NOVEL SIMULTANEOUS SEPARATION AND QUANTITATIVE DETERMINATION OF FOUR SARTANS IN PRESENCE OF HYDROCHLOROTHIAZIDE BY ISOCRATIC RP-HPLC Under the guidance of: Dr. Panchumarthy Ravisankar M. Pharm., Ph.D. Professor & HOD Dept. of Pharmaceutical Analysis and Quality Assurance Vignan Pharmacy College Vadlamudi. Presentation by: K. Manjusha (12AB1R0043) V. Laya Sri (12AB1R0044) B. Vijaya Kumar (12AB1R0009) K. Rajyalakshmi (12AB1R0037) K. Avinash kumar (12AB1R0039)
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NOVEL SIMULTANEOUS SEPARATION AND QUANTITATIVE DETERMINATION OF FOUR SARTANS IN PRESENCE OF HYDROCHLOROTHIAZIDE BY ISOCRATIC RP-HPLC
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NOVEL SIMULTANEOUS SEPARATION AND QUANTITATIVE DETERMINATION OF FOUR SARTANS
IN PRESENCE OF HYDROCHLOROTHIAZIDE BY ISOCRATIC RP-HPLC
Under the guidance of:Dr. Panchumarthy Ravisankar M. Pharm., Ph.D.
Professor & HOD Dept. of Pharmaceutical Analysis and Quality AssuranceVignan Pharmacy CollegeVadlamudi. Presentation by:
K. Manjusha (12AB1R0043)V. Laya Sri (12AB1R0044)B. Vijaya Kumar (12AB1R0009)K. Rajyalakshmi (12AB1R0037)K. Avinash kumar (12AB1R0039)
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CONTENTS: AIM AND OBJECTIVE
INTRODUCTION DRUGS PROFILE LITERATURE SURVEY PLAN OF WORK MATERIALS AND METHODS RESULTS CONCLUSION REFERENCES
Vignan Pharmacy College,Vadlamudi.
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AIM:
The core AIM of the present study is to develop a novel, rapid,
precise and accurate RP-HPLC method for simultaneous
separation and quantification of Hydrochlorothiazide along with
four sartans Telmisartan(TELM), Losartan(LOSA),
Olmesartan(OLME) and Valsartan(VALS).
To develop Novel methods for separation and quantification of
all the above said drugs on single chromatographic system
without any minor changes in detection wavelength and mobile
phase composition.
Vignan Pharmacy College,Vadlamudi.
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OBJECTIVE:
Vignan Pharmacy College,Vadlamudi.
To develop rapid, sensitive and economical analytical methods
based on spectrophotometric & RP-HPLC techniques for estimation
and separtion of Hydrochlorothiazide with Telmisartan, Losartan,
Olmesartan and Valsartan in pharmaceutical dosage forms.
To develop method with shorter run time and better sensitivity.
Reducing the solvent consumption to make it more eco-friendly.
To validate the method for different parameters like Accuracy,
Precision, Linearity, specificity, Robustness according to
International Conference on Harmonization ICH Q2(R1)
guidelines[5] .
To apply the developed RP-HPLC method in the analysis of
pharmaceutical formulations.
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INTRODUCTION
Vignan Pharmacy College,Vadlamudi.
HPLC is a versatile tool for the qualitative and quantitative analysis
of drugs and pharmaceuticals, chemical and biological materials has
become indispensable in pharmacokinetics studies.
Of the many instrumental methods available for quantification RP -
HPLC is selected as a tool for developing novel analytical procedure.
Method development is done for the products as follows:
- New products
- Existing products
Alternate methods for existing products are developed for better
precision and ruggedness.
Method development:
Vignan Pharmacy College,Vadlamudi. 6
DRUG PROFILE
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Drug Name : HYDROCHLOROTHIAZIDE
Structure :
IUPAC : 6-chloro-1,1-dioxo-3,4-dihydro-2H-1,2,4-
benzothiadiazine-7-sulfonamide
Category : Anti hypertensive
Molecular Formula : C7H8ClN3O4S2
Molecular Weight : 297.74 g/mol
Brand names : Apo-Hydro, Aquazide H, Hydrodiuril,
HydroSaluric, Hydrochlorot, Microzide,
Esidrex, and Oretic
Vignan Pharmacy College,Vadlamudi.
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Drug name : VALSARTAN
Structure :
IUPAC Name :(S)-3-methyl-2-(N-{[2'-(2H-1,2,3,4-tetrazol-5- yl)biphenyl-4-yl]methyl}pentanamido)butanoic acid
Molecular weight :435.519 g/mol
Molecular formula : C24H29N5O3
Brand names : Diovan, Valent -H
Vignan Pharmacy College,Vadlamudi.
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Drug name : LOSARTAN
Structure :
IUPAC Name :(2-butyl-4-chloro-1-{[2'-(1H-tetrazol-5-yl)biphenyl-
4-yl]methyl}-1H-imidazol-5-yl)methanol
Molecular weight :422.91 g/mol
Molecular formula :C22H23ClN6O
Brand names : Cozaar, Losagard, Zargo-H
Vignan Pharmacy College,Vadlamudi.
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Drug name : OLMESARTAN
Structure :
IUPAC Name : 2,3-dihydroxy-2,2-butenyl-4(1-hydroxy-1- methylethyl)-2-propyl-1-[p-(o-1H-tetrazol-5- ylphenyl)benzyl]imidazole5-carboxylate2,2-carbonate
Molecular weight : 558.59
Molecular formula : C29H30N6O6
Brand names : Olmicep-H, Olmax
Vignan Pharmacy College,Vadlamudi.
N
NN
HN
Olmesartan
N
N OH
H3C
O
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Drug name : TELMISARTAN
Structure :
IUPAC Name : 2-(4-{[4-Methyl-6-(1-methyl-1H-1,3-benzodiazol- 2-yl)-2-propyl-1H-1,3-benzodiazol-1-yl]methyl} phenyl)benzoic acid
Development and Validation of RP-HPLC Method for the Estimation and Separation ofValsartan, Losartan and Irbesartan in Bulk and Pharmaceutical Formulation.
Separation of 3 sartans using c18 column and its validation according to ICH guidelines.
International Journal of Pharmaceutial Sciences Review and Research.(IJPSRR)
Preparation of reagents and standards: Preparation of phosphate buffer pH 3.3
Phosphate buffer with 10 mM was prepared by dissolving 6.056 g KH2PO4 in
445 mL of HPLC grade water. To this said solution 55 mL of 0.1M H3PO4 was added to
adjust the pH 3.3.
Preparation of mobile phase:
The above stated prepared phosphate buffer (pH 3.3) acetonitrile were mixed
completely in the proportion of 50: 50 v/v and was filtered through 0.45 µm nylon
membrane filter and degassed by sonication.
Preparation of standard stock solution:
Stock standard solutions containing (1.25, 0.4, 0.5, 0.4, 0.8 mg/mL) of
Hydrochlorothiazide, Telmisartan, Losartan, Olmesartan and Valsartan respectively
were prepared by dissolving (12.5, 40, 50, 40, 80 mg) of each in methanol in 100 mL
volumetric flask respectively. It was then sonicated for 15 minutes and the final volume
of solutions was made up to 100 mL with methanol to get stock standard solutions.Vignan Pharmacy College,Vadlamudi.
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Preparation of sample solution: A composite of ten Telmisat-H, Zargo-H, Olmecip-H and Valent-H
tablets are prepared by grinding them to a fine, uniform size powder,
triturated using mortar and pestle. After calculating the average tablet
weight, amounts of powder equivalent to HCTZ, TELM, LOSA, OLME
and VALS respectively are prepared by dissolving (12.5, 40, 50, 40, 80
mg) respectively of each type of tablets were accurately weighed and
transferred separately to 100 mL volumetric flasks respectively.
Solutions were sonicated for 15 min and the solutions were then filtered
through 0.45 µm nylon membrane filters. Aliquots of appropriate volume
(10 mL) were transferred to 100 mL calibrated flasks and diluted to
volume with mobile phase to furnish the mentioned concentration above.
Vignan Pharmacy College,Vadlamudi.
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RESULTS:
Method Development:
Selection of a common detection wavelength:
The ultra violet spectrum of four Sartans and hydrochlorothiazide were
scanned individually in the region between 200 - 400 nm. The UV
overlain spectra of these five drugs showed that they absorbed at 238
nm. So this wavelength was selected for the determination of sartans.
Figure 1 shows UV overlain spectra of four sartans with HCTZ.
Vignan Pharmacy College,Vadlamudi.
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Figure 1. UV overlain spectra of four sartans and HCTZ.
Vignan Pharmacy College,Vadlamudi. 25
Method optimization: Numerous trials were performed to obtain the better separations. From all trials, eventually better reproducibility of the results and good
resolution, good peak shape, short runtime, minimal peak tailing were well identified when mobile phase consisting of phosphate buffer mixture properly adjusted to pH 3.3, acetonitrile in the proportion of 50:50 v/v with satisfactory results.
Table 4. Optimized chromatographic conditions
Parameter Chromatographic conditions for 4 sartans & HydrochlorothiazideInstrument Shimadzu LC-20AT Prominence liquid chromatographColumn Welchrom C18 column (4.6 X 250 mm, 5 µm)