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Assay of Boric Acid
Assay of Boric Acid
PRINCIPLE: Boric acid is a local infective. It is very weak acid and hence it can be titrated
with glycerin, glyceryl-boric acid is formed which behaves as a strong acid and can be
PREPARATION OF REAGENTS:
volumetric flask and dissolved in water and make up to 100 with distilled water.
ASSAY:
1gm of boric acid is taken in conical flask and dissolved in distilled water to it add glycerin
and phenolphthalein indicator titrated against with1M NaoH until a faint pink colour was
produced.
REPORT:
2. ASSAY OF ASPIRIN
CHEMICALS: NaoH solution, Aspirin and Phenolphthalein, sodium carbonate, Methyl red
solution.
PRINCIPLE: Assay of aspirin is acid base titration using phenolphthalein indicator. The
amount of aspirin is determined by neutralizing sample solution with excess standard NaoH
solution to acetoxy group. Cooling and back titrated with standard acid using phenol red as
indicator.
PREPARATION OF REAGENTS:
PREPARATION OF 0.5M HCL:-4.25ml of HCL was diluted with water and the volume
was made up to 1000ml.
ASSAY:
1.5gm of aspirin is weighed taken in conical flask and dissolved in ethanol to it add 50 ml of
NaoH boiled for 10mins. The solution was cooled and titrated against 0.5M HCL using
phenol red as indicator.
REPORT:
3. STANDARDISATION OF DISODIUM EDETATE
volume of HCl and bromine water (0.2ml) to ensure oxidation of iron impurity to iron which
forms a much less stable edetate complex than iron. Gently boil to remove excess bromine
cool and add NaOH (2M) until it become neutral, it was diluted to 250ml and ammonia
buffer PH (10) was added until the precipitate was just dissolved and then 5ml in excess was
added. Then a mixture of mordant black-II and NaCl was added which acts as an indicator.It
was then titrated with 0.05M disodium edetate solution until green colour was appeared.
AIM: To perform the assay of calcium gluconate and repeat its percentage purity.
PROCEDURE:
0.125gms of granulated zinc was accurately weighed and dissolved in few ml of HCl.To this
0.2ml of bromine water was added.solution was gently boiled to remove excess of
bromine.Solution was cooled and NaOH was added until solution becomes neutral.dilute to
250ml Ammonia buffer of PH 10 was added until the precipitate was just dissolved, 5ml
excess was added.Then a mixture of mordant black-II and NaCl was added as indicator and
was titrated with 0.05M disodium edetate until the green colour appears.
ASSAY: Drug equivalent to 0.5gms was taken and dissolved in 50ml of warm water. The
solution was cooled and 5ml of 0.05M MgSO4 and 10ml of NH3 was added. The resultant
mixture was then titrated with 0.5M EDTA using mordant black-II as indicator.
PRINCIPLE: Ferrous sulphate is an example of haematinic agent and also reducing agent. It
is an redox titration.KMno4 is an powerful oxidizing agent in the presence of dilute
H2SO4.During the titration ferrous sulphate is oxidized to ferric sulphate. As soon as the
oxidation of ferrous sulphate is completed addition of a drop of KMno4 produces pink color
which indicates end point.
PREPARATION OF REAGENTS:
PREPARATION OF 0.1N OXALIC ACID:- 0.63gms of oxalic acid was diluted with
water and the volume was made up to 100ml.
STANDARDIZATION OF 0.5M HCL:-10ml of 0.1N Oxalic acid was taken and to this add
10 ml of sulphuric acid .The contents were warmed to 70ºC and titrated with KMno4.The
titration was continued until a faint pink color was produced.
ASSAY: 1gm of ferrous sulphate was weighed accurately and dissolved in 20 ml of dilute
sulfuric acid .Then it was titrated with 0.1N KMno4 until permanent color was produced.
REPORT: The percentage purity of given ascorbic acid tablet was found to be
6. ASSAY OF METRONIDAZOLE
AIM: To perform the assay of Metronidazole by non-aqueous titration.
PRINCIPLE: The substances which are weakly basic and weakly acidic, give sharp end
points in non-aqueous titrations. Non-aqueous titration is the titration of substances dissolved
in non-aqueous solvents. It is the most common titrimetric procedure used in assays. These
titrations provide insoluble organic compounds are soluble.
PROCEDURE:
ASSAY:
20 tablets were weighed, powdered and a quantity equivalent of 0.2gms was taken.
Solution is extracted with 6 quantities each of 10ml of hot acetone and cool.
50ml of acetic anhydride was added.
0.1ml of anhydrous glacial acetic acid was added and 1% solution of brilliant green
which acts as indicator, and titrated against 0.1N perchloric acid.
PRINCIPLE: The substances which are weakly acidic and weakly basic give sharp
dissolved.
PROCEDURE:
20 tablets were weighed and powdered and 0.1gm was taken and dissolved in 70ml of
water and diluted to 100ml and filter.
From above solution, 10ml was taken and diluted to 100ml with water, from 10ml
was taken and further diluted to 100ml and absorbance was measured at 232nm.
PRINCIPLE: Ampicillin is a β-lactum antibiotic used against gram +ve bacteria. The
primary amine present in ampicillin reacts with ninhydrin. This reaction in presence of 0.1N
HCl using ninhydrin reagent, it forms L-amino acid which on reaction with ninhydrin gives
colour complex. Absorbance was seen at 490nm.
PROCEDURE:
For calibration curve: 100mg of drug was dissolved in 100ml of 0.1N HCl to give 1mg/ml.
From this 1, 2, 3, 4, 5ml was taken, to each 2ml of ninhydrin, 1.5ml of 0.1N HCl, 2ml
of methanol was added.
Then it was heated until colour developed. It was then cooled and volume was made
up to 100 ml with methanol and absorbance was measured at 490nm.
ASSAY:
20 tablets were weighed and quantity equivalent to 100mg was dissolved in 100ml of
0.1N HCl.
From this 3 ml was taken, 1.5N HCl, ninhydrin, 2ml methanol were added and heated
for 15min until the colour was developed.
It was then cooled and diluted to 10ml and absorbance was measured at 492nm.
CHEMICALS: Furosemide pure drug and tablets, sodium nitrite, methanol, aniline, HCl,
Water.
carbonyl group with sulphonyl moiety in Meta position (carbon no.5), a halide substitution is
highly potent diuretic related to the thiazides but more potent than them. Furosemide has high
efficiency and rapid onset of action and short duration of action. It acts only on proximal and
distal tubules and allow on the ascending limb of loop of Henley It is indicated in the
treatment of edema associated with congestive heart failure, cirrhosis of liver, renal disease
and hypertension The hydrolysed furosemide is diazotized with nitrite in acid medium and
coupled with aniline to give a water soluble coloured dye having an absorption maximum at
480nm.
PROCEDURE:
Nitrite solution: A 1000 ppm nitrite solution was prepared by dissolving 0.15gms of
Aniline solution: A 10% v/v aniline solution was prepared by mixing 10ml of aniline in
PREPARATION OF STANDARD:
10ppm.
From the above 2, 4, 6, 8and 10ml were pipetted and make up to 10ml with methanol
To the aliquots of drug 2 ml of 3.5M HCl was added and 10ml of NaNO2was added.
The solutions were kept for 5min and allow to diazotization reaction to go to
completion.
The resulting solutions are coupled with aniline by adding 7ml of 10% aniline
The orange coloured azo dye was diluted to 100ml with distilled water and
ASSAY:
10 tablets were taken , weight of drug equivalent to 0.05gms was taken and dissolved
in 50ml CH3OH
1ml of above solution was taken and diluted to 10ml with methanol
From the resulting solution any volume ranging from 2-10ml was pipetted out and
To these solutions, 2ml of 3.5M HCl was added followed by addition of 10ml NaNO2
Then 7ml of 10% aniline was added and was boiled in water bath for 6min,
The resulting orange colour azo dye was diluted to 100ml with distilled water and
CHEMICALS: MBTH reagent, distilled water, cephadroxil pure drug and ceric ammonium
nitrate.
PROCEDURE:
For calibration curve: 10 mg of pure drug was weighed and dissolved in 10ml of water.
From the above solution 0.1, 0.2, 0.3, 0.4 and 0.5 ml are taken and to these solutions 2ml of
MBTH solution was taken and to these solutions 2ml of MBTH solution was added and 2ml
of Ce(IV) solution was added and volume was made up to 1oml with distilled water which
produces concentration of 10, 20, 30, 40 and 50 µg/ml. The absorbance was noted at 410nm.
AIM: To estimate the amount of diclofenac present in given tablet using MBTH.
PRINCIPLE: Diclofenac 2(2, 6-dichlorophenyl) amino benzene acetic acid mono salt. It is
an NSAID advised for use in painful and inflammation, rheumatic and non-rheumatic
condition. The proposed mechanism involves the use of MBTH in presence ofcerric
ammonium as the chromogenic agent for diclofenac. It obeys beers law at 0.2-8 µg/ml and
absorbance was noted at 580nm.
PROCEDURE:
ASSAY:
AIM: To estimate the amount of atorvastatin present in given tablet using MBTH.
CHEMICALS: Atorvastatin (AVS) pure drug and tablets, MBTH, methanol, Ce(SO4)2,
They have potent cholesterol-lowering effects and they could reduce morbidity and mortality
associated with coronary heart disease. Atorvastatin calcium is chemically {[R-(R, R*)]-2-(4-
presence of Ce (SO4)2in an acidic medium to form pink coloured oxidative coupling product
that can be measured at 566nm. Under the reaction conditions on oxidation, MBTH loses two
electrons and one H+ forming an electrophilic intermediate, which is active coupling species.
The coupling of oxidized form of the drug with electrophilic intermediate of MBTH results in
PROCEDURE:
Preparation of stock solutions: stock standard solution of 0.5 mg/ml was prepared by
dissolving the appropriate weight of AVS in 100ml volumetric flask, 5ml of methanol was
added the volume was then diluted to the mark with distilled water. 0.01M MBTH solution
was prepared with doubled distilled water and 1% Ce (SO4)2 solution was prepared with
sulphuric acid of 0.369M, medium. Freshly prepared solutions were always used.
Procedure for calibration curve: Aliquots of standard AVS solution (0.1-1.0ml, 0.5mg/ml)
were transferred into a series of 25ml calibrated volumetric flask. Then 1ml of MBTH
solution was added and kept a side for 3 min. after that, 1 ml of Ce(SO4)2 solution was added.
The volume was made up to mark with distilled water mix well and note the absorbance at
566nm as function of time between 0-20min against reagent blank treated similarly.
ASSAY: 20 tablets containing AVS were weighed and pulverized. An amount of the powder
equivalent to 25mg of the cited drug was dissolved in a 25ml of CH3OHand mixed for about
5min and filtered. The methanol was evaporated to about 5ml. the remaining portion of
solution was diluted to 50ml with double distilled water to achieve a concentration of
0.5mg/ml.Then to this above solution 1ml of MBTH was added and kept aside for 3min. after
that 1ml of Ce(SO4)2was added and the volume was made up to mark (25ml) with double
CHEMICALS: Cephalexin pure drug, FC reagent 10% NaOH, distilled water, tablets,
NaNO2
PROCEDURE:
For calibration curve: 100mg of pure drug was weighed and dissolved in 100ml water.
From this solution 10ml was taken and made to 100ml with water to give 100µg/ml. From
this 1, 1.5, 2, 2.5, 3ml were pipetted out and 2ml of FC-reagent and 2.5ml NaOH was added
volume was made up to 10ml with distilled water. The resultant concentrations were 10, 15,
20, 25 and 30 µg/ml respectively. Absorbance was noted at 753nm and curve was plotted.
ASSAY:
20 tablets were weighed and the drug equivalent to 100mg was taken and dissolved in
100ml of distilled water.
From the above stock solution, 2ml was taken and the volume was made up to 100ml,
from this 1ml was taken and to this solution 0.1 ml of FC reagent and 1ml of NaNO 2
was added and volume was made up to 10ml with distilled water and absorbance was
noted at 753nm.
CHEMICALS: 0.2% FeCl3, ciprofloxacin pure drug and tablet, FC-reagent, distilled water.
drug class. It obeys beer lamberts law in a range of 20-120 µg/ml.Folin ciocalteau reagent
reacts with ciprofloxacin to form chromogen that can be detected spectrophotometrically. The
PROCEDURE
For calibration curve: 0.5 mg/ml of stock solution was prepared by taking 50mg of drug in
100ml of water. Pipette out 2, 3, 4, 5, 6ml and diluted to 25ml and add 1ml FeCl3 and 1ml FC
reagent and then diluted to 25ml. Which gives concentration of 40, 60, 80, 100 and 120
µg/ml. The absorbance was measured and calibration curve was plotted.
ASSAY: Weigh and powder tablets and take the weight equivalent to 50mg and diluted to
100ml with distilled water.From the above solution pipette out 2ml and add 1ml FeCl3 and
1ml folin ciocalteau reagent and make up to the mark with distilled water to 25ml. The
AIM: To estimate the amount of Ranitidine HCl pure drug and tablets, distilled water.
This method is based on the condensation of RNH with PDAB to form a red coloured
product. The absorbance of coloured product measured is a quantitative measure of
concentration of RNH. The absorbance was measured at 503nm.
PDAB solution of 0.025N was prepared in 2N HCl. Ranitidine HCl stock solution containing
1000µg/ml RNH was prepared by dissolving 100mg of pure drug in water and diluting to the
mark in a 100ml calibrated flask.
Aliquots containing 50-350µg/ml RNH were transferred into a series of 10ml standard flask
by means of a micro burette. To each flask 3ml of 0.025N PDAB was added and kept a side
for 20min for colour development. It was then diluted up to the mark with distilled water. The
absorbance was measured against reagent blank at 503nm. The graph of PDAB-RNH
condensation product and corresponding reagent blank was plotted by taking concentration
Vs absorbance.
ASSAY: Three tablets were taken and ground into a fine powder. Powder equivalent to
168mg of RNH was weighed into a 100ml calibrated flask, 60 ml of water was added and
shaken well and filtered. First 10ml portion of the filtrate was discarded. A suitable aliquot
was taken and add 3ml of 0.025N P-Dimethyl amino benzaldehyde was added and kept aside
for 20 min for colour development. It was then diluted up to the mark with distilled water.
The absorbance was measured against the reagent blank at 503nm.