CHAPTER ––– 333 Preformulation...

Chapter – 3 Preformulation Study

JJT University Page 30

CHAPTER CHAPTER CHAPTER CHAPTER –––– 3333

Preformulation StudyPreformulation StudyPreformulation StudyPreformulation Study 3.0 Introduction

As per Sir Arthur Conan Doyle, “It is a capital mistake to theorize before one has data”.

Preformulation work is the foundation for development of any robust formulations (G.

Banker et al., 2000). Preformulation is defined as a stage of development during which

the physicochemical properties of drug substance are characterized (L.J. Ravin et al,

1990). Almost all drugs are marketed as tablets, capsules or both. Prior to the

development of these major dosage forms, it is essential that to pertain fundamental,

physical and chemical properties of the drug molecule and other essential properties of

the drug powder are determined. This information decides many of the subsequent events

and approaches in formation development (H. Brittain et al, 1995; E.F. Fiese et al. 1986).

Preformulation involves the application of biopharmaceutical principles to the

physicochemical parameters of drug substance to characterize the designing of optimum

drug delivery system. Prior to the development of any dosage form new drug, it is

essential that certain fundamental physical & chemical properties such as

characterization, solubility, melting point, compatibility etc of drug powder and

formulation are determined. This information may dictate many of subsequent event &

approaches in formulation development. The wrong approach of Preformulation could

affect drug performance and development of an efficacious dosage form”.

The aim of this section was to investigate a robust prototype formulation of rapid

dispersible tablets. The Preformulation of formulation is investigated by using

characterization of active, and drug excipients compatibility study.

� Characterization of active

� Drug Excipients compatibility

On the basis of current objective, the Preformulation study was designed as,

1. Characterization of Actives such as Tolfenamic Acid and Paracetamol

2. Drug excipients compatibility study of Rapid Dispersible Tablets of Paracetamol



3. Drug excipients compatibility study of Rapid Dispersible Tablets of Tolfenamic

Acid

4. Drug excipients compatibility study for Fixed Dose Combination of Tolfenamic

Acid and Paracetamol

3.1 Materials and Equipments

3.1.1 Materials

List of raw materials used in formulation of rapid dispersible tablets is summarized in

Table 3.1.

Table 3.1: List of Materials Used in Formulation and Development

Excipients Grade Manufacturer

Tolfenamic Acid BP Elder Pharma.

Paracetamol IP/BP Ankur Drugs

Lactose Monohydrate IP DMV Fonterra

Microcrystalline Cellulose IP Sigachi

Mannitol IP Roquette

β-Cyclodextrin (Betadex) BP Emcure Pharma

Polyethylene Glycol IP Elder Pharma

Hypromellose USP Shin-Etsu Chemicals

Eudragit –EPO USP Evonik Industries

Urea IP Cadila Pharma

Povidone (PVP K-30) IP FMC Biopolymer

Croscarmellose sodium (Ac-Di-SolTM) IP FMC Biopolymer

Crospovidone (Kollidon CLTM) IP BASF Limited

Sodium Starch Glycolate Type A(ExplotabTM) IP Cadila Pharma

Polacrillin Potassium (Kyron T-314TM) USNF Corel Pharma Chem

Sodium Lauryl Sulfate IP Parchem

Magnesium Stearate IP Pharma Trans

Purified Talc IP Elder Pharma

Aspartame IP Elder Pharma

Flavor Vanilla IH Flavors International


JJT University 2

Flavor Strawberry IH Flavors International

Ethanol and Methanol IH Shree Chemicals

3.1.2 Equipments

List of manufacturing and analytical equipments used in formulation development of

rapid dispersible tablets is summarized in Table 3.2.

Table 3.2: List of Equipments Used in the Formulation Development.

Name of the Equipment Manufacturer

Electronic weighing balance Sartorius

Rapid Mixer Granulator (H.S.M.G – 10) Kevin Machinery

Air jet Mill Shree Engg.

16 Station compression machine Cadmach Machinery Co. Pvt. Ltd

Hardness tester Erweka

Digital Vernier Calipers Mitutoyo, Japan

Friability Apparatus Electrolab

Disintegration time tester Electrolab

Moisture analyzer Sartorius

Bin Blender Solace Engg.

Vacuum tray dryer Shree Engg.

Fluid Bed Processor Pam Glatt

Tap density tester Electro lab

Electromagnetic sieve shaker Electrolab (USP)

Oscillating granulator Frewitt

Mechanical stirrer Remi Electronics, India

Dissolution Equipment Electrolab

Fourier Transform Infrared Spectroscopy Shimadzu

U V Spectroscopy Shimadzu

High Performance Liquid Chromatography Shimadzu



Equipment

Melting point apparatus Veego

3.2 Characterization of API (British Pharmacopoeia, 2010)

3.2.1 Physical Appearance

Physical appearance of Tolfenamic Acid is white or slightly yellow, crystalline powder

(British Pharmacopoeia, 2010).

Physical appearance of Paracetamol is white or almost white crystalline powder (British

Pharmacopoeia, 2010).

A small sample of API was taken for the visual observation.

3.2.2 Melting Point

Confirmation of melting point for the actives was determined by open capillary method.

The capillary tube with opens from one side was filled with active by repeated tapping to

fill a sufficient amount of drug. The melting point was measured by using digital

melting point apparatus. The temperature at which the drug started melting was recorded.

The triplicate reading was recorded by repeating the same procedure. The melting point

of active was compared with the reported melting point of actives as per British

Pharmacopoeia.

3.2.3 Infra-red (FTIR) Analysis

The identification of actives was also confirmed by using Infra-red analysis of drug

sample. The FTIR spectra were obtained using FTIR spectrometer (FTIR 8001,

Shimadzu). The Drug sample were previously triturated and mixed thoroughly with

potassium bromide in 1:5 (Sample: KBr) ratio, KBr discs were prepared by compressing

the powders at a pressure of 5 tons for 5 min in hydraulic press (Sushma Talegaonkar et.

Al, 2007). Scans were obtained at a resolution of 2 cm 1 from 4500 to 400 cm. It was

carried out to check for functional groups of drug. The spectrum was than compared with

standard spectrum of actives as per the British Pharmacopoeia.

3.2.4 U V Spectrophotometer



The identification of actives was characterized by using UV spectrophotometry. The

method is adopted from British Pharmacopoeia.

Tolfenamic Acid

20 mg of Tolfenamic acid was dissolved in a mixture of 1 volume of 1 M hydrochloric

acid and 99 volumes of methanol R and dilute to 100 ml with the same mixture of

solvents. 5.0 ml of above solution was again diluted to 50 ml with a mixture of 1 volume

of 1 M hydrochloric acid and 99 volumes of methanol R. The diluted solution than

examined between 250 nm and 380 nm, the solution shows 2 absorption maxima, at 286

nm and 345 nm. The ratio of the absorbance measured at the maximum at 286 nm to that

measured at the maximum at 345 nm is 1.2 to 1.4.

Paracetamol

Take 50 mg of sample and dissolve in small quantity of methanol R and make the volume

up to 100 ml. Take 2.0 ml of solution to 0.5 ml of a 10.3 g/lt solution of hydrochloric acid

R and dilute to 100.0 ml with methanol R. Immediately measure the absorbance of test

solution at the absorption maximum at 249 nm. The specific absorbance at the maximum

is 860 to 980.

3.3 Drug Excipients Compatibility Study

For the development of rapid dispersible tablet formulation, the pharmaceutical

development was initiated with drug-Excipients compatibility study. The Preformulation

study was conducted to formulate two types of formulation. Earlier formulation is

formulation of Rapid dispersible tablets of Tolfenamic acid, and Paracetamol while later

is combination of Tolfenamic acid and paracetamol. The selection of excipients for all the

formulation was based on reference innovators and various literature searches of

respective formulations. The Preformulation details of remaining formulation and drug

excipients study was summarized in Table 3.3.



Table 3.3: Preformulation Details of Drugs and Excipients

Drug + Excipients Drug Excipients Ratio

Tolfenamic Acid 1 gms

Paracetamol 1 gms

TA + Mannitol 1:1

TA + Microcrystalline Cellulose 1:1

TA + Sodium Lauryl Sulfate 1:1

TA + PVP K-30 1:1

TA + Polacrillin Potassium 1:1

TA + Croscarmellose Sodium 1:1

TA + Crospovidone 1:1

TA + Sodium Starch Glycolate 1:1

TA +Magnesium Stearate 1:1

TA + Aspartame 10:1

TA + Vanilla Flavor 10:1

TA + Strawberry Flavor 10:1

PA + Mannitol 1:1

PA + Microcrystalline Cellulose 1:1

PA + β-Cyclodextrin 1:1

PA+ Urea 1:1

PA+ Lactose Monohydrate 1:1

PA + Sodium Lauryl Sulfate 1:1

PA + PVP K-30 1:1

PA + Polacrillin Potassium 1:1

PA + Croscarmellose Sodium 1:1

PA + Crospovidone 1:1

PA + Sodium Starch Glycolate 1:1

PA +Magnesium Stearate 1:1

PA + Aspartame 10:1



PA + Vanilla Flavor 10:1

Pa+ Strawberry Flavor 10:1

TA + PA + Mannitol 1:1:1

TA + PA + Microcrystalline Cellulose 1:1:1

TA + PA + β-Cyclodextrin 1:1:1

TA + PA+ Urea 1:1:1

TA + PA+ Lactose Monohydrate 1:1:1

TA + PA + Sodium Lauryl Sulfate 1:1:1

TA + PA + PVP K-30 1:1:1

TA + PA + Polacrillin Potassium 1:1:1

TA + PA + Croscarmellose Sodium 1:1:1

TA + PA + Crospovidone 1:1:1

TA + PA + Sodium Starch Glycolate 1:1:1

TA + PA +Magnesium Stearate 1:1:1

TA + PA + Aspartame 10:10:1

TA + PA + Vanilla Flavor 10:10:1

TA + Pa+ Strawberry Flavor 10:10:1

Blend of TA with all excipients of TA mentioned for individual study to make prototype formulation for Rapid dispersible tablets of Tolfenamic Acid

Approx 5 gms

Blend of PA with all excipients of PA mentioned for individual study to make prototype formulation for Rapid dispersible tablets of Paracetamol

Approx 5 gms

Blend of TA + PA with all excipients of TA + PA mentioned for individual study to make prototype formulation for fixed dose combination

Approx 5 gms

3.3.1 Physical Observation

Physical observation such as color, texture, and appearance during Preformulation study

were evaluated for initial and after 15, 30 days interval as per the scheme of drug

excipients compatibility. The scheme for Drug substance-excipients Compatibility study

is tabulated in the Table 3.4.



Table 3.4: Scheme for Drugs - Excipients Compatibility Studies

Sr. No. Condition Time -point Type of packing

1 Initial Zero day Glass vial

2 Stress Condition 50 0 C Up to 15 days Glass vial

3 400C/ 75% RH Up to 30 days Glass vial

3.3.2 Analytical Observation by Spectrophotometric Evaluation

The drug excipients compatibility study also confirmed with spectrophotometric

evaluation. The comparative evaluation of spectrophotometric spectrum for active such

as Tolfenamic Acid, Paracetamol, blend of Tolfenamic Acid, blend of Paracetamol at

initial stage of Preformulation and after completetion of Preformulation studies were

investigated for any degradation.

The FTIR spectra were obtained using FTIR spectrometer (FTIR 8001, Shimadzu). The

samples were previously triturated and mixed thoroughly with potassium bromide in 1:5

(sample: KBr) ratio, KBr discs were prepared by compressing the powders at a pressure

of 5 tons for 5 min in hydraulic press. Scans were obtained at a resolution of 2 cm -1

from 4500 to 400 cm-1.

3.4 Result and Discussion

3.4.1 Characterization of API

(British Pharmacopoeia, 2010; Handbook of Pharma. Excipients, 2009)

3.4.1.1 Physical Appearance

The physical appearance of Tolfenamic Acid and Paracetamol was compared as per the

specification of British Pharmacopoeia 2010 and it complies as per the standard

description of actives. The observation of description is summarized in Table 3.5.



Table 3.5: Physical Observation of Tolfenamic Acid and Paracetamol

Name of Active Description as per British

Pharmacopoeia, 2010 Observation

Tolfenamic Acid white or slightly yellow,

crystalline powder Complies

Paracetamol white or almost white

crystalline powder Complies

3.4.1.2 Melting Point

The average melting point of Tolfenamic acid and Paracetamol by using digital melting

point apparatus is tabulated in table. The melting point of active (Table 3.6) is confirmed

as per the standard value given in the British pharmacopoeia 2010.

Table 3.6: Determination of Melting Point of Tolfenamic Acid and Paracetamol

(mean ± S.D, n = 6)

Description Standard Melting Point

(BP, 2010) Observation

Tolfenamic Acid About 213 °C 212.5 ± 0.30C

Paracetamol 168 °C to 172 °C 170.2± 0.250C

3.4.1.3 Infra-red (FTIR) Analysis

The identification of Tolfenamic acid and Paracetamol was confirmed by using Infra-red

analysis of drug sample, the spectra of standard reference (British Pharmacopoeia, 2010)

and taste reference (sample of Tolfenamic acid and Paracetamol) are presented in Graph

3.1 to 3.6. On the basis of comparison of spectrum the taste spectrum is confirmed as

Tolfenamic acid and Paracetamol.



A. Infra-red (FTIR) Analysis - Paracetamol

Graph 3.1: IR Spectra of Paracetamol (Test Sample)

Graph 3.2: IR Spectra of Paracetamol (Reference Sample - WS)



Graph 3.3: Comparative IR Spectra of Paracetamol (Test vs. Reference)

B. Infra-red (FTIR) Analysis – Tolfenamic Acid

Graph 3.4: IR Spectra of Tolfenamic Acid (Test Sample)



Graph 3.5: IR Spectra of Tolfenamic Acid (Reference Sample)

Graph 3.6: Comparative IR Spectra of Tolfenamic Acid (Test vs. Reference)



3.4.1.4 U V Spectrophotometer

A - Determination of λmax – Paracetamol

λmax of Paracetamol was observed at 249 nm which was then chosen as wavelength for

detection of drug.

Graph 3.7: UV Spectrum of Paracetamol

Table 3.7: Observation for Calibration Curve of Paracetamol

Con µg/ml Abs in nm

0.0060 0.540

0.0080 0.735

0.0100 0.890

0.0120 1.061

0.0140 1.249



Graph 3.8: Calibration Curve of Paracetamol

B - Determination of λmax – Tolfenamic Acid

λmax of Tolfenamic Acid was observed at 286 nm which was then chosen as wavelength

for detection of drug.

Graph 3.9: UV spectrum of Tolfenamic Acid



Table 3.8: Observation for Calibration Curve of Tolfenamic Acid

Con µg/ml Abs in nm

2.5 0.134

5.0 0.242

7.5 0.366

10.00 0.483

12.50 0.622

15.00 0.753

17.50 0.854

Graph 3.10: Calibration curve of Tolfenamic Acid

Conclusion

On the basis of characterization of API, the Tolfenamic Acid and Paracetamol comply as

per the given specification active in British Pharmacopoeia, 2010.

y = 0.049x + 0.002

R² = 0.998

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

0 5 10 15 20

Series1

Linear (Series1)

Linear (Series1)



3.4.2 Drug Excipients Compatibility Study

3.4.2.1 Physical Observation

Physical observation such as color, texture, and appearance and any significant changes

during Preformulation study were evaluated for initial and after 15, 30 days interval. On

the basis of stress study at 500C for 15 days and accelerated condition at 400C 75% RH

for 30 days, there was no any significant changes observed in Preformulation study. So

the selection of excipients does not have any physical effect on stability. The observation

of Preformulation is also summarized in Table 3.9.

Table 3.9: Preformulation details of drug excipients

Drug + Excipients

Physical Appearance

Initial Observation 15 days 500C

30 days 400C 75%

RH Tolfenamic Acid off – white colored powder NC NC

Paracetamol off – white colored powder NC NC

TA + Mannitol off – white colored powder NC NC

TA + MCC off – white colored powder NC NC

TA + SLS off – white colored powder NC NC

TA + PVP K-30 off – white colored powder NC NC

TA + Polacrillin Potassium off – white colored powder NC NC

TA + CCS off – white colored powder NC NC

TA + Crospovidone off – white colored powder NC NC

TA + SSG off – white colored powder NC NC

TA +Magnesium Stearate off – white colored powder NC NC

TA + Aspartame off – white colored powder NC NC

TA + Vanilla Flavor Sweet off – white colored

powder NC NC

TA + Strawberry Flavor Flavored off – white colored

powder NC NC

PA + Mannitol Flavored off – white colored

powder NC NC

PA + MCC off – white colored powder NC NC



PA + β-Cyclodextrin off – white colored powder NC NC

PA+ Urea off – white colored powder NC NC

PA+ Lactose Monohydrate off – white colored powder NC NC

PA + SLS off – white colored powder NC NC

PA + PVP K-30 off – white colored powder NC NC

PA + Polacrillin Potassium off – white colored powder NC NC

PA + CCS off – white colored powder NC NC

PA + Crospovidone off – white colored powder NC NC

PA + SSG off – white colored powder NC NC

PA +Magnesium Stearate Sweet off – white colored

powder NC NC

PA + Aspartame Sweet off – white colored

powder NC NC

PA + Vanilla Flavor Flavored off – white colored

powder NC NC

Pa+ Strawberry Flavor Sweet flavored off – white

colored Powder NC NC

TA + PA + Mannitol Flavored off – white colored

powder NC NC

TA + PA + MCC off – white colored powder NC NC

TA + PA + β-Cyclodextrin off – white colored powder NC NC

TA + PA+ Urea off – white colored powder NC NC

TA + PA+ Lactose Monohydrate

off – white colored powder NC NC

TA + PA + SLS


TA + PA + PVP K-30 off – white colored powder NC NC

TA + PA + Polacrillin Potassium


TA + PA + CCS off – white colored powder NC NC

TA + PA + Crospovidone off – white colored powder NC NC

TA + PA + SSG off – white colored powder NC NC

TA + PA +Magnesium Stearate

Sweet off – white colored powder

NC NC



TA + PA + Aspartame Sweet off – white colored

powder

NC NC

TA + PA + Vanilla Flavor Flavored off – white colored

powder

NC NC

TA + Pa+ Strawberry Flavor Sweet flavored off – white

colored Powder

NC NC

Blend of TA with all excipients of TA mentioned for individual study to make prototype formulation for Rapid dispersible tablets of Tolfenamic Acid

Sweet flavored off – white colored Powder

NC NC

Blend of PA with all excipients of PA mentioned for individual study to make prototype formulation for Rapid dispersible tablets of Paracetamol


NC NC

Blend of TA + PA with all excipients of TA + PA mentioned for individual study to make prototype formulation for fixed dose combination


NC NC

NC – No Change

3.4.2.2 Analytical Observation by Spectrophotometric Evaluation

The drug excipients compatibility study is also confirmed with spectrophotometric

evaluation. The comparative spectrophotometric spectrum for initial blend and after

completetion of Preformulation studies is presented in the respective Graph 3.11 – 3.14.

Their results indicate that there is no chemical incompatibility between drug and

excipients used in the formulation of rapid dispersible tablets.



Graph 3.11: IR Spectrum of Blend of TA (TA#FA) with all excipients of TA for Rapid

dispersible tablets of Tolfenamic Acid at initial stage of Preformulation study

Graph 3.12 IR Spectrum of Blend of PA with all excipients of PA for Rapid dispersible

tablets of Paracetamol at initial stage of Preformulation study



Graph 3.13: IR Spectrum of Blend of TA (TA#FB) with all excipients of TA for Rapid

dispersible tablets of Tolfenamic Acid at final stage of Preformulation study

Graph 3.14: IR Spectrum of Blend of PA with all excipients of PA for Rapid dispersible

tablets of Paracetamol at Final stage of Preformulation study.



3.5 Conclusion

The selection of excipients used in formulation of dispersible tablets was based on the

available literature and database of innovator products. Preformulation study was

completed to reinsure the compatibility of drug with excipients. On the basis of physical

evaluation such as stress study, accelerated studies, there was no significant effect

observed on the appearance of blend. From the Drug: excipients compatibility it was

found that the selected excipients were found to be compatible with the Active

Pharmaceutical Ingredient at the different stability conditions that is 50°C ±

2°C(Stress Condition) for 15 days , 400C ± 2°C I 75%RH ± 5%Rh for 1 month. So

the excipients selection for the proposed formulation is most suitable for the

proposed formulation of Rapid dispersible formulation of Tolfenamic Acid and

combination of Paracetamol and Tolfenamic acid.

The spectrophotometric evaluation of drug with excipients also performed to confirm

the effect of temperature and humidity, and degradation of active during

preformulation studies. There was no significant variation observed on the profile of

active on the basis of comparative FTIR spectrum of drug excipients during

preformulation study.

The characterization of active of Tolfenamic acid and Paracetamol also complies as

per the given specifications of British Pharmacopoeia.

CHAPTER ––– 333 Preformulation...

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