and below these values, e.g. 10 M HCl, in theory, has a pH of -1. Self-test 2.1 Calculate the pH of the following solution: 0.05 M HCl 0.05 M NaOH 0.05 M H 2 SO 4 Answers: (i) 1.3, (ii) 1.0, since 0.05 M H 2 SO 4 contains 0.1 M H +. 6

7. Weak acids and bases Weak acids are not completely ionized in aqueous solution and are in equilibrium with the dissociated acid, as is the case for water, which is a very weal acid. The dissociation constant Ka is given by the expression below: 7

8. Weak acids and bases For instance, in a 0.1 M solution of acetic acid (Ka = 1.75  10 -5 ), the equilibrium can be written as follows: 8

9. Weak acids and bases The pH can be calculated as follows: Since the dissolution of the acetic acid does not greatly change the concentration of the un-ionised acid, the above expression can be approximated to: 9 In comparison, the pH of 0.1 M HCl is 1.

10. Weak acids and bases The calculation of the pH of a weak base can be considered in the same way. For instance, in a 0.1 M solution of ammonia (Kb = 1.8  10 -5 ), the equilibrium can be written as follows: If the concentration / activity of water is regarded as being 1, then the equilibrium constant is given by the following expression: 10

11. Weak acids and bases The concentration of NH 3 can be regarded as unchanged by a small amount of ionization and the expression can be written as: 11

12. Self-test Calculate the pH of the following solutions: 0.1 M formic acid (Ka = 1.77  10 -4 ) 0.05 M phenol (Ka = 1.3  10 -10 ) 0.15 M ethylamine (Kb = 5.6  10 -4 ) Answers: (i) 2.4, (ii) 5.6, (iii) 11.9 12

13. Acidic and basic strength and pKa The pKa value of a compound is defined as: pKa = -log Ka. A pKa value can be assigned to both acids and bases. For an acid, the higher the [H + ] the stronger the acid, e.g.: In the case of a base, it is the protonated form of the base that acts as a proton donor, e.g.: In this case, the lower the [H + ] the stronger the base. If pKa is used as a measure of acidic or basic strength, for an acid, the smaller the pKa value the stronger the acid; for a base the larger the pKa value the stronger the base. 13

14. Henderson-Hasselbach equation Can be rearranged substituting pH for –log [H + ] and pKa for –log Ka to give: For example, when acetic acid (pKa 4.76) is in solution at pH 4.76. the Henderson-Hsselbalch equation can be written as follows: pH=4.76+log[CH 3 COO - ]/[CH 3 COOH] 14

15. Self-test Determine the percentage of ionization of acetic acid at (i) pH 3.76 and (ii) pH 5.76. Acetic acid is 50% ionized at pH 4.76. in the case of a weak acid, it is the protonated form of the acid that is un-ionised, and, ad the pH falls, the acid becomes less ionized. 15

16. Self-test Calculate the percentage of ionization of ammonia at (i) pH 8.25 and (II) pH 10.25 Answers: (i) 90.9%, (ii) 9.09) An alternative way of writing the expression giving the percentage of ionization for an acid or base of a particular pka value at a particular pH value is: Acid: % ionisation = { 10 (pH-pKa) /1+10 (pH-pKa) } × 100 Base: % ionisation = {10 (pka-pH) /1+10 (pKa-pH) } × 100 16

17. Ionization of drug molecules The ionization of drug molecules is important with regard to their absorption into the circulation and their distribution to different tissues within the body. The pKa value of a drug is also important with regard to its formulation into a medicine and to the design of analytical methods for its determination. Calculation example 2.1 Calculate the percentage of ionization of the drugs at pH 7.0 and pka as shown Diphenhydramine This drug contains one basic nitrogen andpka 9.0, at pH7.0, its percentage of ionisation can be calculated as follows: 17

18. Ionization of drug molecules Ibuprofen This drug contains one acidic group andpKa4.40, at pH 7.0, its percentage of ionization can be calculated as follows: 18

19. Self-test Calculate the percentage of ionisation of the following drugs which contain 1 group that ionizes in the pH range 0-14, at the pH values of (i) 4 and (ii) 9. Answers: Bupivacaine (i) 99.99%, (ii) 11.2%, (iii) 79.9%; Indomethacin (i) 24.0%m (ii) 99.99%; 5-fluorouracil (i) 0.01%, (ii) 90.9%. 19

20. Buffers Solution Buffers can be prepared from any weak acid or base and are used to maintain the pH of a solution in a narrow range. This is important in living systems; for example human plasma is buffered at pH 7.4 by a carbonic acid/ bicarbonate buffer system. Buffers are used in a number of areas of analytical chemistry, such as the preparation of mobile phases for chromatography and the extraction of drugs from aqueous solution. The simplest type of buffer is composed of a weak acid or base in combination with a strong base or acid. A common buffer system is the sodium acetate/ acetic acid buffer system. The most direct way of preparing this buffer is by the addition of sodium hydroxide to a solution of acetic acid until the required pH is reached. The most effective range for a buffer is 1 pH unit either side of he pKa value of the weak acid or base used in the buffer. He pKa value of acetic acid is 4.76; thus its effective buffer range is 3.76-5.76. 20

21. Salt Hydrolysis When the salt of a strong acid and a strong base is dissolved in water it produces a pH of ca 7.0. when salts of a weak acid and a strong base or of a strong acid and a weak base are dissolved in water, they will produce, respectively, alkaline and acidic solutions. When sodium acetate is dissolved in water, the acetate ion behaves as a base removing protons from solution. For a weak electrolyte in Kb × Ka = Kw. If a 0.1 M solution of sodium acetate in water is considered. 21

22. 22 Activity, ionic strength an dielectric constant

23. Partition coefficient An understanding of partition coefficient and the effect of pH on partition coefficient is useful in relation to the extraction and chromatography of drugs. The partition coefficient for a compound (P) can be simply defined as follows: P = C o/ C w Where C o is the concentration of the substance in an organic phase and C w is the concentration of the substance in water. The greater P the more a substance has an affinity for organic media. The value of P for a given substance of course depends on the particular organic solvent used to make the measurement. Many measurements have been made of partition between n-octanol and water, since n-octanol, to some extent, resembles biological membranes and is also quite a good model for reverse-phase chromatographic partitioning. P is often quoted as a log P-value, e.g. a log P of 1 is equivalent to P=10. Where P=10 for a particular compound partitioning into a particular organic solvent, and partitioning is carried out between equal volumes of the organic solvent and water, then ten parts of the compound will be present in the organic layer for each part present in the water layer. 23

24. Effect of pH on partitioning Many drugs contains ionisable groups and their partition coefficient at a given pH may be difficult to predict if more than one ionized group is involved. However, often one group in a molecule may be much more ionized than another at a particular pH, thus governing its portioning 0f 0rganic acids and bases into organic solvent with represent to the pH of the solution that they are dissolved in. From the Henderson – Hasselbalch equation For acid : Papp = P/1+10 pH-pKa For bases : Papp = P/1+10 pka-pH Papp is the apparent partition coefficient which varies with pH. Thus it can be seen that, when a compound, acid or base, is 50% ionized (i.e. pH = pKa), its partition coefficient is half that of the drug in the un-ionised state. Papp =P/1+10 0 = P/2 AS a general rule, for the efficient extraction of a base into an organic medium from an aqueous medium, the pKa of the aqueous medium should be at least 1 pH unit higher than the pKa value of the base, and, in the same situation for an acid, the pH should be 1.pH until lower than the pka value of the acid. 24