ASSAY PPT-md.ppt

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). 

3 steps in processing of ASSAY :

Solution Preparation

Sample & Standard preparation

Instrumentation & Calculation

STEPS INVOLVED

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 methodDiclofenac Sodium

(Spectrophotometic Method)

1. Solution Preparation:

0.01N NaoH

Dissolve 0.4gm of NaoH in Litre of Solution.

ASSAY methodDiclofenac Sodium(Spectrophotometic Method)

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.

2. Standard Preparation:

ASSAY methodDiclofenac Sodium(Spectrophotometic Method)

10 tablets were weighed individually.

# of Tablet Weight (in mg)

1 219.9

2 218.5

3 223.9

4 220.4

5 218.1

6 219.97 218.48

223.69 218.510 220.4Mean = 2201.6 / 10 =

220.16

3. Sample Preparation:

ASSAY methodDiclofenac Sodium(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 methodDiclofenac Sodium(Spectrophotometic Method)

Dilution: Filter the solution. Take 02ml from filtrate in 100ml

vol. flask. Make up the volume with 0.01N

NaoH & sonicate.

ASSAY methodDiclofenac Sodium(Spectrophotometic Method)

Absorbance of resulting solutions (Standard & Sample) measured at 236nm spectrophotometrically.

ASSAY methodDiclofenac Sodium(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