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RESEARCH ARTICLE
Simultaneous RP-HPLC Method for the Estimation of the Emtricitabine, Tenofovir
Disoproxil Fumerate and Efavirenz in Tablet Dosage Forms
N Appala Raju* and Shabana Begum
*
Department of Pharmaceutical Chemistry, Sultan-Ul-Uloom College of Pharmacy, Mount Pleasant, Road-3, Banjara Hills,
Hyderabad-500 034. INDIA
ABSTRACT:
A simultaneous stability indicating RP-HPLC method is developed for the estimation of Emtricitabine, Tenofovir disoproxil
fumerate and Efavirenz in tablet dosage form. Chromatography was carried on an Inertsil ODS 3V column using gradient
composition of 0.02M sodium dihydrogen orthophosphaste as mobile phase A and mixture of Methanol and water in ratio of 85:15
as mobile phase B at a flow rate of 1.5 ml/min with detection at 265 nm. The retention times of the Emtricitabine, Tenofovir
disoproxil fumerate and Efavirenz was about 5.875, 8.800 and 12.020 mins respectively. The detector response is linear from 8-
120µg/ml, 12-180µg/ml, 20-360µg/ml of test concentration for Emtricitabine, Tenofovir and Efavirenz respectively. The respective
linear regression equation being Y=10175x-76883 for Emtricitabine, Y=6280.8x+219800 for Tenofovir disoproxil fumerate and
Y=1883.5x+323060 for Efavirenz. The limit of detection and Limit of quantification was 0.06, 0.07 and 0.08 µg/ml and 0.14, 0.12
and 0.15µg/ml for Emtricitabine, Tenofovir and Efavirenz respectively. The percentage assay of Emtricitabine, Tenofovir disoproxil
fumerate and Efavirenz was 99.31, 99.77 and 100.20 % respectively and percentage recovery for average of three different
concentrations was 100.87%, 100.04% and 99.52% respectively. The method was validated by determining its sensitivity, Linearity,
accuracy and precision. The proposed method is simple, fast, sensitive, Linear, accurate, rugged and precise and hence can be
applied for routine quality control of Emtricitabine, Tenofovir disoproxil fumerate and Efavirenz in bulk and in tablet dosage form.
KEY WORDS: Emtricitabine, Tenofovir disoproxil fumerate and Efavirenz, Shimadzu HPLC with PDA detector,
Inertsil ODS 3V and Tablets.
INTRODUCTION:
Emtricitabine2 is chemically known as 4-amino-5-fluoro-1- Literature survey reveals few Chromatographic methods11-13
[(2R, 5S)-2-(hydroxymethyl)-1, 3-oxathiolan-5-yl] in biological fluids were reported along with other
pyrimidin-2-one. Tenofovir disoproxil fumerate fumerate3 is antiretroviral dugs. So far, only one HPLC method has been
chemically know as 9-[(R)-2-[[bis [[isopropoxycarbonyl] reported in gradient mode for the estimation of
oxy] methoxy] phosphonyl] methoxy] popyl] adenine Emtricitabine, Tenofovir disoproxil fumerate and Efavirenz
fumarate. Efavirenz1 is chemically known as (4S)-6-chloro- from pharmaceutical dosage form14.
4-(cyclopropylethynyl)-1, 4-dihydro-4-(trifluoromethyl) 2-
H-3, 1-benzoxazin 2-one. Emtricitabine, Tenofovir Highly Sensitive, Selective, Rugged stability indicating
disoproxil fumerate, and Efavirenz 4,5,6. are a novel HPLC method will be very useful for the estimation of
formulation combining fixed doses of the nucleoside reverse Emtricitabine, Tenofovir disoproxil fumerate and Efavirenz
transcriptase inhibitors Emtricitabine (200mg) and in pharmaceutical formulations. The purpose of the this
Tenofovir disoproxil fumerate fumarate (300mg) with the study was to develop sensitive, simple, precise, accurate and
non-nucleoside reverse transcriptase inhibitor Efavirenz Rugged Simultaneous Stability Indicating RP HPLC method
(600mg) represents the first once-daily, one-tablet for the estimation of Emtricitabine, Tenofovir disoproxil
antiretroviral regimen.10It is official in Martindale2-The fumerate and Efavirenz in bulk drug samples and in
Extra pharmacopoeia. pharmaceutical dosage form.
EXPERIMENTAL:
Materials and Methods:
Received on 05.01.2009 Modified on 10.01.2009 Emtricitabine, Tenofovir disoproxil fumerate and Efavirenz
Accepted on 11.01.2009 © RJPT All right reserved was obtained as a gift sample from Aurobindo Pharma Ltd,
Research J. Pharm. and Tech. 1(4): Oct.-Dec. 2008;Page 522-525 Hyderabad. HPLC grade Methanol, Acetonitrile (Merck)
522
Research J. Pharm. and Tech. 1(4): Oct.-Dec. 2008,
,
and AR grade sodium dihydrogen orthophosphate (Merck) Fig 1:Typical Chromatogram of Emitricetabine (5.88mins),
was used. Milli-Q water was used in mobile phase Tenofovir (8.796) and Efavirenz (12.015) by HPLC
preparation. Commercially available Emtricitabine,
Tenofovir disoproxil fumerate and Efavirenz dosage forms mV
8.796
1750
(Vireday, Cipla) were purchased from local market.
1500
Instrument: 1250
Shimadzu prominence High Performance Liquid
1000
Chromatograph with PDA detector and Auto injector mode
12.015
5.883
750
was used with LC solutions software.
500
10 3 0 6 0 2 0
4 59 4 74 0
523
Research J. Pharm. and Tech. 1(4): Oct.-Dec. 2008,
,
Table I: Linear Regression Data for Calibration curves:
Drug Emtricitabine Tenofovir Disoprxil Efavirenz
Concentration range (µg/ml) 8 – 120 12 – 180 20 - 360
Slope (m) 10175 6281 1884
Intercept (b) -76883 219800 323060
% Intercept -1.0 2.8 6.7
Residual Sum of Squares 154626 148254 117281
Correlation coefficient 0.9994 0.9994 0.9990
RSQ(r2) 0.999 0.999 0.998
% RSD 0.0 0.1 0.1
524
Research J. Pharm. and Tech. 1(4): Oct.-Dec. 2008,
,
59 4 4 170
4 8 776 0 4
3 8 110 3 8
2 74 4 4 72
16 779 0 6
6 113 4 0
240 605 9 70 13 3 5 170 0 2065 2430 2 79 5 3 16 0 3 52 5
Conc . ( µg/ mL)
525