Chitosan Loaded Mucoadhesive Microspheres of Gliclazide - Journal

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software of the instrument which is directly proportional to the concentration of the standard solutions (Graph.2). The method was applied for the known concentration and was found to be satisfactory for the analysis of tablet formulations. Method C: Area Under Curve (AUC) Shobha Manjunath et al./ A Novel Spectrophotometric Estimation of Pramipexole in Bulk Drug and Formulations This method is applicable when there is no sharp peak or broad spectra are obtained. It involves the calculation of integrated value of absorbance with respect to the wave length between two selected wave lengths λ1 and λ 2. Area calculation processing item calculates the area bound by the curve and the horizontal axis. The horizontal axis is selected by entering the wavelength range over which the area has to be calculated. This wave length range is selected on the basis of repeated observations so as to get the linearity between AUC and concentrations. Suitable dilutions of standard stock -1 solution (100 µg mL ) of the drug were prepared and scanned in the spectrum mode from the wavelength range 400-200 nm (Fig. 3). From the spectrum, AUC in the range of 255-265 nm was selected for analysis. The calibration curve was plotted as -1 concentration (9-45 µg mL ) against AUC (Graph 3). The method was checked by analyzing the samples with known concentration for the AUC. As per the results obtained were satisfactory, the method was applied for pharmaceutical formulations. For the estimation of Pramipexole in tablet formulations by the three methods, fifty tablets of each brand were weighed and triturated to fine powder. Tablet powder equivalent to 5mg of Pramipexole was weighed and dissolved and further diluted with sufficient quantity of distilled ethanol to get stock -1 solution of concentration 100 µg mL . For method A, analysis of Pramipexole in both tablet formulations Parpex (T 1) and Pramipex (T 2 ) was done using various sample solutions at 261.8 nm against reagent blank in quantitation mode for six times. For method B, analysis of T 1 and T2 was done for first order derivative spectra using various sample solutions at 249.4 nm against reagent blank for six times. For method C the sample solutions were scanned in the spectrum mode and AUC calculations were done in the wavelength range of 255- 265 nm for both the tablet formulation T 1 and T 2 for six times and concentrations calculated using the calibration curve. Methods A, B and C were validated for linearity, accuracy. RESULTS AND DISCUSSION: The Optical Characteristics such as Absorption maxima, Beer's law limits, Molar absorptivity are presented in (Table1). The regression analysis using the method of least squares was made for the slope (b), intercept (a) and correlation coefficient (r) obtained from different concentrations and the results are summarized in (Table 1). The percent relative standard deviation and percent range of error (0.05 and 0.01 level of confidence limits) calculated from the eight measurements, ¾ of the upper Beer's law limits of Pramipexole are given in (Table 1). Relative standard deviation for analysis of six replicate samples of two brands T 1 and T 2 in method A, B and C are given in (Table 2). The percentage recovery was found to be within range. The results obtained by proposed method are in good agreement with the label claims (Table 2). Table 1: Optical Characteristics and Other Parameters Parameters Method A Method B Method C λl max (nm) 261.8 249.4 255-265 -1 Beer ’ s Law Limit (µg mL ) (C) 9 – 45 9 - 45 9 – 45 -1 -1 Molar absorptivity (L mole cm ) 3 5.050 x 10 3 4.349 x 10 4 4.7603 x 10 Regression Equation (Y*) Slope (b) 0.0165 0.014 0.001 Intercept (a) 0.0006 0.013 -3.398 Correlation Coefficient ® 0.9998 1.003 1.822 % RSD Range of errors** 0.1983 0.0527 0.0278 Confidence limits with 0.05 level ±0.0007 ±0.0001 ±0.298 Confidence limits with 0.01 level ±0.0011 ±0.0002 ±0.441 -1 Limit of Detection (LOD)( µg mL ) 0.1783 0.0482 0.013 -1 Limit of Quantification (LOQ) ( µg mL ) 0.5404 0.1463 0.041 -1 Y*= bC+a where C is the concentration of Pramipexole in µg mL and Y is the absorbance at the respective λ max. ** For eight measurements. 108 RJPS, Jul - Sep, 2011/ Vol 1/ Issue 2
Shobha Manjunath et al./ A Novel Spectrophotometric Estimation of Pramipexole in Bulk Drug and Formulations Table 2: Evaluation of Pramipexole in Pharmaceutical Preparations Method Brand Labelled amount (mg) Amount obtained (mg)* %Recovery A T1 1 0.986 99.73 T2 1 0.983 99.62 B T1 1 0.989 99.83 T2 1 0.987 99.82 C T1 1 0.991 99.87 T2 1 0.986 99.73 *Average of Six determinations Fig.1: Selection of Wavelength for Pramipexole. Method A Graph.1: Calibration curve of Pramipexole Method A Fig.2: First order derivative spectrum of Pramipexole with n=1. Method B 109 Graph.2: Calibration Curve of Pramipexole Method B Fig.3: Wavelength range selected for AUC. Method C Graph.3: Calibration Curve of Pramipexole Method C RJPS, Jul - Sep, 2011/ Vol 1/ Issue 2
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