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

Dextromethorphan HBr and Cetrizine HCL in Syrup

VERMA S. D., GUPTA K. R., HEMKE A. T.
Shrimati Kishoritai Bhoyar College of Pharmacy Kamptee,Nagpur
Sunitaverma.bltr@gmail.com
ABSTRACT :
The aim of present study was to developed a stabilityindicating HPLC method for the determination of Phenylephrine
HCL, Dextromethorphan HBr and Cetrizine HCL in bulk and its marketed formulation(syrup).HPLC method was
developed on a Xterra C18 (250 x 4.6 mm, 5) stationary phase with a mobile phase consisting of
Acetonitrile:Methenol:Ammonium sulphate buffer (30:15:55) having P H 3.4, effluent flow rate (1.0 mL/min) and detected
at multiwavelength i.e. 272, 230 the column was maintain at temperature 2830 oC. These method precise, accurate, rapid,
reasonable, specific and rugged, hence method may be adopted for routine assay of selected drugs free of interference from
its degradated product in marketed formulation.
INTRODUCTION
Phenylephrine HCL is a selective 1-adrenergic receptor agonist, use as a decongestant, Dextramethorphan HBr used as a
antitussiveand Cetirizine used as allergics agents. There are several analytical methods were reported ondextromethorphan
hydrobromide , 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 B5The
simultaneous determination of phenylephrine hydrochloride, paracetamol, chlorpheniramine maleate and
dextromethorphan hydrobromide in pharmaceutical preparations. The objective of the present study was to develop a
Stability Indicating Assay method for the combination of three drugs because there was several methods develop only for
determination of two drug combination.
MATERIAL And METHODS
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 solution PC1 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)

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 a Xterra C18 (250 x 4.6 mm, 5) stationary phase with a mobile phase consisting of
Acetonitrile:Methenol:Ammonium sulphate buffer (30:15:55) having P H 3.4, effluent flow rate (1.0 mL/min) and detected
at multiwavelength i.e. 272, 230 the column was maintain at temperature 2830 oC.
PROCEDURE :
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. The recorded chromatograms of blank (Fig A)
and sample after 24 h are depicted in Fig B-D in acidic, alkaline and peroxide stress respectivel

RESULTSAND DISSCUTION :
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.
Observations and results of estimation
Agrus-NS
Wt. of

AUC (V)
Std.

syrup
(mL)

5.0

PPH

2357018

DEX

8702983

Amt. estimated (mg)

Sample
CET

5880978

% of Labeled claim

PPH

DEX

CET

PPH

DEX

CET

PPH

DEX

CET

2369752
2358341
2364760

8599511
8670524
8689577

5949972
5910544
5978647

7.53
7.49
7.51

14.93
15.05
15.09

5.00
4.97
5.02

99.30
100.12
100.34

99.60
98.94
100.08

2377597

8582191

5936232

7.55

14.90

4.99

99.68
99.20
99.47
100.0

99.10

99.37

2388058

8587387

5951167

7.58

14.91

5.00

99.16

99.62

2356439

8590850

5926674

7.48

14.91

4.98
Mean

99.20
99.53
0.54

99.21
99.47
0.39

0.55

0.39

SD
%RSD

1
100.4
5
99.12
99.65
0.50
0.51

Summary of estimation
Combined
PPH
dosage form
Mean*
99.65
SD
0.50
%RSD
0.51
*Mean of five/six observations

M5
DEX
99.53
0.54
0.55

CET
99.47
0.39
0.39

VALIDATION OF PROPOSEDV HPLC METHOD:

Accuracy, Precision, Srecision, Linearity,Robustness, Limit of detection (LOD) and limit of quantitation (LOQ)
CONCLUSION:
The method is precise, accurate, rapid, reasonably specific and rugged. The developed method was found to be superior with
respect 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.
REFERANCE :
1

Her Majesty Stationary Office Landon Medicine and Healthcare Products Regulatory Agency, London, (1994)
British Pharmacopoeia, Vol. II: 1300.

Rajan, Sekar, Colaco, Socorrina, Ramesh, N, Meyyanathan, Subramania N, Elango, K. (2014) Development and
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Biological Sciences,; 17(3); 429-433.

Thakkar K, Parmar V, Minal R, Meshram DB.( 2013) UV Spectrophotometric method for estimation of
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pharmaceutical and chemical sciences,; 2: 2277-2283.

Reddy PR, Reddy VK, Goud ES, Reddy YR. (2014) Development and validation of a ultra performance liquid
chromatographic method for assay of cetirizine dihydrochloride. International Journal of Pharmacy and

Pharmaceutical Sciences,; 6(2): 430-432.

Patel BN, Patel CN, Patel NB.(2014) The simultaneous determination of phenylephrine hydrochloride,
paracetamol,

chlorpheniramine

maleate

and

dextromethorphan

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hydrobromide

in

pharmaceutical