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Sabyasachi Mandal
Himalaya Drug Company
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1
Department of Quality Assurance, Acharya and B. M Reddy College of Pharmacy.
Bangalore – 560107.
INTRODUCTION
Common cold related to virus is quite common phenomena in our daily life. Due to week
immunity of young age children become more vulnerable to this common cold virus. The
symptom related common cold like headache, running nose, nasal congestion and sneezing
may sometime become troublesome in our common life. The common cold symptoms
usually resolve in seven to ten days but sometimes it can persist for up to three weeks. The
average duration of cough is around 18 days. So to reduce the symptoms sometimes
physician prescribe three or more combination drugs in the form of tablet or syrup. The most
commonly used combination drugs like Paracetamol, cetirizine and pseudoephedrine are
commonly available in market. Paracetamol are used mainly to reduce the headache and fever
related to the common cold. Cetirizine acts as an antihistaminic drug. Pseudoephedrine
reduces nasal congestion and acts as an effective decongestant.Due to different mechanism of
these drugs, the combination dosage form of these three drugs become one of the best options
for general medical practitioner to prescribe for common cold patients. The simultaneous
analysis of these drugs by RP-HPLC in pharmaceutical industry not only reduces the time of
analysis but also it is most cost effective.
So the need of our present study is to provide a simple, sensitive, precise and accurate
validated method of RP-HPLC analysis which will reduce the run cost of routine analysis.
EXPERIMENTAL
Instrumentation
RP-HPLC instrument (Shimadzu, Japan) equipped with a SPD 20A UV-visible detector and
LC-20AT pump, manual Rheodyne injector with 20 µl loop, Phenomenex Luna C18 column
(250 mm×4.6mm i.d., 5µ particle size) and LC solution software.
Chromatographic conditions
Chromatographic separation was carried out on Phenomenex Luna C18 column (250 mm x
4.6 mm id, 5µ), mobile phase of Acetonitrile: 0.025 mM Potassium dihydrogen
orthophosphate buffer pH 7.5(50:50 v/v), with flow rate of 0.5 ml/min, detection wavelength
at 220 nm,temperature 25 C and Injection volume was 20 μL.
Linearity
Thelinearity studies were performed to ensure that the test results are directly proportional to
the concentration of analyte. 20 µl of each standard solution of Pseudoephedrine (5-40
µg/ml), Paracetamol (75-600 g/ml) and Cetirizine (2.5-20 g/ml) was injected into HPLC
system. The peak area v/s concentration were plotted to get a standard calibration curve.
Precision
Precision of the assay was determined by intra-day and inter-day precision of the developed
method.Intra-day precision was done by three different analysts in same day whereasInter-
day precision was done in three different days. The results are presented in terms of %
relative standard deviation (% RSD)(Table 3& 4).
Accuracy
The accuracy of an analytical method is the closeness of test results obtained by that method
to the true value. The accuracy was performed by addition of known amounts of standard
drug. The accuracy of the method was determined by calculating percentage recovery of
known added amounts of analytes (Table 5, 6 & 7).
Robustness: This parameter was carried out to check the ability of the system to give
unaffected results for small deliberate changes in system parameters and method parameters.
Robustness was performed by changing the flow rate, wavelength and mobile phase ratio for
standard drugs as well as formulations.
Assay: Formulation used was Tablet and Brand Name – Kold time (500 mg of Paracetamol +
30 mg Of Pseudoephedrine + 10 mg of Cetirizine)
2.5 ml from the sample stock solution of 1000 µg/ml containing PCM, CTZ and PSEDN,
transferred into 10 ml volumetric flask and volume adjusted up to 10 ml with mobile phase to
get 250, 5 and 15 µg/ml concentration respectively.
DISCUSSION
The chromatographic parameters were fixed and HPLC system was studied for the suitability
of drug analysis. The system suitability parameters were given in Table 1. The developed
method was validated to make suitable it for drug analysis. Validation of the HPLC method
was performed for linearity, precision, accuracy, specificity, robustness, LOD and LOQ.
CONCLUSION
HPLC method was developed and validated as per ICH guidelines. It can be concluded that
the method is specific for estimation of PSEDN, PCM and CTZ in pharmaceutical dosage
form. The method has linear response in stated range and is accurate and precise.Statistical
analysis proves that the method is suitable for the analysis of PSEDN, PCM and CTZ as bulk
drug and in pharmaceutical formulation without any interference from the excipients. It may
be extended to study the degradation kinetics of PSEDN, PCM and CTZ and also for its
estimation in plasma and other biological fluids andImpurity profiling for both the drugs.
ACKNOWLEDGEMENT
The authors would like to thank Medriech, Micro Labs, Bangalore andAcharya and B M
reddy college of pharmacy, Bangalore for providing sample of Paracetamol, Cetirizine and
Pseudoephedrine bulk drug.
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