Spectrophotometric Method Development of ...

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has also been studied for the treatment of reactive hypoglycemia, the dumping syndrome and certain types of hyperlipoproteinaemia.[2]. Acarbose is official in ...

G. Kumar et al. / Journal of Pharmacy Research 2009, 2(10),1595-1597

Research Article ISSN: 0974-6943

Available online through http://jprsolutions.info Spectrophotometric Method Development of Acarbose From Bulk And In Its Tablet Dosage Form 1

G. Kumar *1,V.Juyal2, P.P.Badoni3, S.Kumar1,M.S.M.Rawat4. Department of Pharmacy, S.G.R.R.I.T.S. Patel Nagar, Dehradun -248001, Uttarakhand, India 2 Department of Pharmaceutical Sciences, Kumaon University, Bheemtal, (UA) 3 Department of Chemistry, H.N.B. Garhwal University, Pauri Campus (UA) 4 Department of Chemistry, H.N.B. Garhwal University, Srinagar (UA) - 246174 Received on: 14-05-2009; Accepted on:15-08-2009

ABSTRACT A simple rapid spectrophotometeric method has been developed for estimation of acarbose from bulk drug and tablet dosage form by using potassium permagnate and sodium hydroxide as oxidizing agent. The method is based on the formation of green colored complex of drug with 0.1 N alkaline potassium permanganate having absorbance maxima at 625 nm. The beer’s law is obeyed in the concentration range of 10-70 µg/ ml of the drug but more precisely it obeys in the range of 10-50 µg/ml. The slope and intercept values are 0.0118 and 0.0071, respectively. Results of analysis of this method were validated statistically and by recovery studies. The method is applied to the marketed tablet formulation. Result of analysis of tablet formation given as percentage of label claim+ standard deviation is 99.78 + 0.1053. The precision and accuracy was examined by performing recovery studies and was found to be 99.59 + 0.180.The developed method is simple, sensitive and reproducible and can be used for routine analysis of acarbose from bulk and tablet dosage form. Keywords: Acarbose, Green complex, Spectrophotometery, Alkaline Potassium permagnate. INTRODUCTION Acarbose(O-4,6-dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy3-(hydroxymethyl)-2-cyclohexen-1-yl]amino]-a-D-glucopyranosyl-(14)-O-a-?D-glucopyranosyl-(1-4)-D-glucose),[1] is an oral alpha-glycosidase inhibitor, especially sucrase and structure shown in figure 1. It is given by mouth in the treatment of type 2 diabetes mellitus. It has also been studied for the treatment of reactive hypoglycemia, the dumping syndrome and certain types of hyperlipoproteinaemia.[2] Acarbose is official in Indian pharmacopoeia. Some analytical methods have been reported for the determination of the studied drugs. The reported methods of acarbose include gas chromatography – mass spectrophotometery (GC-MS),[3] High performance liquid chromatography,[4-5] and capillary electrophoresis.[5-6] To the best of our knowledge, no Spectophotometric method have been reported for the analysis of acarbose till now, The results obtained were promising. Several analytical techniques such as, Spectrophotometery, chromatography, capillary electrophoresis, kinetic flourimetry, flow injection analysis and chemiluminescence were utilized for selective oxidation and determination of many pharmaceutical compounds in formulations using potassium permagnate as reagent viz benzenediol and benzenetriol,[7] perchloroethylene,[8] propanolol.[9] *Corresponding author. Tel.: + 91-9897435971 Telefax: +91-135-2721762 E-mail: [email protected]

The method developed is based on the formation of complex of acarbose with 1% potassium permagnate in 0.1N sodium hydroxide to give green colored chromogen withλmax 625 nm. Reaction conditions were optimized to obtain maximum color intensity. The method is simple, reproducible and requires low cost and method is applied successfully to the analysis of the marketed tablet formulation.[10] MATERIALSAND METHODS: Instrumentation An Elico UV-visible spectrophotometer (model UV-160) with 1cm matched quartz cells was used for all absorbance measurements. Reagents All chemical used were of analytical reagent grade and the solvents were of spectroscopic grade •Potassium permagnate (Merck, Germany) 0.01 N aqueous solution. •Sodium hydroxide (Thomas baker (chemicals) Pvt. Ltd. India)0.1N aqueous solution. Materials The different pharmaceutical preparations were purchased from the commercial source in the local market. they include: •Acarbose was kindly offered as gift sample from Biocon pharma

Journal of Pharmacy Research Vol.2.Issue 10.October 2009

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G. Kumar et al. / Journal of Pharmacy Research 2009, 2(10),1595-1597 Table 1: Optical Parameters and Regression Characteristics of Acarbose S.N. 1 2 3 4 5 6 7

Parameters Beers law limit(µg/ml) molar absorptivity (1 mole-1 cm-1) Sandell’s Sensitivity (mg/cm2 /0.001Absorbance unit) Regression equation (y=a +bc) Slope ( b) Intercept ( a) Correlation coefficient (r2)

Figure.3.

Observations 10-70 14.01 x 103 4.60 x 10-2 Y= .0071+ 0.0118c 0.0118 0.0071 0.9937

Table 2: Summary of validation parameters Validation parameter Linearity and Range (µg/ml) Correlation Coefficients (r) Precision (RSD) Repeatability (n=6) Intraday (n=3) Interday (n=3) % Recovery Specificity Limit of Detection (ppm) Limit of Quantitation (ppm)

Acarbose observation 10 -70 0.9937 0.0634 0.0744 0.2939 100.59 2.5 x 10-3 1.77 17.7 10 -70

Table 3: Results of acarbose in tablet Brand

Acarbose

Label Theoretical Calculated Amount % claim conc. conc. found Assay (mg/tab) (µg/ml) (µg/ml) (mg/tab) 49.822 49.822 99.64 49.944 49.944 99.88 50 50 49.923 49.923 99.84 49.889 49.889 99.77

India. •Glucobay 50 tablets, labeled to contain 50 mg acarbose/tablet batch P-09151 the product of Bayer pharmaceutical Pvt. Ltd. Preparation of standard stock solution of Acarbose. a)Accurately about 100mg of drug was weighed and transferred into 100 ml of distilled water in volumetric flask and 2 ml of this solution Mean was taken in 8 ml of distilled water. Other concentrations were pre% Assay±SD pared by further dilution with distilled water. These solutions also were found stable for at least three days without alteration when kept in refrigerator. 99.78 ± 0.1053 b)5 ml of 0.01 N Potassium Permagnet in 0.1 N Potassium Hydroxide solution were prepared. Determination of wavelength of maximum absorbance (λ max) 0.5 ml of the standard stock solution was pipette out and transferred to a 10 ml of volumetric flask and added 5 ml of 0.01N Potassium paramagnet (coloring agents) in dissolving 0.1N NaOH solution. The volume was made up to the mark with distilled water. This solution contained 10µg/ml of the drug, the absorbance of this solution was scanned by UV-visible spectrophotometery in the range of 300-750. The maximum absorbance of Acarbose was obtained at 625 nm shown in figure2. Preparation of calibration curve for Acarbose at 625 nm Firstly, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5ml of standard stock solution of acarbose separately pipetted out in a series of 10 ml volumetric flask and added 5 ml of 0.01 KMnO4 in 0.1 N NaOH solution remaining up to mark volume was made by distill water. It were make the concentration. Range of 10, 20, 30, 40, 50, 60, 70µg/ml of the drug. The absorbance of these resultant solution were measured at 625 nm by placing 0.01 N alkaline with acarbose in sample cell, and 0.01 N alkaline solution placed in reference cell, and a graph was plotted between absorbance v/s concentrations of the solutions (figure 3). The Lambert-Beer’s law was obeyed in range of 10 to 70 µm/ml at 625 nm. Result of method development and validation were shown in table 1 and table 2. Procedure for the determination of acarbose in dosage form:

An accurately weighed quantity of the mixed contents of 10 Figure 2 : wavelength of maximum absorbance (λ max) Journal of Pharmacy Research Vol.2.Issue 10.October 2009 1595-1597

G. Kumar et al. / Journal of Pharmacy Research 2009, 2(10),1595-1597 powdered tablets equivalent to 100mg of the drug was transferred into 100ml volumetric flask. The content of the flask was completed to 100ml with distilled water then sonicated for 15 minutes and filtered. An aliquots of the cited solutions was taken and the above procedure was applied. The nominal content was calculated either from a previously plotted calibration graph and result shown in table 3. RESULT AND DISCUSSION: Oxidation of the studied drugs with Potassium permagnate was carried out in presence of sodium hydroxide. Trial was made to determine the drug through oxidation with KMnO4 neutral and acidic media, but no apparent reaction product were observed. Potassium permagnate in alkaline medium oxidized the studied drug and yielded the green color of magnate radical, which absorbs maximally at 625nm. Using various normality ranges from 1N to 0.1N the normality of sodium hydroxide was optimized. Sodium hydroxide of normality equivalent to 0.1N was found to yield reproducible results. Beer’s law is obeyed in concentration range of 10-70 µg/ml. The slope and intercept values are 0.011 and 0.0071. The correlation coefficient was found to be 0.9937.To study the recovery of acarbose, drug from the tablet sample was taken to which different quantities of pure drug (reference standard) was added within the analytical concentration range in the proposed method. The added quantity of individual drug was estimated in the method.% concentration + SD and coefficient of variance for acarbose bulk drug and acarbose recovery sample were found 100.59 ± 0.6317, 99.75 ± 0.6279 and 1.013,1.019 respectively. From these values it seems that method is accurate and reproducible for both bulk drug and formulation. The marketed tablet formulations with 50mg of drug claim are used for applying developed method on formulation. % concen-

tration + SD and coefficient of variance for the tablet formulation were found to be 99.78 ± 0.1053 and 0.1090, respectively. These reaction conditions were optimized to obtain maximum color intensity. Proposed method was use for pure bulk drug as well as for marketed formulation. The result obtained compared favorably with labeled amount of drug as well as that of the formulation. None of the usual diluents, lubricant, film formers employed in preparation of tablet dosage form was found to interfere in the proposed procedure. The proposed method hence is specific, precise, accurate and reliable. ACKNOWLEDGEMENTS: Sincere thanks are due to M/s Biocon Pharma for providing gift sample of drug and Prof Vijay Juyal, Principal of Department of pharmacy, Kumaon university, Bheemtal for providing necessary facilities for experimentation. REFERENCES: 1.

Merck Index, An Encyclopaedia of Chemicals, Drugs and Biologicals, 12 Ed.; by Merck & Co. Inc., NJ., White House Station, 1995. 2. Parfitt, K. “Martindalle the Complete Drug Reference” 32nd Ed., the Pharmaceutical Press, Massachusets. 1999; 317-9. 3. Goromaru T, matsuki K, and matsuki Y. bio.pharm.bull.1994; 17:1568. 4. Youssef D, Samir C, Xavier C, Emmanuel V, et al. V. J. Sep. Sci. 2002; 25: 280. 5. Cherkaoui S, Daali Y, Christen P, and Veuthey JL. J Pharm Biomed. Anal.1998;18: 729-31. 6. Rethfeld I, and Blaschke G J. Chromatogr. B. 1997; 700: 249-51. 7. Fan SL, Zhang LK and Lin JM. Talanta. 2006; 68: 646-9. 8. Zhai X, Hua I, Rao PSC, and Lee LS J. Contaminant Hydrology. 2006; 82: 61-4. 9. Marques Santos J L M, and. Lima JLFJ. Pharm. Biomed. Anal. 2005; 39: 886-8. 10. Ibrahim FA Ali, Ahmed FA, Tolba SM. Kinetic Determination of Acarbose and Miglitol in Bulk and Pharmaceutical Formulation using Alkaline Potassium prmanganate I J Bio Sci., 2007;(1):20-2-6

Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.2.Issue 10.October 2009

1595-1597

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