Int J Curr Pharm Res, Vol 12, Issue 2, 68-71Original Article
SPECTROPHOTOMETRIC DETERMINATION OF DASATINIB IN PHARMACEUTICAL FORMULATIONS
YOGESH THORAT1, ANITA SHEGAONKAR1*, SMITA KUMBHAR1, VINOD MATOLE1, AVINASH HOSMANI2
1D. S. T. S. Mandal’s College of Pharmacy, Solapur 413004. Maharashtra, India, 2Government College of Pharmacy, Ratnagiri
Email: anitashegaonkar11@gmail.com
Received: 20 Nov 2019, Revised and Accepted: 19 Jan 2020
ABSTRACT
Objective: A new, simple, sensitive, precise and reproducible bioanalytical method was developed for the determination of Dasatinib in pharmaceutical formulations with Chloranilic acid.
Methods: The method is based on formation of violet colored complex. The UV spectrum of Dasatinib in methanol showed λ max at 521 nm. Beer’s law is valid in the concentration range of 10-60 μg/ml. This method was validated for linearity, accuracy, precision, ruggedness and robustness.
Results: The method has demonstrated excellent linearity over the range of 10-60 μg/ml with regression equation y = 0.021x-0.083 and regression correlation coefficient r2= 0.997. Moreover, the method was found to be highly sensitive with LOD (2.96μg/ml) and LOQ (8.98μg/ml).
Conclusion: Based on results the proposed method can be successfully applied for the assay of Dasatinib in various pharmaceutical dosageforms.
Keywords: Dasatinib, Bioanalytical method, Spectrophotometry, Chloranilic acid, Method development, Validation
© 2020 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
DOI: http://dx.doi.org/10.22159/ijcpr.2020v12i2.37492. Journal homepage: https://innovareacademics.in/journals/index.php/ijcpr
INTRODUCTION
Cancer is an abnormal, continuous multiplying of cells. The cells divide uncontrollably and may grow into adjacent tissue or spread to distant parts of the body [1].
Dasatinib is an anticancer agent which is given orally to treat the chronic myeloid leukemia and acute lymphoblastic leukemia. Specifically it is used to treat cases that are Philadelphia chromosome-positive (Ph+). It is classified as a kinase inhibitor. The main action of kinase inhibitors is to prevent the growth of tumors by the action of reducing the activity of proteins that controls growth, cell division and survival. Dasatinib was approved for medical use in the United States in 2006. .It is on World Health Organization List of Essentials Medicines, the most effective andsafe medicines needed in a health system [2-5].
Fig. 1
Structure of dasatini
The chemical name of Dasatinib is N-(2-Cholro-6-methylphenyl)-2-[[6-[4-(2-hydroxyehyl)-1-piperazinyl]-2-methyl-4-pyrimidinyl] amino]-5-iazolecarboxamide monohydrate. It has molecular formula of C22H26Cl1N7O2Sand molecular weight of 488.01 gm/ml. Dasatinib is white powder and has melting point 280 °-286 °C. The drug substance is soluble in acetonitrile and slightly soluble in methanol [6-8].
The aim of this work is to introduce a simple, precise and rapid procedure for the simultaneous quantitation of the cited drug in pure and pharmaceutical formulation.
MATERIALS AND METHODS
Dasatinib was taken as a gift sample from Microlab, Bengaluru, India. Chloranilic acid, Methanol and Dichloromethane were used were of analytical grade.
Instruments
A UV visible double beam spectrometer [systronics 2201] and Shimadzu 1800-UV spectrophotometer with 1 cm quartz cuvettes was used for all absorbance measurement.
All weights were taken on analytical balance. Sonicator was used for dissolving Dasatinib in methanol.
Experimental
Preparation of chloranilic acid
0.3% (w/v) of Chloranillic acid was dissolved in some amount of methanol (20% of the total volume) and make up the volume using methylene chloride.
Preparation of standard stock solution
Accurately weighed 10 mg of Dasatinib transferred to 100 ml volumetric flask. It was dissolved in methanol and sonicated for 10 min. The volume was made up to mark with methanol to obtain final strength.
Procedure for plotting calibration curve
To a series of 10 ml volumetric flask, 1-6 ml of standard stock solution was pipetted out separately and into each flask 1 ml of 0.3% Chloranilic acid was added. The volume was adjusted using methanol. The produced violet color was measured at wavelength 521 nm against blank solution which was prepared without adding drug in similar manner.
20 Tablet containing Dasatinib were weighed. An accurately weighed portion of the powder equivalent to 10 mg of Dasatinib was dissolve in a 100 ml of methanol and mixed for about for 10 min and sonicated for about 15 min then filtered. From formed solution with concentration of 100μg/ml seven aliquots were pipetted out into a 100 ml of volumetric flask having concentration 10-60μg/ml to each flask and 1 ml of 0.3% Chloranilic acid was added. The volume was made up to mark with methanol. These solutions were analyzed at selected wavelength 521 nm and results were statistically validated.
RESULTS AND DISCUSSION
The absorption spectra shows result of wavelength at 521 nm.
Fig. 2: The proposed method was validated according to ICH Guidelines [9, 10]
Linearity
The linearity was confirmed by taking aliquots of concentration of 10-60 µg/ml and absorbance was measured. It was performed in single day only. The obtained absorbance shows good regression coefficient at wavelength 521 nm. The slope and intercept values were recorded. The linearity was plotted against absorbance of Dasatinib vs concentration of Dasatinib.
Table 1: Results for linearity
S. No. |
Concentration (µg/ml) |
Absorbance |
1 |
10 |
0.107 |
2 |
20 |
0.37 |
3 |
30 |
0.59 |
4 |
40 |
0.8 |
5 |
50 |
1.01 |
6 |
60 |
1.21 |
Fig. 3: Calibration curve for dasatinib
Accuracy
The accuracy is parameter of an analytical method which describes the closeness to the test results obtained by that method to the theoretical value. The standard addition method is used to analyze accuracy which is performed by using previously analyzed standard solutions. The percentage relative standard deviation and percentage recovery were analyzed by using standard solutions.
Range
The range is the analytical parameter of interval between lower and upper concentration limit of an analyte i.e. 10-60 µg/ml.
Precision
The precision is performed as inter-day and intra-day. Intra-day precision was performed in one day and inter-day precision was performed in three days. Dasatinib was evaluated at concentration 30 µg/ml. The percentage RSD for intra-day precision was found to be 0.69% and inter-day precision was found to be 0.86%.
Limit of detection (LOD)
The limit of detection (LOD) or lower limit of detection is the lowest quantity of a substance that can be able to distinguish from the absence of that substance with a stated experimental level.
LOD = 3 Sa/b
Limit of quantitation (LOQ)
The limit of quantitation (LOQ) is the lowest concentration at which the performance of a method or measurement system is acceptable for a specified use.
LOQ =10 Sa/b
Ruggedness
The ruggedness is the study of degree of reproducibility of test results obtained by variety of external conditions like different analysts, laboratories, days and reagents. This study shown that there is no any influence of these conditions on test results.
Table 2: Regression analysis of the calibration curve for proposed method
| Parameters | Method values |
| λ max | 521 nm |
| Beer’s law | 10-60μg/ml |
| Correlation coefficient (r) | 0.997 |
| Regression equation (Y = mx+c) | 0.021x-0.083 |
| Slope (m) | 0.021 |
| Intercept (c) | 0.083 |
| LOD(μg/ml) | 2.96 |
| LOQ(μg/ml) | 8.98 |
Table 3: Result for precision (Intra-day)
| S. No. | Concentration (µg/ml) | Absorbance 1 | Absorbance 2 | Absorbance 3 | %RSD |
| 1 | 30 | 0.590 | 0.589 | 0.592 | |
| 2 | 30 | 0.592 | 0.595 | 0.583 | |
| 3 | 30 | 0.591 | 0.594 | 0.599 | |
| 4 | 30 | 0.595 | 0.592 | 0.595 | |
| 5 | 30 | 0.600 | 0.597 | 0.594 | |
| 6 | 30 | 0.597 | 0.591 | 0.598 | |
| %RSD | 0.64% | 0.48% | 0.96% | 0.69% |
Table 4: Result for precision (Inter day)
| S. No. | Concentration (μg/ml) | Day 1 | Day 2 | Day 3 | %RSD |
| 1 | 30 | 0.590 | 0.592 | 0.598 | |
| 2 | 30 | 0.592 | 0.599 | 0.602 | |
| 3 | 30 | 0.591 | 0.609 | 0.591 | |
| 4 | 30 | 0.595 | 0.594 | 0.593 | |
| 5 | 30 | 0.600 | 0.593 | 0.605 | |
| 6 | 30 | 0.597 | 0.597 | 0.597 | |
| %RSD | 0.65% | 1.05% | 0.88% | 0.86% |
Table 5: Result for Robustness
| Temperature | 30 °C | 25 °C |
| Concentration | 8μg/ml | 8μg/ml |
| Absorbance | 0.590 | 0.601 |
| 0.591 | 0.599 | |
| 0.592 | 0.603 | |
| 0.595 | 0.597 | |
| 0.600 | 0.595 | |
| 0.597 | 0.598 | |
Average SD |
0.594167 0.003859 |
0.598833 0.002858 |
Table 6: Result of ruggedness
| Concentration | Analyst 1 | Analyst 2 |
| 8µg/ml | 0.590 | 0.598 |
| 0.591 | 0.594 | |
| 0.592 | 0.599 | |
| 0.595 | 0.591 | |
| 0.600 | 0.601 | |
| 0.597 | 0.592 | |
Average SD |
0.595833 0.00407 |
Robustness
The robustness is the small but deliberate variations in method parameters such as temperature and stability of analytical solution.
Linearity
Six different concentrations of Dasatinib were prepared and analyzed. Then wavelength was found to be 521 nm. The regression coefficient was found to be 0.997. The absorbance was found in limit i.e. 0-2. Hence the analyzed parameter was found to be validated (table 1).
Precision
Intra-day precision
Intra-day precision was found within limit i.e.30 µg/ml at 521 nm. The relative standard deviation is less than 2%. Hence the parameter was found to be validated (table 3).
Inter-day precision
Inter-day precision was performed in three days and the obtained results of concentration 30 µg/ml at 521 nm shown that the relative standard deviation is less than 2%. Hence the parameter was found to be validated (table 4).
Robustness
The change in concentration i.e. 8 µg/ml and change in temperatures i.e. at 25 °C and 30 °C. And obtained results shown that there is negligible effect on results. The robustness was found to be in limit i.e. the relative standard deviation is less than 2%. Hence the performed parameter was found to be validated (table 5).
Ruggedness
The change in analyst at concentration of 8µg/ml showed that the obtained result does not affected by it (table 6).
Limit of detection
The limit of detection was found to be 2.96 µg/ml (table 2).
Limit of quantification
The limit of quantification was found to be 8.98 µg/ml (table 2).
CONCLUSION
Bioanalytical method was developed and validated thoroughly for quantitative determination of Dasatinib Tablets. The presented method was found to be rugged, simple, accurate, precise, and reproducible and gives an acceptable recovery of the analyte, which can be directly easily applied to the analysis of pharmaceuticalTablets formulation of Dasatinib.
ACKNOWLEDGEMENT
Authors are thankful to the Principal, D. S. T. S. Mandal’s College of Pharmacy, Solapur, for providing the necessary facilities.
FUNDING
Nil
AUTHORS CONTRIBUTIONS
All the authors have contributed equally.
CONFLICT OF INTERESTS
Declare none
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