Murli Krishna Pharma Private Limited

PROTOCOL FOR STABILITY STUDY OF

OMEPRAZOLE PELLETS
8.5 %W/W

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 Murli Krishna Pharma Private Limited

INDEX

SrNo Name Page no

1 Approval page 03
4 Finished product specification 04
5 Method of analysis 05
8 Stability schedule, report tables 09

Approval

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 Murli Krishna Pharma Private Limited

Post approval of this protocol will be the joint responsibility of the following
functional area

Functional area Name Signature Date

Quality assurance Mr. Sunil kumar.N

Production Mr. Doshi

Quality Control Miss.Sushila Kumari

Research and Mr. Srinivas rao

development

Authorized By: Dr.V.K.Shastri, Technical Director

Signature :

Omeprazole Pellets Specification

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01. Description : White to off white color spherical pellets.

02. Identification : Positive test for Omeprazole

03. Water by KF method : NMT 3.0% w/w.

04. Bulk density : Between 0.75 to 0.90 gm/ml

05. Gastric resistance : NLT 85.0% of label claim should retain as

residue after two hours in 0.1 N HCl.

06. Dissolution : NLT 75.0% of label claim should dissolve in

Phosphate buffer pH 6.8 in 45 minutes.

07. Assay : ± 5% w/w of the labeled amount of Omeprazole

08. Pellet mesh size : Retention of 90 % pellets of required mesh size

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Method of analysis of Omeprazole Pellets
01.Description: By physical observation.
02.Identification:
In the assay, the principle peak in the chromatogram obtained with sample solution has the same retention
time as that of the peak due to Omeprazole in the chromatogram obtained with standard solution.
03.Water By KF Method:
Take about 15-20 ml of methanol in titration vessel and neutralize the solvent with Karl Fischer Reagent.
Weigh accurately about 1.0 gm of powdered pellets, transfer to the
Pre-neutralized KF titration vessel, carry out the titration and note the titre value and calculate the
moisture content from the following formula.

Titre Value X KF Factor

Moisture Content = -------------------------------------------------
Weight of sample taken for titration
04. Bulk density:
Take about 10 g of pellets into a dry 25 ml measuring cylinder and apply 100 strokes and measure the
volume calculate gms in ml.
Formula: wt of the sample in gm / ml of the sample
05.Gastric Resistance
Dissolution test apparatus: Apparatus II as per USP
Medium : 0.1 M HCl, 900 ml
RPM : 100
Temp. : 37°C ± 0.5° C
Time : 2 hours
Sample Preparation: Place 900ml of 0.1 N HCl in the vessel, and assemble the apparatus. Allow the
medium to equilibrate to a temperature of 37°C ± 0.5° C. Transfer an accurately weight equivalent to 20
mg of Omeprazole pellets (Calculated based in the assay of the batch to be tested) in the dissolution vessel
and operate the apparatus for exactly for specified time. At the end of two hours drain the medium slowly
without loosing the pellets, transfer to a filter paper and dry the pellets by blotting with filter paper.
Transfer the pellets into 100 ml volumetric flask. Add 40 ml of 0.1 M NaOH and sonicator to dissolve.

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 Murli Krishna Pharma Private Limited
Dilute to the volume with 0.1 M NaOH. Mix and Centrifuge about 15 ml of solution for 5 minutes, dilute
5 ml of the clear supernatant liquid to 50 ml with mobile phase.
Procedure: Inject standard solution in duplicate and sample solution in single by follow the analytical
condition of mobile phase and chromatographic system as mentioned in the assay. Calculate the % content
of Omeprazole in the supernatant liquid as follows:

AT WS 5 100 50
Calculation: -------- X --------- X -------- X --------- X ------ X P
AS 100 50 WT 5
AT = Average peak area of sample preparation
AS = Average peak area of standard replicate injection
WS = Weight of working standard in mg
WT = Weight of sample taken in mg
P = Purity of working standard
Assay after Resistance
% of acid resistance = --------------------------------X 100
Assay of pellets
06.Dissolution: Apparatus required: Same as under gastric resistance
Medium : 900 ml of pH 6.8 phosphate buffer as per IP
RPM : 100
Temperature : 37° C ± 0.5° C
Time : 45 minutes
Standard Preparation: Transfer an accurately weighed quantity of about 20 mg of Omeprazole working
standard to a 100 ml volumetric flask. Add 20 ml of 0.1 M NaOH to dissolve. Make volume up to the
mark with 0.1 M NaOH and mix. Transfer 5 ml of the solution to 50 ml with dissolution medium.
Sample Preparation: Weigh a quantity of pellets equivalent to 20 mg of Omeprazole. Transfer
individually to six dissolution flasks having 900 ml 0.1 N HCl equilibrated to 37° C ± 0.5° C taking care
to exclude air bubbles from the surface of the pellets and start the apparatus at 100 rpm for 2 hours. After
2 hours drain the medium without loosing the pellets, add 900 ml of pre heated buffer solution. Start the

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apparatus at 100 rpm for 45 minutes, collect the sample from zone midway between surface of dissolution
medium and top of rotating paddle, not less than 1.0 cm from the vessel wall. Centrifuge at 5000 rpm for 5
minutes of the above solution, and use clear supernatant liquid.
Procedure: Inject standard preparation in duplicate and sample preparation in single by follow the
analytical conditions of mobile phase and chromatographic system as mentioned in the assay.
Calculate the % content of Omeprazole dissolved in the phosphate buffer in the supernatant liquid.
AT WS 5 900
Calculation: -------- X --------- X -------- X --------- X P
AS 100 50 WT
AT= Average peak area of sample preparation
AS = Average peak area of standard replicate injection
WS = Weight of working standard in mg
WT = Weight of sample taken in mg
P = Purity of working standard

Assay after dissolution

DISSOLUTION % IN BUFFER: = --------------------------------X 100
Assay of pellets

07. Assay: Chromatographic condition

Column: 4.6mm ID x 25cm long. Packaged with C18 with particle size 5µ.
Flow Rate: 1.0 ml/min.
Wave length: 302 nm
Buffer PH: Dissolve 6.8 gm of potassium dihydrogen orthophosphate in 1000ml water. Adjust pH 7.4
with 0.1 M sodium hydroxide.
Mobile Phase: Prepare a suitable quantity of a filtered and degassed mixture of 65 volumes of phosphate
buffer pH 7.4 and 35 volumes of acetonitrile.
Standard preparation: Transfer an accurate weighed quantity of about 20 mg of Omeprazole working
standard to a 100ml volumetric flask. Add 40 ml of 0.1 M Sodium hydroxide and sonicate to dissolve.
Make volume up to the mark with 0.1 M sodium hydroxide and mix. Transfer 5 ml of the solution to 50
ml with mobile phase and mix.

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Sample Preparation: Take about 5 gms of pellets into a mortar and pestle and grind the pellets into a
uniform fine powder. Weigh accurately about a quantity equivalent to 20 mg of Omeprazole into a dry
100 ml volumetric flask, add about 50 ml of 0.1 M NaOH and sonicate to dissolve. Make volume up to the
mark with 0.1 M sodium hydroxide and mix. Transfer 20 to 30 ml of solution into dry stoppered test-tube,
centrifuge at 5000 rpm for 5 minutes. Further, dilute 5 ml of the supernatant solution to 50 ml with mobile
phase.
Procedure: Inject sample preparation in duplicate into the chromatograph and record the chromatograms.
Measure the response for the major peaks. Calculate the quantity in % w/w of Omeprazole by using below
formula.
AT WS 5 100 50
Calculation: -------- X --------- X -------- X --------- X --------X P
AS 100 50 WT 5

AT= Average peak area of sample preparation

AS = Average peak area of standard replicate injection
WS = Weight of working standard in mg
WT = Weight of sample taken in mg
P = Purity of working standard
08. Pellets mesh size:
Place 100 g of sample in a lab sifter containing standard required mesh sieves. And calculate retention
of the pellets

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TABLE I

LONG TERM STABILITY OF OMEPRAZOLE PELLETS AT STORAGE TEMP. (25°C ± 2°C / RH60 % ± 5 %)

Batch No.:
Mfg Date:
Exp Date:
Period (Months)
No Parameter 0 3 6 9 12 18 24 36
1. Description

2. Dissolution

Acid medium: NLT 85%

Buffer Medium: NLT 70 %

3. Assay Limits:
8.075 – 8.925 % w/w

ANALYST CHECKED BY: MANAGER Q.A.

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TABLE II

ACCELERATED STABILITY OF OMEPRAZOLE PELLETS AT TEMP. (40°C ± 2°C / 75 % RH ± 5 %)

Batch No.:
Mfg. Date:
Exp Date:
Period (Months)
No Parameter 0 1 2 3 6
1. Description

2. Dissolution

Acid medium: NLT 85%

Buffer Medium: NLT 70 %

3. Assay Limits:
8.075 – 8.925 % w/w

ANALYST CHECKED BY: MANAGER Q.A.

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