Pharm.

analysis: NON-AQUEOUS TITRATIONS

Its is used for the assay of certain materials which cannot be easily titrated in aqueous systems

Reasons for titration in NON-AQUEOUS solvents:-there are 3 common reasons The reactants or products might be insoluble in water. The reactants or products might react with water. The analyte too weak an acid or base to be titrated in water.

Solvents in NON-AQUEOUS TITRATIONS:1) 2) 3) 4) The solvents are classified into 4 types Protogenic solvents Protophillic solvents Amphiprotic solvents Aprotic solvents

1) Protogenic solvents:-

2) Protophilic solvents:-

They are acidic solvents they enhances the basicity of weak bases. E.g.:- sulfuric acid, formic acid They are basic in nature .they enhances the acidity of weak acids. E.g.:-pyridine, n-butylamine, ethylene-di-amine,

3) Amphiprotic solvents:-

They category of solvents behave as acid or base depending upon the substance dissolved in it. They accept or donate protons. E.g.:- Glacial acetic acid , Dioxon

4)Aprotic solvents:-

In non-aqueous solutions the Selection of solvents based on the following considerations:1) Solubility & nature of sample 2) There should not be any side reactions between the sample or titrant & the solvent 3) The solvent should not effect on the sharpness of the End-point during titrations. 4) The titrant & solubility should be readily miscible with the solvent. 5) The solvent should have high dielectric constant. 6) The solvent should be readily available of low toxicity, easily purified and in expensive.

Solvents in neutral in nature These solvents do not accept or donate protons. These are useful in dissolving the drugs & they act as solvents. E.g.:- Benzene, Carbon tetrachloride.

SOLVENTS
Formic acid Ethylene diamine Acitic acid,glacial acetic acid

Dielectric constant
58.5 12.9 6.13

Example of drugs
Levodop, methyl dopa Phenols Adrenaline

Titrating in NON-AQUEOUS solvents:Acidic titrants Basic titrants

Perchloric acid in Dioxon Perchloric acid in glacial acetic acid P-toluene sulphonic acid 2,4-dinitrobenzene sulphonic acid 2,4,6-trinitrobenzene sulphonic acid

Tetra alkyl ammonium hydroxide Tetra butyl ammonium hydroxide Sodium methoxide Potassium methoxide Lithium methoxide

Among the acidic titrants perchloric acid in glacial acetic acid is mostly used. In basic titrants methoxide of sodium or potassium or lithium & tetrabutylammonium hydroxide are widely used. The titrants are standardized before the assay of any pharmaceutical substances. The primary standards available for the standardization of acidic titrants are potassium hydrogen phthalate, diphenyl guanidine & anhydrous sodium carbonate. Benzoic acid & phenyl cinchonic acid are suitable primary standards for the standardization of the basic titrations.

Detection of end point:-

In non aqueous titrimetry, the end point can be detected by the color change of the indictors or by the potentiometric method. Ex:- crystal violet-violet-yellowish green Orange blue blue pink 2-naphthol benzein blue dark green

Classification of NON-AQUEOUS solvents:1) A. B. 2) They are mainly categorized mainly into two classes. Titration of basic substances Titration of primary, secondary & tertiary amines Titration of halogen acid slts of bases Titration of acidic substances

1) Titration of basic substances:Preparation Mix 8.5 ml Perchloric acid + 500ml glacial acetic acid+ 30 ml of acetic anhydrade Cool & add more glacial acetic acid to makeup 1000ml .Allow tht solution to stand for 24 hours before use. Acetic anhydrade reacts with water in the perchloric acid & acetic anhydrade & readers the mixture virtually anhydrous. Standardization Weight accurately about 0.7g of potassium hydrogen phthalate dissolved in 50ml of glacial acetic acid. Add 2 drops of crystal violet solution & titrate with perchloric acid solution until the violet changes to green. Detect the volume of perchloric acid consumed by 50ml of glacial acetic acid. Each 0.0241g of potassium hydrogen phthalate is equivalent to 1ml of N\10 perchloric acid.

Perchloric acid 0.1 N in glacial acetic acid:-

Titration of 10, 20 & 30 amines Ex:-Adrenaline can be titrated with acetous perchloric acid. Titration of halogen acid salts of bases
Chlorides, Bromides, iodide are very weakly basic & do not react quantitatively with acetous perchloric acid.

 A solution of mercuric acetate is added during the titration. This replaces the halides ion by an equivalent quantity of acetate ion. Which is strong base in acetic acid? 2R.NH2.Hcl (CH3 Coo)2 HG +2CL2R+ NH2 + 2ClHgCl2 + 2CH3Coo4CH3CooH

(un dissolved)

2CH3 CooH2+ +2CH3Coo-

2) Titration of acedic substances: Basic titrants are used. Alkali methoxides or tetrabutyl ammonium hydroxide. Lithium mithoxide is preferred because less quantity of lithium metal is required to prepare the solution & the solution is easy to prepare. Solvents required to dissolves the substances include dimethyl formamide N-butylamine Ethylene diamine Pyridine Preparation

Lithium methoxide 0.1 N

Dissolve 0.7 g of lithium +150ml of methyl alcohol cooling the flask during the
addition of methyl

When the reaction is complete add 850ml of toluene If cloudiness or precipitate occurs add sufficient methyl alcohol to clarity the
solution

Store the solution in a container protected from co2 & moisture.

Standardization Weigh accurately 0.25g of benzoic acid dissolved in 25ml of dimethylformamide & titrates with lithium methoxide solution. Indicator is QUINOLINE RED. Protect the solution from carbon dioxide during the titration. Perform the blank titration using 25ml of dimethyl formamide. Each 0.01221g of benzoic acid is equivalent to 1ml of 0.1 lithium methoxide.

Preparation Dissolve 40g of tetrabutyl ammonium iodide in 90ml of dehydrated methanol in a glass stoppered flask. Place the flask in ice bath add 20g of powdered silver oxide, insert the stopper in the flask and agitate vigorously for one hour. Centrifuge a few minutes & test the super titrant liquid for iodide. If the test is positive add another 2g of silver oxide, and continue to stir for 30 min when all iodide has reacted. Filter through fine sintered glass filter. Rinse the flask & filter with three quantities Each of 50ml anhydrous toluene Add washing to the filtrate and dilute to 1000 ml with anhydrous toluene. Flush the solution for 10min with dry carbon dioxide free nitrogen Store the container protected from carbon dioxide and moisture Standardization Weigh accurately 0.25g of benzoic acid dissolved in 80ml of dimethylformamide.

Tetrabutyl ammonium hydroxide 0.1 N

 Add 3 drops of 1% solution of thymol blue in dimethylformamide & titrate with tetra butyl ammonium hydroxide. End point is blue. Perform blank titration Each 0.01221g of benzoic acid is equivalent to 1ml of 0.1 tetrabutyl ammonium hydroxide.

Pharm.analysis: Complexometric Titrations

A complexometric titration is one in which a soluble, undissociated and stoichiometric complex is-formed during the addition of titrant to the sample solution (usually solution of a metal ion). It is a method of volumetric analysis developed after the introduction of the analytical reagent, commonly known as ethylene diaminetetraacetic acid disodium salt (E.D.T.A). Complexometric titrations are those reactions in which simple metal ion are transformed into complex by the addition of a reagent which is known as ligand or complexing agent. The complex formed is stable and water-soluble. A complexing agent in its widest sense includes any electron-donating system, which by its ability to form a bond or bonds (Covalent or dative), with a metal ion, produces with it a complex which has different properties from that of the free metal ion. The simplest complexing agents are elementary anions, e.g., the fluoride ion in K+ (BF4),the iodide ion in 2K+ (HgI4)--2 and complex anions, e.g., the cyanide ion in potassium ferri-and ferrocyanides. The link with the metal in the complex is covalent involving either the sharing of two electrons, one from metal and one from the ligand or donation of a lone pair of electrons from the ligand. Once the complex is formed, there is hybridization and equalization of all the bonds. There are also neutral groups involving lone pairs of electrons forming a covalent bond, usually from a nitrogen atom (e.g., NH3 in the complex Co(NH3)36+).These are called simple ligands. There are several organic compounds which possess more than one donating group which may be neutral or charged and thus form bi-, tri-, or polydentate ligand. For example, ethylenediamine is a bidentate ligand and E.D.T.A. can act as a hexadentate ligand. The term sequestering agent is generally applied to ligands which form water-soluble complexes with metal ions (e.g., EDTA). The term chelating agent is generally applied to ligands which form waterinsoluble complexes with metal ions (e.g., salicylaldoxime). EDTA disodium salt is the most widely used titrant in complexometric titrations. There are several reasons for the versatility of EDTA. They are:

1. EDTA forms stable, soluble, stoichiometric 1:1 complexes with metal ions. 2. Certain amount of selectivity can be obtained because of differences in stability constants and through the proper control of the pH of the solution. 3. All the metal-EDTA complexes are soluble and most complexes form rapidly. 4. The end-point is readily detected by chemical or instrumental methods. 5. The titration is suitable for a semi micro to macro concentration range. If EDTA (can be generally represented as Na2H2Y) is used, the reaction with a metal ion and the formation constant for the complex formed are represented by

From the above reaction it is evident that complexometric titrations will be very sensitive to pH and all procedures in which EDTA is used as a titrant must include a buffer with sufficient capacity to take care of the hydrogen ion produced during the titration. DETECTION OF END-POINT In complexometric titrations, the end-point is detected by using (i) visual indicators or (ii) instrumental methods. Amongst the visual category are important complexometric indicators (Metal-ion indicators. Metallochromic indicators or indicators). The complexometric indicator is a dye which is capable of acting as ligand to form dye-metal complex. This dye-metal complex is different in colour from the dye itself, and also has a lower stability constant than the chelate-metal complex. The colour of the solution, therefore, remains that of dye-metal complex until the end-point. As soon as there is slight excess of EDTA, the metal-dye complex decomposes to produce free-dye, this is accompanied by a change in colour. It is essential in a complexometric titration to use a buffer solution to maintain the required pH during the titration. Some examples of commonly used complexometric indicators are: 1. Murexide or Ammonium purpurate 2. Solochrome Black T or Eriochrome black T or mordant black II 3. Catechol violet 4. Xylenol organge 5. Methyl thymol blue 6. Alizarin complexone 7. Sodium alizarine sulphonate 8. Diphenyl carbazone 9. Tiron In addition to the visual indicators, instrumental methods like potentiometric titration, photometric titration and amperornetric titration are also employed to detect the end-point. TYPES OF COMPLEXOMETRIC TITRATIONS

Direct Titrations: To the metal ion solution a suitable buffer solution and indicator are added and the solution is titrated with standard EDTA solution until the indicator just changes colour. A blank titration may be performed omitting the sample as a check on the presence of trace of metallic impurities in the reagents. 2. Indirect or Back Titrations : This procedure is necessary for metals which precipitate as hydroxides from solution at the pH required for titration, for insoluble substances (like lead sulfate and calcium oxalate) for substances which do not react instantaneously with EDTA, and for those metal ions which form more stable complexes with EDTA than with the desired indicator. In this titration excess of standard EDTA solution and a suitable buffer solution to added to the metal solution or suspension. The solution is heated to effect complex formation, cooled and the EDTA not consumed by the sample back-titrated with magnesium or zinc chloride (or sulphate) using a suitable indicator. 3. Replacement or Substitution Titrations : When direct or back titration do not give sharp endpoints, the metal may be determined by the displacement of an equivalent amount of magnesium or zinc from a less stable edetate complex according to the following scheme : M2+ + mgx 2 m x 2 mg +2
1.

Calcium, lead and mercury can be estimated satisfactorily by this method using mordant black II as indicator. 4. Alkalimetric Titration : In this method, protons from EDTA are displaced by a heavy metal and titrated with standard alkali according to the following scheme : (m y ),'.-4 2 H + mn+2 H,y2 The titration is carried out in unbuffered solution. A visual pH indicator may be used, but a potentiometric method of locating the end-point is also suggested. MASKING AND DEMASKING AGENTS Masking agents: When it is required to estimate selectively one or more ions in a mixture of cations and to .eliminate the effects of possible impurities which would add to the titre, "masking agents" are used. These act either by precipitation or by formation of complexes more stable than the interferring ion-edetate complex. MASKING BY PRECIPITATION Many heavy metals (e.g., cobalt copper and lead) can be separated either in the form of insoluble sulphides using Na2S or as insoluble complexes using thioacetamide. These are filtered, decomposed and titrated with EDTA. Other common precipitating agents are sulphate for lead and Barium; oxalate for calcium and lead, fluoride for calcium, magnesium and lead, ferrocyanide for zinc and copper, and cupferron and 8- hypdroxyquinoline for many heavy metals. Thioglycerol is used to mask copper by precipitation in the assay of lotions containing copper

and zinc. Dimercaprol forms precipitates with many cations like mercury, cadmium, zinc, arsenic, antimony, tin, lead and bismuth. MASKING BY COMPLEX FORMATION Potassium cyanide reacts with silver, copper, mercury, iron, zinc, cadmium, cobalt and nickel ions to form complexes in alkaline solution which are more stable than corresponding EDTA complexes. Therefore, ions such as lead, magnesiuni and manganese can be determined in their presence. Other examples of masking agents belonging to this category are potassium iodide (He+), Tiron (Aluminium and Titanium) and triethanolamine (Aluminium). Another simple method of masking is pH control. An example is that the alkaline earth metals do not form EDTA complexes below pH 7.0 and transition elements form EDTA complexes stable down to pH 3.0. DEMASKING AGENTS These are substances which release masked metal ion. Formaldehyde or chloral hydrate can be used to release the masked zinc ion by potassium cyanide. These agents are useful in the determination of specifc metal ion from their mixture with other cations. PREPARATION AND STANDARDISATION OF M/20 EDTA M/20 EDTA solution: Dissolve 18.6 g of disodium edetate in sufficient quantity of distilled water to produce 1000 ml. M/20 CaC1 solution: Dissolve 5.005 g of AnalaR CaCO3 in water (25 ml) 2 containing the minimum quantity of dil. HC1. Boil off carbon dioxide, cool and transfer quantitatively to a 1000 ml flask. Dilute with distilled water to 1000 ml. Buffer Solution:Dissolve 67.5 g of ammonium chloride in 570 ml of strong ammonia solution, dilute to 900 ml with water and add a solution of 0.616 g of magnesium sulphate and 0.93 g of sodium edetate in 50 ml of water. Add sufficient water to produce 1000 ml (the small amount of magnesium edetate complex is included in the buffer to render the endpoint sharper in calcium titrations). Method: Pipette 20 ml of standard calcium chloride solution into a conical flask, add 1 ml of buffer (pH 10) solution and 3 drops of solochrome black T indicator and titrate with EDTA solution until the colour changes from winered to blue. A blank titration should be performed on the reagents. APPLICATIONS Many pharmacopoeias drugs and chemicals are assayed by complexometric method. These are given the below Table Drugs Assayed by Complexometry Drug Type of titration

Direct

Calcium aminosalicylate Calcium carbonate Calcium chloride Dibasic calcium phosphate Tribasic calcium phosphate Magnesium chloride Heavy magnesium carbonate Heavy magnesium oxide Magnesium sulphate Zinc chlroide Zinc stearate Zinc Sulphate Zinc undecylenate Aluminium hydroxide gel Aluminium sulphate Alum Calcium gluconate Calcium lactate

Indirect (Back) Indirect (Back) Indirect (Back) Replacement Replacement Replacement