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Reverse Phase-hplc Method Development and Validation for the Simultaneous Estimation of Azilsartan Medoxomil and Chlortalidone in Pharmaceutical Dosage Forms

Reverse Phase-hplc Method Development and Validation for the Simultaneous Estimation of Azilsartan Medoxomil and Chlortalidone in Pharmaceutical Dosage Forms

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Published by: jamonline on Feb 29, 2012
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Jamonline /Jan-Feb 2(1) / 117–126 Madhu Babu K & Bikshal Babu K
All rights reserved© 2011 www.jamonline.in 
Research Article
Journal of Atoms and Molecules
  An International Online Journal An International Online Journal An International Online Journal An International Online Journal
ISSNISSNISSNISSN –– 2277227722772277 –– 1247124712471247
*, Bikshal Babu Kasimala
Faculty in Chemistry, Department of Allied Sciences, College of Marine Science and Technology,Massawa, Eritrea, North East Africa.
QC Department, RV Labs, Guntur, Andhra Pradesh, India.
Received on: 01-01-2012 Revised on: 14-02-2012 Accepted on: 29–02–2012Abstract:
A simple, selective, linear, precise and accurate RP-HPLC method was developed and validated forthe simultaneous estimation of Azilsartan Medoxomil and Chlortalidone in pharmaceutical dosageforms. Isocratic elution at a flow rate of 0.9ml min
was employed on a symmetry C18 column atambient temperature. The mobile phase consisted of Methonal: Water: Acetonitrile : 0.1% Orthophosphoric acid 30:35:15:5(v/v/v/v). The UV detection wavelength was at 251 nm. The retentiontime for Chlortalidone was 3.923min and Azilsartan Medoxomil was 7.208 min. The method wasvalidated as per the ICH guidelines. The proposed method can be successfully applied for theestimation of Azilsartan Medoxomil and Chlortalidone in pharmaceutical dosage forms.
Key Words:
Azilsartan Medoxomil and Chlortalidone, HPLC Development, Validation, 251nm.
Chlortalidone or chlorthalidone is a diureticdrug used to treat hypertension, originallymarketed as Hygroton in the USA. It isdescribed as a thiazide diuretic (or, rather, athiazide-like diuretic because it acts similarlyto the thiazides but does not contain thebenzothiadiazine molecular structure).Compared with other medications of thethiazide class, chlorthalidone has the longest* Corresponding authorMadhu Babu Kasimala,Email: madhu.lucky09
 Tel: 00291 – 7251105
Jamonline /Jan-Feb 2(1) / 11
All rights reserved© 2011duration of action but a similat maximal therapeutic doses.in the management of hoedema.
Figure 1: Stricture of C
Chlortalidone works on the k to treat fluid retention caukidney conditions,
heart failure
(a conpumping action of the heart
diabetes insipidus
(a conditiindividual produces large aurine). It removes excess watby increasing how much andpass urine. This removal oblood pressure and helps redthe heart. How it works in dihowever is not fully understoChlortalidone is used to trpressure (hypertension), heheart is unable to properly puthe body), fluid collection(ascites) due to cirrhosis of retention due to kidney disyndrome) and a type of diabetes insipidus that cauexcessively thirsty and pass lurine. It is a type of diurknows as water tablets. It is
7–126 Madhu Ba
r diuretic effectIt is often usedpertension and
idney. It is useded by liver or
(high blooddition where theis reduced) andn in which anounts of diluter from the bodyhow often youf fluid reducesuce the work of abetes insipidusd.eat high bloodrt failure (thep blood aroundin the stomachthe liver, fluidease (nephroticdiabetes calledses you to bearge amounts of tic, sometimesused to removeexcess fluid from thproduction of urine.your body causes yoIn diabetes insipiduproduction of urine.used to reduce(hypertension) and rbody (oedema) in ceused to treat a typeinsipidus.
is anantagonist usedhypertension that wa
Figure 2: Stri
Azilsartan is anblocker (ARB) thatblocking the actiovasopressor hormontreat high bloodLowering high bloostrokes, heart attack Azilsartan belongs tangiotensin receptorworks by relaxing blcan flow more easily
bu K & Bikshal Babu K
body by increasing yourThe reduction in fluid inr blood pressure to drop., the drug stops the overIn general this drug ishigh blood pressuremove excess fluid in ther
tain conditions. It is alsof diabetes called diabetesangiotensin II receptorin the treatment of s developed by Takeda.
ture of Azilsartan
angiotensin II receptorlowers blood pressure byof angiotensin II, ae. Azilsartan is used topressure (hypertension).d pressure helps prevents, and kidney problems.a class of drugs calledblockers (or ARBs). Itood vessels so that blood.
Jamonline /Jan-Feb 2(1) / 117–126 Madhu Babu K & Bikshal Babu K
All rights reserved© 2011 www.jamonline.in 
Azilsartan has a boxed warning that says theuse of the drug should be avoided in pregnantwomen because use of the drug during thesecond or third trimester can cause injury andeven death in the developing fetus. If awoman becomes pregnant while using thedrug, it should be discontinued as soon aspossible.
ExperimentalChemicals and reagents
All solvents used like Methanol, Water,Acetonitrile, and Ortho Phosphoric Acid(OPA) which are of HPLC grade werepurchased from E.Merck, Mumbai.
Instrumentation and analytical conditions
The analysis of the drug was carried out onShimadzu HPLC model (VP series) containingLC-10AT (VP series) pump, variable wavelength programmable UV/visible detectorSPD-10AVP and Rheodyne injector (7725i)with 20µl fixed loop. Chromatographicanalysis was performed using Gemini C-18column with 250 x 4.6mm internal diameterand 5µm particle size. Shimadzu electronicbalance (AX-200) was used for weighing.Isocratic elution with Methonal : Water :Acetonitrile : 0.1% Ortho phosphoric acid30:35:15:5(v/v/v/v). was selected with a flowrate of 0.9ml min
- .
The detection wavelengthwas set at 251nm with a runtime of 10 min.The mobile phase was prepared freshly and itwas degassed by sonicating for 5 min beforeuse. The column was equilibrated for at least30min with the mobile phase flowing throughthe system. The column and the HPLC systemwere kept at ambient temperature.
Preparation of Stock, working standardsolutions and Sample solutions
100mg of Azilsartan Medoxomil andChlorthalidone was weighed separately andtransferred (working standard) into a 100mlvolumetric flask. The diluent Methanol wasadded and sonicated to dissolve it completelyand made up to the mark with the samesolvent. Further 1ml of the above stock solution was pipetted into a 10ml volumetricflask and diluted up to the mark with diluent.The contents were mixed well and filteredthrough Ultipor N
Nylon 6, 6 membranesample filter paper. The calibration curve wasplotted with the concentrations of the 10 to 80ppm working standard solutions. Calibrationsolutions were prepared and analyzedimmediately after preparation.The formulation tablets of AzilsartanMedoxomil and Chlorthalidone were crushedto give finely powdered material. Powderequivalent to10 mg of drug was taken in 10ml of volumetric flask containing 5 ml of mobile phase and was shaken to dissolve thedrug and then filtered through Ultipor N
 Nylon 6,6 membrane sample filter paper.Volume of the filtrate was adjusted to themark with the same solvent to obtainconcentration of 40 ppm.

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