Gravimetric assays

-Gravimetric analyses is the procedure for isolation & weighing of an element or compound in as pure a form as possible(or pure derivative form). -In case of derivative, equivalent Wt. of the substance serves as the basis for calculating the result. -A definite portion is taken(weight or volume) from bulk substance or sample being examined.

-The element or compound is separated(pure or pure derivative) from the weighed or measured portion. -Separation techniques: Physical(frequently) Chemical -The weight of the constituent element or compound in the sample is calculated from the weight of the product.

The gravimetric assays may be grouped into following categories: 1)Weighing the active ingredient after separation,

2)Precipitation & weighing of derivative of the active ingredient. 3)Weighing of the residue after ignition of the sample.

1)Weighing the active ingredient after separation: -Active principle separated, dried & weighed.

Estrone injection: Determination of Estrone C18H22O2 -An elaborate purifcation procedure is involved.
-The estrone is converted to water soluble derivative.

-Trimethylacethydrazide ammonium chloride is used for derivatisation.(Girards reagent for carbonyl Compd.).

-The aqueous extract contains only ketonic material.

-The aqueous extract is then decomposed with acid to regenerate estrone. -Extracted into chloroform.
-The solvent is removed & residue weighed.

Caffeine & sodium benzoate injection: Determination of Caffeine C8H10N4O2 -The caffeine is extracted into chloroform in alkaline condition. -The chloroform extract is evaporated to dryness. -The residue is weighed.

2)Precipitation & weighing of derivative of the active ingredient: Thiamine hydrochloride: Determination of the percentage purity of C12H17ON4 SCl,HCl -Depends upon the precipitation of insoluble thiamine silicotungstic acid. -In the slightly acidified sample solution, a solution of silicotungstic acid is added.

-The precipitating reagent is a complex silicate with some what variable degree of hydration. SiO2,12WO3 ,xH2O.

-Thiamine silicotungstate is of constant composition. 2C12H17ON4 SCl,HCl + [SiO2 ,12WO3 ] +6H2O (C12H17ON4 SCl,HCl),[SiO2 (OH) 2 ,12WO3 ]4H2O 674.6g C12H17ON4 SCl,HCl 3858g of residue. 0.1929g C12H17ON4 SCl,HCl 1g of residue.

Anticoagulant citrate phosphate dextrose solution: Determination of C6H12O6.H2O . -The sample solution is boiled with Fehling solution. -Cu2O precipates out, washed, dried to constant Wt.
1.0 mg of cuprous oxide(Cu2O) 0.496mg C6H12O6.H2O

Wool alcohol: Determination of cholesterol

-3 hydroxyls(Cholesterol) form an insoluble molecular addition complex with the steroidal saponin, digitonin.
-The Complexes have very low solubilities.

-Are stable & strongly crystalline.

C27 H46O + C56H92O29 C83H138O30

Cholesterol Digitonin Complex

386.3g cholesterol 1616g of complex 0.239g cholesterol 1g of complex.

-The weighted sample is melted on a water-bath, mixed thoroughly & cooled. -Weighed sample is dissolved in warm ethanol(90%), filtered immediately through a No.2 sintered glass crucible.

-Residue on filter is washed with the warm ethanol(90%). -The filtrate is diluted quantitatively in a V.F.
-In to a pipetted amount , digitonin(0.5%) is added.

-For complete complexation, the mixture is warmed to 60. -Then the mixture is filtered through the dried NO.2 sintered glass crucible. -The ppt. is washed with ethanol(90%), acetone & carbon tetrachloride successively. -Then after draining , ppt. is dried to constant weight at 105C.

3)Weighing of the residue after ignition of the sample. Sodium aurothiomalate:Determination of the percentage of Au. -It is of undefined constitution,

-Contains variable proportion of moisture.

-So that a portion of the substance is dried over phosphorous pentoxide at maximum pressure of 5mm of mercury.

-The percentage of gold is calculated W.R.T. dried substance. -The organic matters are oxidised to volatile products by sulphuric acid. -Sulphuric acid is reduced to sulphur dioxide & hydrogen sulphide.
-Hydrogen sulphide reduces gold salts to metallic gold.

-Metallic gold is precipitated.

-Further treatment & digestion with nitric acid.

-The stage ensures complete decomposition of organic matter & oxidation of inorganic matter e.g. sulphur(from theomalic acid). -Finally the sample is filtered, heated & ignited to the constant weight.

Cognate determinations: Sodium aurothiomalate injection, Aurothioglucose.

Zinc oxide & salicylic acid paste:Determination of ZnO. -Weighed amount of the paste is gradually heated in a porcelain crucible. -After thorough charring , the substance is heated to constant Wt. removing all carbonaceous materials. -The Wt. of the residue represent the quantity of ZnO the weigh of the paste taken for the assay.

Cognate determination: Silica in colloidal silicon dioxide, Aluminum oxide in Aluminum monostearate etc.

Alfonso R. Gennaro: Remington: the Science and Practice of Pharmacy, USP, Beckett/Stenlake Practical pharmaceutical chemistry