Determination of Emtricitabine from Pharmaceutical Dosage Form By extractive ion pair complex colorimetric method
Rele Rajan V.*, Prathamesh P. Tiwatane
Central research Laboratory, D.G. Ruparel College Mahim, Mumbai 400 016, India.
*Corresponding Author E-mail: drvinraj@gmail.com
ABSTRACT:
It is a 4-amino-5-fluoro-1-[(2R,5S)-2 (hydroxyl -methyl)-1,3-oxathiolan-5-yl]-1,2-dihydro- pyrimidin-2-one. Emtricitabine is a nucleoside reverse transcriptase inhibitor (NRTI) for the treatment of HIV infection in adults. EMT is an analogue of cytidine. The drug works by inhibiting reverse transcriptase, the enzyme that copies HIV RNA into new viral DNA.
According to the literature review several methods has been developed for drug, like spectroscopy methods1-6. HPLC7-22, HPTLC23,24 and miscellnous25,26. These spectroscopy methods can be used for the routine analysis. In the proposed methods optimization and validation of this method are reported.
Structure of Emtricitabine:
MATERIALS AND METHODS:
A Shimadzu -160 A double beam UV-Visible recording spectrophotometer with pair of 10mm matched quartz cell was used to measure absorbance of solutions. A Shimadzu analytical balance was used.
Cobalt nitrate, Ammonium thiocynate, and chloroform of A.R. grade were used in the study.
Preparation of standard solution and reagents:
Stock solution of Emtricitabine (100μg/ml) was prepared in distilled water. From this stock solution working standard (10μg/ml) was prepared by diluting 10 ml stock solution to 100 ml with distilled water.
Experimental:
Preparation of solutions:
Stock solutions of emtricitabine (100μg/ml) were prepared in distilled water.
From these stock solutions, working standard solutions (10 μg/ml) were prepared by diluting 10ml stock solution to 100 μg/ ml with distilled water.
Preparation of reagents:
Cobalt thiocynate was prepared by dissolving cobalt nitrate and ammonium thiocynate in 1:2 molar proportions in distilled water.
Experimental:
Into a series of separating funnels appropriate amount of the working standard drug solutions were pipetted out. To each funnel 1.6ml of cobalt thiocynate complex solution was added along with 10ml of chloroform to each funnel. The solutions were shaken for thorough mixing of the two phases and were allowed to stand for separation of the layers. The absorbance value of the chloroform layers were measured against their respective reagent blank at the wavelength of the maximum absorbance(λ max 620 nm).
Estimation from tablets:
Twenty tablets of emtricitabine were weighed accurately. Average weight of each tablet of emtricitabine was determined. Tablets were crushed into fine powder. An accurately weighed quantity of powder equivalent to 10mg of emtricitabine was transferred into a beaker and it was shaken with 50ml of distilled water and filtered. The filtrate and the washing were collected in a 100.0 ml volumetric flask. This filtrate and the washing were diluted up to the mark with ethanol to obtain final concentration as 100 μg/ml. A 10ml. of this solution was further diluted to give 10μg /ml. Such solutions were further used for estimation of emtricitabine respectively. Appropriate aliquots of drug solution were taken. The individual assay procedures were carried out for the estimation of drug contents in tablets. The concentration of the drug in the tablets was calculated using calibration curve. Fig. 1
Table 1: Values of results of optical and regression of drug
|
Parameter |
Values |
|
Detection Wavelength (nm) |
620 |
|
Beer Law Limits (µg/ml) |
2-16 |
|
Correlation coefficient(r2) |
0.9999 |
|
Regression equation (y=b+ac) |
|
|
Slope (a) |
0.0071 |
|
Intercept (b) |
0.00006 |
The recovery experiment was carried out by standard addition method. The values of optical and regression terms of analysis are given in table no 1.
Fig. 1: Calibration curve
RESULT:
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 emtricitabine by using chloroform as solvent from its formulations.
The colour ion –pair complexes are formed and 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 optical characteristics such as absorption maxima (nm), correlation coefficient (r) and were calculated and are also summarized in table 1. Assay results of recovery studies are given in Table 2.
Results are in good in agreement with labelled value. The percent recovery obtained indicates noninterference from the common excipient used in the formulation.
Table 2: Results of recovery studies
|
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 |
2.016327 |
100.8163 |
0.009858 |
0.488913 |
|
2 |
2 |
4.036735 |
100.9184 |
0.018177 |
0.4503 |
|
2 |
4 |
6.028571 |
100.4762 |
0.018443 |
0.305923 |
|
2 |
6 |
8.032653 |
100.4082 |
0.019716 |
0.24545 |
|
|
|
|
|
Mean= 0.016549 |
Mean= 0.372646 |
DISCUSSION:
The reproducibility, repeatability and accuracy of these methods were found to be good, which is evidenced by low values of standard deviations. Colorimetric methods suggested in literature were applied in UV region, need costly reagents for development of chromogen and useful in higher concentration. The proposed methods are simple, sensitive, accurate, precise, and reproducible applicable to even very low concentration as compare to previous methods suggested in literature. They are directly applied to drug to form chromogen. Hence they can be successfully applied for the routine estimation of Emtricitabine in bulk and pharmaceutical dosage form even at very low concentration in formulation such as tablets. The strong recommendation is made here for the proposed methods for determination of Emtricitabine from its formulation.
ACKNOWLEDGMENT:
Authors express sincere thanks to the Principal, D. G. Ruparel College for providing necessary facilities and encouragement for research work.
REFERENCES:
1. Dinesh Dhatkar, Sufiyan Ahmad, V. M. Shastry, Development and Validation of UV-Visible Spectrophotometric Method for Estimation of Emtricitabine and Tenofovir in Bulk and Dosage Form. International Journal of Pharmacy and Pharmaceutical Research. 2017; 9(3): 74-84
2. Budagam Lavanya et.al. Method development and validation of combined dosage form of Emtricitabine and Tenofovir disproxil fumarate by ultra violet spectroscopy. International Research Journal of Pharmacy. 2012; 3(12): 104-108.
3. Anindita Behera, Aurobinda Parida1, Amit Kumar Meher, Dannana Gowri Sankar, Swapan Kumar Moitra1, Sudam Chandra Si. Development and Validation of Spectrophotometric method for determination of Emtricitabine and Tenofovir disoproxil Fumarate in Bulk and Tablet dosage form, International Journal of PharmTech Research. 2011; 3(3): 1874-1882.
4. Mohammad H. AbdelHay, Azza A. Gazy, Rasha A. Shaalan, and Heba K. Ashour, Simple Spectrophotometric Methods for Determination of Tenofovir Fumarate and Emtricitabine in Bulk Powder and in Tablets. Journal of Spectroscopy, 2013, Article ID 937409, 7 pages, http://dx.doi.org/10.1155/2013/937409.
5. Rajan V. Rele, Prathamesh P. Tiwatane. Derivative UV Spectrophotometric Method for Validation of Emtricitabine in Bulk and Pharmaceutical Dosage Form. Asian J. Research Chem. 2020; 13(1): 48-51. DOI: 10.5958/0974-4150.2020.00011.5
6. Rajan V. Rele. UV Spectrophotometric Estimation of Emtricitabine by Zero order and area under Curve methods in Bulk and Pharmaceutical Dosage Form. Asian J. Research Chem. 2019; 12(5): 263-267. DOI: 10.5958/0974-4150.2019.00049.X.
7. Rajan V. Rele, Sandip Patil. Development of Analytical Method by RP-HPLC Method for Validation of Emtricitabine in API and Pharmaceutical Dosage Form. Asian J. Research Chem. 2019; 12(3): 143-147. DOI: 10.5958/0974-4150.2019.00029.4
8. Bhushan P. Badgujar, Moreshwar P. Mahajan, Sanjay D. Sawant. development and validation of RP-HPLC method for the simultaneous estimation of tenofovir alafenamide and emtricitabine in bulk and tablet dosage form, International Journal of ChemTech Research. 2017; 10(5): 731-739.
9. Arun Ramaswamy A, Anton Smith Arul Gnana Dhas. Development and validation of analytical method for quantitation of emtricitabine, tenofovir, efavirenz based on HPLC. Arabian Journal of Chemistry. 2018; 11: 275–281.
10. Ajay D. Mali and Uttam B. More. RP-HPLC method for simultaneous estimation of impurities from emtricitabine and tenofovir disoproxil fumarate tablet, International Journal of Pharmaceutical Sciences and Research. 2016; 7(4): 1662-1669.
11. Mallikarjuna rao Nagasarapu1 and Gowri Sankar Dannana. Development and validation of stability-indicating hplc-dad method for simultaneous determination of emtricitabine, elvetegravir, cobicistat and tenofovir in their tablet dosage forms. Indian Journal of Pharmaceutical Education and Research. 2016; 50(1): 205-211.
12. Kalpana Jayapalu, Himaja Malipeddi, Anbarasu Chinnasamy, Chromatographic separation and in vitro dissolution assessment of tenofovir disoproxil fumarate, emtricitabine and nevirapine in a fixed dose combination of antiretrovirals. Journal of Applied Pharmaceutical Science. 2014; 4(11): 076-080.
13. Varma D and A Lakshmana Rao. Stability-Indicating RP-HPLC method for the simultaneous estimation of efavirenz, tenofovir and emtricitabine in pharmaceutical formulations. Indian Journal of Pharmacy and Pharmacology. 2014; 1(1): 1-17.
14. Bala Rami Reddy.Yenumula1, Mutta Reddy. Singampalli and Bala Sekhara Reddy. Challa. Simultaneous estimation of emtricitabine and tenofovir disoproxil fumarate in tablet dosage form by reverse phase high-performance liquid chromatography. International Journal of Analytical Techniques. SOJ Chromatograph Sci. 2015; 1(1): 6.2-6.
15. Prashant S. Devrukhakar, Roshan Borkar, Nalini Shastri, and K. V. Surendranath. A Validated stability-indicating RP-HPLC method for the simultaneous determination of tenofovir, emtricitabine, and a efavirenz and statistical approach to determine the effect of variables. ISRN Chromatography. 2013; Article ID 878295, 8 pages, http://dx.doi.org/10.1155/2013/878295.
16. N.MD. Akram, M. Umamahesh N. A new validated RP-HPLC method for the determination of emtricitabine and tenofovir af in its bulk and pharmaceutical dosage forms. Journal of Chemical and Pharmaceutical Sciences. 2017; 10(4): 54-59.
17. Srinivasa Rao A, Naveen Kumar G, Srilekha K, Aruna Kumari N. Stability indicating method for the simultaneous estimation of tenofovir, emtricitabine and efavirenz in pure and pharmaceutical dosage form by RP-HPLC. 2016; 5(5): 188-200.
18. Umesh Maniyar, M. V. Katariya, Kishor Kumar Koshe, Gopal Karva, Sushil Jaiswal. Stability indicating RP-HPLC method for the simultaneous estimation of efavirenz, emtricitabine and tenofovir disoproxil fumarate in combined pharmaceutical dosage form. World Journal of Pharmacy and Pharmaceutical Sciences. 2016; 5(6): 1180-1198.
19. SK Mastanamma, D Venkata Reddy, P Saidulu and M Varalakhimi. Development and validation of stability indicating RP-HPLC method for the simultaneous estimation of emtricitabine tenofovir alafenamide bulk and their combined dosage form. Journal of Chemical and Pharmaceutical Research. 2017; 9(9): 70-80.
20. Som Shankar Dubey and Mahesh Duggirala. High performance liquid chromatography with PDA detector for combined determination of emtricitabine, tenofovir and efavirenz. Der Pharmacia Lettre. 2015; 7(10): 85-91.
21. P.D. Hamarapurkar, Abhijeet N. Parate. HPLC method for the determination of emtricitabine and related degradation substances. Journal of Chromatographic Science. https://academic.oup.com/ chromsci/article/51/5/419/326539.
22. MD Abdul Sattar and Suneetha Achanta. Analytical Method Development and validation for the determination of emtricitabine and tenofovir disoproxil fumarate using reverse phase hplc method in bulk and tablet dosage form. Journal of Pharmaceutical Science and Research. 2018; 10(5): 1207-1212.
23. J. Saminathan and T. Vetrichelvan. Development and validation of HPTLC method for simultaneous estimation of emtricitabine, rilpivirine and tenofovir disoproxil fumarate in combined dosage form. Bangladesh Pharmaceutical Journal. 2016; 19(1): 114-121.
24. Mahesh S. Wajurkar, Manjusha N. Dole. Development and validation of HPTLC method for determination of emtricitabine in API and tablet dosage form. Journal of Advanced Scientific Research. 2015; 6(3): 51-54.
25. Rajan V. Rele, S.S. Zarapker, Vipul Doshi,V.M. Shah. A simple extractive colorimetric determination of three drug from pharmaceutical preparations. Indian Drug. 1987; 24(12): 560-564.
26. Rajan V. Rele, Pankaj Gurav. A simple extractive spectrophotometric determination of loradatadine desloratadine and rupatadine from pharmaceutical formulation. International Journal of Pharma and Biosciences. 2012; 3(2): 89-92.
|
Received on 12.09.2024 Revised on 15.10.2024 Accepted on 06.11.2024 Published on 25.11.2024 Available online from December 27, 2024 Asian J. Research Chem.2024; 17(6):351-353. DOI: 10.52711/0974-4150.2024.00059 ©AandV Publications All Right Reserved
|