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Indo American Journal of Pharmaceutical Research, 2014

ISSN NO: 2231-6876

DEVELOPMENT AND VALIDATION OF A SIMPLE UV SPECTROPHOTOMETRIC


METHOD FOR THE DETERMINATION OF COBICISTAT IN ITS BULK FORM
Chandni Saha*, Md.Nazeeruddin Ahmed
Dept. of Pharmaceutical Sciences, Bharat Institute of Technology,JNTUH, India.
ARTICLE INFO
Article history
Received 12/12/2014
Available online
24/12/2014

Keywords
Cobicistat,
0.1N HCL,
UV Spectrophotometric
Method Development,
Validation.

ABSTRACT
Cobicistat is a cytochrome P450 3A (CYP3A) inhibitor. It boosts blood levels of the HIV
protease inhibitors Atazanavir and Darunavir by suppressing CYP3A, an enzyme that
metabolizes these drugs in the body. Cobicistat acts only as a pharmacokinetic enhancer and
has no antiviral activity. In the present basic work a rapid, simple and economic UV
spectrophotometric method has been developed for Cobicistat in its bulk form using 0.1N
HCl. The max was determined to be 246.2nm. The method was validated and proved to be
linear in the range of 10-150ug/ml, exhibited good correlation coefficient (R2=0.9998). The
validated method was found to be precise and robust. Thus this simple basic method can be
used for the determination of cobicistat.

Copy right 2014 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical
Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Please cite this article in press as Chandni Saha et al. Development and Validation of a simple UV Spectrophotometric Method
for the Determination of Cobicistat in its Bulk Form. Indo American Journal of Pharm Research.2014:4(12).

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Corresponding author
Chandni Saha
Assistant professor,
Dept. of Pharmaceutical Sciences,
Bharat Institute of Technology,JNTUH, India
chandnisaha7@gmail.com
+91 9966776152

Vol 4, Issue 12, 2014.

Chandni Saha et al.

ISSN NO: 2231-6876

INTRODUCTION
Cobicistat is a cytochrome P450 3A (CYP3A) inhibitor. It boosts blood levels of the HIV protease inhibitors Atazanavir and
Darunavir by suppressing CYP3A, an enzyme that metabolizes these drugs in the body. Cobicistat acts only as a pharmacokinetic
enhancer and has no antiviral activity [1-2]. In contrast to Ritonavir, it is devoid of anti-HIV activity and is thus more suitable for use in
boosting anti-HIV drugs without risking selection of potential drug-resistant HIV variants. Drug shows reduced liability for drug
interactions and may have potential improvements in tolerability over ritonavir. In addition, it has high aqueous solubility and can be
readily co formulated with other agents [3].
Literature review reveals that not many methods are developed for determination of Cobicistat in its bulk form [4]. Hence the
purpose of this work was to develop a simple basic rapid and economic method for the determination of Cobicistat in its bulk form so
as to provide better scope for further research on the drug.

Fig1-Structure of Cobicistat.
MATERIALS AND METHOD
Materials
All chemical and reagents used were of analytical grade. The bulk drug of Cobicistat was obtained as a gift sample from
Indian Institute of Chemical Technology (IICT), Hyderabad, India.
Instrument
Shimadzu uv-visible spectrophotometer (UV-1800) was used for all absorbance measurements with matched quartz cells.
Standard stock Solution:
Accurately weighed 0.1gm of Cobicistat (Bulk Drug) was dissolved in 0.1N HCl to obtain a concentration of 200g/ml
(STOCK SOLUTION).
Determination of max:
Pipette out 2.5ml from above stock solution into a 10ml calibrated volumetric flask and volume was made up to the mark
with 0.1N HCl. The resultant solution was scanned using UV-Visible spectrophotometer along 200 400 nm using 0.1N HCl as
blank.

Intermediate precision:
It was performed by preparing 3 different samples of concentration 50ug/ml and observed the absorbance for 3 different days.
LOD and LOQ
Limit of detection: it was calculated by using the following formulae: - LOD = 3.3 x S.D/Slope

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Intra-Day Precision (Repeatability) and Inter-Day Precision (Intermediate precision)


Repeatability:
8 different samples of same concentration 50ug/ml were prepared and the absorbance was observed under the same
experimental condition over short period of time.

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Linearity and Range:


For linearity study, fifteen solutions at different concentrations (10ug/ml 150ug/ml) were prepared by pipetting out 0.5ml
7.5ml from stock solution and making up to the mark of 10ml using 0.1N HCl. The absorbance was read at absorbtion maxima and the
data obtained were used for linearity calibration curve.

Vol 4, Issue 12, 2014.

Chandni Saha et al.

ISSN NO: 2231-6876

Limit of quantization: it was calculated by using the following formulae: - LOQ = 10 x S.D/Slope
Ruggedness:
Ruggedness of the Cobicistat was performed by preparing three different samples of concentration 50ug/ml by three different
Analysts and observed.
RESULTS AND DISCUSSION
Determination of max: The absorption maximum for Cobicistat was observed to be 246.2 nm.

Fig2: UV Absorbance Spectrum of Cobicistat in 0.1N HCl.


Linearity and Range
The calibration curve was evaluated by its correlation coefficient. The absorbance of the samples in the range of 10 150
ug/ml was linear with a correlation coefficient (R2) of 0.999. The LOD and LOQ were calculated.
Table-1: Optical characteristic and precision.
Parameters
max (nm)
Beers law limits (g/ml)
Molar absorptivity (lit. mol-1 cm-1)
Limit of Detection (LOD/ mcgml-1)
Limit of Quantification (LOQ/ mcgml-1)
Slope[b]
Intercept (a)
Correlation coefficient (r)**

UV Method
246.2
10-150
5x105
2.9357
8.8963
3.8 x 10-3
1.08 x 10-1
0.99952

Absorbance
0.064
0.126
0.190
0.256
0.320
0.381
0.446
0.512
0.576
0.640
0.702
0.765
0.846
0.910
0.952

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Concentration[g/ml]
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150

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Table-2: Calibration values of Cobicistat in 0.1N Hcl.

Chandni Saha et al.

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ISSN NO: 2231-6876

Fig3: Calibration curve of Cobicistat in 0.1NHCl.


Intra-Day Precision (Repeatability) and Inter-Day Precision (Intermediate precision)
The repeatability and intermediate precision study of the developed method confirmed adequate method stability where all
the RSDs where <2%.
Repeatability:
S.D = 0.0033
%RSD = 1.0698

Fig4: Repeatability graph of Cobicistat in 0.1N Hcl.


Inter-Day precision Results.
Table3:- Inter-Day precision Results.
DAY1
S.D = 0.007
% RSD = 1.53

DAY 2
S.D = 0.01
% RSD = 2.22

DAY 3
S.D = 0.01
% RSD = 2.22

CONFLICT OF INTEREST
The research was conducted in the absence of any commercial or financial relationships that could be construed as a potential
conflict of interest.

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CONCLUSIONS
The results and statistical parameters of the proposed basic method demonstrate method is linear, precise, robust, simple,
rapid and economic. As not much research has been carried on Cobicistat, this method can be used for future research works on
Cobicistat in various fields.

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S.D= standard deviation


RSD= Relative standard deviation

Vol 4, Issue 12, 2014.

Chandni Saha et al.

ISSN NO: 2231-6876

REFERENCES
[1] Gilead Sciences New Drug Applications for Cobicistat and Elvitegravir for HIV Therapy Accepted by U.S. FDA, April 21,
2014, http://www.gilead.com/news/pressreleases/2014/4/gilead-sciences-new-drug-applications-for Cobicistat and elvitegravirfor-hiv-therapy-accepted-by-us-fda (22-10-2014).
[2] Tybost Facilitates Once-Daily Dosing of the Protease Inhibitors Atazanavir and Darunavir. September 25, 2013.
http://investors.gilead.com/phoenix.zhtml?c=69964&p=irol-newsArticle&ID=1858868 (20 Dec. 2014)
[3] Eve-Irene Lepist Et al., Cobicistat Boosts the Intestinal Absorption of Transport Substrates, Including HIV Protease Inhibitors
and
GS-7340, In
Vitr,
Antimicrobial
Agents
Chemother.,
Vol
56(10),
pg
5409-5413.
oct
2012.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3457391/ (22-10-2014).
[4] Kavitha, K, Y.Geetha, G.Hariprasad, R.Venkatnarayana, R.Subramanian. (Jan2013) Development and validation of rp-hplc
analytical method for simultaneous estimation of emtricitabine, rilpivirine, tenofovir disoproxil fumarate and its pharmaceutical
dosage forms, Academic Journal Pharmacie Globale, [Online] Vol. 4 Issue 1, p1 January 2013. Available from:
http://connection.ebscohost.com/c/articles/87977558/development-validation-rp-hplc-analytical-method-simultaneous-estimationemtricitabine-rilpivirine-tenofovir-disoproxil-fumarate-pharmaceutical-dosage-forms (22-10-2014).

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