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International Journal of Pharmaceutical InvestigationNOW IN
Article

Method Development and Validation of Olmesartan Medoxomil by Using RP-HPLC

Anchal Kushwaha1,2, Kandasamy Nagarajan3★, Richa Goel4, Garima Kapoor5, Snigdha Bhardwaj6, Pankaj Bhatt7★ Corresponding author

  1. 1Department of Pharmaceutical Quality Assurance, KIET School of Pharmacy, KIET Group of Institutions, Ghaziabad, Uttar Pradesh, INDIA.
  2. 2Dr. A.P.J. Abdul Kalam Technical University, Lucknow, Uttar Pradesh, INDIA.
  3. 3Department of Pharmaceutical Quality Assurance, KIET School of Pharmacy, KIET Group of Institutions, Ghaziabad, Uttar Pradesh, INDIA.
  4. 4Department of Pharmacognosy, KIET School of Pharmacy, KIET Group of Institutions, Ghaziabad, Uttar Pradesh, INDIA.
  5. 5Department of Pharmaceutical Chemistry, KIET School of Pharmacy, KIET Group of Institutions, Ghaziabad, Uttar Pradesh, INDIA.
  6. 6Department of Pharmaceutics, KIET School of Pharmacy, KIET Group of Institutions, Ghaziabad, Uttar Pradesh, INDIA.
  7. 7Department of Pharmaceutics, Lloyd Institute of Management and Technology, Greater Noida, Uttar Pradesh, INDIA.

CORRESPONDENCE

Kandasamy Nagarajan

Principal and Professor, Department of Pharmaceutical Quality Assurance, KIET School of Pharmacy, KIET Group of Institutions, Ghaziabad-201206, Uttar Pradesh, INDIA.

k.nagarajan@kiet.edu

Received: 27-03-2025; Revised: 19-05-2025; Accepted: 03-07-2025.

Volume 15, Issue 4 · pp. 1320–1331 · PUBLISHED Oct-Dec 2025 · DOI: 10.5530/ijpi.20251665

View on IJPI original site ↗

ABSTRACT

Background Olmesartan (OLM) is a drug used to treat hypertension, or elevated blood pressure. Controlling high blood pressure lowers the chance of further strokes, renal problems and heart attacks. Materials and Methods The article presents a comprehensive overview of the drug’s pharmacological characteristics and therapeutic effectiveness. A UV spectrophotometric technique was used to determine the solubility of Olmesartan medoxomil in a range of solvent ratios, including ethanol, methanol, acetonitrile, chloroform, pyridine, acetone, ethyl ether and distilled water. This was carried out since parameter analysis, drug validation and HPLC technique development all heavily depend on the solubility of the drug. The dissolution rates of several compounds were measured in a number of solvents, including distilled water, ethanol, methanol, acetonitrile, acetone, chloroform and pyridine. For every solvent, the UV spectra were measured between 200 and 400 nm. The λmax values for several solvents were analyzed and recorded. Results and Discussion The study revealed that Olmesartan medoxomil is very soluble in methanol and ethanol. A wavelength of 256 nm for methanol and an absorbance of 0.291 were used. The C18 (4.6×250 mm, 5 µ) particle size column in our approach was filled with water (containing 5% ammonium acetate) and methanol (5%:95%) at room temperature and 1.0/min flow rate using 10-min runtime. Olmesartan medoxomil eluted with a retention time of exactly 2.3 min. Conclusion Using 10 min runtime, Olmesartan medoxomil eluted with a retention time of exactly 2.3 min.

KEYWORDS

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FULL TEXT

INTRODUCTION

Olmesartan is used to treat hypertension, or elevated blood pressure. Reducing hypertension can help avert heart attacks, strokes and renal issues. One drug that belongs to the Angiotensin Receptor Blocker (ARB) class is olmesartan. In order to facilitate easier blood flow, it relaxes blood vessels. The molecular formula for olmesartan medoxomil is 2, 3-dihydroxy-2-butenyl 4-(1-hydroxy-1-methylethyl).2-propyl-1-[benzyl p-(o-1H-tetrazol-5-ylphenyl)] imidazole cyclic 2, 3-carbonate, 5-carboxylate. Chemical structure of Olmesartan medoxomil is shown in Figure 1.

Figure 1: Structure of Olmesartan medoxomil.

Drug Profile

Angiotensin II receptor antagonist olmesartan medoxomil is mostly used to treat hypertension, or high blood pressure. The following is a comprehensive medication profile that includes important details about olmesartan medoxomil:

Olmesartan medoxomil belongs to the class of medications known as Angiotensin II Receptor Blockers (ARBs) or Angiotensin Receptor Blockers (ARBs). It works by blocking the action of angiotensin II, a hormone that causes blood vessels to narrow. By inhibiting angiotensin II, olmesartan medoxomil relaxes blood vessels, thereby reducing blood pressure (Arora et al., 2020; Gopani et al., 2017; Ghante et al., 2019; Celebier et al., 2014). The principal indication for Olmesartan medoxomil is the treatment of hypertension (high blood pressure) in adults and children six years of age and older (Dhole et al., 2013). The recommended starting dose for most adults is usually 20 mg once daily. The dosage can be increased to 40 mg based on the individual patient's response (Beg et al., 2016). For paediatric patients (6-17 years old), the starting dose is weight-dependent (Jain et al., 2011). It is typically administered orally in the form of tablets. Olmesartan medoxomil is a prodrug and once ingested, it is rapidly converted to its active form, olmesartan, in the gastrointestinal tract (Thomas et al., 2022). The peak plasma concentration is reached about 1-2 hr after oral administration (Biagini et al., 1998). Olmesartan is primarily eliminated via the hepatic (liver) route. It is excreted in faeces (about 35%) and urine (about 60%) (Rump et al., 2004). Common side effects may include dizziness, light headedness, diarrhoea, or flu-like symptoms. Serious side effects are rare but may include kidney problems, allergic reactions and low blood pressure (Fogari et al., 2010). It is important to monitor blood pressure regularly. Patients with impaired liver function may require dose adjustments. Caution is advised in patients with renal impairment (Wabkhede et al., 2007). Olmesartan medoxomil may interact with certain medications, including potassium-sparing diuretics, Nonsteroidal Anti-Inflammatory Drugs (NSAIDs) and lithium (Brunner, 2002). Olmesartan medoxomil is contraindicated in pregnancy (especially in the second and third trimesters) due to the risk of harm to the foetus and its use should be avoided in breastfeeding women (Laeis et al., 2001).

Importantance of Analytical Method and its Validation

The process of confirming that the method works as intended is called validation. Validation requires a series of experiments to check the performance of the method. The method should be able to produce reliable and accurate results after it is developed. The only way to assess the validity of a method is to perform laboratory tests. Validation experiments involve measuring different aspects of the method to evaluate its suitability. The key parameters of Validation are as follows:

Specificity

To make sure the analytical technique can measure the analyte properly in the presence of other factors that might interfere with the analysis, specificity tests are carried out. For example, excipients should not affect the analytical process when it comes to specificity investigations (Srikanth et al., 2023).

Linearity

The concept of linearity of analytical method refers to a methods ability to show a linear relationship between observed responses and various concentrations (Sandhu et al., 2016). To investigate linearity, a response vs concentration graph is employed. Regression analysis is one of the statistical approaches that must be utilized to establish whether the analytical process is linear or not (Dhole et al., 2012).

Precision

Precision investigations assess how closely the observed values match up at the same concentrations and under the same circumstances (Dubey and Ghosh, 2015). The standard deviation and relative and standard deviation of the observed value are calculated for this research (Shah et al., 2022).

Limit of detection

The Limit of Detection (LOD) in analytical method is the smallest amount of the analyte that can be accurately and precisely detected under ideal circumstances by the proposed analytical method (Al-Masri and Al-Mardini, 2005).

Limit of quantitation

The Limit of Quantification (LOQ) is the lowest analyte concentration that can be quantitatively detected with a stated accuracy and precision (Amim, 2012).

Robustness

These parameters, which involve a little adjustment to the ideal chromatographic conditions such column temperature, flow rate and pH, are verified on various instruments and operators. In this method flow rate parameter was used (Mahaparale et al., 2013).

Ruggedness

Ruggedness test is determined between two different days and two different analysts and instruments (Bharti et al., 2022).

Accuracy

A threefold precision is employed: 80%, 100% and 120% of the intended concentration (Singh, 2015). A low % recovery and relative standard deviation number show how accurate the spectrophotometric and HPLC methods (McDowall, 2005).

MATERIALS AND METHODS

Instruments Condition

Ultraviolet-visible (UV-vis) spectrophotometer: Model UV-1800 (Shimadzu).

HPLC system

The Waters-HPLC (Model-2489) was equipped with a manual injector port with 10 µL fixed loop, a Waters 515 HPLC Pump and a Waters 2489 UV/visible detector. The Waters C-18 (4.6×250 mm, 5 µm) column was utilized and data were collected using the Empower-3 programming software.

Additional equipment used in the study included a sonicator (Remi Instrument Ltd., Mumbai, 10A) for sample dissolution, an analytical balance from Mettler-Toledo International Inc. Greifensee (Switzerland) for precise weighing and a highly precise hot air oven (Labman Scientific). Samples were injected into the HPLC system using a Hamilton micro syringe.

Chromatographic parameters

Column: Waters C-18 (4.6×250 mm, 5 µm).

Detector: Waters 2489 UV/visible detector.

Wavelength: 256 nm.

Flow Rate: 1.00 mL/min-1.

Mobile Phase: Ammonium acetate buffer: methanol (5%: 95%).

Reagent: (For API).

Olmesartan medoxomil (API) was obtained as generous gift sample. Acetonitrile (ACN), Methanol (MeOH), Ammonium acetate and water was used. All reagents were HPLC analytical grade and were procured from Thermo Fisher Scientific India Pvt. Ltd.

Reagent: (For Market Formulation)

The tablet of Olmesartan Medoxomil (OlmetimeTM Batch No. J5IAV005, Mfg. Date. 10/2022, Exp. Date. 09/2024) containing 40 mg Olmesartan Medoxomil, with Colour: Titanium Dioxide IP was taken for studies. They were procured from Medical Store.

Mobile phase selection and optimization (pilot scale-up study)

Optimization Method 1

Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. Acetonitrile (100%) was added and sonicated for better dissolution. The volume was made up to mark with diluent and final concentration was made and Acetonitrile (100%) was chosen as a mobile phase.

Optimization Method 2

Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. ACN: Methanol (90%:10%) was added and sonicated for better dissolution. The volume was made up to mark with diluent and final concentration was made and ACN: DW (70:30) was chosen as a mobile phase.

Optimization Method 3

Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. Methanol (100%) was added and sonicated for better dissolution. The volume was made up to mark with diluent and final concentration was made and Acetonitrile: Ammonium acetate buffer (95%:5%) was chosen as a mobile phase.

Optimization Method 4

Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. Methanol (100%) was added and sonicated for better dissolution. The volume was made up to mark with diluent and final concentration was made and Methanol: Distilled water (95%:5%) was chosen as a mobile phase.

Final optimization

Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. ACN: Methanol (9:1) was added and sonicated for better dissolution. The volume was made up to mark with diluent and final concentration was made and Ammonium acetate buffer: methanol (5%: 95%) was chosen. We check the peaks (chromatograms) and finalise the solvent mobile phase based on the peak in the chromatogram with its retention time.

Preparation of buffer

The buffer was prepared freshly; the accurately weighed 0.577 g ammonium acetate was transferred in 250 mL volumetric flask, followed by volume make-up to the mark with Methanol and ACN (90:10). After the preparation of 0.1N ammonium acetate, the filtration of the buffer solution was done by using A0.22 µ/mL pour size membrane filter.

Preparation of Standard solution

Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. ACN: Methanol (9:1) was added and sonicated for better dissolution. The volume was made up to mark with diluent and final concentration was made.

Preparation of Sample solution

To analyse the medicament in tablet, 1 tablet 0.2894 g was precisely weighed and the coarse powder was triturated in the mortar to get a fine powder. The equivalent of 0.2768 g tablet powder was weighed, transferred to a 100 mL volumetric flask and diluent 10% Acetonitrile and 90% Methanol was added to make up the mark and mixture was sonicated for 15 min. The tablet solution was diluted to a final concentration. Olmesartan medoxomil tablet shown in Figure 2 and crushed tablet shown in Figure 3.

Figure 2: Olmesartan medoxomil tablet.
Figure 3: Olmesartan medoxomil crushed tablet.

Analytical Method Validation of HPLC

The procedures provided above were validated in accordance with the ICH Q2 standard (R1). Robustness, analyte stability in solution, linearity, precision, limit of detection, limit of quantitation, accuracy/recovery, selectivity/specificity and ruggedness were among the many qualities that were examined for method validation. The ICH criteria were followed in terms of approach (ICH guideline Q2 (R1 and R2).

System Suitability

An evaluation of the systems suitability is necessary for best results of analytical procedures. In order to find out whether the system was suitable, six standard analyte solutions of pure drug Olmesartan medoxomil were injected at a concentration of 200 ppm. Several parameters were assessed, including the USP tailing factor, theoretical plate count, peak asymmetry factor and percentage Relative Standard Deviation (%RSD). Based on the results obtained, the suitability of the system was assessed. Six duplicate injections were used to acquire the %RSD of the region. A comparison was made between the recommendation and the observed outcomes.

Specificity/selectivity

An injectable sample must be able to distinguish the target analyte from any other drugs or adulterants in order to demonstrate specificity. By injecting a blank and a placebo, the HPLC technique was used to analyze these parameters.

Linearity

A fresh 200 ppm stock of pure drug Olmesartan medoxomil was used to dilute the compound to eight various concentrations using a 90%:20% mixture of methanol and acetonitrile. We used least squares regression to examine the collected data.

System precision

In order to assess the repeatability, six repeats were performed on the same day using three distinct Olmesartan medoxomil concentrations (2 ppm, 4 ppm, 8 ppm, 16 ppm, 32 ppm and 64 ppm) in a 9:1 combination of methanol and acetonitrile. We obtained an intermediate level of accuracy by measuring the same concentration with six separate samples on same day but different times. For three times, the dilution was determined to have remained stable in terms of accuracy. To ascertain the repeatability, three distinct Olmesartan medoxomil concentrations (24 ppm, 32 ppm and 40 ppm) were examined using an RP-HPLC. The concentration estimates derived from the standard curve were accompanied by the following statistics for each dataset: confidence interval, standard deviation and percentage Relative Standard Deviation (%RSD).

Accuracy

A threefold precision was employed: 80%, 100% and 120% of the intended concentration. A low % recovery and relative standard deviation number exhibited how accurate the spectrophotometric and HPLC methods are.

Robustness

The robustness of the approach validation was determined by seeing no significant difference between flow rates. The relative standard deviation as a percentage was less than 2, which is within the allowed range.

Ruggedness

The Ruggedness of the approach validation was determined by seeing no significant difference between analyst A and analyst B. The relative standard deviation as a percentage was less than 2, which is within the allowed range.

Limit of detection

In order to determine the limit of detection, the sample was diluted to a known concentration of 0.00001 ppm. The computed area percentage RSD was less than 2%, which is below the limit.

Limit of quantitation

In order to determine the limit of quantitation, the sample was diluted to a known concentration of 0.00003 ppm. The computed area percentage RSD was less than 2%, which is below the limit.

RESULTS

Optimization method

The mobile phase of the final solvent is chosen by taking into account the retention period of the peak in the chromatogram. The outcomes of the optimization procedure are shown in Figures 4-7. The final optimization method result for API is presented in Figure 8.

Figure 4: Optimization Method 1: Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. Acetonitrile (100%) was added and sonicated for better dissolution.
Figure 5: Optimization Method 2: Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. ACN: Methanol (90%:10%) was added and sonicated for better dissolution.
Figure 6: Optimization Method 3: Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. Methanol (100%) was added and sonicated for better dissolution.
Figure 7: Optimization Method 4: Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. Methanol (100%) was added and sonicated for better dissolution.
Figure 8: Final Optimization Method : Standard stock solution of 200 ppm, 20 mg of olmesartan medoxomil was prepared in a 100 mL volumetric flask. ACN: Methanol (9:1) was added and sonicated for better dissolution. The volume was made up to mark with diluent and final concentration was made and Ammonium acetate buffer: methanol (5%: 95%) was chosen.

System suitability

The system suitability is determined by the several parameters. Tailing factor, retention time and AUC were studied for the drug Olmesartan medoxomil. The observations obtained were analysed after administering six injections of 100 ppm. The tailing factor was under the limit, as less than 2 and retention time and AUC %RSD was less than 2%. Results of System suitability of API and market formulation are presented in Tables 1 and 2 respectively.

Table 1: Results of System suitability, Linearity, LOD, LOQ and Accuracy % Recovery for chosen Active Pharmaceutical Ingredients and (API) Olmetime-40 Market Formulation.
System Suitability
Sl. No.CONC. (PPM)Chosen Active Pharmaceutical Ingredients (API)Olmetime-40 (market formulation)
RTAUCHeightTailingPlate CountRTAUCHeightTailingPlate Count
11002.37648735437677521.6864952918.0249352.37351925548849861.8323494330.702933
21002.38548971967441661.4614833116.828222.39251169368159441.8923172427.847396
31002.38348044716903681.5478092224.2884152.35650900158068831.7114172577.520328
41002.38949501147863621.5019712808.2614512.38449963737836031.5745452737.726276
51002.38848632867307811.3866252973.385692.35251493187132731.7703581621.540456
61002.39247486856743491.3791832504.1597492.38450799077375551.7643711332.271637
SUM14.3132913729543937788.96356616544.9484614.24130625103474224410.54535715027.60903
MEAN2.38554856215.833732296.33331.4939276672757.491412.37355104183.8337903741.75755952504.601504
SD0.0056124970833.4181343449.429020.114702161332.43473260.016318766966.1009361086.921560.109166561053.978754
RSD0.235275041.4586134675.9333123827.67789252912.055694220.687537431.3119845047.7288627366.21125826442.08169451
Linearity
Sl. No.CONC. (PPM)Chosen Active Pharmaceutical Ingredients (API)Olmetime-40 (market formulation)
RTAUCHeightTailingPlate CountRTAUCHeightTailingPlate Count
122.373114435172031.7837763597.8691522.354112758145281.7653392167.812283
242.389212905291771.8448632576.4602492.353187852243551.7332752117.375404
382.401447689580181.5097852096.2780672.363411105563881.6344082630.673969
4162.4147764821190591.3607683183.1312462.3637506731024681.6553232542.207064
5322.41315170462096311.2538172222.8567212.35615554872025471.6632662320.672767
6642.3929821255063721.7253364736.8361512.35531157373962661.6940852128.391062
SUM14.3860506829394609.47834518413.4315914.144613361279655210.14569613907.13255
MEAN2.396666671008447156576.66671.5797241673068.9052642.357333331022268.667132758.66671.6909493332317.855425
SD0.015807171091281.78185444.66430.241738424997.91259210.004501851152187.269146141.28430.050135101222.1706393
RSD0.65954818108.2140935118.436972915.3025717632.516891410.19097221112.7088511110.08040982.9649085479.585181067
LOD
Sl. No.CONC. (PPM)Chosen Active Pharmaceutical Ingredients (API)Olmetime-40 (market formulation)
RTAUCHeightTailingPlate CountRTAUCHeightTailingPlate Count
10.000012.3413490850151.6886632507.0826672.3772470229461.9707141556.136891
20.000012.3482697037572.0338372526.8006432.3583590944751.9075631923.47954
30.000012.352113326851.849992293.93952.3582956235571.8122891384.493297
SUM7.03983011114575.572497327.822817.09390173109785.6905664864.109728
MEAN2.3463327670.3333338191.8574966672442.6076032.3643333330057.666673659.3333331.8968553331621.369909
SD0.004736914.1526111166.2366830.172709395129.12727580.010969665619.917823769.6196030.079753437275.3507809
RSD0.2014124.9876014430.537750279.2979652695.286451890.4639639818.6971193921.031688914.20450815816.98260091
LOQ
Sl. No.CONC. (PPM)Chosen Active Pharmaceutical Ingredients (API)Olmetime-40 (market formulation)
RTAUCHeightTailingPlate CountRTAUCHeightTailingPlate Count
10.000032.3525482074791.6505562590.9835812.3691435116831.6906911374.430763
20.000032.352286130741.9501212615.4318692.3551753220261.896505772.690756
30.000032.3491767722451.886782479.9229792.3591793717131.717447597.864041
SUM7.05195358127985.4874577686.3384297.0834982054225.3046432744.98556
MEAN2.350333178642661.8291523332562.112812.36116606.666671807.3333331.768214333914.9951867
SD0.0015320115.723482813.2431460.15787814472.220536520.00721111963.932365189.96403170.111905506407.3719206
RSD0.0649963.2848533265.945690258.6312190022.8187883160.3054257811.8261683910.51073586.32872971244.52175558
Accuracy% Recovery
Sl. No.% CONC (CONC. (PPM)Chosen Active Pharmaceutical Ingredients (API)Olmetime-40 (market formulation)
RTAUCHeightTailingPlate CountRTAUCHeightTailingPlate Count
80% (24)2.35213720971786401.5284562438.774952.37812685292181731.6563633815.99845
2.36213758211814751.3176942809.8226272.37912524592150061.7243294706.88849
2.3613621091835621.3724683012.2657442.37812396662098351.6752584028.9473
SUM7.07441100277.1353760654
MEAN2.35813700092.3781253551.333
SD0.0052915037090.4544280.0005773514462.47165
%RSD0.2244063880.5175480180.0242754141.153719937
1100% (32)2.36817411992480061.5959193371.7093122.34515784442393081.8705732496.08201
22.35417553632571851.4855113436.7415412.36815520292858102.0980046612.28417
32.38417502672328131.3538272590.3267792.37415796012853121.8889325646.38485
SUM7.10652468297.0874710074
MEAN2.36917489432.3621570024.667
SD0.0150111077174.221630.0153079515595.43768
%RSD0.6337365750.4102032850.648001270.993324373
1120% (40)2.37922042643024991.4759031114.6173672.36718681693724431.9711757825.28124
22.38322269102967201.368281896.4789262.3818432813591281.9502977719.7097
32.38321725052678221.3111812105.0087822.38118304813182911.7818653883.77944
SUM7.14566036797.1285541931
MEAN2.3822201226.3332.3761847310.333
SD0.00230940127329.408530.0078102519164.36749
%RSD0.0969657551.241553770.3287142121.037420034
Table 2: Results of Ruggedness of chosen Active Pharmaceutical Ingredients (API) for 32 ppm & Olmetime - 40 Market Formulation.
Chosen Active Pharmaceutical Ingredients (API)Olmetime-40 (market formulation)
Sl. No.CONC. (PPM)Interday 1Interday 2Interday 3Interday 1Interday 2Interday 3
Mean±SD%RSDMean±SD%RSDMean±SD%RSDMean±SD%RSDMean±SD%RSDMean±SD%RSD
12177332.5± 5335.1203.008174937±748.1180.427175670±393.1510.223174335±4147.8882.379178422±3054.7011.712176360.5±658.3160.373
24356996±2805.7990.785361581±2345.4730.648361825.5±4629.4281.279261441±2296.6820.878259440±4505.6841.736264873.5±89.8020.033
38549230.5±7128.34341.297539168.5±11986.1672.223538356±1035.2040.192518390.5±1750.0890.337517627±1673.0140.323500628±8198.1961.637
4161019060.5±2822.0630.2761027466.5±344.3610.033998769±15832.1201.585888573±5293.4010.595889726±13873.4351.559887560.5±12659.3321.426
5322032484.5±8313.4540.4092028810±1639.0730.081966491.5±4359.3130.2211669611±15225.4230.9111649432±12331.9420.7471677359±12310.7290.733
6644035070±66014.0741.6364044463.5±50937.8511.2594067503.5±91334.8612.2453416847.5±44077.5011.293450709±26763.9910.7753335549.5±24547.2110.735

Linearity

The concentration range of 2ppm to 64ppm was shown to be linear in the standard calibration curve. In calibration curve chart peak area was plotted on the X-axis and concentration on the Y-axis. After performing a linear regression analysis, the correlation coefficients (R2) value was found to be 0.9997. The equation for the calibration curve-based peak area and concentration ppm was y=45992x-42605. Linearity data of API and market formulation are shown in Table 1 and calibration curve chart is shown in Figure 3 and Figure 4 respectively. The market formulation (R2) value was found to be 0.9998. The equation for the calibration curve-based peak area and concentration ppm was y=48562x-2466.9.

Table 3: Results of robustness of chosen Active Pharmaceutical Ingredients (API) and Olmetime-40 Market Formulation.
Robustness Flow
Chosen Active Pharmaceutical Ingredients (API)Olmetime-40 (market formulation)
Sl. No.CONC. (PPM)RTAUCHeightTailingPlate CountFlow RateRTAUCHeightTailingPlate CountFlow Rate
1642.57933973854613481.6351492454.2306130.92.36118048512854671.620232229.7483270.9
2642.59433665714687961.547372560.2725272.36118015592828571.5552572191.542183
3642.58233630634869091.6177922970.4463112.3617943772815681.4344812491.739229
4SUM5.17667296349557053.1651625530.7188384.72135959365644252.9897384683.281412
5MEAN2.5883364817477852.51.5825812765.3594192.36051797968282212.51.4948692341.640706
6SD0.008492480.5312807.825130.049795874290.03666410.0007071075078.4409911.46064090.085401529212.2713669
7RSD0.327870.073722.68028843.14649763510.488208590.0299558050.28245450.3229696215.7129774329.065069905
8642.32531742745074851.6192163128.10208412.36817483552811881.528412720.2244911
9642.3331376485192191.624143646.3961212.36517915343045751.4856342516.825068
10642.33431148614809901.5613662826.6329172.3617894452808261.5061272693.688829
11SUM6.989942678315076944.8047229601.1311227.09353293348665894.5201717930.738388
12MEAN2.329673142261502564.66671.6015743200.3770412.3643333331776444.672888631.5067236672643.579463
13SD0.0045129973.919583.701650.034908078414.63318690.00404145224348.778413608.194920.021394241110.5714374
14RSD0.193560.95393.8967525872.17961066712.955760580.1709340991.37064664.7109511851.4199180364.182640959
15642.13427622144486331.6477812670.3053631.12.11816701192717441.5246611927.4395041.1
16642.1627252174503091.574822904.5091932.16916455762730061.5692522888.527126
17642.13727356794932301.5832163637.6278512.14916883362844431.5376832779.269798
18SUM6.431822311013921724.8058179212.4424076.43650040318291934.6315967595.236428
19MEAN2.143672741037464057.33331.6019393070.8141362.1453333331668010.33276397.66671.5438653332531.745476
20SD0.0142219071.525278.164580.039921672504.64958080.02569695221457.84846995.9775820.022929354526.1877724
21RSD0.663550.695785.4472072222.4920844316.433739021.1978069531.286433782.5311275841.48519129620.78359682

Limit of detection

In order to determine the limit of detection, the sample was diluted to a known concentration of 0.00001 ppm. The computed area percentage RSD is less than 2%, which is below the limit. The results of LOD of API and market formulation are presented in Table 1.

Limit of quantitation

In order to determine the limit of quantitation, the sample was diluted to a known concentration of 0.00003 ppm. The computed area percentage RSD is less than 2%, which is below the limit. The result of LOQ of API and market formulation are presented in Table 1.

Accuracy

To evaluate the techniques' dependability, we conducted recovery experiments. Standards are added to the sample solution at concentrations of 24 ppm, 32 ppm and 40 ppm to guarantee accuracy. Results of Accuracy of API and market formulation are presented in Table 1.

Precision

Precision determines random errors on reproducibility and repeatability. This method is quantified using the terms %RSD (Relative Standard Deviation). % RSD of less than 2% is acceptable. The precision investigations for the developed analytical techniques included assessing both intra-day and inter-day precision. To determine accuracy, experiments were conducted in terms of repeatability, interday and intraday precision. The results were found to have %RSD in acceptable limit, as not more than 2%. These results for API and market formulation are presented in Table 2.

Robustness

The robustness of the approach validation was determined by seeing no significant difference between flow rates. The relative standard deviation as a percentage is less than 2, which is within the allowed range. The results of robustness of API and market formulation are presented in Table 3.

Ruggedness

The Ruggedness of the approach validation was determined by seeing no significant difference between analyst A and analyst B. The relative standard deviation as a percentage is less than 2, which is within the allowed range. The ruggedness result of API and market formulation are presented in Tables 4 for 32 ppm and 64 ppm.

Table 4: Results of Ruggedness of chosen Active Pharmaceutical Ingredients (API) for 32 ppm.
Ruggedness 32PPM
Sl. No.CONC. (PPM)Chosen Active Pharmaceutical Ingredients (API)Olmetime-40 (market formulation)
RTAUCHeightTailingPlate CountRTAUCHeightTailingPlate Count
ana11322.37918086852590591.4322051163.3321462.36718196292814281.5611943287.591355
2322.40317998862414161.3868682047.2048682.36417705402729361.486172752.023114
3322.3818432472454031.3261692202.5261432.36917799032754471.4853822526.712694
Ana2 Ishu4322.3718158142718821.5175272716.313412.40618067442310221.3981021764.157253
5322.38218121452097181.3644831738.156272.3717946712682021.7352132964.642854
6322.36617937652603701.9058573713.5602492.39517744732633981.6752562639.359673
SUM14.281087354214878488.93310913581.0930914.2711074596015924339.34131715934.48694
MEAN2.381812257247974.66671.48885152263.5155142.37851790993.333265405.51.5568861672655.747824
SD0.01288409917209.2985421731.403280.214596576876.87757930.01751285219472.369617935.084050.127402094512.984152
RSD0.5413486860.9496058538.76355781614.4135647938.739631950.7362981891.0872385316.7576158188.18313483119.31599632
Ruggedness 64PPM
Sl. No.CONC. (PPM)Chosen Active Pharmaceutical Ingredients (API)Olmetime-40 (market formulation)
RTAUCHeightTailingPlate CountRTAUCHeightTailingPlate Count
ana11642.37931190434279361.3722961566.1842832.34828227474531271.6567673015.543632
2642.37631385524239931.4068691158.9496242.36127966994893841.5669194263.653623
3642.377317163931716391.3731772286.9969592.37229087524927931.543883863.286109
Ana2 Ishu4642.36231220374595291.3928971668.1752562.41528189924812061.7497864373.364064
5642.37331572694458931.3009782294.4106342.37929216015178521.7894514469.110563
6642.37231650294545841.4179032816.4656692.3728940104922291.7085493550.611427
SUM14.2391887356953835748.2641211791.1824314.24517162801292659110.01535223535.56942
MEAN2.3731673145594.83897262.33331.3773533331965.1970712.3741672860466.83487765.16671.6692253333922.594903
SD0.00604722368.9271114302.3960.041553016605.65057140.02267553671.727920956.470770.098816832562.6410048
RSD0.2548090.71111914124.18914233.01687407530.81882120.9550821.876327574.2964262725.91992164214.3435919

DISCUSSION

I have completed my work by following the ICH guidelines and all the results are mentioned above. The study revealed that Olmesartan medoxomil is very soluble in methanol and ethanol. A wavelength of 256 nm for methanol and an absorbance of 0.291 were used. The C18 (4.6×250 mm, 5 µ) particle size column in our approach was filled with water (containing 5% ammonium acetate) and methanol (5%:95%) at room temperature and 1.0/min flow rate using 10-min runtime.

CONCLUSION

The C18 (4.6×250 mm, 5 µ) particle size column in our approach was filled with water (containing 5% ammonium acetate) and methanol (5%:95%) at room temperature and 1.0mi/min flow rate. Using 10 min runtime, Olmesartan medoxomil eluted with a retention time of exactly 2.3 min. The developed approach was reliable and reproducible based on the ICH guidelines with all relevant parameter’s studies such as linearity, accuracy, robustness, ruggedness, limit of detection, limit of quantitation. Hence the same methodology of approach for analytical method development and validation may be adopted for Olmesartan formulation in near future as the retention time was lower (Rt=2.3 min) as compared with the previous reported literature having retention time of 7 min.

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Kushwaha, A., Nagarajan, K., Goel, R., Kapoor, G., Bhardwaj, S., & Bhatt, P. (2025). Method Development and Validation of Olmesartan Medoxomil by Using RP-HPLC. International Journal of Pharmaceutical Investigation, 15(4), 1320–1331. https://doi.org/10.5530/ijpi.20251665