Development of Stability Indicating Assay Method for Simultaneous Estimation of Phenylephrine HCl,

Dextromethorphan HBr and Cetrizine HCl in Syrup

VERMA S. D. HEMKE A. T and GUPTA K. R
Shrimati Kishoritai Bhoyar College of Pharmacy Kamptee, Nagpur
sunitaverma.bltr@gmail.com
ABSTRACT
The aim of present study was to developed a stability-indicating RP-HPLC method for the determination of Phenylephrine
HCl, Dextromethorphan HBr and Cetrizine HCl in bulk and its marketed formulation. The method was developed using
Xterra C18 (250 x 4.6 mm, 5) as stationary phase with a mobile phase consisting of Acetonitrile: Methanol: Ammonium
sulphate buffer (30:15:55%v/v/v) pH-3.4, at flow rate 1.0 mL/min. The detection of analytes was carried out at 272 and 230
nm. The stability of the drugs and formulation were evaluated by exposing to various stress conditions like acidic, alkaline,
peroxide, thermal and UV light. The drugs were estimated in presence of its degradation products (if any) without
interference. The % recovery by the proposed method was found equal to 100% .The method was validated as per ICH
guidelines and the results were found to be satisfactory. Hence the method can be adopted for routine analysis of drug in its
syrup formulation.
INTRODUCTION
Phenylephrine HCl (PPH) is a selective 1-adrenergic receptor agonist used as a decongestant, while Dextramethorphan HBr
(DEX) as a antitussive and Cetrizine (CET) as antiallergic agent.. The numbers of analytical method were located in
literature on selected drugs under study. Meyyanathan N2 reported development and validation of dissolution study of
sustained release dextromethorphan hydrobromide tablets. Thakkar K3 reported UV Spectrophotometric method for
estimation of dextromethorphan in bulk and syrup formulation by area under curve method. Reddy P 4 reported development
and validation of a ultra performance liquid chromatographic method for assay of cetirizine dihydrochloride. Patel B5
reported simultaneous determination of phenylephrine hydrochloride, paracetamol, chlorpheniramine maleate and
dextromethorphan hydrobromide in pharmaceutical preparations. No method was found in literature for determination of
selected drugs in their combined dosage form. Hence, objective of the present study was to develop a stability indicating
assay method for the estimation of these drugs.
MATERIALS AND METHOD
Preparation of Mix stock standard solution:

75.0 mg PPH, 150.0 mg DEX and 50.0 mg CET were transferred to a 50.0 mL volumetric flask and diluted upto
mark with mobile phase. (Conc. 1500g/mL of PPH , 3000g/mL of DEX and 1000g/mL of CET)

Preparation of Working mix standard solution

A 5.0 mL of mix stock standard solution was diluted to 50.0 mL in a volumetric flask with mobile phase. (Conc.
150g/mL of PPH, 300g/mL of DEX and 100g/mL of CET)

Preparation of Buffer solution- Ammonium Sulphate solution (pH-3.4)

Accurately weighed quantities about 460 mg of Ammonium Sulphate and 1000 mg of Octane 1-sulphonic acid
sodium salt were dissolved in 1000.0 mL of double distill water and pH was adjusted to 3.4 with 10% glacial
acetic acid.

HPLC method was developed on Jasco LC-Net II/ADC using Xterra C18 (250 x 4.6 mm, 5) stationary phase with a mobile
phase containing Acetonitrile: Methanol: Ammonium sulphate buffer (30:15:55%v/v/v) of pH 3.4.The reasonable retention
time achieved using flow rate 1.0 mL/min at detection wavelengths 272, 230 nm. AR and GR grade chemicals used
throughout the experimentation. The HPLC chromatogram of Std. PPH, DEX and CET using optimized chromatographic
conditions is shown in Fig. 1.

Fig. 1: HPLC Chromatogram of Std. PPH, DEX and CET

FORCED DEGRADATION STUDY
This study was carried out via: i) Solution state analysis ii) Solid state analysis
Preparation of stress sample solutions

Accurately weighed quantities of 5.0 mL of syrup equivalent to 7.5 mg of PPH (15.0 mg DEX, 5.0 mg CET) were
transferred to series of 50.0 mL volumetric flasks. To each flask 5.0 mL of reagent (0.1N HCl/0.1N NaOH/ 3%
H2O2) was added and kept at RT for a period of 24 h. After 24 h, the content of each flask was diluted with mobile
phase, sonicated for 15 min and samples were filtered separately. In solid state analysis sample exposed to heat,
humidiy and UV light. After specified period stress samples were diluted, filtered and injected to system.

RESULTS AND DISCUSSION

The stability of the drugs was carried out for solution state stability and solid state stability .The results of solution state
analysis indicate that in sample PPH, DEX and CET were degraded to about 4.55%, 8.08% and 4.67% in acid, 4.44%, 6.88%
and 4.07% in alkali, 2.34%, 6.37% and 2.66% in peroxide when compared to unexposed sample. The results of solid state
stability after 48 h indicate that in sample PPH, DEX and CET were degraded to 1.83%, 3.88% and 3.63% when exposed to
humidity, 1.7%, 4.28% and 2.83% when exposed to thermal, 3.99%, 6.43% and 3.45% when exposed to UV radiation as
compared to unexposed sample.

VALIDATION OF PROPOSED METHOD

Validation parameters such as system suitability, linearity, precision, accuracy, limit of detection (LOD), limit of
quantification (LOQ), Stability of sample and standard stock solutions and robustness were studied. Summary of validation
parameters is given in Table 1.
Table 1: Summary of validation parameters
Drugs
PPH
DEX
CET

CONCLUSION

Accuracy
Mean
%RSD
99.74
0.94
99.66
0.85
99.78
0.41

VALIDATION PARAMETERS
Precision
Intra-day study
Mean
%RSD
Mean
%RSD
99.65
0.51
99.93
0.42
99.53
0.55
99.76
0.40
99.47
0.39
99.76
0.46

Inter-day study
Mean
%RSD
98.27
1.01
98.60
1.04
99.05
0.68

Robustness
Mean
%RSD
below less
than 2

The method is precise, accurate, rapid, reasonably specific and rugged. The developed method was found to be superior with
srespect to resolution of drug from its degradation products under applied stress conditions. Hence, method may be adopted
for routine assay of selected drugs free of interferences from its degradation products in bulk and marketed formulations.
REFERENCES
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in

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