Development of a UV Spectrophotometric Method and Validation of Paroxetine Hydrochloride

 

Ch. Bhavani¹, V. Padmabhushana Chary¹*, P.Mahitha¹, R. Mahesh¹, R. Chenchulaxmi¹, M. Chinnaeswaraiah²

¹Department of Pharmaceutical Analysis, Anurag Pharmacy College,

Kodad-508206, Suryapeta (Dist), Telangana, India.

²Department of Pharmacognosy, Anurag Pharmacy College, Kodad-508206, Suryapeta (Dist), Telangana, India.

 

ABSTRACT:

For the development and validation of the Paroxetine Hydrochloride method, a simple, reliable, cost-effective, and repeatable UV Spectrophotometric method was used. With a correlation value (R2 = 0.999), paroxetine hydrochloride exhibits maximum absorbance (λmax) in methanol at 294 nm and follows Beer's law at concentrations between 10 - 60 µg/ml. By using the standard addition approach, the proposed method's validity was evaluated. The additional standard's mean percentage recovery was found to be between 98% and 102%. The determined values for the limits of detection and quantification were 0.206µg/ml and 0.625µg/ml, respectively. The International Conference for Harmonisation (ICH) Q2 (R1) guidelines have been followed in the validation of the method. The suggested approach is rapid, simple to use, precise, sensitive, and cost-effective, making it suitable for the regular quantification of paroxetine in pharmaceutical formulations and bulk form.

 

 

 

INTRODUCTION:

The chemical structure of paroxetine is known as (3S, 4R) - 3-[(2H-1, 3-methyl 3-benzodioxol-5-yloxy)]piperidine (4-(4-fluoro phenyl))^[1]. The problems for which paroxetine was designed specifically are premenstrual dysphonic disorder (PMDD), depression, panic disorders, categorised anxiety disorder (GAD), and trauma-related stress disorder.

 

 

The first antidepressant used to treat panic disorders was paroxetine. As an inhibitor of selective serotonin reuptake (SSRI), paroxetine works by preventing the reuptake of specific serotonin neurotransmitters. Paroxetine prolongs the activity of the neurotransmitter at at the synaptic receptor places and stimulates 5-HT in the central nervous system (CNS) by perhaps preventing serotonin from being reabsorbed at the neuronal membrane and enhancing serotonergic neurotransmission by delaying neurotransmitter turnover^{1, 2,4}.

 

According to literature reviews, recently there were just two UV methods ^{1, 2,} as well as one colorimetric method ³ reported for paroxetine hydrochloride estimation. However it was found that the current work found to be accurate and sensitive.

 

Figure 1:Structure of Paroxetine Hydrochloride.

 

MATERIALS AND METHODS:

Instrument:

To validate paroxetine hydrochloride in the bulk, the instruments used were one centimetre matched of quartz cells and a UV/Visible spectrophotometer (Lab India).

 

Materials:

The supplier of paroxetine hydrochloride API was Yarrow Chemicals Private Limited in India. Yarrow Chemicals Private Limited supplied all of the solvents, including methanol, needed for this investigation.

 

Solvent selection for analysis

Several trials were conducted to determine the best solvent system for the drug's dissolving. Various solvents, including methanol, 0.1N NaOH, DMF and distilled water, were tested with regard to the drug's solubility. The drug's maximum absorption in methanol was determined to be 294 nm. Accordingly, methanol was chosen as the best solvent for this spectrophotometric method.

 

Standard stock solution preparation:^5-8

Precisely weighed and transferred exactly 25 milligrams of paroxetine hydrochloride into a 100 ml dry volumetric flask, followed by dissolving and adding methanol to make up the volume, and then sonicating for five minutes to designate it as stock-1 (250μg/ml).The above solution  dilutedproperly by using methanol to get a final concentrations40μg/ml. For the purpose of studying precision, robustness, and roughness, this solution was considered as the working standard solution.

 

Wavelength selection for λmax determination:

The ultraviolet spectrum was used to determine the wavelength for the examination of paroxetine hydrochloride. A solution containing 10 µg/ml of paroxetine hydrochloride was made, and it was examined in the 200–400 nm UV region to determine the λmax. The absorbance maxima (λmax) of paroxetine hydrochloride in relation to methanol were found at 294 nm.

 

 

 

RESULTS AND DISCUSSION:

Linearity:

Aliquots of the prepared standard solution (0.4, 0.8, 1.2, 1.6, 2.0, and 2.4 millilitres) were put into a series of 10-milliliter volumetric flasks and diluted using methanol to provide a concentration ranges of 10–60 μg/ml. The a fore mentioned solutions were scanned within 200 - 400 nm using a reagent blank. Each solution's absorbance at 294 nm was compared to methanol as a reference. Absorbance versus concentration was plotted to generate a calibration curve ^9-14. Calibration curve and overlay UV spectrum of different concentration of paroxetine hydrochloride was shown in Figure 2, 3 respectively. Table 1 presented the results.

 

Table 1: Results of the linearity of paroxetine hydrochloride

Concentration (µg/ml)	Absorbance (n=3)
10	0.166
20	0.328
30	0.482
40	0.635
50	0.833
60	0.981

n= number of replicate of each concentration.

 

Figure 2: Linearity curve of Paroxetine Hydrochloride.

 

Figure 3: Overlay UV spectra of different concentrations of Paroxetine Hydrochloride.

 

Limit of Detection and Quantification:

Limit of detection (LOD) and limit of quantification (LOQ) measurements were used to characterise the method's sensitivity ^15-19. It was computed by extracting the response's slope and intercept from the analyte's calibration curve, which were used to assess linearity. Utilising the subsequent formulas, it was found that the suggested method was sensitive because the results were less than 1 (Table 2).

 

LOD = 10(σ/S) and LOQ = 10(σ/S)

 

Where σ = the standard deviation and S is the slope of the calibration curve.

 

Table-2:Results of sensitivity

API	Limit of detection (LOD)	Limit of quantification (LOQ)
Paroxetine Hydrochloride	0.206µg/ml	0.625µg/ml

 

Precision:

In the intra-day study, the drug replicates' concentrations were determined twice in the same day. The drug concentration in the inter-day study was determined on two consecutive days, expressing the variance in the laboratory on various days ^20-25. % RSD was determined for the methods in the intra- day and inter-day precision study and the findings are displayed in Tables 3, 4.

 

Accuracy:

The standard addition method was used to conduct recovery parameter at three different levels (50%, 100%, and 150%). Recovery percentage and mean recovery percentage are used to calculate  accuracy ^26-36. It was discovered to be in the 98–102% range, as indicated in Table 5. The formula below is used for determining the recovery percentage.

Amount found

% Recovery= -------------  X 100

Amount added

 

Robustness: Six replicates of the working standard solution were made, and absorbance were measured under variable method conditions, such as applying various wavelengths (max ± 2nm), to demonstrate the method's resilience. Table 6 showed that the % RSD values were less than 2.

 

Table-3: Results for intra-day precision

Intraday precision(n=3)
Concentration (µg/ml)	Morning	Evening
40	0.647	0.659
40	0.637	0.669
40	0.638	0.669
40	0.650	0.664
40	0.643	0.665
40	0.641	0.668
Mean	0.642	0.665
Standard Deviation (SD)	0.005	0.003
% Relative standard deviation (%RSD)	0.778	0.451

n= number of replicate of each concentration.

 

Table-4: Results of Inter day precision

Inter day precision (n=3)
Concentration (µg/ml)	Day 1	Day 2	Day 3
40	0.650	0.606	0.658
40	0.653	0.618	0.652
40	0.656	0.612	0.656
40	0.658	0.614	0.662
40	0.650	0.610	0.650
40	0.649	0.616	0.653
Mean	0.652	0.612	0.655
Standard Deviation (SD)	0.003	0.004	0.004
%RSD	0.460	0.653	0.610

n= number of replicate of each concentration.

 

Table-5: Recovery results of proposed method

% Level	Test concentration added (Tablet powder - µg/ml)	Added Amount (API- µg/ml)	Amount found (µg/ml)	%Recovery	%Mean recovery
50%	40	20	19.64	98.20	100.03
50%	40	20	20.01	100.03
50%	40	20	20.37	101.86
100%	40	40	39.27	98.19	99.35
100%	40	40	39.82	99.56
100%	40	40	40.13	100.32
150%	40	60	58.97	98.28	99.33
150%	40	60	59.76	99.60
150%	40	60	60.07	100.11

 

Table 6: Results of robustness

n= number of replicate of each concentration i.e n=3.

 

Ruggedness:

For ruggedness investigations, six replicas of the working standard solution were used to investigate analyst variation. Table 7 showed that the percentage RSD values were less than 2.

 

Table-7: Results of ruggedness

Analyst-1(n=3)		Analyst-2(n=3)
Concentrations (µg/ml)	Absorbance	Concentrations (µg/ml)	Absorbance
40	0.645	40	0.643
40	0.648	40	0.640
40	0.648	40	0.637
40	0.653	40	0.640
40	0.658	40	0.630
40	0.655	40	0.644
Mean	0.651	Mean	0.651
SD	0.005	SD	0.005
%RSD	0.77%	% RSD	0.78%

 n= number of replicate of each concentration i.e n=3.

 

CONCLUSION:

A simple, accurate, and precise UV spectrophotometric method was successfully developed and validated for the estimation of Paroxetine Hydrochloride using methanol as the solvent. The drug exhibited a maximum absorbance (λ_max) at 294 nm. The method showed excellent linearity in the concentration range of 10–60 μg/mL with a correlation coefficient (R²) of 0.9991. The limits of detection (LOD) and quantification (LOQ) were found to be 0.206 μg/mL and 0.625 μg/mL, respectively, indicating the method's sensitivity. Precision studies, including both intraday and interday variations, demonstrated %RSD values of less than 1%, and confirming the method's repeatability and reproducibility. Robustness studies performed by varying the wavelength ±2 nm and ruggedness assessments using two different analysts showed %RSD values below 1%, reflecting the method's reliability under varied conditions. Recovery studies yielded results within the acceptable range of 99.33% to 100.03%, establishing the method's accuracy. Overall, the proposed UV spectrophotometric method is robust, rugged, and suitable for routine analysis of Paroxetine Hydrochloride in pharmaceutical formulations.

 

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Received on 27.07.2024      Revised on 03.03.2025 Accepted on 24.07.2025      Published on 01.10.2025 Available online from October 04, 2025 Research J. Pharmacy and Technology. 2025;18(10):4716-4720. DOI: 10.52711/0974-360X.2025.00678 © RJPT All right reserved
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