Publication Ref No.

: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-9/NOV/002

ISSN 0974 9446

PREFORMULATION PARAMETERS CHARACTERIZATION TO DESIGN, DEVELOPMENT AND FORMULATION OF VANCOMYCIN HYDROCHLORIDE TABLETS FOR PSUDOMEMBRANOUS COLITIS M. M. Gupta*, 1, T.R. Saini 2 Department of Pharmaceutics, Jaipur College of Pharmacy, ISI-15, RIICO Institutional Area, Sitapura, Tonk Road, Jaipur 302015, 2 Industrial Pharmacy Research Lab., Department of Pharmacy, S.G.S.I.T.S., Park Road Indore, M.P., India, E-Mail: mmingupta@gmail.com
1

ABSTRACT Tablets1 of Vancomycin hydrochloride2, 3 were prepared by direct compression4, 5 method using Lactose (Direct Compression Lactose-21)6, 7 as diluent and Avicel 2006,7 as filler disintegrating agent, whereas Kollidon- CL (Crosslinked polyvinyl pyrollidone)8 was used as superdisintegrant. So before selection of all these excipients as well as direct compression method, the Preformulation study of drug Vancomycin hydrochloride is completed for successful formulation of tablet. The % compressibility was calculated of this drug which is required in the selection of direct compression method and was found 17.5% which suits for this direct compression method. The stability of drug was studied in distilled water and as well as in 0.1N HCl which was required for drug analysis to check release profile without degradation of drug in these solvent and found as such no degradation of drug in water as well as in the 0.1N HCl. The drug -excipient compatibility study is carried by simple physical mixing as well as by Infrared Spectroscopy (IR), and as such no major interaction was found between Vancomycin hydrochloride and all selected excipients (lactose, Avicel, Kollidon, Magnessium stearate and Aerosil). The drug release from tablet was studied by UV spectrophometer so interference of additives during analysis was also checked and as such no interference was found by any excipient in the estimation of drug. The solubility as well as partition coefficient of Vancomycin hydrochloride was also calculated and found 99210 g / ml and 0.2142 respectively. Key Words: Vancomycin hydrochloride, % compressibility, compatibility studies, partition coefficient, stability of drug, water, absorbance

International Journal of Pharma Research and Development Online

www.ijprd.com

Publication Ref No.: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-9/NOV/002

ISSN 0974 9446

INTRODUCTION Preformulation9 commences when a newly synthesized drug shows sufficient pharmacologic promise in animal models to warrants evaluation in man. These studies should focus on those physicochemical properties of the new compound that could affect drug performance and development of an efficacious dosage form. A through understanding of these of these properties may ultimately provide a rational for formulation design, or support the need for molecular modification. Vancomycin hydrochloride2,3 is amphoteric glycopeptide antimicrobial substance produced by the growth of certain strains of Streptomyces orientalis used in the treatment of enterocolitis10-12 caused by Staphylococcus aureus, antibiotic associated pseudomembranous colitis10-12 caused by C.difficile. Drug is poorly absorbed from the gastrointestinal tract, although absorption may be somewhat greater when the gastrointestinal tract is inflamed2, 3, 10-12. So this drug having almost zero oral bio availability but the drug is freely soluble in water which is good for dissolution study. The % compressibility was calculated because we want to prepare tablet by direct compression method due to cost effective factor as well as due moisture sensitive property of Vancomycin hydrochloride. METHOD AND MATERIALS Vancomycin hydrochloride USP, lactose (DCL-21), Avicel-200, Kollidon-CL were obtained as a gift samples from Alkem lab Mumbai (India), Aerosil 200 was obtained from Ranbaxy lab. Ltd. Dewas (India) as a gift sample, magnesium stearate was collected from local pharmaceutical industry of pharmaceutical grade. All other chemical were from reagent grade. STABILITY OF VANCOMYCIN HYDROCHLORIDE IN SOLVENTS:

Stability of Vancomycin Hydrochloride in distilled water Stability of vancomycin hydrochloride in distilled water was determined by keeping the known concentration (40 g/ ml), in kinetic mode of UV Visible spectrophotometer (UV 160 A) for 30 minute. The data obtained is shown in table 1. Table1: Stability of Vancomycin Hydrochloride in Distilled Water in Kinetic Mode of UV- Visible Spectrophotometer at max 281.0 nm (Absorbance of 40 g/ ml drug solution is 0.201) Time (Second) Absorbance dA 0 0.201 0.000 200 0.201 0.000 400 0.200 0.001 600 0.197 0.003 800 0202 -0.001 1000 0.201 0.000 1200 0.200 0.001 1400 0.198 0.003 1600 0.204 -0.002 1800 0.203 -0.002

International Journal of Pharma Research and Development Online

www.ijprd.com

Publication Ref No.: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-9/NOV/002

ISSN 0974 9446

Stability of Vancomycin Hydrochloride in 0.1N HCl This was determined by keeping the known concentration (20 g/ ml) solution, in kinetic mode of UV Visible double beam spectrophotometer (UV 160 A) for 30 minute.The data obtained is shown in table no 2 Table2: Stability of Vancomycin Hydrochloride in 0.1N HCl in Kinetic Mode of UV Visible Spectrophotometer at max 281.0 nm (Absorbance of 20 g/ ml of drug solution is 0.112) Time Absorbance dA (Second) 0 0.112 0.000 200 0.111 0.001 400 0.114 -0.002 600 0.108 0.004 800 0.110 0.002 1000 0.112 0.000 1200 0.112 0.000 1400 0.113 -0.001 1600 0.116 -0.004 1800 0.115 -0.003 Preparation of Calibration Curve13 of Vancomycin Hydrochloride in Distilled Water: A stock solution (200 g / ml) of vancomycin hydrochloride prepared and from this different -different concentration solutions were prepared and then the absorbance of dilutions was measured on a UV- Visible spectrometer (UV- 160 A) at 281.0 nm. The absorbance data are given in the table 3 and the calibration curve is plotted is shown in figure 1. Table 3: Absorbance Data for Calibration Curve of Vancomycin HCl in Distilled Water at 281.0 nm Concentration Absorbance Set 1 Set 2 Set 3 (g / ml) Average SD 0 0.000 0.000 0.000 0.000 40 0.200 0.202 0.201 0.201 0.001 60 0.304 0.300 0.308 0.304 0.004 80 0.409 0.416 0.402 0.409 0.007 100 0.510 0.516 0.504 0.510 0.006 120 0.605 0.600 0.610 0.605 0.005

International Journal of Pharma Research and Development Online

www.ijprd.com

Publication Ref No.: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-9/NOV/002

ISSN 0974 9446

Concentration mcg / ml

150 100 50 0 -50 0

y = 197.15x - 0.0043 R2 = 0.9999

0.2

0.4
Absorbance

0.6

0.8

Fig. 1: Calibration Curve of Vancomycin Hydrochloride in Distilled Water at 281.0 nm Determination of Solubility14 of Vancomycin HCl Excess of drug was placed in distilled water and this solution was occasionally stirred for 24 hours at room temperature. After 24 hours sample was filtered and filtrate was suitably diluted and absorbance was taken at 281.0 nm against distilled water as blank on UV Visible spectrophotometer (UV 160 A). The result is shown in table 4.

Observation: Absorbance of Vancomycin HCl in distilled water: 0.506 A Table 4: Solubility of Vancomycin HCl in Distilled Water Solvent Solubility Inference Distilled water Freely soluble 99210 g / ml

Determination of interference of additives in the estimation of Vancomycin HCl The highest concentration of the additives that would be present in 900 ml of dilution media was estimated on the basis of amount present in per tablet of different batches and the same quantities of different additives were taken in 10 ml volumetric flasks containing 10 ml of 40 g / ml concentration of vancomycin HCl solution. The flasks were kept for 24 hours with occasional shaking and filtration was done. The absorbance of filtrate were measured at 281.0 nm on UV Visible spectrophotometer (UV 160 A) and compared with the absorbance of control drug sample of 40 g / ml concentration without additives.Observations are shown in table 5:

International Journal of Pharma Research and Development Online

www.ijprd.com

Publication Ref No.: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-9/NOV/002

ISSN 0974 9446

Table 5: Interference of Additives in the Estimation of Vancomycin HCl in Distilled Water at 281.0 nm (Absorbance of plain drug solution (40g / ml): 0.200 A) S.No. Additives Absorbance Interference 1 Lactose (DCL 21) 0.198 Nil 2 Avicel 200 0.202 Nil 3 Kollidon CL 0.200 Nil 4 Ac-di-Sol 0.199 Nil 5 Magnessium Sterate 0.201 Nil 6 Aerosil 200 0.197 Nil 7 Dicalcium Phosphate 0.310 Significant Determination of % Compressibility14 I = Dt - Db 100 Dt Bulk Density (Db) =0.33 (Calculated) Tapped Bulk Density (Dt) = 0.40 (Calculated) I = (0.40 0.33 / 0.40) 100 = 17.5 Determination of Partition Coefficient14 10 mg drug was added in 50 ml of n-Octanol (pre saturated with water) and it was shaked and then 50 ml of distilled water (pre saturated with n- Octanol ) was added and was shaked the mixture by mechanical shaker for 24 hours. After 24 hour both phases are separated. Absorbance was taken of both the phases and calculated the concentration in each phases. Po / w = Coil /cwater So Po / w = 17.64706 / 82.35294 = 0.2142 Drug Excipient Compatibility Studies A small amount of drug substance with excipients that is, physical mixture of the drug and excipients (in 1:1 ratio were prepared to have maximum likelihood interaction between them) was placed in a vial, and rubber stopper was placed on the vial and sealed properly. A storage period of 2 weeks at 60C (except for dicalcium phosphate and stearic acid for which 40C is used), and the same sample was retained for 2 months at 40C. After storage the sample were observed physically for liquefaction, caking, odour or gas formation, discolouration. The compatibility study is also carried out by IR Spectroscopy curves are shown below. Observations are recorded in table 6 and 7. Table 6: Drug -Excipient Compatibility Observations Additives (50 mg each) with Observation at 60C Observation at 40C drug for 2 weeks for 2 month Drug (Vancomycin HCl) No Change No Change Drug + Lactose (DCL 21) No Change No Change Drug + Avicel 200 No Change No Change Drug + Kollidon CL No Change No Change Drug + Ac- di Sol No Change No Change Drug + Magnesium Stearate No Change at 40C No Change at 40C Drug + Aerosil 200 No Change No Change International Journal of Pharma Research and Development Online

Remarks Accepted Accepted Accepted Accepted Accepted Accepted Accepted 5

www.ijprd.com

Publication Ref No.: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-9/NOV/002

ISSN 0974 9446

IR Spectroscopic Studies15, 16 IR Spectra of pure Vancomycin HCl and in combination with tablet excipients (physical mixture) were recorded between 4000-450cm-1. The spectra are shown in figure 2. The characteristics spectral bands of pure vancomycin HCl are sumerzed in table7 Table 7: IR Spectral Bands of Vancomycin HCl Wave Number ( cm-1) Characteristics 3401.18 Phenolic OH ( stretching) 1651.87 Aromatic C=C ( stretching) 1505.44 C=O (stretching) associated with secondary amide function 1396.30 C-O (stretching) due to phenolic OH group 1231.41 Ar-O-Ar (stretching) 1061.56 OH (deformation)

RESULTS AND DISCUSSION: There was no significant change in the absorbance value of vancomycin hydrochloride up to 30 minute, which indicates that vancomycin hydrochloride is stable in distilled water as well as in 0.1N HCl. The calibration curve shows a linear relation between concentration (0 120 g / ml) and absorbance. It shows that Beer Lambert law is obeyed in the concentration range of 40 120 g / ml. There was no significant change of absorbance was absorbed other than in Di Calcium Phosphate between additives and drug sample solutions. This indicates there was no interference with vancomycin hydrochloride during analysis by Uv spectrophotometer. Calculated value of % compressibility (17.5) indicates that vancomycin hydrochloride is suitable for direct compression as well as having good flow property so almost negligible chances of weight variation during filling of dye by the blend. Compatibility studies performed on the physical mix of vancomycin HCl and different tablet excipients at temperature 400C and 600C shows no physical changes The characteristics spectral bands of vancomycin HCl were not significantly affected in the physical mixture of drug and excipients. All the characteristics bands of the drug were retained at their respective positions in the IR spectra of drug-excipient physical mixtures. No significant shift in the position of the characteristics bands observed this shows that there was no interaction between vancomycin HCl and the selected tablet excipients in the physical mixtures.

CONCLUSION In the light of this research work, it may be concluded that tablets of vancomycin hydrochloride can be made by direct compression method by using lactose (DCL-21), Kollidon-Cl, Aerosil -200, magnesium stearate and Avicel-200 as suitable excipients as all they showed the compatibility with the Vancomycin hydrochloride.

International Journal of Pharma Research and Development Online

www.ijprd.com

Publication Ref No.: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-9/NOV/002

ISSN 0974 9446

REFERENCES 1. Ansel,H.C., Allen, L.V., and Popvich, N.G., Pharmaceutical Dosage Forms and Drug Delivery systems, Seventh Edition(1999), Lippincott Williams and wilkins, Philadelphia, USA, 179-228. 2. www.lilly.com 3. www.viropharma.com 4. Khan,K.A., and Rhodes, C.T., The Production of Tablets by Direct Compression, Can. J. Pharm, Sci., 1973, 8: 1-5. 5. Shangraw, R.F., Compressed Tablets by Direct Compression Granulation Pharmaceutical Dosage Forms: Tablets, Vol.-1, Marcel Dekkar, USA, Second Edition, (1989), 195-246. 6. Handbook of Pharmaceutical Excipients, Second Edition (1994), ALPA Pharm. Sc.Great Britain, AphA, Washington DC, 89-86. 7. Armstrong, N.A., Selection of Excipients for Direct Compression Tablet Formulation, Pharm. Technol. Eur., 1997, 9: 24-30. 8. Antonio Moronim, A Novel Copovidone Binder for Dry Granulation and Direct Compression Tableting, Pharm. Technol., 1986, 24: 8 12. 9. Lachman,L., Liberman, H.A., and Kanig, J.L., The Theory and Practice of Industrial Pharmacy, Lea & Febiger, Philadelphia, 1986,171-195. 10. www.health.allrefer.com/health/pseudomembranous-colitis-info.html 11. www.cdc.gov/ncidod/hip/gastro/clostridiumDifficileGen.htm 12. www.ascp.com/public/pubs 13. Backett, A.H., and Stenlake, J.B., Practical Pharmaceutical Chemistry, First Edition, Reprint, 2004, CBS Publishers and Distributors, New Delhi, 275 325. 14. Carstensen, J.T., Pharmaceutical Preformulation, 1998, Technomic Publishing Company, Inc., New Holland Avenue, Lancaster, Pennysylvania, USA, 13 24, 41 48, 259 274. 15. Vogals Textbook of Quantitative Chemical Analysis, Fifth Edition, reprint, 1996, English Language Book Society, London, 645 651, 752 755. 16. Pavia, D.L., Lampman, G.M., and Kriz, G.S., Introduction to Spectroscopy, Third Edition, 2001, Harcourt College Publishers, Orlando, Florida, USA, 353 359.

International Journal of Pharma Research and Development Online

www.ijprd.com