Development And Validation Of RP-HPLC Method For Determination Of Fimasartan In Bulk And Their Dosage Form – A Review

An example of a typical antihypertensive drug is fimasartan. When renin is released from the kidney, the kidney's rennin-angiotensin cascade begins, breaking down angiotensinogen into angiotensin I. This is how fimasartan acts on the kidney. Vasoconstriction is reduced and vasodilation is encouraged when fimasartan binds to the AT1 receptor. 2-(2-Butyl-4-methyl-6-oxo-1-{[2'-(1H-tetrazol-5-yl)-4-biphenylyl] methyl}-1,6-dihydro-5-pyrimidinyl)-N, N-dimethylethanethioamide is chemical name of fimasartan as shown in fig. 1.1

Few methods, such as ultraviolet (UV), high-performance liquid chromatography (HPLC), and LC-MS, appear to be available for biological fluids and pharmaceutical dosage forms, according to a study of the literature [1–16]. Using a UV detector at 265 nm, the current method offers precise information about the development and validation of Fimasartan in pharmaceutical dosage form and bulk. It is easy to use, precise, quick, repeatable, and sensitive, meeting acceptance criteria based on ICH and FDA guidelines.

METHODS:

Fimasartan Potassium Trihydrate was obtained as sample from pharmaceutical company in Maharashtra, India. The pharmaceutical tablet formulation is obtained from the pharmaceutical firm. HPLC grade water, orthophosphoric acid, and acetonitrile of HPLC grade were used. Analytical grade chemical reagents were the other chemicals used.

Selecting a wavelength

Fimasartan was dissolved in an appropriate solvent (acetonitrile) to determine the wavelength, and the resulting solution was scanned in the ultraviolet (UV) area, or between 200 and 400 nm. According to Fimasartan, the greatest absorption at 265 nm was chosen, as seen in Fig. 2.

Conditions for chromatography

An HPLC from Shimadzu was used to examine the entire process. Acetonitrile:0.1% OPA (80:20, v/v) at a flow rate of 0.8 ml/min was used as the mobile phase and a Primacel C18 (150 mm × 4.6 mm i.d., 5 ?m) column as the stationary phase. A rheodyne injector with a 25 ?l loop was used to inject the samples. Finally, the sample was analysed at 265 nm after the mobile phase had been sonicated, filtered, and degassed. Acetonitrile and 0.1% OPA were used in the mobile phase at an 80:20 v/v ratio to prepare it for the current experiment. This was accomplished by dissolving in a reservoir 20 millilitres of 0.1% OPA and 80 millilitres of acetonitrile. 0.1% orthophosphoric acid was produced by carefully measuring 0.1 ml of OPA and dissolving it in 100 ml of HPLC-grade water. The mobile phase that was prepared earlier is sonicated and degassed.

Preparation of the standard solution

100 mg of fimasartan were precisely weighed and then transferred into a 100 ml volumetric flask to create the standard solution. Add a few milliliters of acetonitrile to this to fully dissolve the medication. Lastly, add acetonitrile (1000 ?g/ml) to the solution to make it up to 100 ml.

Preparation of a sample solution

The tablet powder to 100 mg of fimasartan was carefully weighed, then put into a 100 ml volumetric flask, with the volume being raised to 100 ml using acetonitrile. Following around fifteen minutes of sonication, pour the mixture into a second 100 ml volumetric flask (1000 ?g/ml) and strain it using Whatman filter paper (No. 1). The solution was further diluted with the diluent until it reached the working concentration range of the calibration curve.

Curve of calibration

A 100 milliliter volumetric flask was filled with ten milliliters of the standard solution. The remaining volume was increased to 100 milliliters by adding 100 ?g/ml of acetonitrile. From this solution, several dilutions with concentrations ranging from 5 ?g/ml to 30 ?g/ml were made. Following three more injections of these concentrations into the system, the calibration curve was made by plotting the concentration on the X-axis and the peak area on the Y-axis.

How the commercial formulation is assayed

Fimasartan, a film-coated tablet containing a similar amount of potassium trihydrate, is sold. Locally, fimasartan is available under the trade name FIMANTA (120 mg, Ajanta Pharma Limited, India). A sample solution was prepared and diluted to 10 ?g/ml, and then 1000 ?g/ml was injected into the HPLC apparatus. After determining the percentage assay, the outcomes are shown in Table 1.

END RESULTS AND TALK

Specificity

The system was injected with the blank after the formulation excipients were prepared. The specificity of the developed procedure is shown by an absence of peaks in the retention period that match the analyte peak, suggesting that the excipients in the formulation are not interfering. A blank and standard chromatogram are displayed in Fig. 3

A 100 ml volumetric flask was filled with ten ml of the standard solution. The remaining volume was increased to 100 ml by adding 100 ?g/ml of acetonitrile. From this solution, several dilutions with concentrations ranging from 5 ?g/ml to 30 ?g/ml were made. These concentrations were injected into the system three more times, and the calibration curve was made by graphing the concentration of fimasartan on the X-axis and the mean peak area on the Y-axis. The method has strong linearity (r2 = 0.9995). The linearity graph and table were supplied by Table 2 and Fig.

Determination limit (LOD)

This represents the lowest detectable analyte concentration. The formula below can be used to compute this:

LOD = 3.3 × SD/slope.

It was calculated and determined to 1.3 ?g/ml.

Quantification limit (LOQ)

This represents the lowest quantifiable analyte concentration. The formula below can be used to compute this:

LOD = 10 × SD/slope.

It was calculated and determined to be 4 ?g/ml.

 Accuracy

There is a consensus between the measured and true values. Three injections of 50%, 100%, and 150% levels into the HPLC apparatus are required to achieve accuracy. As a conclusion, Table 3 provides a clear representation of the computed amounts found, added, recovered percentage, mean recovery, percentage SD, and percentage RSD, all of which indicate good agreement between the findings.

Repeatability

System Precision

The procedure is as follows: an HPLC system is filled with 10 ?g/ml of standard solution six times; the peak regions are noted, and the average, standard deviation, and percent RSD are computed, as indicated in Table 4 and Figure 5.

Method precision

This involves injecting 10 ?g/ml of sample solution six times into an HPLC machine, noting peak regions, and calculating % assay, average, SD, and %RSD, as shown in Table 5 and Fig. 6.

Intermediate Precision/Ruggedness

It is conducted in two different laboratories under two different analysts' conditions on two different days, as shown in Fig. 7. The two laboratories are kept similar.

Analyst-1:

Inject the sample six times into the HPLC system at a concentration of 100% (10 ?g/ml). Take note of the area after every injection and compute the SD and % RSD values, as indicated in Table 6.

Analyst 2:

Inject the sample six times into the HPLC system at a concentration of 100% (10 ?g/ml). Take note of the area after every injection and compute the SD and % RSD values, as indicated in Table 6.

Day 1:

Inject 100% concentration of sample (i.e., 10 ?g/ml) into HPLC system 6 times. Note the area for each injection and calculate the SD and% RSD values, as given in Table 6.  On Day 2, inject the sample at 100% concentration (10 ?g/ml) into the HPLC system six times. Record the area for each injection and calculate the SD and percentage RSD values, as shown in Table 6.

Reproducibility

This is generally considered as the laboratories' precision. In two different laboratories, inject 100% of the sample (10 ?g/ml) six times into separate HPLC systems to carry out the test. As indicated in Table 7 and Figure 8, note the area for each injection and calculate the SD and% RSD values.

A purposeful or accidental change of a parameter shouldn't impact any method. In order to accomplish this, adjust the mobile phase (± 10%) and flow rate (± 10%).

Change in flow rate

The flow rate can be adjusted by ± 10% (0.7 ml/min and 0.9 ml/min) to achieve this. Six injections at 0.7 and 0.9 ml/min flow rates should be made with the 100% concentration (10 ?g/ml). Table 8 and Figures 9 and 10 illustrate this process. Take note of the peak location and compute the percentage assay, average, SD, and % RSD for both flow rates.

The organic phase percentage (88:12 v/v and 72:28 v/v) is adjusted by ± 10% to achieve this. Using both mobile phase proportions (88:12 v/v and 72:28 v/v), inject the 100% concentration (10 ?g/ml) six times. Table 9 and Figures 11 and 12 illustrate how to compute the % Assay, Average, SD, and % RSD for both mobile phase proportions while noting the peak area.

This is carried out for 0, 24, and 48 hours. Inject 100% concentration (10 ?g/ml) 6 times during 0, 24, and 48 hours. Table 10 and Figures 13–15 illustrate how to compute the percentage assay, average, SD, and %RSD for both mobile phase proportions while noting the peak area.  

Force Degradation Studies

This includes acid and alkali degradation, oxidative deterioration, photolytic degradation, and thermal degradation, all of which are well illustrated in Table 11 and Figures 16-20.

Preparation of Solutions