Estimation of Imatinib Mesylate by Extractive Colorimetric Methods in Bulk and Pharmaceutical Dosage Form
Rajan V. Rele*, Prathamesh P. Tiwatane
Central Research Laboratory, D.G. Ruparel College, Matunga, Mumbai 400016.
*Corresponding Author E-mail: drvinraj@gmail.com
ABSTRACT:
Simple sensitive and accurate extractive spectrophotometric methods have developed for the estimation of Imatinib mesylate in pharmaceutical dosage form. The methods are based on the formation of coloured complexes by the drug with reagents like bromophenol blue, solochrome dark blue and bromocresol green in acidic medium. The ion associated complexes were formed and quantitatively extracted under the experimental condition in chloroform. The absorbance values were measured at 420 nm, 495 nm and 430 nm respectively. The proposed methods were validated statistically. Recoveries of methods were carried out by standard addition methods. The linearity was found to be 1-8 μg/ml, 2 -16μg/ml, 1-16 μg/ ml for methods 1, 2 and 3 respectively. The low values of standard deviation and percentage RSD indicate high precision of methods. Hence these methods are useful for routine estimation of Imatinib mesylate in tablets.
KEYWORDS: Imatinib mesylate, Bromophenol blue, Solochrome dark blue, Bromocresol green, Chloroform.
INTRODUCTION
It is a 4-[(4-methylpiperazin-1-yl)methyl]-N-(4-methyl-3-{[4-(pyridin-3-yl)pyrimidin-2-yl]amino}phenyl) benzamide, imatinib mesylate. It is known for chronic myelogenous leukemia and acute lymphocytic leukemia that are philadephia chromosome positive, certain types of gastrointestinal stromal tumors, hypersinoiphilic syndrome, chronic eosinophilic leukemia, systemic mastocytosis and myelodysplastic syndrome.
According to the literature review several methods has been developed for drug, like spectroscopy methods1-7. HPLC8-22, and HPTLC23. This extractive colorimetric can be used for the routine analysis. In the proposed methods optimization and validation of this method are reported.
Structure of imatinib mesylate:
MATERIALS AND METHODS:
A Shimadzu -160 A double beam UV-Visible recording spectrophotometer with pair of 10mmmatched quartz cell was used to measure absorbance of solutions. A Shimadzu analytical balance was used.
Bromophenol blue, solochrome dark blue, bromocresol green, hydrochloric acid, potassium hydrogen phthalate and chloroform of A.R. grade were used in the study.
Preparation of standard solution and reagents:
Stock solution of Imatinib mesylate (100 μg/ml) was prepared in distilled water. From this stock solution working standard (10 μg/ml) was prepared by diluting 10 ml stock solution to 100mlwith distilled water. 0.6%w/v solution of bromophenol blue, 0.25 %w/v solochrome dark blue and 0.2 %w/v bromocresol green were prepared in distilled water respectively.
Potassium hydrogen phthalate buffer solution of pH 4.01was prepared in distilled water. Dilute hydrochloric acid was used to adjust desired pH of buffer solution.
EXPERIMENTAL:
Method 1 (with bromophenol blue):
Into a series of separating funnels appropriate amount of the working standard drug solutions were pipetted out. To each funnel 1.0ml of buffer (pH= 4.2) and 6 ml of 0.6 %w/v bromophenol blue were added. 10 ml of chloroform was added to each funnel. The solutions were shaken for thorough mixing of the two phases and were allowed to stand for clear separation of the layers. The absorbance values of the chloroform layers were measured against their respective reagent blank at the wavelength of the maximum absorbance (λ max 420 nm).
Method 2 (with solochrome dark blue):
Into a series of separating funnels appropriate amount of the working standard drug solutions were pipetted out. To each funnel 4.0 ml of buffer (pH = 1.10) and 4 ml of 0.25% w/v solochrome dark blue were added. 10ml of chloroform was added to each funnel. The solutions were shaken for thorough mixing of the two phases and were allowed to stand for clear separation of the layers. The absorbance values of the chloroform layers were measured against their respective reagent blank at the wavelength of the maximum absorbance (λ max=495 nm).
Method 3 (with bromocresol green):
Into a series of separating funnels appropriate amount of the working standard drug solutions were pipetted out. To each funnel 1.0ml of buffer (pH= 3.7) and 5.0 ml of 0.02% w/v bromocresol green were added. 10 ml of chloroform was added to each funnel. The solutions were shaken for thorough mixing of the two phases and were allowed to stand for clear separation of the layers. The absorbance values of the chloroform layers were measured against their respective reagent blank at the wavelength of the maximum absorbance (λmax =430 nm).
Estimation from tablets:
Twenty tablets were weighed accurately and average weight of each tablet was determined. Powder equivalent to 10 mg of imatinib mesylate was weighed and transferred in 100 ml of volumetric flask. A 30 ml of distilled water was added and sonicated for 15 minutes and filtered. The filtrate and washing were diluted up to the mark with distilled water to give concentration as 100 μg /ml. Such solution was used for analysis.
Table 1: Values of results of optical and regression of drug
|
Parameter |
bromophenol blue |
Solochrome dark blue |
Bromocresol green |
|
Detection Wavelength (nm) |
420 |
495 |
430 |
|
Beer Law Limits (µg/ml) |
1-8 |
2-14 |
1-16 |
|
Correlation coefficient(r2) |
0.9999 |
0.9999 |
0.9999 |
|
Regression equation (y=b+ac) |
|
|
|
|
Slope (a) |
0.0301 |
0.02020 |
0.0254 |
|
Intercept (b) |
0.0009 |
-0.00014 |
-0.0003 |
RESULTS:
The extractive spectrophotometric methods are popular due to their sensitivity in assay of the drug and hence ion pair extractive spectrophotometric methods have gain considerable attention for quantitative determination of many pharmaceutical preparations. These proposed methods are extractive spectrophotometric methods for the determination of Imatinib mesylate by using chloroform as solvent from its formulations i.e. tablets. The colour ion pair complexes formed are very stable. The working conditions of these methods were established by varying one parameter at time and keeping the other parameters fixed by observing the effect produced on the absorbance of the colour species. The various parameters involved for maximum colour development for these methods were optimized. The proposed methods were validated statistically and by recovery studies. The molar absorptivity show the sensitivity of methods while the precision was confirmed by %RSD (relative standard deviation). The optical characteristics such as absorption maxima (nm), molar absorptivity (l -mole-1 cm-1), co-relation coefficient (r) were calculated and are also summarized. Assay results of recovery studies are given in table 2 (A, B, C).
Table no 2: A (Bromophenol blue)
|
Amount of Sample Added in (µg/ml) |
Amount of Standard Added in (µg/ml) |
Total amount recovered |
Percentage recovery (%) |
Standard deviation |
Percentage of relative standard deviation (C.O.V.) |
|
1 |
0 |
1.008571 |
100.8571 |
0.011524 |
1.142613 |
|
2 |
1 |
2.020952 |
101.0476 |
0.024699 |
1.222159 |
|
3 |
2 |
3.070952 |
102.3651 |
0.0046 |
0.149805 |
|
4 |
3 |
4.042857 |
101.0714 |
0.03709 |
0.917418 |
|
|
|
|
|
Mean= 0.019478 |
Mean= 0.8579 |
Table no 2: B (Solochrome dark blue)
|
Amount of Sample Added in (µg/ml) |
Amount of Standard Added in (µg/ml) |
Total amount recovered |
Percentage recovery (%) |
Standard deviation |
Percentage of relative standard deviation (C.O.V.) |
|
2 |
0 |
2.005575 |
100.2787 |
0.004389 |
0.218838 |
|
2 |
2 |
3.990383 |
99.75958 |
0.033397 |
0.836932 |
|
2 |
4 |
5.95122 |
99.18699 |
0.028163 |
0.473238 |
|
2 |
6 |
7.993031 |
99.91289 |
0.086479 |
1.081924 |
|
|
|
|
|
Mean= 0.038107 |
Mean= 0.652733 |
Table no 2: C (Bromocresol green)
|
Amount of Sample Added in (µg/ml) |
Amount of Standard Added in (µg/ml) |
Total amount recovered |
Percentage recovery (%) |
Standard deviation |
Percentage of relative standard deviation (C.O.V.) |
|
2 |
0 |
2.005602 |
100.2801 |
0.003827 |
0.190818 |
|
2 |
2 |
3.989356 |
99.73389 |
0.043828 |
1.098618 |
|
2 |
4 |
5.979272 |
99.65453 |
0.038803 |
0.648953 |
|
2 |
6 |
7.955182 |
99.43978 |
0.035284 |
0.443532 |
|
|
|
|
|
Mean= 0.030435 |
Mean= 0.59548 |
Results are in good in agreement with labelled value.
DISCUSSION:
The percent recovery obtained indicates non interference from the common excipients used in the formulation. The reproducibility, repeatability and accuracy of these methods were found to be good, which is evidenced by low standard deviation. The proposed methods are simple, sensitive, accurate, precise and reproducible. They are directly applied to drug to form chromogen. Hence they can be successfully applied for the routine estimation of Imatinib mesylate, in bulk and pharmaceutical dosage form even at very low concentration and determination of stability of drug in formulation such as tablets. The strong recommendation is made here for the proposed methods for determination of Imatinib mesylate from its formulation.
ACKNOWLEDGMENT:
Authors express sincere thanks to the Principal, of D. G. Ruparel college.
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Received on 03.06.2024 Modified on 18.06.2024
Accepted on 01.07.2024 ©AJRC All right reserved
Asian J. Research Chem. 2024; 17(3):145-148.