Design and Validation of Dissolution
Profile of Rivaroxaban by Using
RP-HPLC Method in Dosage Form
Pinaz A. Kasad*,
K.S. Muralikrishna
Shree Dhanvantary
Pharmacy College, Department of Quality assurance, Kim, Surat
(India)
*Corresponding Author E-mail: pinazkasad@yahoo.in
Rivaroxaban,
anti-coagulant is a novel drug for the prevention of venous thromboembolism (VTE) in patients who have undergone
elective total hip replacement or total knee replacement surgery and no dissolution study for
its estimation has been reported yet. The aim of this
work was to develop and validate a dissolution profile for Rivaroxaban
in a tablet dosage form using RP-HPLC method. The conditions established for
dissolution were: 900 mL of acetate buffer pH=4.5 +
0.2 % sodium lauryl sulphate
(SLS) as dissolution medium, using a
paddle type dissolution apparatus at a stirring rate of 75 rpm which was able
to give % drug release of 98.86% . The drug release was evaluated by RP-HPLC
method at 250 nm and a good linearity was observed in the concentration range
of 20-60 μg/mL with
correlation coefficient of 0.9989. The percentage recovery of Rivaroxaban was found to be 99.63 and % CV (0.22 %; n=6)
indicated a good precision of the analytical method..
Robustness of the method was performed by using different rotation speeds and
Temperatures. The validation parameters included linearity, accuracy, precision
and robustness. Analytical method validation was found to be within the
acceptance criteria of the guidelines of ICH Q2 R1, FDA and FIP. The proposed
method can be applied for routine quality control analysis of Rivaroxaban
KEYWORDS: Rivaroxaban, Validation, Dissolution apparatus,
RP-HPLC Method, 250 nm.
INTRODUCTION (1-10)
Rivaroxaban is an oral anticoagulant invented and
manufactured by Bayer; in a number of countries it is marketed as Xarelto.(1) In the United States, it is marketed
by Janssen Pharmaceutical.(2) It is the first available orally
active direct factor Xa inhibitor.In September
2008, Health Canada and European
Commission granted marketing authorization for Rivaroxaban
as one 10 mg tablet taken once daily for the prevention of venous thromboembolism
(VTE) in patients who have undergone elective total hip replacement or total knee replacement surgery. It was approved by CDSCO on 30 January 2010. In December
2011
Rivaroxaban has been approved by the European
Commission for use in two new indications: prevention of stroke and systemic
embolism in adult patients with non-valvular atrial fibrillation (AF) with one or more risk factors and
treatment of deep vein thrombosis (DVT) and prevention of recurrent DVT and
pulmonary embolism (PE) following an acute DVT in adults.
On July 1, 2011, the U.S. Food and Drug Administration (FDA) approved Rivaroxaban
for prophylaxis of deep vein
thrombosis (DVT),
which may lead to pulmonary embolism (PE), in adults undergoing hip and
knee replacement surgery.(3) On November 4, 2011, the U.S. FDA approved
Rivaroxaban for stroke prophylaxis in patients with
non-valvular atrial fibrillation
.(4)
Figure 1- structural formula of Rivaroxaban
In vitro dissolution study is an official test of pharmacopoeias for evaluation of
drug release from solid and semisolid dosage forms and to establish in vitro/in vivo correlation. It is also routinely
used tool in Quality Control (QC) to ensure batch to batch consistency and in
Research and Development (R&D) to provide some predictive estimate of the
drug release in respect to the In vivo performance.
When dissolution test is not defined in the monograph of
the dosage form, or if the monograph is not available, comparison of drug
dissolution profiles is recommended in different dissolution media, in the pH
range of 1.2–7.5. The selection of a dissolution medium depends upon the
solubility data and dosage range of the drug product. Literature survey reveals
Colorimetric method (7-8), RP-HPLC method(9) and bio analytical method(10) for estimation
of Rivaroxaban. The present investigation was taken
up to develop a suitable dissolution profile for the Rivaroxaban
in its dosage form and validate it by using RP-HPLC methods.
MATERIAL AND METHODS:
Instruments and Reagents:
The analysis of the drug was carried
out on HPLC Model: LC-2010CHT, Shimadzu,
Japan with UV-Visible and photodiode array detector and injector with
20μl fixed loop. Chromatographic analysis was performed using Column: Phenomenex C18 (250mm×4.6mm i.d.)
5μm. Analytical balance: Sartorious CP225D was
used for weighing. Water purifier system used was Sartorious
Arium 611VF. The mobile phase was prepared freshly
and it was degassed by Sonicator-Trans-O-Sonic sonicating for 15 min before use. The column was
equilibrated for at least 30min with the mobile phase flowing through the system.All the chemicals used were of analytical grade.
Pure Rivaroxaban was procured as a gift sample from
Mega Fine Pharma, Mumbai, India.
Preparation of
Standard Solutions:
Preparation of
Dissolution Medium (acetate buffer pH=4.5 + 0.2 % sodium lauryl sulphate (SLS)):
Accurately measured 2.9 gm of Sodium acetate was taken in 1000 ml
measuring cylinder and was dissolved with distilled water.14 ml of acetic
acid(to make pH=4.5) and 2 gm of sodium
lauryl sulphate (SLS) was added, make up to 1000 ml with distilled
water.
Preparation of Rivaroxaban standard stock solution (100 μg/mL):
Stock solution (100μg/mL)
was prepared by weighing accurately 10 mg of Rivaroxaban
and dissolving in 100 mL of Dissolution Medium
(acetate buffer pH=4.5 + 0.2 % sodium lauryl sulphate (SLS)). This stock solution was further diluted
serially (20-60 μg/mL),
for the linearity study.
Dissolution Test Conditions:
Selection of suitable dissolution medium was made on the basis of
solubility study. Precisely, excess quantity of Rivaroxaban
was added to 10 mL of various solvents and buffers
with pH range of 1 to 7.4 in a shaking water bath; set at 100 rpm for 24 h at
room temperature. The solution was then passed through Whatmann
(No. 1) filter paper and the amount of the drug dissolved was analyzed after
suitable dilutions using calibration curve.
In vitro drug release study was carried out
according to dissolution procedures recommended for conventional single-entity
tablet formulations(11-13), in
a 900 mL of selected solvent using USP type-II (paddle
method) dissolution apparatus at 37.0 ± 0.5 oC
at 50,75 and 100 rpm. Sample aliquots of 5 mL were
withdrawn at time intervals of 10, 20, 30, 40, 50 and 60 minutes and replaced
with an equal volume of the fresh medium to maintain the sink condition. After
the end of each test time, sample aliquots were filtered, and quantified. The
Peak Area was measured and from the regression equation of linearity curve, the
percentage release on each time of dissolution profile was calculated. The
percentage of drug released was plotted against time, in order to obtain the
release profile and to calculate the in vitro dissolution data.
Figure-2 Blank Solution (acetate buffer pH=4.5 + 0.2 %
sodium lauryl sulphate
(SLS)
Figure 3-Standard Solution of Dissolution Method.
Figure 4 - % Drug Release Profile at Different RPM of Rivaroxaban
Method Validation:
Rivaroxaban analyzed in
acetate buffer pH=4.5 + 0.2 % sodium lauryl sulphate (SLS) and validated for its specificity,
linearity, precision and accuracy, according to ICH guideline[14], FIP guidelines [15] and US
Pharmacopoeia [16].
RESULTS AND DISCUSSION:
Solubility study of Rivaroxaban in different pH
media showed that it is highly soluble in acetate buffer pH=4.5 with adding 0.2
% sodium lauryl sulphate
(SLS) as surfactant. Moreover, in this media drug was found stable for 24 h.
Thus acetate buffer pH=4.5 + 0.2 % sodium lauryl sulphate (SLS) was chosen as the dissolution medium for
evaluation of Rivaroxaban.
Study of absorbance maxima of solution of Rivaroxaban
(40μg/mL) in acetate buffer pH=4.5 + 0.2 %
sodium lauryl sulphate
(SLS) has shown peak at 250 nm and was chosen owing to good linearity.
Dissolution study of formulation strength 10 mg carried in 900 mL of acetate buffer pH=4.5 + 0.2 % sodium lauryl sulphate (SLS) shown that
at 75 rpm more than 90% of the drug was released in 60 min, which is found
within the acceptance criteria of pharmacopoeia and hence adopted for the study
compared to <75% drug release at 75 rpm (Figure
4).
Validation of developed RP-HPLC method was carried out and the
observations are tabulated (Table
5).
Figure 5 Linearity
curve of Rivaroxaban in for dissolution method
Linearity:
The linearity of Rivaroxaban was studied at
concentration range of 20 – 60 μg/mL and showed a good correlation coefficient at 250 nm
Precision:
The precision of the method was determined by measuring the repeatability
(intra-day precision) and expressed as RSD (%). Tablets (n=6) were subjected to
dissolution test conditions (900 mL of dissolution
medium pre-heated at 37ºC±0.5, paddle with stirring rate of 75 rpm, 60
minutes). All the data (Table 1) are within the acceptance criteria.
Table 1 Precision data for dissolution method
Accuracy:
The accuracy was evaluated by applying the proposed method to the
analysis of the tablet with known amounts of the Rivaroxaban,
corresponding to the concentrations of 80, 100 and 120%, which were subjected
to dissolution test conditions described above. The accuracy was calculated as
the percentage of the drug recovered from the formulation matrix. The percent
recoveries obtained [Table 2] were considered acceptable [14].
Table 5: Regression analysis data and summary of
validation parameters for dissolution method:
Robustness:
It was performed by using different rotation speeds [75
rpm and 100 rpm] and Temperature [35 and 390C]. Results
were expressed in % RSD [Table 3].
CONCLUSION:
The dissolution test developed and validated for Rivaroxaban
tablets was considered satisfactory. The conditions that allowed the
dissolution determination were 900 mL of acetate
buffer pH=4.5 + 0.2 % sodium laurayl sulphate (SLS) at 37.0 ± 0.5 ºC, paddle apparatus, 75 rpm
stirring speed and filtration with quantitative filter. The % drug delivery was
higher than 90% in 60 minutes for all evaluated products; fulfilling the pharmacopeial guidelines for conventional solid oral
tablet dosage form. Moreover, these conditions satisfied the criteria of drug
stability during dissolution study. The method was validated to ensure that proposed
method is linear, precise, accurate, sensitive, robust and cost effective which
can be effectively and successfully employed for the dissolution study of Rivaroxaban in its dosage forms in routine quality control.
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