International Journal of Research and Applications
Oct - Dec © 2015 Transactions 2(8): 376-381
CHEMISTRY
eISSN : 2349 – 0020
pISSN : 2394 - 4544
ORIGINAL A RTICLE
VALIDATION OF DEVELOPED METHOD FOR ZOLMITRIPTAN TABELETS
IN PHARMACEUTICAL DOSAGEFORM BY RP-HPLC
Jagadeesh Kumar Ega , Kavitha Siddoju*
Department of Chemistry,
Chaitanya Postgraduate College,(Autonomous) Warangal, Telangana - 506001.
ABSTRACT
Zolmitriptan acts as Anti-inflammatory Agents, Anti-migraine Agents, Selective Serotonin Agonists and
Vasoconstrictor Agents. A new RP-HPLC method was developed for the determination of Zolmitriptan
((4R)-4-[[3-(2-dimethylaminoethyl)-1H-indol-5-yl] methyl]-1, 3- oxazolidin-2-one) in tablet dosage
form. The HPLC method was then validated to indicate that the analytical procedure used is suitable for
intended use by using various parameters like specificity, linearity, and precision, and accuracy,
stability in analytical solution, system suitability and filter interference. A new, specific stability
indicating method was developed. This Validation describes the procedure for assay of Zolmitriptan
(MF: C16H21N3O2) tablets 5 mg by HPLC as per ICH Guidelines and can be applicable for the analysis of
commercial dosage forms. X-TerraRP-18, 250x4.6mm,5µ
temperature 30°C, run time 8 minutes
column, injection volume20 µl , Column
with a hydrophilic linkage between silica particles and
hydrophobic alkyl chains. Using isocratic elution with UV detection at 225 nm.
Keywords: Zolmitriptan, RP- HPLC, ICH.
1. INTRODUCTION
Zolmitriptan binds with high affinity to human
Venous anastomoses) and sensory nerves of the
5-HT1B and 5-HT1D receptors leading to cranial
trigeminal system which result in cranial vessel
blood vessel constriction. Current theories
constriction and inhibition of pro-inflammatory
proposed to explain the etiology of migraine
neuropeptide release.
headache suggest that symptoms are due to
local cranial vasodilatation and/or to the release
of sensory neuropeptides (vasoactive intestinal
2. MATERIALS
Table No: 1 list of materials
peptide, substance P and calcitonin gene-related
peptide)
through
nerve
endings
in
the
trigeminal system. The therapeutic activity of
S.
Name of
No.
Instrument
Model
Make
CPA225D
Sartorius
zolmitriptan for the treatment of migraine
headache can most likely be attributed to the
agonist effects at the 5HT1B/1D receptors on
intracranial blood vessels (including the arterio-
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Semi micro
balance
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Micro balance
CPA2P
Sartorius
Merck
4
3
4
Precision
balance
pH
HPLC
6
HPLC
7
Column
Column
CHT
Shimadzu
Wakosil
6),5µ
6
Hydrochloric acid
Merck
(GR – Grade)
7
Hydrogen peroxide Merck
Working/reference standards – Zolmitriptan
Filter 0.45µm GHP membrane filter (Manufactured
C18(250X4.
Waters
6),5µ
by PALL) and 0.45µm PVDF
UV-1800
Shimadzu
UCB 70
Spectralab
Test sample
Zolmitriptan tablets 5 mg and
Zolmitriptan tablets Placebo.
3.
Sonicator
membrane filter
(Manufactured by PALL).
ter
10
TKA water
Agilent
C18(250X4.
Spectrophotome
Water
Orion
UV9
5
Thermo
1200 series
LC-2010
(GR grade)
Sartorius
3 Star
Meter
5
8
GPA5202
Triethylamine
METHODOLOGY
Method Development by RP-HPLC
Preparation of buffer solution (Mobile Phase-A)
Reagents and chemicals:
Weighed accurately about 6.85358g of sodium
Table No: 2 list of Reagents and chemicals
dihydrogen orthophosphate and 1.75023g of
disodium hydrogen phosphate dihydrate and
Chemicals/
SS.No.
transferred into 5 liter bottle. Added 5 liter
Make/Grade
Reagents
until the material dissolved completely. Mixed
Merck,
1
(HPLC-Grade)
2
0.45 µm nylon membrane filter and degassed by
sonicating for 5 minutes.
Merck
dihydrogen
orthophosphate
well, checked the pH and adjusted to 7.8 ±0.05
with triethylamine. Filter the solution through
Acetonitrile
Potassium
water and mixed well using magnetic stir bar
Mobile phase B Acetonitrile.
(GR-Grade)
Preparation of Diluent
Merck
3
Transferred 750ml of buffer and 250 ml of
Sodium Hydroxide
(GR-Grade)
Acetonitrile into 1 liter bottle, Mixed well and
degassed by sonicating for 5 minutes.
1.
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Oct – Dec © 2015 Transactions
Standard preparation
Weighed
accurately
Trial No. 3
about
52.54
mg
of
Zolmitriptan working standard and transferred
into a 100 ml volumetric flask. Added about 50
ml of diluent to dissolve by sonicating for 5
minutes and completed to volume with diluent.
Sample Preparation for Assay
Weighed accurately 10 tablets and transferred
to mortar, and crushed the sample and weigh
equivalent
to
(3988.38mg)
80
and
mg
of
Zolmitriptan
transferred
to
100
ml
volumetric flask. To this added about 50 ml of
diluents
and
sonicated
for
20
Figure: 7 HPLC chromatogram of trial 3
minutes.
Completed to volume with diluent and filtered
through 0.45µ GHP filter.Further pipetted 3ml
to 25ml and completed to volume with diluent.
.Trial No. 1
HPLC chromatogram for optimized chromatographic
parameters
Figure: 9 HPLC Chromatogram of blank
Figure: 5 HPLC chromatogram of trial 1
Trial No.2
Figure:
10
HPLC
chromatogram
Zolmitriptan
Figure: 6 HPLC chromatogram of trial 2
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standard
International Journal of Research and Applications
Oct – Dec © 2015 Transactions
Figure: 11 HPLC chromatogram of sample drug of
Zolmitriptan
Figure:15 HPLC chromatogram of Zolmitriptan linearity
Figure: 12 HPLC chromatogram of Zolmitriptan
100%
linearity 25%
Figure: 13 HPLC Chromatogram of
Zolmitriptan
linearity 50%
Figure: 14 HPLC chromatogram of Zolmitriptan
Figure: 16 HPLC chromatogram of Zolmitriptan
linearity 125%
Figure: 17 HPLC chromatogram Zolmitriptan 150%
linearity 75%
1.
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A
4. SUMMARY
HPLC method
Analysis – II, by Nirali Prakashan, 13th Edn,
for
Zolmitriptan
was
2005, pg.47-56
developed and validated in tablet dosage form
as per ICH Guide lines. Agilent 1200 series with
[4] Beckett A.H. and Stanlake J.B. Practical
DAD Detector and Waters X-terra(250 × 4.6 mm,
Pharmaceutical
5 μl) column, injection of 10 µl is injected and
Publishers and Distributors; 4
eluted with the mobile phase of sodium
pg.157-174.
Chemistry,
Part
th
2,
CBS
Edition.2002;
dihydrogen orthophosphate buffer with 0.2M of
7.8 pH and acetonitrile in the ratio 80:20v/v,
which was pumped at a flow rate of 1.0 ml at
225 nm. The peak of Zolmitriptan was found
well separated at 4.2 min. The analytical method
validation of Zolmitriptan by RP HPLC method
[5] Remington. The Science & Practice of a
th
Pharmacy Vol. I, 20 Edn. pp 587-613.
[6] Snyder L.R., Kirkland J.J. and Glajch L.J.,
Practical HPLC Method Development. John
was found to be satisfactory and could be used
Wiley and Sons, INC; 2
for the routine pharmaceutical analysis of
102.
Zolmitriptan.
nd
Edition. 1997, pg. 98-
[7] L.R. Snyder and J.J. Kirkland. Introduction to
5. CONCLUSION
The proposed RP-HPLC method is suitable for
Modern Liquid Chromatography. 2 nd edition;
John Wiley & Sons, New York, 1979; pg.453-482.
the estimation of Zolmitriptan in formulation.
All the validation parameters for Zolmitriptan
[8] L.R. Snyder and J.J. Kirkland, and J.L. Glajch,
meet the criteria as per ICH guidelines. The
Practical
analytical method was found to be simple,
edition, Wiley-interscience, New York, 1997;
sensitive, accurate and precise. The developed
pg.135-138.
methods may be recommended for routine and
QC analysis of the investigated drug to provide
reproducible
determination
quantitative
of
analysis
Zolmitriptan
for
in
the
tablet
formulation.
HPLC
Method
Development,
2nd
[9] Michael W. Dong. Modern hplc for practicing
scientists. John Wiley & Sons. New Jersey. 2006;
pg.193-196.
[10] Hokanson GC. A life cycle approach to the
Validation
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