9.Methodology

8/6/2019 9.Methodology

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Chapter.5 Methodology

NIMS Institute Of Pharmacy, NIM S Un iversity Jaipur Page 51

5. METHODOLOGY

5.1 Characterization of API

1. Official monograph: In house

2. Description: A white to off-white powder

3. Identification:

By HPLC: The retention time of the principal peak in the sample preparation for

assay should corresponds with the retention time of the principal peak in the standard

preparation for assay1.

4. Related substances:

Unknown Impurities: Not more than 0.1%

Total Impurities: Not more than 0.5%

5. Assay by HPLC: 99.87% (98% - 102%w/w)

6. Loss of drying: 0.67% (Not more than 1.0%)

7. Heavy metals: Not more than 0.001% w/w

8. Sulphated ash: Not more than 0.2% w/w

9. pH: Observed – 4.0 Specification: 3.5 – 5.0

10. Melting point: 142~146c

11. Half-life:13 hour

5.2 Selection and justification of excipients2

Diluents: In view of the low or medium dose of drug it is essential to add bulking agents or

diluents to increase the weight of the tablet. Dibasic calcium phosphate was used as diluents

because of its good compaction property and good flow property of the coarse grade material

and also it is insoluble in water. So used as diluents in SR preparation.

Matrix-forming polymers: HPMC which is most widely used matrix-forming polymer in

these days because of its excellent compatibility, multifunctional property. It is available in

different grades depending upon its attached group. It enables control time release of active

ingredient by pH independent swelling and permeability.





Lubricants: Magnesium Stearate, talc &colloidal silicon dioxide as lubricating agent.

5.2 Preformulation study

Preformulation can be defined as investigation of physical and chemical properties of drug

substance alone and when combined with excipients.

Preformulation studies are the first step in the rational development of dosage form of a drug

substance. The objectives of preformulation studies are to develop a portfolio of information

about the drug substance, so that this information is useful to develop formulation.

Preformulation investigations are designed to identify those physicochemical properties and

excipients that may influence the formulation design, method of manufacture, and

pharmacokinetic-biopharmaceutical properties of the resulting product. Followings studies

performed for in the preformulation study.3-7

5.3 Physical appearance:-

5.3.1 Description

Colour: - A white to off-white powder

Table 5.1 Ingredients and their function

Sr.

NO

Ingredients Function

1. AED API

2. LACTOSE MONOHYDRETE Diluent

3. HPMC K4M Thickening agent; viscosity-increasing

agent.

3. HPMC K100M Matrix-forming polymer

4. HPMC K15M Matrix-forming polymer

5. IPA Binder solution

6. Microcrystalline

cellulose(MCC)

Diluent

7. Colloidal silicon dioxide Glidant (0.25-3%)

8. TALC Glidant

8. Magnesium stearate Lubricant (0.25-4%)







5.3.4 Carr’s index

The Compressibility Index of the powder blend was determined by Carr’s compressibility

index. It is a simple test to evaluate the BD and TD of a powder and the rate at which it

packed down. The formula for Carr’s Index is

as below:

Carr’s index (%) = [(TD – BD)*100] / TD…..

5.3.5 Hausner’s ratio

The Hausner’s ratio is a number that is correlated to the flowability of a powder or granular

material.

Hausner’s ratio = TD / BD…………..

Table 5.2 Effect of carr’s index and hausner’s ratio on flow property

Carr’s index (%) Flow character Hausner’s ratio

< 10 Excellent 1.00 – 1.11

11 – 15 Good 1.12 – 1.18

16 – 20 Fair 1.19 – 1.25

21 – 25 Passable 1.26 – 1.34

26 – 31 Poor 1.35 – 1.45

32 – 37 Very poor 1.46 – 1.59

>38 Very, very poor >1.60

5.3.6 Angle of repose

The angle of repose of API powder was determined by the funnel method. The accurately

weight powder blend were taken in the funnel. The height of the funnel was adjusted in such a

way the tip of the funnel just touched the apex of the powder blend. The powder blend was

allowed to flow through the funnel freely on to the surface.





The diameter of the powder cone was measured and angle of repose was calculated using the

following equation.

Where, h and r are the height and radius of the powder cone respectively.

5.3.7 Particle size analysis: ( By malvern master seizer, dry method)

For many active substances, particle size has an impact on powder flow; content uniformity

and drug dissolution. In order to assure consistent product quality, the particle size of the API

has been characterized. From the results obtained, the limits will be derived which will be

routinely applied by the API manufacturer during analysis of drug.

Particle size of drug was determined by Malvern particle size analyser.

D (0.10) = 2.39 µ, D (0.50) = 7.37 µ , D (0.90) = 19.46 µ.

5.3.8: Assay

Standard solution

Parameters for Preparation of Standard Solution

Column: Inertsil ODS 3r (250×4.6)

Colum Id: AD3/Lc-074/10

Wavelength: 220 nm

Flow rate: 1.0ml/min

Injection volume: 10µl

Column temp. : 45°C

Diluent: Water: CAN

First prepare buffer solution by 2.7gm KHPO4in 2 litre purified water. Then prepare mobile

phase by addition of ACN with the ratio of 40:60.Then prepare diluent with addition of water:

methanol (40:60). Finally 50 mg API is added in to the 25 ml diluent. The λmax of API in the

above media was determined by scanning a suitable dilution of the stock. From the stock





solution, various dilutions were made to obtain solutions of 1, 5, 10, 15 and 20 _g/ml, and

absorbance was measured for each dilution.

5.3.9 Drug excipients compatibility study:

API and excipients were been thoroughly mixed in predetermined ratio given in below table

and passed through the 40# sieve. The blend was to be filled in transparent glass vials and

were closed with gray coloured rubber stoppers and further sealed with aluminium seal and

charged in to stress condition at above condition. Similarly API should also be kept at all

condition as for the samples. Samples were withdrawn for analysis within two day of

sampling date as per the compatibility study plan. Physical observation should be done at

every week up to 1 month and DSC studies were carried out to determine the compatibility of

excipients with the drug.8

5.3.10In –Vitro Release study of theoretical and Hypothetical Release profile

Dissolution parameter:

Medium: 0.1N Hydrochloric acid

Volume: 900ml

Apparatus: IP (Paddle)

RPM: 50 rpm

Time point: 1, 2, 4, 6, 8, 10, 12 hrs.

Temperature: 37°C ± 0.5°C

Table 5.3In – Vitro Release study of Innovator and Hypothetical Release profile

Dissolution 900ml, USP - II (Paddle) Apparatus, 50 RPM

Time (hrs.) Theoretical % Drug Hypothetical % Drug Release

0.1N HCL

1 23.9 15-25%

2 36 30-35%

PH6.8 Phosphate buffer

4 53.5 50-55%

8 89.6 85-90%

12 97.6 NLT 90%





5.3.11 Drug- Excipient Compatibility Study

Figure 5.5 Drug- Excipient Compatibility Study

No:

Ingredients

Ratio

of Dry

mix

Drug:

Excipi

ent

Initial 1 week 2 week 3 week 4 week

1 API ----- White

crystalline

powder

No

change

No

change

No

change

No change

2 LACTOSE

MONOHY

DRETE

1:1 White

crystalline

powder

no

change

no

change

no

change

No change

3 HPMC

K4M

1:1 White

crystalline

powder

No

change

No

change

No

change

No change

4 HPMC

K100M

1:1 White

crystalline

powder

No

change

No

change

No

change

No change

5 HPMC

K15M

1:1 White

crystalline

powder

No

change

No

change

No

change

No change

6 Microcrys

talline

cellulose(

MCC)

1:1 White

crystalline

powder

No

change

No

change

No

change

No change

7 Colloidal

silicon

dioxide

1:1 White

crystalline

powder

No

change

No

change

No

change

No change

8 TALC 1:1 White

crystalline

powder

No

change

No

change

No

change

No change

9 Magnesiu

m

stearate

1:1 White

crystalline

powder

No

change

No

change

No

change

No change





5.4. Formulation of trials:

5.4.1 (A) Method of preparation of Extended release tablets:

Method: API SR Tablets were prepared by Wet granulation technique.

For F/001 to F/007:

Step 1: Sifting: API was passed through 16 #sieve, and pass HPMC K4M, HPMC K15M, LACTOSE

MONOHYDRETE through 40#seive

Step 2: Wet Granulation: by ISOPROPAYL ALCOHOL

5.6.FORMULATION OF TRIALSF001 TO F003

SR

NO

INGREDIENT F-1 F-2 F-3

1 QUANTITY OF DRUG 300 300 300

2 LACTOSE MONOHYDRATE 177 177 169.5

3 HPMC K4M - - -

4 HPMC K15M 40 - -

5 HPMC K100M 40 50

6 IPA Q.S Q.S Q.S

7 MCC 59 59 56.5

8 COLLOIDAL SILICONE

DIOXIDE

6 6 6

9 TALC 12 12 12

10 MG STEARATE 6 6 6

11 TOTAL 600 600 600





Granulation:

Table 5.8 Wet granulation parameters

BINDER Time Impeller

speed

Chopper speed

Dry

mixing

10minute 150RPM Off

Binder

addition

150ml 2minute 150RPM Off

Extra

IPA

addition

40ml 2 minute 150RPM Off

Kneading 2minute 150RPM 1500RPM

Step 3: Drying:

Granules were dried in Fluidized bed dryer.

Inlet temperature: 45°C

Outlet temperature: 35 °C

LOD: 0.62 to 1.03 %

Step 4: Sifting:

Granules were passed through 20 # by manually.

And retained 20# grenules passed through co-mill 1905 sieve

Step 5: calculation

Calculation of extra granular part

Step 6: shifting

MCC (Microcrystalline cellulose), colloidal silicone dioxide and TALC were passed through

40 # and Magnesium stearate through 60 #.





Step 5: Blending and Lubrication:

Blending time for MCC, TALC & Colloidal silicon dioxide was 10 minutes and lubricated

with magnesium stearate for 3 minutes.

Step 6: Compression:

The prepared blend was compressed (16 × 8.0” mm oval shape standard shape with concave,)

using 16 station tablet compression machine (Cadmach, Ahmedabad, India).

5.4.1 (B) Evaluation of powder blend:

Evaluation parameters of powder blend were done as in Preformulation study.

5.4.1 (C) Evaluation of tablets:

1. Appearance:

Twenty tablets of each formulation were taken to check any discoloration or surface ruffness

in the tablet formulation.

2. Weight variation test:

To study weight variation twenty tablets of the formulation were weighed using a Mettler

Toledo electronic balance and the test was performed according to the official method.9

3. Hardness:

The hardness of five tablets was determined using the Dr.schleunizer type hardness tester and

the average values were calculated.

4. Thickness and diameter:

The Thickness and Diameter of the tables was determined by using Digital vernier calipers.

Five tablets were used, and average values were calculated.

5. Friability:

The friability of twenty tablets was measured by Roche friabilator for 4min at 25rpm for 100

revolutions. Accurately weigh twenty tablets placed into Roche friabilator for 100 revolutions

than dedust the tablets and weigh.

% Friability = (W0 - W)* 100 / W…..





5.5. Formulation of trialsF004 TO F008:-

5.5.1 (A) Method of preparation of Extended release tablets:

Method: API SR Tablets were prepared by Wet granulation technique.

For F/001 to F/007:

Step 1: Sifting: API was passed through 16 # sieve, 0and pass HPMC K4M, HPMC K15M,

LACTOSE MONOHYDRETE through 40#seive

Step 2: Wet Granulation: by ISOPROPAYL ALCOHOL

Table 5.9 Formulation of trialsF004 TO F008

Sr

no

INGREDIENT F/004 F/005 F/006 F/007

1 QUANTITY OF DRUG 300 300 300 300

2 LACTOSE

MONOHYDRATE

162 162 162 162

3 HPMC K4M - 10 20 30

4 HPMC K15M - - - -

5 HPMC K100M 60 50 40 30

5 IPA Q.S Q.S Q.S Q.S

6 MCC(MICROCRYSTE

LLAIYINE

CELLULOSE)

54 54 54 54

7 COLLOIDAL SILICONE

DIOXIDE

6 6 6 6

8 TALC 12 12 12 12

9 MGNESIUM

STEARATE

6 6 6 6

10 TOTAL 600 600 600 600





Granulation:

Table 5.10 Wet granulation parameters

BINDER Time Impeller

speed

hopper speed

Dry

mixing

10minute 150RPM Off

Binder

addition

150ml 2minute 150RPM Off

Extra

IPA

addition

40ml 2 minute 150RPM 1500RPM

Kneading 2minute 150RPM 1500RPM

Step 3: Drying:

Granules were dried in Fluidized bed dryer.

Inlet temperature: 45°C

Outlet temperature: 35 °C

LOD: 0.62 to 1.03 %

Step 4: Sifting:

Granules were passed through 20 # by manually.

And retained 20# grenules passed through co-mill 1905 sieve

Step 5: calculation

Calculation of extra granular part

Step 6: shifting

MCC (Microcrystalline cellulose), colloidal silicone dioxide and TALC were passed

through 40 # and Magnesium stearate through 60 #.

Step 5: Blending and Lubrication:

Blending time for MCC, TALC & Colloidal silicon dioxide was 10 minutes and lubricatedwith magnesium stearate for 3 minutes.







5.5.2 (D).4 Selection of packaging material:

Tablets will be packed in a blister formed by the following primary packaging components.

Base foil: PVC/PVDC blister pack.

Lidding Material: 0.025 mm aluminum foil

Justification: PVC/PVDC Blister pack provides complete protection against light, water vapour, gases

etc.

5.6Conclusion

From the result, concluded that batch taken in combination with HPMC K4M&HPMC

K100M (30+30%) in has good sustained property. Batch F007 was charged for Accelerated

stability study.

5.7. Stability studies of optimized tablets

The ICH Guidelines have established that long term stability testing should be done at

25°C±2°C / 60%±5% RH; stress testing should be done at 40°C±2°C / 75%±5% RH for 6

months. If significant change occurs at these stress conditions, then the formulation should be

tested at an intermediate condition at 30°C±2°C /75%±5% RH. Table 6.7 shows different

temperatures and period of stability testing.

Table 5.11 ICH guidelines for stability study

Study Storage condition

Minimum time period

covered by data at

submission

Long term 25°C±2°C / 60%±5% RH 12 months

Intermediate 30°C±2°C / 65%±5% RH 6 months

Accelerated 40°C±2°C / 75%±5% RH 6 months

The stability studies of the optimized tablets were carried out at 40○C temperature and 75 %

relative humidity (accelerated stability) in stability chamber for three months .Tablets were

withdrawn at 1, 2, 3 months intervals and evaluated for disintegration time, hardness, drugcontent and in vitro release.

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9.Methodology