DRUG ASSAY

Presenter:
Madiha Moid
(Pharmacist)
(I.B.B.P.S,DUHS,OJHA

ASSAY (DEFINITION)
Assay is analytical
procedure for assessing
the presence or amount
of a target entity (the
Analyte).

STEPS INVOLVED
3 steps in processing of ASSAY :

Solution Preparation

Sample & Standard preparation

Instrumentation & Calculation

DIFFERENT TECHNIQUES
Chemical Assays:
Coulimetry,
Spectrophotometry,
Spectrofluorimetry,
Chromatography,
Immunoassays
Microbiological assays

DIFFERENT TECHNIQUES
ASSAY METHOD

ADVANTAGES

LIMITATION

COULIMETRIC

Simple

Semi-quantitative

Rapid

Sample
Pretreatment

Inexpensive
SPECTROPHOTOMETRIC

Simple

Semi-quantitative

Rapid

Sample
Pretreatment

Inexpensive
HPLC

Reliable

Expensive

High resolution

Training of
personnel

Accurate

Sample
Pretreatment

Methodology Sources
1. Official Compendia
2. Petition Methods (NDAs and ANDAs)
3. Pharmacopeial Forum
4. Code of Federal Regulations(Title 21 of the
Code of Federal Regulations (CFR)
5. Journals and Other Literature
Laboratory Information Bulletin, Journal of Pharmaceutical Sciences

6. Links to Scientific Sites

CDER Medical Library web site

ASSAY METHOD
Diclofenac Sodium
(Spectrophotometic Method)

50

Spectrophotometry (UV ,VIS &

IR)

PRINCIPAL
Instrument

used to measure the absorbance of

light by compounds within a liquid

Based

on the BEER-LAMBERT LAW

absorbance is proportional to concentration

UV Spectroscopy

Sample dissolved in liquid; placed

in cuvette.

A range of UV light wavelengths

are passed through solution.

Some light will pass through; some

light will be absorbed.

ASSAY method
Diclofenac Sodium
(Spectrophotometic Method)

1. Solution Preparation:
0.01N NaoH
Dissolve 0.4gm of NaoH in Litre of
Solution.

ASSAY method
2.

Diclofenac Sodium
(Spectrophotometic Method)

Standard Preparation:

Weigh 50mg of standard in

100 vol. flask.
Make up vol with 0.01N
NaoH & sonicate.
Filter above solution.
Take 02ml of filtrate in 100ml
vol. flask.
Make up volume with 0.01N
NaoH & sonicate.

ASSAY method
3.

Diclofenac Sodium
(Spectrophotometic Method)

Sample Preparation:

10 tablets were weighed

individually.

# of Tablet

Weight (in mg)

219.9

218.5

223.9

220.4

218.1

219.9

218.4

223.6

218.5

10

220.4
= 2201.6 / 10 =
220.16

Mean

ASSAY

Diclofenac Sodium
method (Spectrophotometic Method)

Powder tablets. Powder material equivalent to 50

mg disclofenac sodium was taken in 100 vol.
flask and make up the vol with 0.01N NaoH &
sonicate.
= Avg wt of tab x req. quantity
label claim
= 220.16mg x 50mg
50mg
= 220.16mg

ASSAY

Diclofenac Sodium
method (Spectrophotometic Method)

Dilution:
Filter the solution.
Take 02ml from filtrate in 100ml
vol. flask.
Make up the volume with 0.01N
NaoH & sonicate.

ASSAY

Diclofenac Sodium
method (Spectrophotometic Method)

Absorbance of resulting solutions (Standard &

Sample)
measured
at
236nm
spectrophotometrically.

ASSAY

Diclofenac Sodium
method (Spectrophotometic Method)

Absorbance:
=

Standard: 0.3058
Sample: 0.2970

Abs of Sample x wt. of STD x Avg. wt. of tablets x Potency of STD x 100
Abs of Standard x wt. of sample x C: wt. X 100

= 0.2970 x 50.4 x 2 x 100 x 100 x 220.16 x 99.5 x 100

0.3058 100 100 220.4 2
50
100
= 96.40 %
For mg/tablet:
%age x label claim / 100
96.04 x 50 /100 = 48.02mg/tablet