RP-HPLC Method Development and Validation for The Estimation of Citalopram in Bulk and Pharmaceutical Dosage Form

Citalopram is a medication used to treat depression. Citalopram hydrobromide increases serotonergic activity in the central nervous system to produce its antidepressant effects. Citalopram hydrobromide has been shown in numerous trials to be a selective serotonin reuptake inhibitor (SSRI) with negligible effects on dopamine and norepinephrine neuronal reuptake. ^[1-3] Citalopram is chemically (RS)-1- [3-(Di methylamino) propyl]-1-(4-fluorophenyl)-1,3-dihydroisobenzofuran-5-carbonitrile.

The chemical structure of Citalopram is ^[4,5]

R)-(−)-citalopram (top),
(S)-(+)-citalopram (bottom)

Figure 1: Chemical Structure of Citalopram

MATEIALS AND METHODS

Chemicals and Reagents

Citalopram, the working standard medication, was acquired from MSN Laboratories Pvt Ltd in Hyderabad, Telangana. The local pharmacy provided the formulation dose form citalopram 80. We bought HPLC-grade acetonitrile, water, and methanol from Merk Chemicals Private Limited in Mumbai. The AR Grade buffer solutions used in the study were acquired from Merck Specialities Private Limited in Mumbai, India.

Preparation of standard drug solution

20 mg of the bulk medication Citalopram should be precisely weighed before being put into a 20 ml volumetric flask. To ensure that the medicine is completely dissolved in the diluent, add 10 ml of it and sonicate for 15 minutes. Now add diluent to bring the volume up to 20 ml. The solution has a 1000µg/ml concentration. To obtain a 100µg/ml solution, take 1 ml of this solution and dilute it to 10 ml. A working standard solution of 20µg/ml is obtained by diluting 1 ml of this solution to 10 ml.

Preparation of formulation solution

A clean, dry mortar and pestle were used to grind 10 Citopam 20 tablets, each containing 20 mg of citalopram, to a fine, consistent powder. An airtight 20 mL volumetric flask was filled with a precisely weighed tablet powder containing 20 mg of citalopram. 10 mL of diluent was used to dissolve the tablet powder, resulting in a final volume of 20 ml. In the described method, formulation analysis was conducted using the same solution. Take 1 ml of this solution and dilute it in a 10 ml volumetric flask to make 10 ml. Take 1 ml of this solution and dilute it in a 10 ml volumetric flask to make 10 ml. The formulation analysis is conducted using this solution.

METHOD DEVELOPMENT

Selection of Wavelength

A standard solution of 10 μg/mL was made and scanned in a UV detector between 200 and 400 nm in order to choose an appropriate wavelength. The maximum wavelength that was acquired was chosen as the ideal wavelength for the detection.

Optimized Chromatographic Conditions

METHOD VALIDATION

In accordance with the ICH requirements, the method was validated in terms of linearity, accuracy, precision, robustness, LOD & LOQ, and system appropriateness. Replica injections of the sample and standard solutions into the column were used to conduct validation investigations. ^[6]

RESULTS AND DISCUSSION

Method Development

Selection of Wavelength

Figure 2: UV Spectra of Citalopram

Table 1: Results for Optimized Chromatogram

Figure 3: Optimized Chromatogram of Citalopram

METHOD VALIDATION

System suitability

Figure 4: Chromatogram of Blank

LOD & LOQ

Citalopram's LOD was found to be 1.213µg/mL, while its LOQ was determined to be 3.129µg/mL. The method's sensitivity and potential utility for drug detection and analysis at extremely low concentrations were validated by the results.

Linearity

The concentration range of 10–35 µg/mL showed linearity, with a correlation coefficient of 0.9998.

Table 2: Results for Linearity

Figure 5: Linearity graph for Citalopram

Precision

Citalopram's percentage RSD was determined to be 0.42 for intraday precision and 0.45 for interday precision. For both intraday and interday precision, the percentage RSD was found to be within the acceptable level of less than 2. Thus, it was discovered that the developed procedure was accurate.

Table 3: Results for Intraday Precision

Table 4: Results for Interday Precision

The recovery percentage was found to be between 98.10 and 98.95%. At 50%, 100%, and 150% spiking levels, the percentage RSD was found to be within the permissible range for Citalopram. The results showed that the suggested approach was accurate, with a % RSD of less than two and an acceptability limit of 98-102.

Table 5: Accuracy results for Citalopram

The percentage RSD used to express ruggedness had to be less than 2. Citalopram's percentage RSD in the developed technique was 0.45. The method's robustness is confirmed by results that fall within the acceptable range.

Robustness

The percentage change for Citalopram in the developed technique was found to be within the permissible level of less than 2. As a consequence, the suggested approach was determined to be appropriate for the analysis of Citalopram when there was a slight alteration in the analytical conditions. This confirms that a slight change in the analytical conditions has no effect on the results.

Assay

Citalopram's assay percentage in formulation analysis was 98.95%. As a result, the technique was determined to be appropriate for the regular examination of Citalopram in both formulations and bulk drug.

Figure 6: Chromatogram of Formulation

Table 6: Results for Formulation