A NOVEL HPLC METHOD FOR THE SIMULTANEOUS DETERMINATION OF CHLORPHENIRAMINE MALEATE AND DEXTROMETHORPHAN IN BULK AND PHARMACEUTICAL FORMULATION

A NOVEL HPLC METHOD FOR THE SIMULTANEOUS DETERMINATION OF CHLORPHENIRAMINE MALEATE AND DEXTROMETHORPHAN IN BULK AND PHARMACEUTICAL FORMULATION

Blessy Sirigiri, Prasanthi Chengalva^*, S. Angala Parameswari and G. Aruna

Department of Pharmaceutical Analysis, Krishna Teja Pharmacy College, Tirupati - 517506, Andhra Pradesh, India.

ABSTRACT: Chlorpheniramine maleate and Dextromethorphan are used in treatment of cough caused by allergies and the common cold by decreasing intra ocular pressure. The present study was designed to develop and validate a simple, sensitive, precise and accurate reverse phase high performance liquid chromatographic (HPLC) method for simultaneous determination of Chlorpheniramine maleate and Dextromethorphan in tablet dosage form. Chromatographic separation of these two drugs was achieved on Discovery C18 column (250 x 4.6 mm, 5 μm) as stationary phase with a mobile phase of Water (pH 2 adjusted with OPA): Acetonitrile (60:40v/v)  at a flow rate of 1 ml/min and PDA detection at 218 nm. The retention times of Chlorpheniramine maleate and Dextromethorphan were found to be 3.0 min and 3.6 min respectively. The proposed method was validated for system suitability, linearity, accuracy, precision, LOD, LOQ and robustness. The calibration curves were linear in the concentration range of 50% to 150% of the working concentration (r² = 0.999) for both the drugs in binary mixture. The LOD was found to be 0.04 μg/ml and 0.11 μg/ml and LOQ was found to be 0.14 μg/ml and 0.43 μg/ml for Chlorpheniramine maleate and Dextromethorphan respectively. Hence the proposed RP-HPLC method can be used in routine analysis of tablets containing Chlorpheniramine maleate and Dextromethorphan.

Chlorpheniramine maleate, Dextromethorphan, RP- HPLC, Method development and validation

INTRODUCTION: Chlorpheneramine maleate is chemically 2-[p-Chloro - a - [2-(dimethylamino) ethyl] benzyl] pyridine maleate, is a histamine H1 antagonist used in allergic reactions, hay fever, rhinitis, urticaria, and asthma. Dextromethorphan is chemically (4bS, 8aR, 9S) -3- Methoxy-11-methyl-6,7,8,8a,9,10-hexahydro-5H-9,4b-(epiminoethano) phenanthrene, is an N-methyl-D-aspartate receptor antagonist used as cough suppressant.

Extensive literature survey revealed that there were analytical methods for the estimation of specified drugs with other combinations ^{1 - 14}. There was no method has been reported for the simultaneous estimation of Chlorpheniramine maleate and Dextromethorphan. Hence we presented a simple analytical method for simultaneous determination of Chlorpheniramine maleate and Dextromethorphan in bulk and pharmaceutical preparation.

MATERIALS AND METHODS:

Materials: Chlorpheniramine maleate and Dextromethorphan were obtained from Spectrum Labs, Hyderabad. Cheston meltees tablets were taken for study which contain 2 mg Chlorpheniramine maleate and 10 mg Dextromethorphan.  Acetonitrile, Ortho phosphoric acid (OPA) and HPLC grade water were used of analytical grade from Merck.

Diluent: Water: Acetonitrile has taken in the ratio 50:50 %v/v.

Standard stock solution preparation: 2 mg of Chlorpheniramine maleate and 10 mg of Dextromethorphan standards were accurately weighed and transferred into a 10 ml clean dry volumetric flask, 5 ml of diluent was added, sonicated for 5 minutes and made up to the final volume with diluent. Further 1ml from the above stock solution was taken into a 10ml volumetric flask and made up to 10ml with diluent.

Sample solution preparation: 20 tablets were weighed and crushed. A powder equivalent to 2 mg of Chlorpheniramine maleate and 10 mg of Dextromethorphan was taken and then transferred into a 10 ml clean dry volumetric flask, 5 ml of diluent was added, sonicated for 5 minutes and made up to the final volume with diluent. Further 1 ml from the above stock solution was taken into a 10 ml volumetric flask and made up to 10 ml with diluent.

Chromatographic Conditions: The criteria employed for selecting the mobile phase for the analysis of the drugs were cost involved, time required for the analysis and better separation of drugs. Chromatographic separation was performed on reverse phase Waters Discovery C18 (250 x 4.6 mm, 5m) column. The mobile phase consisted of water (pH 2 adjusted with OPA): acetonitrile (60:40v/v) with PDA detection at 218 nm. The flow rate was set at 1.0 ml/min for simultaneous determination of Chlorpheniramine maleate and Dextromethorphan.

Method Development: Different chromatographic conditions were tried for separation and resolution. Waters symmetry Discovery column was found satisfactory. Peak purity of Chlorpheniramine maleate and Dextromethorphan was checked using photo diode array detector and 218 nm was considered satisfactory for detecting both the drugs with adequate sensitivity. A typical RP-HPLC chromatogram for simultaneous determination of Chlorpheniramine maleate and Dextromethorphan from standard preparation and from pharmaceutical formulation was shown in Fig. 1 and 2.

FIG. 1: CHROMATOGRAM OF CHLORPHENIRAMINE MALEATE AND DEXTROMETHORPHAN IN STANDARD PREPARATION

FIG. 2: CHROMATOGRAM OF CHLORPHENIRAMINE MALEATE AND DEXTROMETHORPHAN IN SAMPLE PREPARATION

RESULTS AND DISCUSSION:

Method Validation: The developed RP-HPLC method was validated for parameters like system suitability, linearity, accuracy, precision, LOD, LOQ and robustness according to ICH guidelines¹⁵.

System Suitability: Standard solutions were prepared as per the test method and injected into the chromatographic system. The system suitability parameters like theoretical plates, resolution and asymmetric factor were evaluated. The system suitability parameters were tabulated in Table 1. All the parameters were found to be within the limits.

Precision:

Method Precision: The precision of the analytical method was verified by method precision studies. The sample solution was prepared at working concentration and analysis was carried out at replicates. The sample solutions of Chlorpheniramine maleate and Dextromethorphan were prepared as per the test method and injected 6 times into the column. The results of precision were tabulated in Table 2. The average was taken; % RSD was calculated and reported. % RSD values were found within the limits, indicating the developed method was precise.

TABLE 1: RESULTS OF SYSTEM SUITABILITY

# min: minutes

 TABLE 2: RESULTS OF METHOD PRECISION

# Rt: Retention times, # n: number of injections=6

Linearity: The linearity of the test solutions for the assay method was prepared from Chlorpheniramine maleate and Dextromethorphan standard stock solution at five concentration levels from 50% to 150% of assay concentration. The peak area versus concentration data was treated by least-squares linear regression analysis (Fig. 3 and 4).

FIG. 3: LINEARITY CHART OF CHLORPHENIRAMINE MALEATE

FIG. 4: LINEARITY CHART OF DEXTROMETHORPHAN

The results tabulated in Table 3 have shown an excellent correlation between peak areas and concentration within the concentration range of 10 - 30 μg/ml for Chlorpheniramine maleate, 50 - 100 μg/ml for Dextromethorphan. The correlation coefficients were found to be 0.999 for both the drugs, which meet the method validation acceptance criteria and hence the method was said to be linear for both the drugs.

Accuracy: To ensure the reliability and accuracy of the method recovery studies were carried out by standard addition method. A known quantity of pure drug was added to pre-analyzed sample and contents were reanalyzed by the proposed method and the percent recovery was reported. The results were given in Table 4.

Limit of Detection and Limit of Quantitation: The limit of detection (LOD) and limit of quantitation (LOQ) were established at signal-to-noise ratio of 3:1 and 10:1 respectively. The LOD and LOQ of Chlorpheniramine maleate and Dextromethorphan were experimentally determined by injecting six injections of each drug. The LOD of Chlorpheniramine maleate and Dextromethorphan was found to be 0.04 µg/ml and 0.11 µg/ml respectively. The LOQ of Chlorpheniramine maleate and Dextromethorphan was found to be 0.14 µg/ml and 0.43 µg/ml respectively.

Robustness: Robustness of the method was verified by altering the chromatographic conditions like mobile phase composition, wavelength detection, flow rate, etc. and the % RSD should be reported. Small changes in the operational conditions were allowed and the extent to which the method was robust was determined. A deviation of ±2 ^oC in the column temperature and ±0.2ml/min in the flow rate, were tried individually. A solution of 100 % test concentration with the specified changes in the operational conditions was injected to the instrument in triplicate. % RSD was reported in the Table 5.

TABLE 3: RESULTS OF LINEARITY

TABLE 4: RESULTS OF ACCURACY

TABLE 5:  RESULTS OF ROBUSTNESS

CONCLUSION:  The proposed RP-HPLC method was found to be simple, accurate, precise, robust, rapid and economical. This method gives good resolution between two compounds with a short analysis time. Hence this method can be used in quality control departments with respect to routine analysis for the assay of the tablets containing Chlorpheniramine maleate and Dextromethorphan.

ACKNOWLEDGEMENT: Author expresses sincere thanks to the Principal and Head of Pharmaceutical Analysis department, Krishna Teja Pharmacy College for providing facilities and great support to carry out the research work.

CONFLICTS OF INTEREST: The authors declare that there is no conflict of interest

REFERENCES:

1. Gyanendra P, Palled MS, Bhat AR, Gyanendra PS , Aruna M: Analytical method development and validation of Acetaminophen, Caffeine, Phenylephrine Hydrochloride, Dextromethorphan Hydrobromide and Chlorpheniramine Maleate in tablet dosage form by RP- HPLC. International Journal of Pharmaceutical Research 2017; 1:10-13.
2. Rushikesh B, Nikam A and Sanjay S: Analytical method development and validation of Phenylephrine hydrochloride, Chlorpheniramine maleate, Paracetamol and caffeine in bulk drug and tablet dosage form by RP-HPLC. Indo American journal of pharmaceutical Research 2016; 1:1-7.

3. Akwasi A, Wilfred OG, Godfred D and Sylvester AA: Validated RP-HPLC method for simultaneous determination and quantification of Chlorpheniramine maleate, Paracetamol and Caffeine in tablet formulation. Springer Plus 2016; 5:625-629.
4. Vishal J and Mukesh CS: Validated RP-HPLC method for determining the levels of Bromhexine HCl, Chlorpheniramine maleate, Dextromethorphan HBr and Guaiphenesin in their pharmaceutical dosage forms. Journal of Taibah University of science 2016; 10:38-48.
5. Kapildev RJ: Analytical method development and validaton of RP-HPLC for simultaneously estimation of Chlorpheniramine maleate, Dextromethorphen hydro-bromide, Phenylephrine hydrochloride in cough syrup. Institute for Research and Academic Journal 2016; 4: 53-58.
6. Ali A, Ahmed M, Mahmud T, Qadir MA, Nadeem K and Saleem A: Stability indicating High Performance Liquid Chromatography method for simultaneous determination of Aminophylline and Chlorpheniramine Maleate in Pharmaceutical Formulations. Indian Journal of Pharmaceutical Sciences 2015; 77: 515-521.
7. Bhaskar D, Reddy GS and Kamarapu SK: Development and validation of RP-HPLC method for simultaneous estimation of Chlorpheniramine maleate and Diethyl carbamazine citrate in pharmaceutical dosage forms. Asian Journal of Pharmaceutical and Clinical Research 2014; 7:98-102.
8. Alagar RM, Samatha MDB, Rao KNV, Sujitha, Vinod J and Selva KD: Analytical method development and validation of Acetamenophen, Dextromethorphan hydro bromide, Doxylamine succinate in soft gel capsule dosage form by using RP-HPLC. World Journal of Pharmacy and Pharmaceutical Sciences 2013; 2:5852-5862.
9. Redasani VK, GorleAP, Badhan RA, Jain PS and Surana SJ: Simultaneous determination of Chlorpheniramine maleate, Phenylephrine hydrochloride, Paracetamol and Caffeine in pharmaceutical preparation by RP-HPLC. Chemical Industry & Chemical Engineering Quarterly 2013; 19:57−65.
10. Pinak MS, Falgun AM andNirav BU: Development and Validation of an RP-HPLC Method for Estimation of Chlorpheniramine Maleate, Ibuprofen, and Phenylephrine Hydrochloride in Combined Pharmaceutical Dosage Form. Chromatography Research International 2013; 6:42-48.
11. Renu S, Khata M, Mukesha M and Sunil K: Simultaneous determination of Chlorpheniramine maleate, Paracetamol and Phenylephirine Hydrochloride in Tablet dosage form by High Performance Liquid Chromatography. International journal of drug development and research 2013; 5:258-263.
12. Sandeep R: Simultaneous determination of Chlorpheniramine maleate, Paracetamol and Pseudoephedrine hydrochloride in pharmaceutical preparations by HPLC. International Journal of Life Sciences and Pharma Research 2011; 1:28-32.

13. Mukesh M, Richa R, Vertika G, Dharmendra K, Amrendra KC, Anand G and Ranjit S: Development and validation of a RP-HPLC method for the determination of Chlorpheniramine maleate and Phenylephrine in pharmaceutical dosage form. International Journal of Comprehensive Pharmacy 2010; 1:1-4.
14. Heydari R: A New HPLC method for the simultaneous determination of Acetaminophen, Phenylephrine, Dextromethorphan, Chlorpheniramine in Pharmaceutical formulation. Analytical Latters 2008; 41: 965-976.
15. International conference on harmonization of technical requirements for registration of pharmaceuticals for human use. Validation of analytical procedures; Text and methodology ICH Q2 (R1); 2005.
    

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    How to cite this article:

    Sirigiri B, Chengalva P, Parameswari SA and Aruna G: A novel HPLC method for the simultaneous determination of chlorpheniramine maleate and dextromethorphan in bulk and pharmaceutical formulation. Int J Pharm Sci Res 2018; 9(3): 1147-51. doi: 10.13040/ IJPSR. 0975-8232.9(3).1147-51.

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