Sirigiri et al., IJPSR, 2018; Vol. 9(3): 1147-1151.

E-ISSN: 0975-8232; P-ISSN: 2320-5148

IJPSR (2018), Volume 9, Issue 3 (Research Article)

Received on 15 June, 2017; received in revised form, 10 August, 2017; accepted, 17 August, 2017; published 01 March, 2018

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.
Keywords: ABSTRACT: Chlorpheniramine maleate and Dextromethorphan are used in
Chlorpheniramine maleate,
treatment of cough caused by allergies and the common cold by decreasing intra
Dextromethorphan, RP- HPLC, ocular pressure. The present study was designed to develop and validate a
Method development and validation simple, sensitive, precise and accurate reverse phase high performance liquid
Correspondence to Author: chromatographic (HPLC) method for simultaneous determination of
Mrs. C. Prasanthi Chlorpheniramine maleate and Dextromethorphan in tablet dosage form.
Chromatographic separation of these two drugs was achieved on Discovery C18
Assistant Professor, column (250 x 4.6 mm, 5 μm) as stationary phase with a mobile phase of Water
Department of Pharmaceutical (pH 2 adjusted with OPA): Acetonitrile (60:40v/v) at a flow rate of 1 ml/min
Analysis, Krishna Teja Pharmacy and PDA detection at 218 nm. The retention times of Chlorpheniramine maleate
College, Chadalawada Nagar, and Dextromethorphan were found to be 3.0 min and 3.6 min respectively. The
Renigunta Road, Tirupati - 517506,
proposed method was validated for system suitability, linearity, accuracy,
Chittoor Andhra Pradesh, India.
precision, LOD, LOQ and robustness. The calibration curves were linear in the
E-mail: [email protected] 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.
INTRODUCTION: Chlorpheneramine maleate is Extensive literature survey revealed that there were
chemically 2-[p-Chloro - a - [2-(dimethylamino) analytical methods for the estimation of specified
ethyl] benzyl] pyridine maleate, is a histamine H1 drugs with other combinations 1 - 14. There was no
antagonist used in allergic reactions, hay fever, method has been reported for the simultaneous
rhinitis, urticaria, and asthma. Dextromethorphan is estimation of Chlorpheniramine maleate and
chemically (4bS, 8aR, 9S) -3- Methoxy-11-methyl- Dextromethorphan. Hence we presented a simple
6,7,8,8a,9,10-hexahydro-5H-9,4b-(epiminoethano) analytical method for simultaneous determination
phenanthrene, is an N-methyl-D-aspartate receptor of Chlorpheniramine maleate and Dextromethorphan
antagonist used as cough suppressant. in bulk and pharmaceutical preparation.
QUICK RESPONSE CODE MATERIALS AND METHODS:
DOI:
10.13040/IJPSR.0975-8232.9(3).1147-51
Materials: Chlorpheniramine maleate and
Dextromethorphan were obtained from Spectrum
Article can be accessed online on:
Labs, Hyderabad. Cheston meltees tablets were
www.ijpsr.com taken for study which contain 2 mg
Chlorpheniramine maleate and 10 mg
DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.9(3).1147-51
Dextromethorphan. Acetonitrile, Ortho phosphoric

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Sirigiri et al., IJPSR, 2018; Vol. 9(3): 1147-1151. E-ISSN: 0975-8232; P-ISSN: 2320-5148

acid (OPA) and HPLC grade water were used of Chromatographic Conditions: The criteria
analytical grade from Merck. employed for selecting the mobile phase for the
analysis of the drugs were cost involved, time
Diluent: Water: Acetonitrile has taken in the ratio required for the analysis and better separation of
50:50 %v/v. drugs. Chromatographic separation was performed
on reverse phase Waters Discovery C18 (250 x 4.6
Standard stock solution preparation: 2 mg of
Chlorpheniramine maleate and 10 mg of mm, 5) column. The mobile phase consisted of
Dextromethorphan standards were accurately water (pH 2 adjusted with OPA): acetonitrile
weighed and transferred into a 10 ml clean dry (60:40v/v) with PDA detection at 218 nm. The
volumetric flask, 5 ml of diluent was added, flow rate was set at 1.0 ml/min for simultaneous
sonicated for 5 minutes and made up to the final determination of Chlorpheniramine maleate and
volume with diluent. Further 1ml from the above Dextromethorphan.
stock solution was taken into a 10ml volumetric Method Development: Different chromatographic
flask and made up to 10ml with diluent. conditions were tried for separation and resolution.
Sample solution preparation: 20 tablets were Waters symmetry Discovery column was found
weighed and crushed. A powder equivalent to 2 mg satisfactory. Peak purity of Chlorpheniramine
of Chlorpheniramine maleate and 10 mg of maleate and Dextromethorphan was checked using
Dextromethorphan was taken and then transferred photo diode array detector and 218 nm was
into a 10 ml clean dry volumetric flask, 5 ml of considered satisfactory for detecting both the drugs
diluent was added, sonicated for 5 minutes and with adequate sensitivity. A typical RP-HPLC
made up to the final volume with diluent. Further 1 chromatogram for simultaneous determination of
ml from the above stock solution was taken into a Chlorpheniramine maleate and Dextromethorphan
10 ml volumetric flask and made up to 10 ml with from standard preparation and from pharmaceutical
diluent. 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

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Sirigiri et al., IJPSR, 2018; Vol. 9(3): 1147-1151. E-ISSN: 0975-8232; P-ISSN: 2320-5148

RESULTS AND DISCUSSION: Precision:

Method Validation: The developed RP-HPLC Method Precision: The precision of the analytical
method was validated for parameters like system method was verified by method precision studies.
suitability, linearity, accuracy, precision, LOD, The sample solution was prepared at working
LOQ and robustness according to ICH guidelines15. concentration and analysis was carried out at
replicates. The sample solutions of Chlorpheniramine
System Suitability: Standard solutions were maleate and Dextromethorphan were prepared as
prepared as per the test method and injected into per the test method and injected 6 times into the
the chromatographic system. The system suitability column. The results of precision were tabulated in
parameters like theoretical plates, resolution and Table 2. The average was taken; % RSD was
asymmetric factor were evaluated. The system calculated and reported. % RSD values were found
suitability parameters were tabulated in Table 1. within the limits, indicating the developed method
All the parameters were found to be within the was precise.
limits.
TABLE 1: RESULTS OF SYSTEM SUITABILITY
Analytes Retention times Resolution Theoretical plates Tailing factor
Chlorpheniramine maleate 3.0 min - 5751 1.5
Dextromethorphan 3.6 min 4.1 5304 1.0
# min: minutes

TABLE 2: RESULTS OF METHOD PRECISION

N Chlorpheniramine maleate Dextromethorphan
Rt (min) Peak area Rt (min) Peak area
Injection 1 3.005 457727 3.665 1920019
Injection 2 3.005 455312 3.666 1931638
Injection 3 3.006 457351 3.665 1927814
Injection 4 3.005 458951 3.674 1919839
Injection 5 3.006 451597 3.670 1910973
Injection 6 3.005 458744 3.666 1925603
Average 456614 1922648
SD 2780.1 7312.3
%RSD 0.6 0.4
# 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. 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
FIG. 3: LINEARITY CHART OF CHLORPHENIRAMINE drugs, which meet the method validation
MALEATE

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Sirigiri et al., IJPSR, 2018; Vol. 9(3): 1147-1151. E-ISSN: 0975-8232; P-ISSN: 2320-5148

acceptance criteria and hence the method was said was found to be 0.04 µg/ml and 0.11 µg/ml
to be linear for both the drugs. respectively. The LOQ of Chlorpheniramine
maleate and Dextromethorphan was found to be
Accuracy: To ensure the reliability and accuracy 0.14 µg/ml and 0.43 µg/ml respectively.
of the method recovery studies were carried out by
standard addition method. A known quantity of Robustness: Robustness of the method was
pure drug was added to pre-analyzed sample and verified by altering the chromatographic conditions
contents were reanalyzed by the proposed method like mobile phase composition, wavelength
and the percent recovery was reported. The results detection, flow rate, etc. and the % RSD should be
were given in Table 4. reported. Small changes in the operational
conditions were allowed and the extent to which
Limit of Detection and Limit of Quantitation: the method was robust was determined. A deviation
The limit of detection (LOD) and limit of of ±2 oC in the column temperature and ±0.2ml/min
quantitation (LOQ) were established at signal-to- in the flow rate, were tried individually. A solution
noise ratio of 3:1 and 10:1 respectively. The LOD of 100 % test concentration with the specified
and LOQ of Chlorpheniramine maleate and changes in the operational conditions was injected
Dextromethorphan were experimentally determined to the instrument in triplicate. % RSD was reported
by injecting six injections of each drug. The LOD in the Table 5.
of Chlorpheniramine maleate and Dextromethorphan

TABLE 3: RESULTS OF LINEARITY

Analytes Correlation coefficient (r2)
Chlorpheniramine maleate 0.999
Dextromethorphan 0.999

TABLE 4: RESULTS OF ACCURACY

Analytes Pre-analysed Amount added Amount found Recovery
sample conc. (µg/ml) (µg/ml) (µg/ml) (%)
Chlorpheniramine maleate 20 10 9.88 98.83
20 20 19.65 98.28
20 30 29.89 99.57
Dextromethorphan 100 50 50.16 100.33
100 100 98.01 98.01
100 150 149.61 99.74

TABLE 5: RESULTS OF ROBUSTNESS

Analytes Flow rate (ml) % RSD Column temperature (oC) % RSD
Chlorpheniramine maleate 0.8 1.2 28 1.4
1.2 1.1 32 1.3
Dextromethorphan 0.8 0.6 28 1.2
1.2 0.8 32 1.5

CONCLUSION: The proposed RP-HPLC method Pharmacy College for providing facilities and great
was found to be simple, accurate, precise, robust, support to carry out the research work.
rapid and economical. This method gives good
resolution between two compounds with a short CONFLICTS OF INTEREST: The authors
analysis time. Hence this method can be used in declare that there is no conflict of interest
quality control departments with respect to routine
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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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