Novel Validated UV Spectroscopic Method for the Analysis of Ramipril and Olmesartan medoxomil in Drug Substance as Fixed Dosage Form

 

Navin Kumar J¹, K. Sonia²*, K. Manikandan³

¹Department of Pharmaceutical Chemistry, Sri Ramachandra Faculty of  Pharmacy,

SRIHER (DU), Porur, Chennai-600116. Tamil Nadu, India.

²Department of Pharmaceutical Analysis, SRM College of Pharmacy,

SRM Institute of Science and Technology, Kattankulathur, Chengalpattu-603203, Tamil Nadu, India.

 

ABSTRACT:

The current study focuses on the development and analytical validation of a unique, precise, and accurate UV-Visible Spectrophotometric method for estimating Ramipril and Olmesartan medoxomil simultaneously. The regression strength of Ramipril and Olmesartan medoxomil over its absorbances were obtained as y=0.0378x-0.05347 and y=0.0397x-0.0042 respectively with a correlation coefficient (r²) of 0.9917 for Ramipril and 0.9999 for Olmesartan medoxomil. The intra-day precision in addition inter-day precision for Ramipril and its % RSD were obtained as 1.46% and 1.40% respectively. The intra-day precision in addition inter-day precision for Olmesartan medoxomil and % RSD were obtained as 2.28% and 1.46% respectively.

 

 

 

INTRODUCTION: 

New chemical entities (NCE) are discovered or synthesized for pharmaceutical applications every day in today’s world. NCE cannot be used as a human drug or medicine. A few NCE which are approved by concerned regulatory authorities do not automatically become inclusive in Pharmacopoeias. Regulatory approval need not automatically grant permission to be included in the pharmacopoeia. They are manufactured and are used. This necessitates the development of analytical methods using the latest available equipment^1-5. A formulation of two or more active components together in a single dosage form, in specific quantities, is known as a combination of several pharmaceuticals or a fixed-dose combination. By reducing the number of doses required, the fixed-dose combination of medications aids in therapy and improves compliance. Fixed-dose medication combinations are used to treat a variety of cardiovascular disorders in a variety of dosage forms, including tablets, capsules, modified release formulations,

 

controlled-release formulations, and prolonged-release combinations^6-9. Ramipril chemically is 1-[(2S,3aS,6aS) -1-[(2S)-2-[[(1S)-1-(Ethoxycarbonyl)-3-phenylpropyl] amino] [1-oxopropyl] octahydrocyclopenta [b] pyrrole-2-carboxylate] |A-D Glucopyranuronic Acid. It's an antihypertensive prodrug and a nonsulfhydryl angiotensin-converting enzyme(ACE) inhibitor. Ramipril is converted to its active form in the liver by de-esterification, which inhibits ACE and hence prevents the conversion of angiotensin I to angiotensin II. Angiotensin II's powerful vasoconstrictive effects are blocked, resulting in vasodilation. This drug also promotes diuresis and natriuresis by increasing bradykinin levels and decreasing angiotensin II-induced aldosterone production by the adrenal cortex. Ramipril has the appearance of a solid powder. Its melting point is 109°C and the refractive index is 1.731. It is soluble^10-15. The structure is given in Fig.-1.

 

 

Fig. 1: Structure of Ramipril

Olmesartan Medoxomil Chemically, it is 5-Methyl-2-oxo-1,3-dioxol-4-yl) methyl 4-(2-hydroxy-2-propanyl)2-propyl-1- [2'-(1H-tetrazol-5-yl)-4-biphenylyl] methyl -1H-imidazole-5-carboxylate. Olmesartan Medoxomil is an antihypertensive prodrug made from a synthetic imidazole derivative. Olmesartan medoxomil is transformed into olmesartan after hydrolysis. By attaching to the AT1 receptor of angiotensin II in the smooth muscle of the vascular smooth muscle and the adrenal gland, olmesartan competes with angiotensin II binding to the receptor. This reduces aldosterone production and prevents angiotensin II-induced vasoconstriction, decreasing sodium retention and potassium excretion caused by aldosterone. The appearance of Olmesartan Medoxomil is a solid powder. Its melting point is 109°C and the refractive index is 1.731 and it is soluble^16-18. The structure is given in Fig. 2.

 

Fig. 2: Structure of Olmesartan Medoxomil

 

EXPERIMENITAL:

Materials and Reagents:

Ramipril, Amilodipine besylate, Olmesartan medoxomil and Valsartan were Active pharmaceutical ingredient were procured from Yarrow chemicals, Mumbai, India. Were procured from the local pharmacy. Solvents like Acetonitrile, Methyl alcohol, Acetic acid, Ammonium acetate, Water (Milli Q grade) were obtained from Distilled water are used for the experiment.

 

Equipment used:

S. No.	Name of the equipment	Company
1.	UV 3092 UV/Visible spectrophotometer with 1 cm matched Quartz cells.	LABINDIA
3.	Electronic Balance	Shimadzu BL-220H, Japan
4.	LI 120 PH Meter	Elico India.
5.	R8c Laboratory Centrifuge	Remi motors Ltd, India
6.	Vortex Mixer	Remi motors Ltd, India.
7.	Ultra-Sonicator	ILE

 

Method Validation ^19-22:

Standard stock solution Preparation:

Meticulously weighed and transferred into 100-milliliter standard flasks of 100milligram of each drug Olmesartan medoxomil and Ramipril in two separate standard flasks and dissolved in methyl alcohol to get the strength of 1milligram/milliliter of each. From the stock, 1milliliter is pipette out and reduced the strength of working standard solution is 10microgram per milliliter of both drugs with methyl alcohol.

 

λ max determination:

Both Olmesartan medoxomil and Ramipril standard concentrations were prepared of dilutions 10microgram per milliliter both the solutions were scanned in UV range against blank solvent. The wavelength spectra of Olmesartan medoxomil and Ramipril in methyl alcohol are shown. The representative of each spectrum revealed that Olmesartan medoxomil shows a λ max at 230 nanometer and Ramipril shows at 210nanometer.

 

Preparation of calibration curve:

Olmesartan medoxomil and Ramipril working standard solutions were prepared of dilution 100µg/milliliter. Different dilution was taken from the standard stock solution and diluted with methyl alcohol in the concentration of 2microgram/milliliter to 12microgram/ milliliter solutions at 2microgram/milliliter interval. The working solutions of Olmesartan medoxomil and Ramipril were prepared and scanned at 230nanometer and 210nanometer respectively. The absorbances were recorded and are outlined against the strength to obtain the respective calibration curves (Fig. 3 and Fig. 4).

 

Sample preparation:

Twenty tablets weight were taken and finely grinded in a mortar. The equivalent amount of 20milligram of Olmesartan medoxomil and 5milligram of Ramipril was meticulously weighed and relocated into a 100milliliter tidy volumetric flask in which 70milliliter methyl alcohol and sonicated for 5min in which drug go into solution completely. It is made up to the mark with methyl alcohol to get a stock concentration of Olmesartan medoxomil another Ramipril. The solution was separated by using Whatman filter paper, discarding the first few milliliters. Further dilutions were made to get a strength of 5 microgram/milliliter of Ramipril and 20 microgram/milliliter of Olmesartan medoxomil.

 

Linearity:

For the estimation of Olmesartan medoxomil and Ramipril lambda max were found to be 260 nanometer and for Ramipril was found to be 210 nanometers in a methyl alcohol solvent. The linearity for both Olmesartan medoxomil and Ramipril in the strength range of 5-30 microgram/milliliter. (Table 1 and Figures 5 and 6).

 

Precision:

For the intra-day calculation of precision 0-10 hours with the interval of every two hours and interday precision 1-6 days were chosen and readings were taken for every day for Olmesartan medoxomil and Ramipril tabulated in table 2.

 

Stability parameter:

The precise amount of tablet formulation which is equal to 20milligram of Olmesartan medoxomil and 5milligram of Ramipril was transferred into a 100-milliliter standard flask and maintained under the subsequent conditions which hold Alkaline 0.1 N NaoH, Acidic 0.1 N HCL reflux for 3 hours, 3% Oxydol at 50ºC, heat (60ºC), humidity (75 percentage Relative humidity) for 24 hour and after the particular time quantity was diluted to the mark with distilled water, separated using Filter paper. From this stock solution, a 5-milliliter portion of the filtrate was pipetted out and further diluted with distilled water in a 100-milliliter standard flask (10 micrograms/milliliter). The standard stock solution of two drugs was prepared and compared against a label claim and results were tabulated in table 3.

 

Limit of detection and Limit of quantification:

The limit of detection and limit of quantification for Olmesartan medoxomil were verified to be 0.15 microgram/milliliter and 0.32 microgram/milliliter approximately. The LOD and LOQ for Ramipril was examined to be 0.35 microgram/milliliter and 0.95 microgram/milliliter (Table 4) respectively.

 

Accuracy:

 Accuracy was determined for drugs by spiking with 80, 100 and 120 percentage of pure drug and the mean recovery of the Olmesartan medoxomil and Ramipril were to be 99.19% and 99.2% respectively (Table 5).

 

Assay:

The assay of Olmesartan medoxomil and Ramipril was done and its percentage purity was found to be 99.7% and 100.02% respectively.

 

RESULTS AND DISCUSSION:

The extent of Ramipril another Olmesaratan medoxomil bulk samples and their tablet forms were determined by simultaneous equation method by using UV Spectrophotometer. The regression strength of Ramipril and Olmesaratan medoxomil over its absorbances were obtained as y=0.0378x-0.05347 and y=0.0397x-0.0042 respectively with a correlation coefficient (r2) of 0.9917 for Ramipril and 0.9999 for Olmesaratan medoxomil. The intra-day precision in addition to inter-day precision for Ramipril its % RSD was found to be 1.46% and 1.40 % respectively. The intra-day precision in addition to inter-day precision for Olmesaratan medoxomil and % RSD was obtained as2.28% and 1.46% respectively. This confirms the procedure is precise. Accuracy is determined for both drugs by spiking with 80, 100 and 120% of additional pure drug and the % mean recovery of the Ramipril and Olmesaratan medoxomil were obtained as 99.2 and 99.19 respectively (Table 5). The percentage purity for the assay of Ramipril and Olmesaratan medoxomil were obtained as100.02% and 99.7% respectively (Table 6). The assay result shows that the methodology was selective for the evaluation of Ramipril and Olmesaratan medoxomil without hindering the inactive substance used in the tablet dosage form.

 

Figure 3: Chromatogram of Olmesartan medoxomil

 

Figure 4: Chromatogram of Ramipril

 

Figure 5: Linearity graph of Ramipril

 

Figure 6: Linearity graph of Olmesartan medoxomil

 

Table 1: Linearity data for Ramipril and Olmesartan medoxomil

 

Table 2: Intra-day and Inter-day precision for Olmesartan medoxomil and Ramipril

Intra-day precision				Inter-day precision
S. No.	Time (Hours)	Olmesartan Absorbance	Ramipril Absorbance	Time (Days)	Olmesartan Absorbance	Ramipril Absorbance
1	0	0.3133	0.5045	1	0.3754	0.3745
2	2	0.3162	0.5142	2	0.3504	0.3643
3	4	0.3019	0.5240	3	0.3802	0.3744
4	6	0.3102	0.5045	4	0.3501	0.3646
5	8	0.3202	0.5085	5	0.3601	0.3746
6	10	0.3213	0.5146	6	0.3802	0.3747
Mean		0.31385	0.51171	Mean	0.3660	0.37118
SD		0.007180	0.00748	SD	0.0143	0.005217
%RSD		2.2877	1.4617	%RSD	3.9071	1.4055

 

Table 3: Stability studies for Olmesartan medoxomil and Ramipril

 

Table 4: LOD and LOQ for Olmesartan medoxomil and Ramipril

 

Table 5: Recovery studies for Olmesartan medoxomil and Ramipril

 

Table 6: Assay of Telmisartan and Ramipril formulations

 

CONCLUSION:

There were many procedures arrived for the determination of double dose combination there is no any stability indicating, simultaneous resolution, so the recent work is useful for stability indicating, simultaneous determination of Telmisartan another Ramipril in the dosage regimen. This procedure can be unique gives higher and speedy analysis throughput. Stress degradation was carried out by different conditions such as acidic, alkaline, oxidative, hydrolytic, thermal and photolytic for specificity. The results obtained from this analytical method are reliable, accurate in assessing the ability of the component and robust as per the ICH guideline.

 

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Accepted on 09.04.2022           © RJPT All right reserved

Research J. Pharm. and Tech 2023; 16(3):1442-1446.