Analytical Method Development and Validation of Chlorpheniramine Maleate by Uv Spectroscopy

Abstract

A UV Spectrophotometric method for the determination of Chlorpheniramine Maleate in Bulk and its pharmaceutical dosage form. The wavelength was selected using water, 0.1 N HCl and 0.1 N NaOH as solvents (later HCL was selected as final solvent). The method was found to be specific with the appearance of the corresponding maxima at 268 nm. Linearity was observed at concentration of 10µg/ml – 50µg/ml and a correlation coefficient (R2) of 0.997. Limits of detection and quantification were respectively 0.62 and 0.187. The tolerance limits for the different concentration levels (80, 100 and 120%) were respectively: 100.2, 100.5 and 99.5%. The results of method validation were in the acceptable range as per ICH guidelines.

Keywords

Introduction

Figure 1: Structure of Chlorpheniramine Maleate

Unfortunately, due to lack of resources, the latter could not check the quality of all pharmaceutical products circulating in the countries [6, 7, 8, 9]. In the treatment of allergy, Chlorpheniramine Maleate, as an essential drug to its effectiveness and lower cost and is distributed worldwide as a generic compound [10, 11]. The present study describes the development and validation of novel, alternative and simplified analytical method for the quantification of antihistamine drug Chlorpheniramine Maleate that meets the requirements of the International Conference on Harmonization, which shows the practicality, reliability, safety and low cost of using this method routinely in pharmaceutical industry. Analytical method performance characteristic are defined to meet the need of analytical target profile. According to USP & ICH guidelines there are many validation parameters for separations, which are considered as method performance characteristics.

MATERIAL AND METHOD:

• Instrument: UV- Visible spectrophotometer (Double beam) having matched quartz cells of light path 1 cm of ELICO - Model (SL-159).
• Apparatus: Electronic analytical weighing balance 
• Glassware: Volumetric flask (Borosilicate), Pipettes, etc.
• Reagents and chemicals: Chlorpheniramine Maleate (Jai Radhe Sales), hydrochloric acid (Sigma Aldrich), Chlorpheniramine Maleate Tablets (Dr. Reddy’s).

Preparation of Standard and Working Solutions:

1. Chlorpheniramine Maleate Standard Stock Solution (1000 µg/ml):
   • Weigh 10 mg of Chlorpheniramine maleate and transfer it to a 10 ml volumetric flask.
   • Add 5 ml of hydrochloric acid (HCl) to dissolve the drug.
   • Finally, add more HCl to bring the solution up to the 10 ml mark.
2. Chlorpheniramine Maleate Working Stock Solution (100 µg/ml):
   • Take 1.0 ml of the above stock solution and transfer it to a 10 ml volumetric flask.
   • Fill the flask with HCl up to the 10 ml mark.
   • This solution can then be further diluted to prepare concentrations between 10-50 µg/ml.

Analysis of Tablets:

1. Finely grind a tablet that contains 5 mg of Chlorpheniramine maleate.
2. Transfer the powdered tablet into a 50 ml volumetric flask and add HCl to reach the 50 ml mark.
3. Filter the solution using Whatman filter paper.
4. Dilute the filtered solution further to get a concentration of 30 µg/ml.
5. The drug content in this solution is determined using the calibration curve of the standard Chlorpheniramine maleate solution.

METHOD DEVELOPMENT:

The method was validated by evaluating the following parameters: selectivity, linearity and range, accuracy, trueness (recovery), limit of detection (LOD) and limit of quantification (LOQ) as specified in the International Conference on Harmonization protocol [12]. The method has been validated using the accuracy profile as previously described [13,14,15,16].

Selection of solvent system:

The selection of solvent as made after determining the solvent of Chlorpheniramine maleate in different solvents like water, 0.1N HCl, 0.1N NaOH, 0.1N H₂SO₄. Chlorpheniramine maleate is insoluble in water and soluble in HCl. So HCl is selected as a solvent system for this UV-method development.

Preparation of standard stock solution:

Chlorpheniramine maleate standard stock solution (100µg/ml):

A 10mg of Chlorpheniramine maleate was weighed accurately and transferred to a 100ml volumetric flask. Now add 10ml of HCl to this volumetric flask to make up the volume with 0.1N HCl up to 100ml.

Preparation of 0.1N HCl solution:

2.2ml of 0.1N HCl is transferred in a 250ml volumetric flask and makes up the volume up to 250ml with distilled water.

Selection of Wavelength:

The drug is soluble in 0.1N HCl, prepared with different concentrations of solution. These solutions recorded the spectrum between 200 to 400 nm. In UV-Spectrophotometric method wavelength 268nm was selected for determination of Chlorpheniramine maleate.

Method Validation: According to ICH Q2 (R1) Guideline the method was validated [17,18,19,20].

Linearity:

The solution was prepared by pipetting 1, 2, 3, 4, 5 ml from working stock solution into 10ml volumetric flask and the volume was adjusted to mark with HCl to produce 10 - 50 μg/ml respectively. The absorbance of solutions was measured at 268nm. Calibration curve was generated by taking the absorbance verses concentration.

Accuracy:

The accuracy of the method was determined by calculating recovery of Chlorpheniramine maleate by the standard addition method. Reference standard solution of each drug was added to samples at three different concentrations level (80,100 and 120%). At each level, samples were prepared in triplicate and the mean percentage recoveries and % RSD value was calculated.

Precision (Repeatability):

Aliquots of 0.4 ml of working standard solution of Chlorpheniramine maleate (100μg/ml) were transferred to 10ml volumetric flask and volume was adjusted to HCl to get concentration of 4μg/ml. The absorbance of solution was measured spectrophotometry six times to calculate % RSD. Intraday aliquots of 0.2, 0.3, 0.4 ml of working standard solution of Chlorpheniramine maleate (100μg/ml) were transferred to 10ml volumetric flask and volume was adjusted to 0.1N HCl to get concentration of 2, 3 and 4 μg/ml. The absorbance of solution was measured by spectrophotometry three times and % RSD was calculated.

Ruggedness:

Ruggedness is a measure of reproducibility of test results under various conditions.

Robustness:

The robustness of an analytical procedure is a measure of its capacity to remain unaffected. It provides an indication of reliability during normal usage.

Limit of detection (LOD)

The LOD is estimated from the set of 5 calibration curves used to determine method linearity. The LOD may be calculated as,

LOD=3.3XS.DSlope

 

Where, SD = Standard deviation of the Y- intercepts of the 5 calibration curves.

Slope = Mean of the 5 calibration curves.

Limit of Quantitation (LOQ)

The LOQ is estimated from the set of 5 calibration curves used to determine method linearity. The LOQ may be calculated as,

LOQ=10XS.DSlope

 

Where, SD = Standard deviation of the Y- intercepts of the 5calibration curves.

Slope = Mean of the 5 calibration curves

RESULTS AND DISCUSSION:

The present work describes UV Spectrophotometric method for analysis of Chlorpheniramine Maleate in tablets. Chlorpheniramine Maleate was soluble in 0.1N HCl. Standard Chlorpheniramine Maleate solution showed absorption maximum at 268nm was selected as the detection wavelength.

Figure 2 Absorption Maxima in 0.1N HCl

Figure 2: Calibration curve of Chlorpheniramine maleate

CONCLUSION:

The proposed method development and validation of UV-spectrophotometric method was to determine Chlorpheniramine maleate. The developed method was validated in 0.1N HCl according to ICH guidelines. It has shown to be accurate, precise and cost effective. It doesn't require expensive and enlightened and chemicals in contrast with chromatographic method. It can be used for quality control analysis and quantification of the drug in the formulations The Chlorpheniramine maleate maximum absorption is 268 nm. The linearity occurs in the range 10-50µg/ml. The correlation coefficients should be near to 0.997. Precision was determined thrice in a day with 2 concentrations taken 3 times with % RSD 0.329,0.370 for 2 µg/ ml and 4µg/ml respectively. Accuracy for Chlorpheniramine maleate was evaluated for 80,100 and 120%. % recovery of accuracy was found to be 100.2%, 100.5%, 99.5% respectively. Ruggedness was evaluated RSD was found to be less than 2%. For robustness absorbance taken at 266, 268 and 270 nm and RSD was found to be less than 2%. The limit of Detection and Quantitation were respectively 0.62 µg/ml and 1.87µg/ml

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