UPLC SEPARATION ANALYSIS OF EMTRICITABINE, TENOFOVIR, COBICISTAT AND ELVITEGRAVIR FROM THEIR DEGRADATION PRODUCTS
Objective: A simple, rapid, accurate and precise stability-indicating UPLC analytical method has been developed and validated for the quantitative analysis of Emtricitabine, Tenofovir, Cobicistat and Elvitegravir in bulk drugs and combined dosage forms.
Methods: ACE C18 (50 mm x 3 mm, 2Âµ). The column temperature was maintained at 30oC and run time 8 min. The mobile phase was a mixture of Mobile Phase: Aâ€“0.1% TFA in Acetonitrile, Bâ€“0.1% TFA in Milli-Q-water. The injection volume of samples was 20Î¼l. UV detection was carried out using a UV-PDA detector at 240 nm. The validation of this method was done as per ICH guidelines.
Results: The retention times were observed as 1.46, 3.59, 4.13, 4.64 min for Emtricitabine, Tenofovir disoproxyl fumarate, Cobicistat, and Elvitegravir respectively. Linearity ranges were observed 150-275 Âµg/ml Emtricitabine, 250-375 Âµg/ml Tenofovir, 100-225 Âµg/ml Cobicistat and 100-225 Âµg/ml Elvitegravir. Relative Standard Deviation did not exceed 2.
Conclusion: The newly developed UPLC method for separation of different degradation products along with the pure drugs were found to be capable of giving faster retention times while still maintaining good resolution than that achieved with conventional HPLC. The decreased flow rate 0.4 ml/min, in UPLC indicate more economical. This method exhibited an excellent performance in terms of sensitivity and speed. The results of stress testing undertaken according to the ICH guidelines reveal that the method is specific and stability-indicating. The proposed method has the ability to separate these drugs from their degradation products in tablet dosage forms and hence can be applied to the analysis of routine quality control samples and samples obtained from stability studies.
Keywords: Stability indicating assay, RP-UPLC, Emtricitabine, Tenofovir, Cobicistat, Elvitegravir, Forced degradation studies
http://www.drugbank.ca/drugs/DB00879. [Last accessed on 10 Nov 2015].
http://www.drugbank.ca/drugs/DB00300. [Last accessed on 10 Nov 2015].
Lepist EI, Phan TK, Roy A, Tong L, MacLennan K, Murray B, et al. "cobicistat boosts the intestinal absorption of transport substrates, including hiv protease inhibitors and gs-7340, in vitro". Antimicrob Agents Chemother 2012;56:5409-13.
Thaczuk, Derek, Carter, Michael. ICAAC: "Best response to elvitegravir seen when used with T-20 and other active agents". Aidsmap; 2007. p. 19.
ICH guidelines. "Validation of analytical procedures: Text and Methodology, Q2 (R1)"; 2005.
Ilango K, Sunitha PG. "Development and validation of UV spectrophotometric methods for the simultaneous determination of emtricitabine and tenofovir disoproxyl fumarate in combined dosage form". J Pharm Res 2012;5:873-5.
Vishnu P Choudhari, Snehal Ingale, Sacchidanand R Gite, Dipali D Tajane, Vikram G Modak, Archana Ambekar. "Spectrophotometric simultaneous determination of Tenofovir disoproxyl fumarate disoproxil fumarate and Emtricitabine in combined tablet dosage form by ratio derivative, first order derivative and absorbance corrected methods and its application to dissolution study." Pharm Methods 2011;2:47-52.
Budawari S. The Merck Index. Whitehouse Station, NJ: Merck and Co. Inc; 2006. p. 927, 1573.
Sweetman SC, Martindale. The complete drug reference. London; Pharmaceutical Press; 2005. p. 635-42.
Raghu Ram Jampala, V Kiran Kumar, Appala Raju Nemala. Development and application of the liquid chromatographic method for simultaneous determination of elvitegravir, tenofovir disoproxil fumarate, emtricitabine, and cobicistat in fixed dosage formâ€. Pharm Methods 2014;5:7-13.
Putchakayala Purnachandra Rao, Dondeti Mogili Reddy, D Ramachandran. Stability indicating HPLC method for simultaneous estimation of emtricitabine, tenofovir disoproxyl fumarate, cobicistat and elvitegravir in pharmaceutical dosage formâ€. World J Pharm Sci 2014;2:1822-9.
Swati S Pardhi, Kiran B Aher, Girija B Bhavar, SR Chaudhari. Development and validation of a UV-Spectrophotometric method for the estimation of elvitegravir in bulk and pharmaceutical formulationsâ€. Int J Pharm Sci Res 2015;6:213-8.