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PART-[B]
Simultaneous reverse phase
high performance liquid
chromatographic [RP-HPLC]
determination of salicylamide,
salicylic acid and deferasirox in
the bulk API dosage forms
SECTION-[III]
HPLC method development and
validation of deferasirox
Section- [III] Deferasirox
PART-B Page 95
1.0 Introduction of deferasirox
Deferasirox is chemically 4-(3, 5-bis (2-hydroxyphenyl) benzoic acid. Its
molecular formula is C21H15N3O4 and molecular is weight 373.36 gm/mole.
Deferasirox is an oral iron chelator. Its main use is to reduce chronic iron
overload in patients who are receiving long term blood transfusions for
conditions such as beta-thalassemia and other chronic anemia [1-5].
Deferasirox is an oral iron chelator.
Deferasirox is a white to slightly yellow powder. Inactive Ingredients:
Lactose monohydrate, crospovidone, povidone (K3O), sodium lauryl sulphate,
microcrystalline cellulose, silicon dioxide, and magnesium stearate. Its main
use is to reduce chronic iron overload in patients who are receiving long-term
blood transfusions for conditions such as beta-thalassemia and other chronic
anemias [6-8]. It is the first oral medication approved in the USA for this
purpose. It was approved by the United States Food and Drug Administration
(FDA) in November 2005 [9]. According to FDA (May 2007), renal failure and
cytopenias have been reported in patients receiving deferasirox oral suspension
tablets. It is approved in the European Union by the European Medicines
Agency (EMA) for children 6 years and older for chronic iron overload from
repeated blood transfusions [10-12]. Its low molecular weight and high
lipophilicity allows the drug to be taken orally unlike deferoxamine which has to
be administered by IV route (intravenous infusion). Together with deferiprone,
deferasirox seems to be capable of removing iron from cells (cardiac myocytes
and hepatocytes) as well as removing iron from the blood.
[2.0] Chemical structure, IUPAC and common name of deferasirox
Common name
20kg, Exjade, ICL-670, CGP 72670, Deferasirox, Deferasirox (CGP-72670),
Deferasirox for research, 4-[3,5-bis(2-hydroxyphenyl),4-[3,5-bis(2-
Section- [III] Deferasirox
PART-B Page 96
hydroxyphenyl)-1,2,4-triazol-1-yl]benzoic acid 4-[3,5-bis(2-hydroxyphenyl)-
1,2,4-triazol-1-yl]benzoic acid.
3.0 Brief overview of synthetic pathway of deferasirox
Deferasirox can be prepared from simple commercially available starting
materials (salicylic acid, salicylamide and 4-hydrazinobenzoic acid) in the
following two-step synthetic sequence.
4.0 Description
[4.1] Pharmacodynamics
Deferasirox is an orally active chelator that is selective for iron (as Fe3+).
It is a tridentate ligand that binds iron with high affinity in a 2 : 1 ratio.
Although deferasirox has very low affinity for zinc and copper there are variable
decreases in the serum concentration of these trace metals after the
administration of deferasirox. The clinical significance of these decreases is
uncertain.
Section- [III] Deferasirox
PART-B Page 97
[4.2] Mechanism of action
Two molecules of deferasirox are capable of binding to 1 atom of iron.
Deferasirox works in treating iron toxicity by binding trivalent (ferric) iron (for
which it has a strong affinity), forming a stable complex which is eliminated via
the kidneys.
[4.3] Pharmacokinetics
Pharmacokinetic data
Bioavailability 70 %
Protein binding 99 %
Metabolism Hepatic glucuronidation
Half-life 8 to 16 hours
Metabolism
Hepatic. CYP450-catalyzed (oxidative) metabolism of deferasirox appears
to be minor in humans (about 8 %). Glucuronidation is the main metabolic
pathway for deferasirox, with subsequent biliary excretion. Indication for the
treatment of chronic iron overload due to blood transfusions (transfusional
hemosiderosis) in patients 2 years of age and older.
Table 1: Physical and chemical properties
Physical and chemical properties
Property Value
Molecular Weight 373.3615 g/mol
Molecular Formula C21H15N3O4
Molecular mass 373.362 g/mol
Exact Mass 373.106256 g/mol
Density 1.5 g/cm3
Solubility Soluble in water (< 1 mg/mL at 25°C), water
solubility (0.038 mg/mL at 37°C), soluble in
acetonitrile and methanol
CAS number 201530-41-8
Physical state White to Off-White Solid
Melting point 116-117°C
Boiling point 260-2620°C
Stability Less stable up to 45°C
Categories oral iron chelator
Section- [III] Deferasirox
PART-B Page 98
[5.0] Survey of analytical method/Literature reviews
The literature reviews regarding deferasirox suggest that various
analytical methods were reported for its determination in pharmaceutical
formulation and in various biological fluids. As per discussion in the literature
reviews UV, LC-MS, HPLC methods for the determination of deferasirox in
pharmaceutical dosage forms are reported. Most of the reported methods do not
include stress degradation studies or are not completely optimized, validated
and they are cumbersome, time-consuming and expensive. The literature
reviews for analysis of deferasirox are as under:
[1] VS. Rao Somisetty, D. Dhachinamoorthi, SA. Rahaman, CH MM Prasada
Rao et al, have developed a simple, sensitive, specific UV-spectroscopic and
RP-HPLC methods are developed for the estimation of deferasirox in bulk and
pharmaceutical formulation. The first method was based on UV–spectroscopic
determination of the drug. It involves absorbance measurements at 319 nm in
0.1 M sodium hydroxide. Calibration curve was linear with the correlation
coefficient was 0.9997 % over a concentration range of 5 to 30 μg/mL for the
drug. The second method was based on HPLC separation of the drug in reverse
phase mode using C18 column (150 mm × 4.6 mm i.d. 5μ). The mobile phase
constituted of acetonitrile: water pH 3.5 adjusted with orthophosphoric acid (70
: 30 v/v) and flow rate 1.0 mL/min. Detection was performed at 248nm.
Separation completed within 5 minutes. Calibration curve was linear with the
correlation coefficient was 0.9996 % over a concentration range of 1 to 6 μg/mL
for the drug. The relative standard deviation (R.S.D) was found < 2.0 % for UV–
spectroscopic and RP-HPLC methods [13].
[2] PK. Tiwari, Padmakar Sathe, Navin Devadiga et al, have developed HPLC
method using Zorbax SB C18 column (250 4.6 mm i.d.), 5 μm, with UV
Detector was used. The proposed method was cost effective, specific, linear,
accurate, rugged and precise. The calibration curves showed good linearity over
the concentration range of 0.5 μg/mL-1.5 μg/mL with respect to the sample.
The correlation coefficient was 0.999 %. Excellent recoveries of 102 % were
obtained at the level 0.5 μg/mL [14].
[3] S. Saravanan, R. Swetha et al, have developed and validated a simple and
isocratic reverse phase high performance liquid chromatography (RP-HPLC)
method for determination of deferasirox and its impurities. The method was
Section- [III] Deferasirox
PART-B Page 99
validated as per International Conference on Harmonization (ICHQ2A (R1))
guidelines for system suitability, precision, linearity, specificity, solution
stability, robustness and ruggedness, limit of detection and limit of
quantitation. Deferasirox was analyzed by using Inertsil ODS-3V (150 mm X 4.6
mm, 5 μm) at 35°C column temperature, with isocratic elution. The analysis
was performed at a wavelength of 245 nm using dual λ absorbance detector.
Efficient UV detection at 245 nm enabled determination of deferasirox without
any interference of impurities salicylic acid & salicylamide. The retention time
(RT) for deferasirox was around 8.7 min [15].
[4] S. Vattikuti, RS. Phani, R. Seetharaman, KS. Lakshmi et al, have
developed a simple rapid and precise reversed phase high performance liquid
chromatography method for the analysis of deferasirox. Chromatography
separation of deferasirox was performed by using a kromosil C18 column (250 x
4.6 mm, 5 μm) as stationary phase with a mobile phase comprising of
Methanol: O-phosphoric acid : 0.01 % Tetrahydrofuran [90 : 7 : 3 (v/v)] at a
flow rate of 1.5 mL/min and UV detection at 245 nm. The linearity of
deferasirox in the range of 0.01 mg/mL to 0.09 mg/mL. The limit of detection
for deferasirox was found to be 40 nanograms. The recovery was calculated by
standard addition method [16].
[5] M. Khan, S. Sinha, M. Todkar, V. Parashar, KS. Reddy, U. Kulkarni et
al, have developed a reverse phase high performance liquid chromatography
(RP-HPLC) method was developed and validated for the assay and process and
degradation related impurities of deferasirox. The method involves quaternary
gradient pumps and variable wavelength UV detector. Chromatographic
conditions include a 20 μL injection volume Xterra RP-18 (150 X 4.6 mm, 5 μm)
column (column temperature: 25°C), flow rate of 1.0 mL/min and detection at
250 nm. The separation of drug from its impurities was achieved by a gradient
elution method, using the mobile phase composed of solution A (water adjusted
to pH 2.5 with perchloric acid) and solution B (methanol). The retention times of
deferasirox, impurity A and impurity B are 37.7 min, 43.7 min and 49.3 min,
respectively. The method has been validated both for assay and impurity
profiling as per ICH guideline [17].
[6] VK. Chakravarthy, DG. Sankar et al, have developed an isocratic reverse
phase liquid chromatography (RP-HPLC) method has been developed and
Section- [III] Deferasirox
PART-B Page 100
subsequently validated for the determination of deferasirox in pharmaceutical
formulation. Separation was achieved with a Develosil ODS HG-5 (150 mm
4.6 mm I.D; particle size 5 μm) and Sodium dihydrogen phosphate
monohydrate Buffer (pH adjusted to 3.0 with dilute orthophosphoric acid):
acetonitrile (55 : 45) as eluent at flow rate 2.0 mL/min. UV detection was
performed at 245 nm. The method is simple, rapid, and selective. The described
method of Deferasirox is linear over a range of 11.999 μg/mL was 35.997
μg/mL. The method precision for the determination of assay was below 2.0 %
RSD. The percentage recoveries of active pharmaceutical ingredient (API) from
dosage forms ranged from 100.5 to 101.0 [18].
[7] N. Padmaja, N. Ramathilangam et al, have developed a simple, accurate,
precise and linear isocratic RP-HPLC has been developed and subsequently
simultaneous for determination of deferasirox in pharmaceutical formulations.
Develosil ODS HG-S (150 X 4.6 mm) 5 μ with flow rate of 2 mL/min. By using
HPLC water PU-2695 pump and photodiode array detector-2996 at 245 nm.
The separation was carried out using a mobile phase consisting of mixture of
sodium dihydrogen phosphate monohydrate buffer (pH = 3.0) and acetonitrile in
the ratio of 55 : 45. The retention time of Deferasirox was found to be 6.31 min.
The mean percentage recovery was found to be 99.55 %. The correlation
coefficient was found to be 0.998 %. The percentage estimation of the drug was
found near to 100 % representing the accuracy of the method [19].
[8] JR. Kothawade, KM. Agrawal, VS. Lad et al, have a simple stability
indicating RP-HPLC assay method has been developed and validated for the
determination of deferasirox from tablet dosage form. Drugs were determined on
a Hypresil BDS 150 × 4.6 mm column packed with 5μm particles. The
optimized mobile phase was a 50 : 50 (v/v) Buffer (1 mL OPA in 2 lit. Milli-Q
Water) and Acetonitrile, pumped at a flow rate of 1 mL/min. UV detection was
performed at 245 nm. The method was validated in the concentration ranges of
50 ppm to 150 ppm where it demonstrated good linearity with R2 = 0.998 %.
The method was found to be robust, resisting to small deliberate changes in
flow rate, column temperature and composition of the mobile phase [20].
Section- [III] Deferasirox
PART-B Page 101
[6.0] Aim and scope of present work
The primary objective of the present work was thus to develop and
validate a RP-HPLC method for the assay of deferasirox from API dosage forms.
Hence, the method is useful for routine quality control analysis and also for
determination of stability. Purpose of the present study was to develop and
validate a RP-HPLC Method for determination of deferasirox in API
pharmaceutical dosage forms. The aim and scopes of the proposed work are as
under:
1. To select suitable mobile phase (solvent buffer ratio)
2. To optimize RP-HPLC conditions
3. To develop suitable HPLC method for deferasirox
4. Perform the validation for the developed method
[7.0] Experimental
[7.1] Materials
Deferasirox obtained received from Cadila pharma dholka Ahmedabad.
Orthophosphoric acid was obtained from s d Fine Chemical Limited. Acetonitrile
(fisher Qualigens, HPLC grade) were obtained from Thermo Fisher Scientific
India Pvt. Ltd. and potassium dihydrogen phosphate was obtained from (Merck
Specialties Private Limited).
[7.2] Equipment
Equipment Apparatus
HPLC System Dionex ultimate 3000 (Germany) High performance liquid chromatographic system equipped with ultimate 3000 Pump, Auto Sampler, Column Compartment and RS Diode Array Detector
Software Dionex Chromeleon ® 7 (Version 7.1, Simply Intelligent)
Column oven Ambient
Column Waters symmetry C18 (4.6 x 250 mm, 5 µm, 110 Å)
[7.3] Preparation of stock and sample solutions
[7.3.1] Preparation of buffer
Dissolved 1.36 g of potassium dihydrogen phosphate in 1000 mL high
purity demonized Milli-Q water [Millipore, Milli-Q, Bedford, MA, USA,
purification system] and pH was adjusted 3.2 with ortho-phosphoric acid
and filtered through 0.22 μ size nylon filter under vacuum.
Section- [III] Deferasirox
PART-B Page 102
[7.3.2] Preparation of mobile phase
The mobile phase was prepared by mixing 400 mL phosphate buffer pH
3.2 and 600 mL of acetonitrile [HPLC Grade]. The mixture was sonicated
in Expo-Hi Tech sonicator for 5 minutes.
[7.3.3] Preparation of diluent
Diluent for deferasirox was 99.5 mL acetonitrile and 0.5 mL water
[7.3.4] Preparation of standard and sample solution
The Standard stock solutions were prepared by accurately weighing 100
mg of deferasirox in 100 mL volumetric flask (1000 µg/mL) in
acetonitrile. Sample solutions were prepared by appropriate dilution of
the standard solutions with the diluent.
[8] Method development and optimization of chromatographic conditions
(UV graph/chromatograms)
To develop a precise, accurate and suitable RP-HPLC method for the
simultaneous estimation of salicylamide different mobile phases, solvent-buffer
ratios and pH were tried to proposed final chromatographic conditions.
Deferasirox is soluble in methanol and acetonitrile : water mixture. The peak
shape, resolution and symmetry of deferasirox were good with above gradient
elution at a 1.0 mL/min flow rate. The method developed was unique in
determining the impurities even at low levels than that of specifications. The
developed method was successfully applied to estimate the amount of
deferasirox.
Optimized chromatographic conditions
Parameter Optimized condition
Flow rate 1.0 mL/min
Mobile phase 40 : 60 v/v (Buffer : ACN)
Buffer pH Potassium phosphate buffer pH 3.2 adjusted by OPA
Wavelength 245 nm
Injection volume 10 µL
Run time 15 min
Column and column oven temperature
30ºC
Section- [III] Deferasirox
PART-B Page 103
Figure 1: Chromatogram and UV calibration curve for deferasirox standard
(Mobile phase: ACN : buffer, 60 : 40 v/v, pH 3.2 with OPA).
Figure 2: Chromatogram and UV calibration curve for deferasirox sample
(Mobile phase: ACN : buffer, 60 : 40 v/v, pH 3.2 with OPA).
Section- [III] Deferasirox
PART-B Page 104
Figure 3: Chromatogram and UV calibration curve for deferasirox sample
(Mobile phase: ACN : buffer, 60 : 40 v/v, pH 3.2 with OPA).
Section- [III] Deferasirox
PART-B Page 105
Figure 4: Chromatogram and UV calibration curve for deferasirox sample
(Mobile phase: ACN : buffer, 60 : 40 v/v, pH 3.2 with OPA).
[9] Analytical method development/validation and its result and
discussion
The optimized RP-HPLC assay method was validated for specificity, linearity,
accuracy, precision (repeatability and intermediate precision), recovery and
system suitability according to International Conference on Harmonization
(ICH) guidelines for the validation of bioanalytical method [21] and the US Food
and Drug Administration (FDA) [22].
[9.1] System suitability
System suitability was performed by using 100 µg/mL of salicylamide by
making six replicate injections. Chromatographic parameters calculated from
experimental data, such as Number of theoretical plates, % RSD of peak area
and resolution factors (Rs) are given in table-2. The system was deemed to be
Section- [III] Deferasirox
PART-B Page 106
suitable for use if the capacity factors were in the range of 2-20 (2 < K’ < 20),
lower than 2 for tailing factor, more than 2 for resolution (Rs), greater than
6988 number of theoretical plates (N), resolution between salicylamide of at
least two and less than 2 % relative standard deviation (% RSD) for peak area.
Table 2: System suitability parameters
System suitability parameters
Sr. No. Parameters Salicylic acid
1 Linearity range (µg/mL) 10.0-100.0 µg/mL
2 Retention time (Min.) 7.030
3 Theoretical plates (N) 6988
4 Peak Asymmetry (T) 2.46
5 Resolution (Rs) 2.4
6 Accuracy 99.99 %
7 Precision 99.70 %
8 % RSD (For peak area) 0.089 %
[9.2] Precision
The precision of the assay was studied with respect to both intra-day
(Repeatability) and Inter-day (Intermediated) precisions. Repeatability was
calculated from five replicate injections of three different concentrations of
deferasirox in the same equipment on the same day. Inter day precision was
checked with the same concentrations as intra-day assay and the
determination of each compound was repeated day by day during three days.
The method was found to be precise with RSD values within for intra-day and
inter day assay. Evaluation of the intra-day and inter-day precision for the
determination of deferasirox by the proposed HPLC method according to ICH
guidelines.
Intra Day (Repeatability) precision
Repeatability can be defined as the precision of the procedure when repeated by
same analyst under the same operating conditions over a short interval of time
or same day. It is normally expected that at least six replicates be carried out
and individual result provided from mean, standard deviation and coefficient of
variation should be calculated for set of n value. The RSD values are important
for showing degree of variation expected when the analytical procedure is
repeated several time in a standard situation (RSD below 2 % for assays in
finished product).
Section- [III] Deferasirox
PART-B Page 107
Inter day (Intermediate) precision
Repeatability can be defined as the precision of the procedure when repeated by
same analyst under the same operating conditions and the determination of
each compound was repeated day by day during three days or study repeat
three days over a long interval of time.
Table 4: Intra-day and Inter-day precision data for deferasirox standard
Standard con. (µg/mL)
Intra day Inter day
Area (mAU*min) Area (mAU*min)
1000 596.335 596.337
596.252 596.389
596.399 596.299
596.288 596.249
596.372 596.399
Average 596.3292 596.3346
% SD 0.059989 0.062664
% RSD 0.01006 0.010508
Standard potency 99.77 %
Table 3: Intra-day and Inter-day precision data for deferasirox sample Sample
Con.
(µg/mL)
Intra day Inter day
Area
(mAU*m
in)
Mean
area
% SD %
RSD
Area
(mAU*m
in)
Mean
area
% SD %
RSD
100 60.359 60.323 0.064 0.106 60.358 60.342 0.053 0.089
60.288 60.252
60.345 60.375
60.232 60.339
60.395 60.389
500 291.159 291.301 0.139 0.048 291.157 291.201 0.056 0.019
291.219 291.198
291.374 291.217
291.245 291.148
291.508 291.289
1000 596.335 596.362 0.064 0.010 596.338 596.381 0.065 0.010
596.399 596.399
596.427 596.332
596.388 596.489
596.265 596.351
[1] % Assay =
P
AT = Average area of obtained in sample preparation
AS = Average area of obtained in standard preparation
W1 = Weight taken of reference standard (mg)
Section- [III] Deferasirox
PART-B Page 108
W2 = Weight taken of test sample (mg)
P = Potency of reference standard (%)
[2]
[1] Intra day
1. % Assay =
99.77
= 1.00 0.9993 99.77 = 99.70 %
2.
=
= 0.010 %
[2] Inter day
1. % Assay =
99.77
= 1.00 0.9995 99.77 = 99.72 %
2.
=
= 0.010 %
[9.3] Limit of detection (LOD) and Limit of Quantification (LOQ)
The limit of detection (LOD) is defined as the lowest concentration of an analyte
that can reliably be differentiated from background levels. The standard
solutions of the compounds for LOD were prepared by diluting them
sequentially. Limit of quantification (LOQ) of an individual analytical procedure
is the lowest amount of analyte that can be quantitatively determined with
suitable precision and accuracy (ICH Guideline Q2B, 2005). LOD and LOQ were
determined calculating the signal-to-noise ratio of each compound by injecting
a series of solution until the S/N ratio 3 for LOD and 10 for LOQ. where S is the
standard deviation of y-intercepts of regression.
[9.4] Specificity
Specificity of method can be absence of any interference at retention
times of samples. The specificity of the method was demonstrated by injection
of standard solution of deferasirox at concentration of 100 µg/mL. The elution
peaks of salicylamide, salicylic acid and deferasirox presented in representative
chromatograms. The representative chromatogram for simultaneous
determination of the studied drugs in API pharmaceutical dosages forms.
Section- [III] Deferasirox
PART-B Page 109
Table 5: Specificity study of deferasirox
Con. (µg/mL) Sample Standard
Area (mAU*min) Area (mAU*min)
100 59.978 59.99
59.992 59.98
59.957 59.97
59.983 59.99
59.978 59.95
Average 59.9776 59.976
% SD 0.012856905 0.016733201
% RSD 0.021436177 0.027899827
Standard potency 99.77 %
[1] % Assay =
99.66
= 1.00 0.9998 99.66 = 99.64 %
[2]
=
= 0.01018 %
[9.5] Linearity
The linearity of deferasirox was studied by preparing standard solution at
five different concentrations ranging from 10.0-100.0 µg/mL. Each
concentration was injected in a five replicates and mean value of peak area was
taken for calibration curve.
Construction of the calibration curves
Working solutions containing (10.0-100.00) μg/mL were prepared by serial
dilution of standard solution with the acetonitrile. In all cases, 10 µL aliquots
were injected (triplicate) and eluted with the mobile phase under the following
chromatographic conditions. The average peak area ratio of each drug and the
internal standard were plotted versus the final concentration of the drug in
μg/mL to get the calibration graph. Alternatively, the corresponding regression
equation was derived.
Sr. No. Concentration (µg/mL) Area (mAU*min)
1 1 0.968
2 10 6.51
3 20 10.12
4 30 14.29
5 40 21.277
6 50 28.353
7 100 59.977
Section- [III] Deferasirox
PART-B Page 110
Figure 5: Linearity curve
Table 6: Summary of linearity data
Summary of linearity data
Sr. No. Parameters Deferasirox
1 Linearity range (µg/mL) 1.0-100.0 µg/mL
2 Slope ± Standard error 1.2427
3 Intercept ± Standard error 0.5984x
4 Linearity equation 0.5984x - 1.2427
5 r2 0.99 %
[9.6] Accuracy
Accuracy of the assay method was calculated for deferasirox by recovery studies
at three concentrations of 50 %, 100 %, 150 % and 200 % levels by standard
addition method. The mean % recovery for deferasirox was found is given in the
follow table 7.
Section- [III] Deferasirox
PART-B Page 111
Table 7: Accuracy data for deferasirox
Sr. No.
Deferasirox Added (µg/mL)
Found (µg/mL)
% Recovery
% Mean Recovery
SD % RSD
1 50% 4.754 4.754 99.99 99.99 0.026 0.026
4.755 4.754 100.02
4.752 4.754 99.96
4.755 4.755 100.00
4.753 4.754 99.96
2 100% 10.057 10.057 99.99 99.99 0.028 0.028
10.059 10.056 100.03
10.054 10.057 99.96
10.057 10.058 99.98
10.052 10.056 99.96
3 150% 14.664 14.664 99.99 99.98 0.088 0.088
14.671 14.664 100.04
14.647 14.665 99.88
14.679 14.665 100.09
14.652 14.664 99.91
4 200% 20.388 20.388 100.00 100.01 0.081 0.081
20.378 20.388 99.95
20.371 20.387 99.91
20.413 20.388 100.12
20.399 20.387 100.05
Table 8: Sample and standard area for deferasirox
Sample Area (mAU*min)
Standard
Area (mAU*min)
50% 100% 150% 200% 1000%
28.353 59.977 87.45 121.58 596.335
28.359 59.989 87.49 121.52 596.252
28.343 59.959 87.35 121.48 596.399
28.358 59.973 87.54 121.73 596.288
28.345 59.948 87.38 121.65 596.372
Average 28.351 59.969 87.442 121.592 596.329
% SD 0.0073 0.0159 0.0779 0.0897 0.0599
% RSD 0.0258 0.0266 0.0890 0.0738 0.0100
Standard potency
99.77 %
[1] Amount added (µg/mL) =
=
= 4.754 µg/mL
[2] Amount found (µg/mL) =
=
= 4.754 µg/mL
Section- [III] Deferasirox
PART-B Page 112
[3] % Recovery =
= 99.99 %
[10.0] Summary and Conclusion
A new RP-HPLC-DAD method has been developed for the estimation of
deferasirox in bulk API dosages forms. The developed method was
validated as per ICH guidelines and was found to be simple, precise,
accurate, and sensitive. The proposed method is rapid, where the total
analytical run time for deferasirox (Rt = 7.030 min.) and the internal standard is
less than 10 min and can be used for routine analysis of deferasirox in
bulk API dosages forms.
[11.0] References
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Journal of pharmacy research. 2011, 4, 2998-3000.
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Satyanarayana. Chromatographia. 2010, 72, 441-446.
[4] VK. Chakravarthy, D. Gowrisankar. Journal of Global Trends in
Pharmaceutical Sciences. 2010, 1, 37-45.
[5] V. Sambasivarao, RS. Phani, R. Seetharaman, KS. Lakshmi. IJPI’s
Journal of Analytical Chemistry. 2011, 1, 31-35.
[6] Official manufacturer website including information for health care
professionals about indications, dosing, safety and more:
http://www.exjade.com.
[7] VP. Choudhry, R. Naithani. Current status of iron overload and chelation
with deferasirox. Indian J Pediatr. 2007, 74, 759-64.
[8] LP. Yang, SJ. Keam, GM. Keating . Deferasirox: a review of its use in the
management of transfusional chronic iron overload. Drugs. 2007, 67,
2211-2230.
[9] FDA Approves First Oral Drug for Chronic Iron Overload (Press release).
United States Food and Drug Administration. November 9, 2005.
[10] http://www.ema.europa.eu/docs/en_GB/document_library/EPAR
Summary for the public/human/000670/WC500033927.pdf.
Section- [III] Deferasirox
PART-B Page 113
[11] http://www.lancet.com/journals/lancet/article/PIIS01406736%2813%
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