11. Annexures

11. ANNEXURES Annexure I

Drug Profiles

I-A. MEBENDAZOLE(1-7)

Generic Name: Mebendazole

Chemical IUPAC Name: Methyl 5-benzoyl-2-benzimidazolecarbamate

Empirical Formula:C16H13N3O3

Molecular weight: 295.3 g/mol

Structural Formula:

Appearance: White to slightly yellow amorphous powder almost colorless

Solubility: Insoluble in water and most of the organic solvents; freely soluble in formic

acid, dimethylsulphoxide Melting point: 288.5OC

Log P/Hydrophicity: 2.8

Mechanism of action: Although the exact mechanism of anthelmintic activity of

mebendazole has not been fully elucidated, the drug appears to cause selective and

irreversible inhibition of the uptake of glucose and other low molecular weight nutrients

in susceptible helminths; inhibition of glucose uptake appears to result in endogenous

depletion of glycogen stores in the helminth. Mebendazole does not inhibit glucose

uptake in mammals. Mebendazole appears to cause degenerative changes in the intestine

of nematodes and in the absorptive cells of cestodes. The principal anthelmintic effect of

the drug appears to be degeneration of cytoplasmic microtubules within these intestinal

and absorptive cells. Microtubular deterioration results in inhibition of organelle

movement and interferes with the absorptive and secretory function. As a result of

excessive accumulation of intracellular transport secretory granules, hydrolytic and

proteolytic enzymes are released and cause cellular autolysis. This irreversible damage

leads to death of the parasite.

Pharmacokinetics:

Absorption: Mebendazole appears to be minimally absorbed from the GI tract following

oral administration. Limited data indicate that about 2–10% of an oral dose is absorbed.

Peak plasma concentrations of mebendazole occur approximately 0.5–7 hours after oral

administration of the drug and exhibit wide interpatient variation. Following oral

administration of multiple doses of mebendazole (40 mg/kg daily) to 2 adults with

hydatid cysts, mean peak plasma concentrations of about 0.08 mcg/mL occurred at 0.5–2

hours. Following oral administration of a single 10-mg/kg dose of mebendazole to

patients with hydatid cysts in another study, peak plasma concentrations of about 0.02–

0.5 mcg/mL occurred at 1.5–7.25 hours. Following oral administration of multiple doses

of mebendazole (100 mg 2 times daily for 3 days) to several children, peak plasma

mebendazole concentrations did not exceed 0.03 mcg/mL and peak plasma

concentrations of the 2-amino metabolite of the drug (the major metabolite) did not

exceed 0.09 mcg/ml.

Distribution: Mebendazole is highly bound to plasma proteins.

Metabolism: Although the exact metabolic fate of mebendazole has not been fully

determined, the drug is metabolized via decarboxylation to 2-amino-5(6)-benzimidazolyl

phenylketone; this metabolite does not have anthelmintic activity.

Elimination: The elimination half-life of mebendazole has been reported to be about

2.8–9 hours. Approximately 2–10% of an oral dose of mebendazole is excreted in urine

within 24–48 hours of administration, principally as unchanged drug and the 2-amino

metabolite. The metabolic fate and rate of excretion of unabsorbed mebendazole have not

been determined.

Category: Anthelmintic

Indications: For the treatment of a variety of nematode (roundworm) infections,

including trichuriasis (whipworm infection), enterobiasis (pinworm infection), ascariasis

(roundworm infection), hookworm infections and hydatid disease

The drug’s broad spectrum of activity makes it useful in the treatment of mixed

helminthic infections.

Dose: The dose vary depending upon the type of infection and age of the patient.

Toxicity: Overdosage of mebendazole may result in GI symptoms lasting up to a few

hours. If acute overdosage of mebendazole occurs, vomiting and purging should be

induced.

Adverse effects: Mebendazole is well tolerated even by patients in poor health.

Diarrhoea, nausea and abdominal pain have attended its use in heavy infestation. Allergic

reactions, loss of hair and granulocytopenia have been reported with high doses. Safety of

Mebendazole during pregnancy is not known, but it is contraindicated on the basis of

animal data.

Drug Interactions: Reduced plasma levels with enzyme inducres e.g. phenytoin,

carbamazepine. Increased plasma levels with cimetidine

Dosage Forms: Tablets (100 mg), Suspension (100mg/5ml)

Table-11.1 Marketed Formulations of Mebendazole

Sr. No. Product Name Dosage form Name of company

1 Helmintol Tablet, Suspension Medley

2 Lupimeb Tablet Lupin

3 Mebazole Tablet Ranbaxy

4 Mebex Tablet, Suspension Cipla

5 Wormin Tablet, Suspension Cadila

6 Mendazole Tablet Glaxo Smith Kline

I-B LOVASTATIN [1-4,8,9]

Generic Name: Lovastatin

Chemical IUPAC Name: 1S,3R,7S,8S,8aR)-8-{2-[(2R,4R)-4-hydroxy-6-

oxooxan-2-yl]ethyl}-3,7-dimethyl-1,2,3,7,8,8a-hexahydronaphthalen-1-yl

(2S)-2-methylbutanoate

Empirical Formula: C24H36O5

Molecular weight: 404.55 g/mol

Structural Formula:

Appearance: White, nonhygroscopic crystalline powder

Solubility: Insoluble in water and sparingly soluble in ethanol, methanol, and

acetonitrile Melting point: 174.5OC

Log P/Hydrophicity: 4.5

Mechanism of action: Lovastatin is a lactone that is readily hydrolyzed in vivo to

the corresponding b-hydroxyacid, a potent inhibitor of HMG-CoA reductase, the enzyme

that catalyzes the conversion of HMG-CoA to mevalonate. The conversion of HMG-CoA

to mevalonate is an early step in the biosynthetic pathway for cholesterol.

Pharmacokinetics:

Absorption: It is incompletely absorbed for g.i.t. and its oral bioavailabiity is 30%.

Distribution: It is highly bound to proteins

Metabolism: Lovastatin undergoes extensive first-pass extraction in the liver.

Elimination: Metabolites are mainly excreted in bile and elimination half-life is 5.3

hours.

Category: Hypolipidaemic drug

Indications: The first choice drugs for primary hyperlipidaemias with raised LDL and

total CH levels, with or without raised TG levels, as well as for secondary

hypercholesterolaemia. For primary propylaxis of coronary artery disease

Dose: 10-40 mg/day (max. 80 mg)

Toxicity:

Adverse effects: Increased creatine phosphokinase; flatulence, nausea, dyspepsia,

constipation or diarrhoea, abdominal pain;muscle cramps, myalgia, weakness; blurred

vision; headache, dizziness; rash.

Drug Interactions: Reduced absorption with cholestryamine, isradipine. It may increase

warfarin effect. Increase risk of myopahty & rhabdomyolysis with amiodarone, fibrates,

danazol, niacin, verapamil, protease inhibitors. Increased levels with diclofenac,

doxycycline, isoniazid, quinidine, diltiazem.

Dosage Forms: Tablets (10mg, 20mg)

Table-11.2 Marketed Formulations of Lovastatin

Sr. No. Product Name Dosage form Name of company

1 Aztatin Tablet Sun

2 Elstatin Tablet Glenmark

3 Lestric Tablet Ranbaxy

4 Lotin Tablet Intas

References:

1. Goodman and Gilman’s Pharmacological Basis of Therapuetics/ Ed by Joel G.

Hardman and Lee E. Limbird, 10th edition, Mc-Graw Hill, Inc., New York, 2001.

2. Essentials of Medical Pharmacology Ed by K. D. Tripathi, 6th edition, Jaypee,

India, 2008, 807-809.

3. Rang and Dale’s Pharmacology Churchill Livingstone/Ed by Rang H P.; Dale

M.M.; Ritter J.M.; Flower R.J., 6th ed.,2007,714.

4. The Merck Index 11th ed., Merck Research Laboratories,1989,904.

5. Van den Bossche H. Rochette F. Horig C.,1982. Mebendazole and related

anthelmintics. Adv.Pharmaco Chemother.19,287-296

6. Kumar A.,Chattopadhyay T.K.,1992. Management of hydatid disease

of the liver. Postgrad.Med.J.68:853-856.

7. Erdincler P.; Kaynar M. Y.; Babuna O.; Canbaz B.,1997. The role of

mebendazole in surgical treatment of central nervous system hydatid disease. Br.

J. Neurosurg. 11(2):116-120.

8. W. Jacobsen et. al (1999) Small Intestinal Metabolism of the 3-Hydroxy-3-

methylglutaryl-Coenzyme A Reductase Inhibitor Lovastatin and Comparison with

Pravastatin . The Journal of Pharmacology and experimental therapeutics 29(1):

131-139.

9. Henwood JM and Heel RC (1988) Lovastatin: A preliminary review of its

Pharmacodynamic properties and therapeutic use in hyperlipidemia. Drugs

36:429–454.

Annexure II

Identification and Estimation of Drugs

II-A. MEBENDAZOLE

IDENTIFICATION:

1. Melting Point Determination: Melting point is the temperature at which the pure liquid and solid exist in equilibrium.

The Thiel’s tube method of melting point determination in liquid paraffin was used in the

present study. The melting point of Mebendazole was found to be 289oC. This matches

with the standard melting point (288.5oC) indicating the identity of the drug. [1]

2. UV Spectrum: UV scanning was done for pure drug from 200-400nm in 0.1M HCl by Shimazdu UV-

1601, Japan. The λmax was found at 245 nm.

3. FTIR Spectra: FTIR spectra of drug in KBr pellets at moderate scanning speed between 4000-600 cm-1

was made. The spectrum is shown in figure 11.1. The peak related to the functional group

present in standard drug and procured drug are given in Table 10.3.

Figure 11.1 FTIR Spectra of Procured Mebendazole

Table 11.3 Results of FTIR Study of Mebendazole

Functional Group present Standard drug Procured drug[2]

NH stretching 3415 3420

C=O (amide) 1720 1720

C=O 1650 1650

CH3-O 1410-1460 1410-1460

The procured drug shows similarity with the reported value of functional group present in

standard drug, which indicates purity and identity of Mebendazole.

4. Selection of dissolution Medium: Mebendazole is insoluble in water. Thus, proper selection of dissolution medium for

Mebendazole was found essential as these drugs attain saturation solubility quickly in the

dissolution medium which may affect the release behavior of the drug loading to the poor

release behavior. MBZ has three polymorphic forms (A, B and C) that have different

solubility and therapeutic effects.5 In 0.1 M HCl polymorph C dissolves faster when

compared to polymorph B and polymorph A.5 Polymorphic form C is pharmaceutically

favoured.5

5. Estimation of drug: UV spectroscopic method was selected for the estimation of the drug.

6. Preparation of dissolution Medium: 8.5 ml of concentrated hydrocholoric acid was taken in 1000 ml volumetric flask and

diluted with distilled deionized water upto the mark. The pH was adjusted to 1.2.

7. Preparation of Standard Curve: Mebendazole (100mg) was accurately weighed and transferred into a 100 ml volumetric

flask. Volume was made up to the mark by using 0.1M HCl. This standard stock solution

is having concentration of 1000mcg/ml. From the standard stock solution, a series of

dilution were made to get 10 to 50mcg/ml solution using dissolution medium. The

absorbance of these solutions was measured at 245 nm against 0.1M HCl as a blank using

UV/VIS double beam spectrophotometer. The experiment was performed in triplicate and

based on average absorbance; the equation for the best line fit was generated. The results

of standard curve preparation are shown in Table 10.4 and Figure 11.2.

Table 11.4 Standard Curve of Mebendazole in 0.1 M HCl

Concentration

(mcg/ml)

Absorbance

1

Absorbance

1

Absorbance

1

Average

Absorbance

0 0 0 0 0

10 0.159 0.155 0.157 0.157

20 0.321 0.325 0.317 0.321

30 0.480 0.476 0.472 0.476

40 0.637 0.640 0.634 0.637

50 0.790 0.794 0.792 0.792

SUMMARY OUTPUT

Regression statistics Standard Error 0.002683

Multiple R 0.9999 Slope 0.01586

R square 0.9999 Intercept 0.0008

Adjusted R

square

0.9998

Observations 6

Absorbance = Slope * Concentration + Intercept

Absorbance = 0.01586 * Concentration + 0.0008

Standard curve of Mebendazole in 0.1 M HCl

0

0.2

0.4

0.6

0.8

1

0 10 20 30 40 50

Concentration (mcg/ml)

Abs

orba

nce

Figure 11.2: Standard Curve of Mebendazole in 0.1 M HCl

II-B. LOVASTATIN

IDENTIFICATION:

1. Melting Point Determination: Melting point is the temperature at which the pure liquid and solid exist in equilibrium.

The Thiel’s tube method of melting point determination in liquid paraffin was used in the

present study. The melting point of Lovastatin was found to be 175oC. This matches with

the standard melting point (174.5oC) indicating the identity of the drug. [1]

2. UV Spectrum: UV scanning was done for pure drug from 200-400nm in 0.05M Phosphate buffer pH-7

ctg. 0.25% SLS by Shimazdu UV-1601, Japan. The λmax was found at 239 nm.

3. FTIR Spectra: FTIR spectra of drug in KBr pellets at moderate scanning speed between 4000-600 cm-1

was made. The spectrum is shown in figure 11.3. The peak related to the functional group

present in standard drug and procured drug are given in Table 10.5.

Figure 11.3 FTIR Spectra of Procured Lovastatin

Table 11.5 Results of FTIR Study of Lovastatin

Functional Group present Standard drug Procured drug[3]

Lactone and ester carbonyl 1725 1730.88

Methyl asymmetric bond 1460 1457.33

Lactone C-O-C asymmetric 1260 1263.16

bend

Ester C-O-C asymmetric

stretch

1222 1227.87

The procured drug shows similarity with the reported value of functional group present in

standard drug, which indicates purity and identity of Lovastatin.

4. Selection of dissolution Medium: Water solubility of LVS is 0.0004 mg/ml. 0.05M Phosphate buffer pH-7 ctg. 0.25% SLS

was selected as a dissolution medium.6

5. Estimation of drug: UV spectroscopic method was selected for the estimation of the drug.

6. Preparation of dissolution Medium: 756 ml of 0.1 M disodium hydrogen phosphate and 244 ml of 0.1 M HCl was mixed

together to make the volume 1000 ml. The pH of above solution was adjusted to 7. In the

above solution, 2.5 gm of Sodium Lauryl Sulphate was added and dissolved.

7. Preparation of Standard Curve: Lovastatin (10mg) was accurately weighed and transferred into a 250 ml volumetric

flask. Volume was made up to the mark by using 0.05M Phosphate buffer pH-7 ctg.

0.25% SLS. This standard stock solution is having a concentration of 40mcg/ml. From

the standard stock solution, a series of dilution were made to get 20 to 40mcg/ml solution

using dissolution medium. The absorbance of these solutions was measured at 239 nm

against 0.05M Phosphate buffer pH-7 ctg. 0.25% SLS as a blank using UV/VIS double

beam spectrophotometer. The experiment was performed in triplicate and based on

average absorbance; the equation for the best line fit was generated. The results of

standard curve preparation are shown in Table 10.6 and Figure 6.

Table 11.6 Standard Curve of Lovastatin in 0.05M Phosphate buffer pH-7 ctg. 0.25%

SLS

Concentration

(mcg/ml)

Absorbance

1

Absorbance

1

Absorbance

1

Average

Absorbance

0 0 0 0 0

20 0.3038 0.3048 0.3044 0.3043

25 0.349 0.351 0.3503 0.3501

30 0.4198 0.4195 0.4201 0.4198

35 0.5133 0.5177 0.5187 0.5166

40 0.6024 0.6038 0.601 0.6024

SUMMARY OUTPUT

Regression statistics Standard Error 0.01612

Multiple R 0.9976 Slope 0.0148

R square 0.9953 Intercept 0.0062

Adjusted R

square

0.9941

Observations 6

Absorbance = Slope * Concentration + Intercept

Absorbance = 0.0148 * Concentration -0.0062

Standard Curve of Lovastatin in 0.05M Phosphate buffer pH-7 ctg. 0.25% SLS

0.30.350.4

0.450.5

0.550.6

0.65

20 25 30 35 40

Concentration (mcg/ml)

Abs

orba

nce

Figure 11.4 Standard Curve of Lovastatin in 0.05M Phosphate buffer pH-7 ctg. 0.25%

SLS

REFERENCES:

1. The Merck Index By M.J.O’Neil, Merck & Co., USA, 2001

2. Analytical Profile of Drug substance and excipients Hary G Brittain volume 16,

291

3. Analytical Profile of Drug substance and excipients. Hary G Brittain volume 21,

277

4. Indian Pharmacopoeia By Government of India, New Delhi,1996,2007

5. Swanepoel E; Liebenberg W; Devarakonda B; de villiers M M Developing a

discriminating dissolution test for three mebendazole polymorphs based on

solubility differences Die Pharmazie 2003;58(2):117-121

6. Dissolution methods database, available at

www.accessdata.fda.gov/scripts/cder/dissolution/dsp_SearchResults_Dissolutions

.cfm?PrintAll=1 - 300k Accessed on 20th March, 2011

Annexure III

Excipient Profiles III-A. POLY VINYL ALCOHOL [1]

1 Nonproprietary Names PhEur: Poly(Vinyl Alcohol)

USP: Polyvinyl Alcohol

2 Synonyms

Airvol; Alcotex; Celvol; Elvanol; Gelvatol; Gohsenol; Lemol; Mowiol; poly(alcohol

vinylicus); Polyvinol; PVA; vinyl alcohol polymer.

Chemical Name and CAS Registry Number

Ethenol, homopolymer [9002-89-5]

Empirical Formula and Molecular Weight

(C2H4O)n 20 000–200 000

Polyvinyl alcohol is a water-soluble synthetic polymer represented by the formula

(C2H4O)n. The value of n for commercially available materials lies between 500 and

5000, equivalent to a molecular weight range of approximately 20 000–200 000; see

Table I.

Table I: Commercially available grades of polyvinyl alcohol

5 Structural Formula

6 Functional Category

Coating agent; lubricant; stabilizing agent; viscosity-increasing agent.

7 Applications in Pharmaceutical Formulation or Technology

Polyvinyl alcohol is used primarily in topical pharmaceutical and ophthalmic

formulations; see Table II. It is used as a stabilizing agent for emulsions (0.25–3.0%

w/v). Polyvinyl alcohol is also used as a viscosity-increasing agent for viscous

formulations such as ophthalmic products. It is used in artificial tears and contact lens

solutions for lubrication purposes, in sustained-release formulations for oral

administration,(4) and in transdermal patches. Polyvinyl alcohol may be made into

microspheres when mixed with a glutaraldehyde solution.

8 Description

Polyvinyl alcohol occurs as an odorless, white to cream-colored granular powder.

9 Pharmacopeial Specifications

10 Typical Properties

Melting point

2280C for fully hydrolyzed grades;

180–190oC for partially hydrolyzed grades.

Refractive index nD

25 = 1.49–1.53

Solubility

Soluble in water; slightly soluble in ethanol (95%); insoluble in organic

solvents. Dissolution requires dispersion (wetting) of the solid in water at

room temperature followed by heating the mixture to about 908oC for

approximately 5 minutes. Mixing should be continued while the heated

solution is cooled to room temperature.

Specific gravity

1.19–1.31 for solid at 258C;

1.02 for 10% w/v aqueous solution at 258C.

Specific heat 1.67 J/g (0.4 cal/g)

Viscosity (dynamic) see Table IV.

11 Stability and Storage Conditions

Polyvinyl alcohol is stable when stored in a tightly sealed container in a cool, dry place.

Aqueous solutions are stable in corrosionresistant sealed containers. Preservatives may be

added to the solution if extended storage is required. Polyvinyl alcohol undergoes slow

degradation at 1008C and rapid degradation at 2008C; it is stable on exposure to light.

12 Incompatibilities

Polyvinyl alcohol undergoes reactions typical of a compound with secondary hydroxy

groups, such as esterification. It decomposes in strong acids, and softens or dissolves in

weak acids and alkalis. It is incompatible at high concentration with inorganic salts,

especially sulfates and phosphates; precipitation of polyvinyl alcohol 5% w/v can be

caused by phosphates. Gelling of polyvinyl alcohol solution may occur if borax is

present.

13 Safety

Polyvinyl alcohol is generally considered a nontoxic material. It is nonirritant to the skin

and eyes at concentrations up to 10%; concentrations up to 7% are used in cosmetics.

Studies in rats have shown that polyvinyl alcohol 5% w/v aqueous solution injected

subcutaneously can cause anemia and infiltrate various organs and tissues.

LD50 (mouse, oral): 14.7 g/kg

LD50 (rat, oral): >20 g/kg

14 Handling Precautions

Observe normal precautions appropriate to the circumstances and quantity of material

handled. Eye protection and gloves are recommended. Polyvinyl alcohol dust may be an

irritant on inhalation. Handle in a well-ventilated environment.

15 Regulatory Status

Included in the FDA Inactive Ingredients Database (ophthalmic preparations and oral

tablets). Included in nonparenteral medicines licensed in the UK. Included in the

Canadian List of Acceptable Non-medicinal Ingredients.

16 Comments

Various grades of polyvinyl alcohol are commercially available. The degree of

polymerization and the degree of hydrolysis are the two determinants of their physical

properties. Pharmaceutical grades are partially hydrolyzed materials and are named

according to a coding system. The first number following a trade name refers to the

degree of hydrolysis and the second set of numbers indicates the approximate viscosity

(dynamic), in mPa s, of a 4% w/v aqueous solution at 200C.

III-B. GLYCERIN [1]

1 Nonproprietary Names BP: Glycerol

JP: Concentrated Glycerin

PhEur: Glycerol

USP: Glycerin

2 Synonyms

Croderol; E422; glicerol; glycerine; glycerolum; Glycon G-100; Kemstrene; Optim;

Pricerine; 1,2,3-propanetriol; trihydroxypropane glycerol.

3 Chemical Name and CAS Registry Number

Propane-1,2,3-triol [56-81-5]

4 Empirical Formula and Molecular Weight

C3H8O3 92.09

5 Structural Formula

6 Functional Category

Antimicrobial preservative; cosolvent; emollient; humectant; plasticizer; solvent;

sweetening agent; tonicity agent.

7 Applications in Pharmaceutical Formulation or Technology

Glycerin is used in a wide variety of pharmaceutical formulations including oral, otic,

ophthalmic, topical, and parenteral preparations; see Table I.

In topical pharmaceutical formulations and cosmetics, glycerin is used primarily for its

humectant and emollient properties. Glycerin is used as a solvent or cosolvent in creams

and emulsions. Glycerin is additionally used in aqueous and nonaqueous gels and also as

an additive in patch applications. In parenteral solvent. formulations, glycerin is used

mainly as a solvent and cosolvent.

In oral solutions, glycerin is used as a solvent, sweetening agent, antimicrobial

preservative, and viscosity-increasing agent. It is also used as a plasticizer and in film

coatings. Glycerin is used as a plasticizer of gelatin in the production of soft-gelatin

capsules and gelatin suppositories. Glycerin is employed as a therapeutic agent in a

variety of clinical applications, and is also used as a food additive.

8 Description

Glycerin is a clear, colorless, odorless, viscous, hygroscopic liquid; it has a sweet taste,

approximately 0.6 times as sweet as sucrose.

9 Pharmacopeial Specifications

10. Typical Properties

Boiling point 290oC (with decomposition)

Density 1.2656 g/cm3 at 15oC;

1.2636 g/cm3 at 20oC;

1.2620 g/cm3 at 25oC.

Flash point 176oC (open cup)

Hygroscopicity Hygroscopic.

Melting point 17.8oC

Osmolarity A 2.6% v/v aqueous solution is isoosmotic with serum.

Refractive index

n D15 = 1.4758;

n D20 = 1.4746;

n D25 = 1.4730.

Solubility

.

Specific gravity

Surface tension : 63.4mN/m (63.4 dynes/cm) at 20oC.

Vapor density (relative) : 3.17 (air = 1)

Viscosity (dynamic)

11. Stability and Storage Conditions

Glycerin is hygroscopic. Pure glycerin is not prone to oxidation by the atmosphere under

ordinary storage conditions, but it decomposes on heating with the evolution of toxic

acrolein. Mixtures of glycerin with water, ethanol (95%), and propylene glycol are

chemically stable. Glycerin may crystallize if stored at low temperatures; the crystals do

not melt until warmed to 208C. Glycerin should be stored in an airtight container, in a

cool, dry place.

12 Incompatibilities

Glycerin may explode if mixed with strong oxidizing agents such as chromium trioxide,

potassium chlorate, or potassium permanganate. In dilute solution, the reaction proceeds

at a slower rate with several oxidation products being formed. Black discoloration of

glycerin occurs in the presence of light, or on contact with zinc oxide or basic bismuth

nitrate. An iron contaminant in glycerin is responsible for the darkening in color of

mixtures containing phenols, salicylates, and tannin. Glycerin forms a boric acid

complex, glyceroboric acid, that is a stronger acid than boric acid.

13. Regulatory Status

GRAS listed. Accepted for use as a food additive in Europe. Included in the FDA

Inactive Ingredients Database (dental pastes; buccal preparations; inhalations; injections;

nasal and ophthalmic preparations; oral capsules, solutions, suspensions and tablets; otic,

rectal, topical, transdermal, and vaginal preparations). Included in nonparenteral and

parenteral medicines licensed in the UK. Included in the Canadian List of Acceptable

Non-medicinal Ingredients.

III-C. FORMIC ACID [1]

IUPAC name: Formic acid

Synonyms: formylic acid, hydrogen carboxylic acid,

Methanoic acid, aminic acid, methanoic acid

Other names Aminic acid

Formylic acid

Hydrogen carboxylic acid

Hydroxymethanone

Hydroxy(oxo)methane

Metacarbonoic acid

Oxocarbinic acid

Oxomethanol

Molecular formula CH2O2

Structural Formula

CAS number 64-18-6

Description

Formic acid is a colorless, fuming liquid having a highly pungent, penetrating odor at

room temperature. It is miscible with water and most polar organic solvents, and

somewhat soluble in hydrocarbons. In hydrocarbons and in the vapor phase, it consists of

hydrogen-bonded dimers rather than individual molecules. Owing to its tendency to

hydrogen-bond, gaseous formic acid does not obey the ideal gas law. Solid formic acid

(two polymorphs) consists of an effectively endless network of hydrogen-bonded formic

acid molecules. This relatively complicated compound also forms a low-boiling

azeotrope with water (22.4%) and liquid formic acid also tends to supercool.

Physical and chemical properties Appearance Liquid

Colour Colourless to pale yellow

Odour Pungent

Molar mass 46.03 g mol−1

Solubility Miscible

Boiling point (°c) 107.3

Relative density 1.195 20

Vapour pressure <4.4 kPa 20

pH-value, conc. Solution <1

pH-value, diluted solution 2.2 1

Viscosity 1.57 cP at 26 °C

Flash point (°c) 65

Auto ignition Temperature (°C) 500

Density 1.22 g/mL,

Acidity (pKa) 3.77

Functional Category

Preservative and antibacterial agent

Toxicological information

Toxic dose 1 - LD 50 730 mg/kg (oral rat)

Toxic conc. - LC 50 7.4 mg/l/4h (inh-rat)

Ingestion May cause severe internal injury

Skin contact Causes burns.

Eye contact Causes burns.

Handling and storage

Usage precautions

Avoid contact with skin and eyes. Provide good ventilation. Eliminate all sources of

ignition.

Storage precautions

Keep containers tightly closed. Keep in original container.

Storage class

Corrosive storage.

III-D. DICHLOROMETHANE [1]

IUPAC name Dichloromethane

Synonyms: DCM; Methylene chloride (MC); Methylene dichloride; Methylene

bichloride; Methane dichloride

Other names Methylene chloride, methylene dichloride, Solmethine, Narkotil,

Solaesthin, Di-clo, Freon 30, R-30, DCM, UN 1593, MD

CAS number 75-09-2

Molecular Weight: 84.93

Chemical Formula: CH2Cl2

Structural Formula

Uses

Dichloromethane's volatility and ability to dissolve a wide range of organic compounds

makes it a useful solvent for many chemical processes. Concerns about its health effects

have led to a search for alternatives in many of these applications.

Physical and Chemical Properties

Appearance: Clear, colorless liquid.

Odor: Chloroform-like odor.

Solubility: 1.32 gm/100 gm water @ 20C.

Specific Gravity: 1.318 @ 25C

% Volatiles by volume 100

@ 21C (70F):

Boiling Point: 39.8C (104F)

Melting Point: -97C (-143F)

Vapor Density (Air=1): 2.9

Vapor Pressure (mm Hg): 400 @ 24C (75F)

Handling and Storage

Store in a cool dry well ventilated area. Keep away from heat and flame. Do not get in

eyes, on skin, or on clothing.

REFERENCES: 1. Handbook of Pharmaceutical Excipients Sixth edition By Raymond C Rowe, Paul

J Sheskey and Arian E Quinn. Pharmaceutical Press Publishers

Annexure IV

Experimental Design

The aim of pharmaceutical formulation and development is to develop an acceptable

formulation in the shortest possible time, using minimum number of working hours and

raw materials.

The formula developed by the formulation and development center is first tried at the

pilot scale and then manufacture scale. Only minor changes are to be made during scale-

up. Thus, it is very ideal to study the formulation from all perspectives at laboratory

levels.

In addition to the art of formulation, a statistical technique is available that can aid in the

pharmacist’s choice of formulation components, which can optimize one or more

formulation additives.

A very efficient way to enhance the value of research and to minimize the process

development time is through the experiment. The need of develop this design because

traditional experiments involve a good deal of efforts and time, especially where complex

formulations are to be developed.

The statistical problem solving approach uses a series of small carefully designed

experiments. We sometimes call the statistical approach ‘strategic experimentation’ or

iterative problem solving strategy. We also call this the ‘stop look and listen’ approach to

experimentation. Analyze the results of few experiments and then plan the next

experiments. Any statistical design consist of the small and efficient experiments, namely

a screening experiments where from many factors affecting the process few important

factors are identified, then an optimization experiment where a predictive model is build

for the few factors in the region of optimum and finally a verification experiment where

the results is confirmed at the predicted setting. In the present work factorial design was

used for the development of effective, functional and perfect dosage form. The help of

systematic formulation approach is taken to get detailed knowledge on the formulation.

In the present study, 24, 32and 33 factorial design and Plackett-burman design were used.

Hence, only these designs are discussed in details.

IV-A. FACTORIAL DESIGNS (1-3)

Factorial designs are used in experiments when the effects of different factors or

conditions, on experiment results are to be elucidated. Factorial designs are the design of

choice of simultaneous determination of the effects of several factors and their

interaction. Factors may be qualitative or quantitative. The levels on each factor are the

values or designations assigned to combinations, of all levels, of all factors. The effects

of a factor are the change in response caused by varying the level(s) of the factor.

The important objective of a factorial experimentation is to characterize the effect of

changing the levels of the factor or combination of factors on the response variable.

Predictions based on results of an undersigned experiment will be more variable than

those, which could be obtained in a designed experiment, in particular factorial design.

The optimization procedure is facilitated by construction of an equation that described the

experimental results as a function of the factor levels. A polynomial equation can be

constructed, where the coefficients in the equation are related to the effects and

interaction of the factors. The goal of pharmaceutical formulation is in the shortest

possible time using minimum time and raw materials.

Optimization by experimental design leads to the evolution of a statistically valid model

to understand the relationship between independent and dependent variables.

The equation constructed from 2n factorial experiment is in the following form.

Y=Bo + B1X1+ B2X2+B3X3+B12X1X2+B13X1X3+B23X2X3+B123X1X2X3

Where,

Y= the measured response

Xi= level of ith factor (independent variable)

Bi= the regression coefficient for the ith independent variable

B0= intercept

The magnitudes of the coefficients represent the relative importance of each factor. Once

the polynomial equation has been established, an optimum formulation can be found out

by grid analysis. A computer can calculate the response based on equation at many

combinations of factor levels. The formulation whose response has optimal

characteristics based on the experimenter’s specification is then chosen.

Advantages of Factorial designs:

In absence of interaction, they have maximum efficiency in estimating mail

effects

Maximum use is made of the data, since all main effects and interaction are

calculated from the data

Since factors effects are measured over varying levels of other factors,

conclusions apply to wide range of conditions

Then are orthogonal; all estimated effect and interaction are independent of the

effect of the other factors

If interaction occur; they are necessary to reveal and identify the interaction

More information is obtained with less work

The effects are measured with maximum precision.

Applications of Factorial designs:

It helps and interprets the mechanism of experimental system

It provides guidance for further experiment

It also useful for the drug-excipient compatibility study

It is very useful in an industrial manufacturing operation because it recommends

or implements, a practical procedure or a set of condition

Factorial designs are either full or fractional. Full factorial design is a design in which

every setting of every factor appears with every setting of every other factor is called as a

full factorial design. When experiments are with a large number of factors and/or a large

number of levels for the factors, the number of factors needed to complete factorial

design is also large.

Full Factorial Design:

24 Factorial Design:

A common experimental design is one with all input factors set at two levels each, these

levels are called ‘high’ and ‘low’ or ‘+1’ and ‘-1’, respectively. A design with all possible

high/low combinations of all the input factors is called a full factorial design in two

levels.

In 24 factorial designs four factors and two levels are used to achieve the proper result.

This implies sixteen runs.

The design of is given in Table 1.

Table 1: Design of 24 factorial design

Trial X1 X2 X3 X4

1 -1 -1 -1 -1

2 -1 -1 -1 1

3 -1 -1 1 1

4 -1 1 1 1

5 -1 -1 1 -1

6 -1 1 -1 -1

7 -1 1 1 -1

8 -1 1 -1 1

9 1 -1 -1 -1

10 1 -1 -1 1

11 1 -1 1 1

12 1 1 1 1

13 1 -1 1 -1

14 1 1 -1 -1

15 1 1 1 -1

16 1 1 -1 1

A statistical model incorporating interactive and polynomial terms is used to evaluate the

response. Yi = B0 + B1X1 + B2X2 + B3X3 + B4X4 + B12X1X2 + B13X1X3 + B14X1X4 + B23X2X3 + B24X2X4 + B34X3X4 + B1234X1X2X3X4

Where, Yi is the dependent variable; b0 is the arithmetic mean of the 16 terms; bi is the

estimated coefficient for the factor Xi.

32 Factorial Design:

In 32 full factorial designs two factors and three levels are used. Total 9 trials are made if

this design is employed.

The design of 32 factorial is as given in Table 2.

Table 2: Design of 32 factorial design

Trial X1 X2

1 -1 -1

2 -1 0

3 -1 1

4 0 -1

5 0 0

6 0 1

7 1 -1

8 1 0

9 1 1

A statistical model incorporating interactive and polynomial terms is used to evaluate the

response. Yi = b0 + b1X1 + b2X2 + b11X1

2 + b22X22 + b12X1X2

Where, Yi is the dependent variable; b0 is the arithmetic mean of the 9 terms; bi is the

estimated coefficient for the factor Xi.

33 Factorial Design:

In 33 full factorial designs three factors and three levels are used. Total 27 trials are made

if this design is employed.

The design of 33 factorial is as given in Table 3.

Table 3: Design of 33 factorial design

Trial X1 X2 X3

1 -1 -1 -1

2 -1 -1 0

3 -1 -1 1

4 -1 0 -1

5 -1 0 0

6 -1 0 1

7 -1 1 -1

8 -1 1 0

9 -1 1 1

10 0 -1 -1

11 0 -1 0

12 0 -1 1

13 0 0 -1

14 0 0 0

15 0 0 1

16 0 1 -1

17 0 1 0

18 0 1 1

19 1 -1 -1

20 1 -1 0

21 1 -1 1

22 1 0 -1

23 1 0 0

24 1 0 1

25 1 1 -1

26 1 1 0

27 1 1 1

A statistical model incorporating interactive and polynomial terms is used to evaluate the

responses. Yi = b0 + b1X1 + b2X2 + b3X3 + b11X1

2 + b22X22 + b33X3

2 + b12X1X2 + b23X2X3 + b13X1X3 Where, Yi is the dependent variable; b0 is the arithmetic mean of the 27 terms; bi is the

estimated coefficient for the factor Xi.

IV-B. PLACKETT-BURMAN DESIGN (4-7)

A popular class of screening designs is the Plackett-Burman design (PBD), developed by

R.L. Plackett and J.P. Burman in 1946.It was designed to improve the quality control

process that could be used to study the effects of design parameters on the system state so

that intelligent decisions can be made. Plackett and Burman (PB) devised orthogonal

arrays are useful for screening, which yield unbiased estimates of all main effects in the

smallest design possible. Various number or ‘n’ factors can be screened in an ‘n + 1’ run

PB design.

For the seven factors, the following PBD having eight runs is used for screening.

Batch

no

X1 X2 X3 X4 X5 X6 X7

1 + + + - + - -

2 - + + + - + -

3 - - + + + - +

4 + - - + + + -

5 - + - - + + +

6 + - + - - + +

7 + + - + - - +

8 - - - - - - -

REFERENCES:

1. Bolton S.,Bon C., Pharmaceutical Statistics: Practical and clinical application, 2nd

Ed., Marcel Dekker Inc., NY, 1990:265-280; 506-538

2. Franz R.M., Browne J.E., and Lewis A.R.; Experimental design, modeling, an

optimization strategies for product and process development: In Livermann,

H.A.Riger, M.M.Banker, G.S., Pharmaceutical dosage form: Disperse system

(volume1), Marcel Dekker, NY, 1988: 427-519

3. Lewis, GA, Mathieu D, Phan-Tan-Luu R. Pharmaceutical Experimental Design

Marcel Dekker, NY, 1999: 185-246

4. R. H. Shobha Rani, K.Vanaja. Design of Experiments: Concept and applications

of Plackett-Burman Design: Clinical Research and Regulatory Affairs,

2007,24(1): 1–23

5. Plackett, R. L. & Burman J. P. (1946) Biometrika 33, 305- 325

6. Vander Heyden, Y., Nijhuis, A., Smeyers-Verbeke, J., Vandeginste, B. G. &

Massart, D.L. (2001) J Pharm BiomedAnal 24, 723-753

7. Draper N.R., “Plackett Burman Designs”, Encyclopedia of Statistical Sciences

Volume 6, Ed Johnson Kotz, 9 volumes; Wiley, 1982-1988

Annexure V

Uniformity Index

Uniformity index (UI) is calculated from the data of particle size distribution by using the

following formula.

UI = Dw/Dn

where Dw and Dn are weight average diameter and number average diameter respectively,

and are calculated as follows:

Dw = ∑NiDi4/∑NiDi3

Dn = ∑NiDi/ ∑Ni

where Ni is the number of particles with Di diameter.

As per Shukla et al, values of UI ranging from 1.0 to 1.1 and 1.1 to 1.2 indicate

monodisperse and nearly monodisperse particles. In the present case, values higher than

1.2 have been regarded as indicative of particles with broad particle size distribution.

REFERENCE:

Shukla P G, Kalidhass B, Shah A, Palashkar D V. Preparation and characterization of

microcapsules of water soluble pesticide monocrotophs using polyurethane as carrier

material. J Microencapsul. 2002; 19(3): 293-304

Annexure VI

Desirability Function Approach

The desirability function1,2,3 is one of the most widely used methods in industry for the

optimization of multiple response processes. It is based on the idea that the "quality" of a

product or process that has multiple quality characteristics, with one of them outside of

some "desired" limits, is completely unacceptable.

During optimization, the responses have to be combined in order to produce a product of

desired characteristics. The application of the desirability function combines all the

responses in one measurement and gives the possibility to predict the optimum levels for

the independent variables. The combination of the responses in one desirability function

requires the calculation of the individual functions.

Individual desirability for each response (ID1) is calculated from the following equation.

Q

ID1 = ----------------------

Rmax – Rmin

Q = Rmax – R OR Q = R – Rmin

Where, Q = difference obtained by substracting of an individual response from the

maximum or minimum value of the response

Rmax = maximum value of the response from the all response values

Rmin = minimum value of the response from the all response values

R = value of the response experimentally determined

Similarly individual desirability ID2 and ID3 with respect to other responses are

determined.

Overall desirability (OD) is calculated from the following equation.

OD = (ID1ID2ID3)1/3

The value of OD near to 1 indicates the batch or product having all the different desired

characteristics.

REFERENCES:

1. Swanepoel E, Liebenberg W, Devarakonda B, Villiers M M D. Developing a

discriminatory dissolution test for three mebendazole polymorphs based on solubility

differences. Pharmazie. 2003; 58(2):117–121

2. Sutariya V B, Mashru R C, Sankalia M G, Sankalia J M. Preparation of rapidly

disintegrating tablets of ondansetron hydrochloride by direct compression method

Ars Pharm 2006; 47(3): 293-311

3. Lewis G, Mathieu D, Phan-Than-Luu R. Optimization: Pharmaceutical process

optimization and validation. In Pharmaceutical Experimental Design, 1st Ed;

Swarbrick, James, Boylan, James C., Eds.; Marcel Dekker, Inc.: New York, 1999;

Vol. 92, 265-276

Appendix VII

Acknowledgement This work was funded (Rs.8.675 lac) by All India Council for Technical

Education (AICTE), New Delhi under Research Promotion Scheme vide letter ref.

no. 220-62/FIN/04/05/1333/319 dated 18/04/2007. Title of the project sanctioned

by AICTE is “Preparation and characterization of spherical agglomerates of some

drugs by novel particle engineering technology”.

Publications Review article on Novel particle engineering technology in Drug Discovery

Technology 2006 Dharmesh M. Modi, Megha Barot, Jolly R. Parikh