Ibandronate
Ibandronate
Ibandronate
ISSN: 0976-4607
Research Article
St. John Institute of Pharmacy and Research, Palghar (E), M.S India.
R.C. Patel Institute of Pharmaceutical Education and Research, Shirpur (M.S.) India.
ABSTRACT
A simple, accurate and sensitive liquid chromatographic method has been developed for the assay of ibandronate
sodium drug substance in tablet dosage form. The separation was achieved on Hypersil BDS C18 (250mm X 4.6mm), 5m
column. The mobile phase consisted of Buffer: ACN (95:05) v/v; flow rate 1.0 ml min1 at ambient temperature. The
analytes were monitored by PDA detector. The drug substance was subjected to stress conditions of hydrolysis, oxidation,
photolytic, thermal and humidity degradation. Considerable degradation was achieved under thermal condition. Mass
balance was demonstrated in all stress conditions. The method was validated for specificity, precision, linearity, solution
stability and accuracy. The average recoveries for ibandronate were in the range of 99.0102.0% and the method can be
successfully applied for the routine analysis of ibandronate sodium drug substance.
Key words: RP- HPLC, Stability indicating, Ibandronate Sodium.
INTRODUCTION
Ibandronate sodium [(1-hydroxy-3-(methyl
chromatographic method with PDA detection for the
pentyl amino) propylidene bisphosphonic acid
determination of ibandronate sodium.
monosodium monohydrate)] is the sodium salt of
ibandronic acid, a synthetic nitrogen-containing
Materials and Methods
bisphosphonate drug. This new third generation
Chemicals and Reagents:
bisphosphonate is used to treat patients with bone disease
The standard sample of ibandronate sodium
like Pagets disease, malignant hypercalcemia and
drug substance was procured from Aarti Drugs Ltd,
postmenopausal osteoporosis (Sankar Babu VR et al.,
Boisar-Thane. Analytical reagent (AR grade) Disodium
2010). For quantification of impurities and assay of
Edetate, Thomas Baker, Pentanesulfonic acid sodium salt
ibandronate sodium, few analytical methods have been
HPLC Grade, Merck, Triethylamine HPLC Grade,
reported. Indirect fluorescence detection was used in a
Rankem, Orthophosphoric acid (OPA) HPLC Grade,
high performance ion exchange chromatographic method
Rankem, Water HPLC grade procured from Milli-Q
based on the formation of the non-fluorescent Al3+system.
ibandronate complex after post-column addition of the
fluorescent Al3+-morin reagent (Narendra kumar M et al.,
High Performance Liquid Chromatography
2011). Ibandronate was determined by high performance
Agilent HPLC 1200 series chromatograph
ion exchange chromatography with UV detection at
equipped with binary pump, 2695 Photodiode Array
240nm after complex formation with Cu2+ ion.
Detector with data processing capacity was used. A
Ibandronate sodium was determined by capillary zone
Hypersil BDS column C18 (250 mm x 4.6 mm, 5 mm)
electrophoretic method within direct detection at 254nm,
was used. The pH measurement was performed by using
the limit of detection (LOD) values reported for
LAB INDIA- PICO controlled pH analyzer equipped
ibandronate was 3521760gml1. The aim of this study
with pH electrode. Mobile phase filtration was performed
by vacuum pump using 0.45 m filter paper. As a
was to develop a simple, sensitive, precise liquid
degasser, PCI Analytics Pathak ultrasonicator was used.
*Corrosponding author
Typical operating conditions include flow rate, 1 ml/min;
injection volume, 10 l; wavelength, 200nm; column
Jineetkumar B. Gawad
compartment temperature, 350C; and operating condition,
Email id: gawadjinit@yahoo.com
room temperature. The retention times of the ibandronate
220
Chromatogram
Sample
Chromatogram
of
of
Sample Preparation
Weigh accurately and transfer 8 intact tablets to
500 mL of volumetric flask. Add about 50 mL of diluent
and sonicate to disperse. Add about 300 mL of diluent
and sonicate for 25 min with intermittent shaking. Allow
to equilibrate to room temperature and dilute to volume
with diluent, mix. Centrifuge the solution at 2500 RPM
for 5 min. dilute 5 mL of supernatant to 20 mL with
diluent, mix. Filter through 0.45 nylon filter discarding
first 3 mL of the filtrate (Figure.2).
Method validation
Linearity study
In order to prepare stock solution, 256 mg
Ibandronate sodium was accurately weighed, dissolved in
Accuracy
The accuracy of an analytical procedure
expresses the closeness of agreement between the value
which is accepted either as a conventional true value or
an accepted reference value and the value found. This is
also termed as trueness. It was done by recovery study.
Sample solutions were prepared with 100% in triplicate.
System Precision (Repeatability)
Repeatability expresses the precision under the
same operating conditions over a short interval of time.
221
Range
The range of an analytical procedure is the
interval between the upper and lower concentration
(amounts) of analyte in the sample (including these
concentrations) for which it has been demonstrated that
the analytical procedure has a suitable level of precision,
accuracy and linearity. Range to be inferred from the data
of linearity, recovery and precision experiments.
Specificity and selectivity
Specificity is the ability to asses unequivocally
the analyte in the presence of components which maybe
expected to present. The analytes should have no
interference from other extraneous components and be
well resolved from them. Specificity is the procedure to
detect quantitatively the analyte in presence of
component that may be expected to be present in the
sample matrix, while selectivity is the procedure to detect
qualitatively the analyte in presence of components that
may be expected to be present in the sample. The method
is quite selective. There was no other interfering peak
around the retention time of Ibandronate Sodium also the
baseline did not show any significant noise.
Forced Degradation Studies
Acid degradation studies
10 tablets were weighed. The average weight
was determined. Sample powder equivalent to 100 mg of
Ibandronate sodium was transferred in to a 100 ml
volumetric flask. About 50 ml diluent was added and
sonicated for 10mins. Added 5 ml of 5M HCl and kept at
80C on water bath for 5 hrs. Then solution was allowed
to cool at room temperature, 5 ml of 5M NaOH solution
was added, for neutralization, and volume was made up
to the mark with diluent and mixed properly.
These solutions were centrifuged and
subsequent solutions were collected. A 10 l of these
solutions were injected into LC, under optimized
chromatographic conditions (Figure.4).
Figure.6 Overlaid
Stressed Sample
Chromatogram
of
Peroxide
222
Figure.8
Sample
Overlaid
Chromatogram
of
Humidity
Humidity degradation
Tablet was kept in 40C/75% RH chamber for
24 hrs. Weighed and transferred sample powder
equivalent to 100 mg of Ibandronate sodium in to a 100
ml volumetric flask, about 75 ml of diluent was added
and sonicated for 10mins and made up to volume with
diluent and mixed. This solution was centrifuged and
supernatant solution was collected. A 10 l of this
solution was injected into LC, under optimized
chromatographic conditions (Figure.8).
Photostability studies
Crushed tablet powder was kept in the photo
stability chamber and exposed to, as per ICH guidelines
(An overall illumination of not less 1.2 million lux hrs
and an integrated near ultraviolet energy of not less than
200 watt hrs/sq m). Weighed and transferred sample
powder equivalent to 100 mg of Ibandronate sodium in to
a 100 ml volumetric flask, about 75 ml of diluent was
added and sonicated for 10mins and made up to volume
with diluent and mixed. This solution was centrifuged
and supernatant solution was collected. A 10 l of this
solution was injected into LC, under optimized
chromatographic conditions (Figure 9).
RESULTS AND DISCUSSION
Method Development and optimization
As there is no chromophore present in
ibandronate sodium, there was no possibility for UV or
fluorescence detection and no suitable groups are present
for derivatization. Ibandronate sodium; for this reason
water was chosen as diluents. Preliminary experiments
were carried out Using Hypersil BDS C18 Column with
Buffer: ACN (70:30) v/v (adjusted to pH 2 using OPA)
Ibandronate was lost peak shape while on Inertsil ODS
Column, with Buffer: ACN (40:60) v/v peak was tailed
with asymmetry 2.41.Ibandronate was determine on
Hypersil BDS C18 column, peak was separated using
phosphate buffer with pH 7.0 (1.75gm pentanesulfonic
acid sodium salt+100mg EDTA in 900ml of water +6ml
TEA, dilute upto 1000ml, adjust pH with OPA); Buffer:
ACN (95:05) v/v. Better resolution obtained using
acetonitrile as organic modifier. Satisfactory separation
and good peak shapes were achieved within a reasonable
time using a mobile phase of 95:5% (v/v) mixture of
buffer and ACN with a flow rate of 1.0 ml min1. The
effect of column temperature on separation was studied at
different temperatures ranging from 35 0C to 650C.
Ambient temperature was found to be optimal from the
point of view of both resolution and peak shape.
Method validation
The proposed method was validated as per ICH
guidelines. The drug solutions were prepared as per the
earlier adopted procedure given in the experiment.
Linearity
The linearity of photodiode array detector
response of ibandronate sodium at different
concentrations was studied in the range 300-900 gml1
223
Area III
204955
327244
370846
411834
451345
487680
609448
Area III
366299
364985
365375
Accuracy
Accuracy of method was determined by
recovery experiments using standard addition technique.
Recoveries were determined by adding the ibandronate
sodium in triplicate, i.e.100% recovery values are given
in Table.4.
Mean
204669
327769
368510
411580
452737
488306
608669
Mean
366299
364985
365375
SD
1456
455
2409
235
4607
1085
709
Recovery
574.36
572.30
572.92
%RSD
0.71
0.14
0.65
0.06
1.02
0.22
0.12
% Recovery
100.0
99.7
99.8
Table.5a
Results
of
System
Precision
(Reproducibility)
System Precision
Injection
Area of Ibandronate
1
443650
2
432008
3
447833
4
438202
5
450637
6
445838
SD
98.7 1
%RSD
1.30
Area
443650
441588
440750
440823
440482
439141
439305
440194
Cumulative SD
18.07
Ibandronate Sample
% Diff w.r.t. Initial
0.5
0.7
0.6
0.7
1.0
1.0
0.8
Cumulative % RSD
0.52
224
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