Kullai Reddy Ulavapalli et al.

/ Journal of Pharmacy Research 2011,4(4),1219-1221

Research Article
ISSN: 0974-6943 Available online through
www.jpronline.info
Simultaneous determination of psuedoephedrine, fexofenadine and loratadine in pharmaceutical
products using high resolution RP-HPLC method
Kullai Reddy Ulavapalli 1, J. Sriramulu 1, Useni Reddy Mallu 1 and Varaprasad Bobbarala2*.
1
Dept of Chemistry, Sri Krishnadevaraya University, Anantapur, AP, India-515003.
2
Translational Research Institute of Molecular Sciences (TRIMS), 2-35-72, Sai Narasimha Towers, MVP Colony, Sector-10, Visakhapatnam – 530017, AP, India
Received on: 05-12-2010; Revised on: 14-01-2011; Accepted on:09-03-2011
ABSTRACT
High resolution RP-HPLC method has been developed for the simultaneous estimation of Psuedoephedrine, Fexofenadine and Loratadine in pharmaceutical products. Separation was achieved
by using C18, 150 x 4.6 mm, 5µm column at a temperature of 350C with a gradient mobile phase composed of sol-A: 0.01M pH 3.0 phosphate (KH2PO4) buffer, sol-B: Acetonitrile and gradient
program was 0-5min, sol-A: 80-60; 5-10min- sol-A: 60-20; 10-15min- sol-A: 20-20; 15-17min- sol-A: 20-80 and 17-20min- sol-A: 80-80. Flow rate was 0.60 ml per min and measured the
absorbance at 210nm. The retention time of Psuedoephedrine, Fexofenadine and Loratadine are 4.3min, 10.4min and 17.0min, respectively. The linearity of the proposed method was evaluated
from 5µg per mL to 100µg per mL and the correlation coefficient results were within the limit (not less than 0.999). The developed method has wide applicable for the quantification of
Psuedoephedrine, Fexofenadine and Loratadine in pharmaceutical dosage forms.

Key words: Psuedoephedrine, Fexofenadine, Loratadine, high resolution and RP-HPLC method.

INTRODUCTION
Fexofenadine (1-8) is indicated to relieve symptoms that are associated with seasonal allergic
rhinitis, such as sneezing, rhino rhea, nose and throat, itchy eyes and red, watery eyes.
Fexofenadine and pseudoephedrine combination is used an antihistamine and decongestant
which provides relief of seasonal allergy symptoms. In cases of seasonal allergic rhinitis the
recommended dosage of fexofenadine is 120 mg daily in a single dose or in two divided doses
for patients 12 years of age and older. In cases of chronic idiopathic urticaria once-daily doses
of 180 mg fexofenadine is recommended. Loratadine (9-12) a class of medications called antihis-
tamines and used to temporarily relieve the symptoms of hay fever (allergy to pollen, dust, or
other substances in the air) and other allergies. These symptoms include sneezing, runny nose,
and itchy eyes, nose, or throat. Loratadine is also used to treat itching and redness caused by
hives. Loratadine is also available in combination with pseudoephedrine. Pseudoephedrine (13-
16) reduces tissue hyperemia, edema and nasal congestion, colds or allergies. Adverse effects are
hypertension, sweating and anxiety. Pseudoephedrine is useful for suppressing of cough.
Chemical structures of Psuedoephedrine, Fexofenadine, Loratadine are represented in figure-1.
Some of available pharmaceutical dosage forms are listed in table-1. Psuedoephedrine,
Fexofenadine and Loratadine have chemical and individual methods (17-21). In the present study
developed a single and high resolution RP-HPLC method for three active ingredients includ-
ing Psuedoephedrine, Fexofenadine, Loratadine in pharmaceutical products and validated the
method with specificity, linearity, accuracy and reproducibility.
MATERIALS AND METHODS
Materials & reagents:
AR grade of Potassium di-hydrogen phosphate (Merck India Pvt.Limited), acetonitrile, metha-
nol, Orthophosphoric acid and Millipore purified water were used for analysis. All purified
standard materials (not less than 98.5%) were used. All market samples were analysed with this
method.
Equipments:
Waters Alliance HPLC system equipped with 2695 separation module connected to 2996-
Photo diode array detector and Empower-2 software and Agilent 1200 series HPLC with dual
absorbance detector were used. Mettler-Toledo analytical balance, sonicator and rotatory shaker
were used for this study.
Columns:
Waters, Agilent, Thermo-scientific, Merck and GL-life sciences make analytical columns were
used for this study.
Chromatographic conditions:
Mobile phase composed of Sol-A: 0.01M Potassium di-hydrogen phosphate buffer (adjusted
the pH to 3.0 with diluted ortho-phosphoric acid) and Sol-B: acetonitrile. 10µl injection
volume, 0.60ml per minute flow rate with a simple gradient elution (0-5min, sol-A: 80-60; 5-
10min- sol-A:60-20; 10-15min- sol-A:20-20; 15-17min- sol-A:20-80 and 17-20min- sol-
A:80-80). Detection of the all three actives was achieved by using a photo diode array (PDA)
detector at 210nm. Inertsil ODS-3V (GL Sciences) 150mm x 4.6mm, 5µm column is used for
analysis and column oven temperature maintained at 350C. Methanol is used as diluent.

Standard preparation:
Standard stock solution (500µg/ml) was prepared in methanol which is used as diluent.
Weighed accurately 50mg of each standard (Pseudoephedrine, Fexofenadine and Loratadine)
Pseudoephedrine Fexofenadine Loratadine into 100ml volumetric flask and dissolved in diluent with intermediate sonication, diluted to
Figure-1: Chemical structure of active ingredients volume with diluent and mixed. Further diluted the resulting solution to achieve a concentra-
tion of 50µg per ml
.
Table-1: Available dosage forms in market.
Sample Preparation:
Dosage form Composition All market samples (combination and individual formulations) were analyzed with the devel-
Tablets Fexofenadine-120mg and Pseudoephedrine (ER)-180mg oped method by preparing 50µg per ml with diluent.
Tablets Fexofenadine-30mg, 120mg and 180mg
Syrup Fexofenadine-30mg per 5mL Calculation:
Tablets Loratidine-5mg and 10mg All active ingredients are quantified with the following calculation.
Syrup Loratadine-5mg per 5mL
Suspension Loratadine-1mg per 1mL Sample area x standard dilution factor
Tablets Loratadine-5mg and pseudoephedrine-120mg ————————————————————————x 100
Loratadine-10mg and pseudoephedrine-180mg Standard area x sample dilution factor

*Corresponding author. RESULTS AND DISCUSSION

Dr. Varaprasad Bobbarala Initial stage of method development trials were performed with acetate buffer and acetonitrile by
Chief Scientist, using waters make C18, 150mm, 4.6 mm ID, 5µm column. 210nm of UV absorbance was
Translational Research Institute of Molecular Sciences, selected for three active ingredients from their UV spectrum. Three active ingredients spectrums
2-35-72, Sai Narasimha Towers, MVP Colony, Sector-10, represented in figure-2. All three active components were eluted but the retention times of all
Visakhapatnam – 530017, AP, India. three components found close to each other and poor baseline. Finally the separation was
Tel.+ 91-9949129539, achieved with phosphate buffer and Acetonitrile with gradient program. The retention time of
E-mail:phytodrugs@gmail.com

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Table-4: System suitability (USP Tailing)
Active Ingredient Tailing factor
Name Injection-1 Injection-2 Injection-3 Injection-4 Injection-5 Average

Psuedophedrine 1.4 1.4 1.4 1.4 1.4 1.4

Fexofenadine 1.5 1.5 1.5 1.5 1.5 1.5
Loratadine 1.1 1.1 1.1 1.1 1.1 1.1

Figure-2: UV spectrum of three active ingredients

Figure-5: System suitability Solution

METHOD VALIDATION
Validated the method as per ICH and FDA (22- 26) guidelines with parameters like specificity,
precision, accuracy, linearity and range, ruggedness and robustness etc.
Figure-3: Blank Chromatogram
Specificity:
Specificity of the method has evaluated with diluent and placebo interference. System suitabil-
ity and precision results found to be good and no interference was observed with diluent and
placebo.

Precision:
Precision of the method was evaluated by carrying out six different sample preparations for all
individual and combination products. Percentage relative standard deviation (% RSD) was
less than 1.5% for within a day and day to day variations, which proves that the method is
precise and results were tabulated in Table-5.

Table-5: Precision Results.

Active Ingredient Name Sample preparations Average (%)

Prep-1 Prep-2 Prep-3 Prep-4 Prep-5 Prep-6

Psuedophedrine 100.01 99.91 99.13 99.65 99.85 98.90 99.71

Figure-4: Standard chromatogram Fexofenadine 99.88 98.90 99.74 99.09 100.10 99.80 99.54
Loratadine 100.40 99.50 100.18 99.18 99.81 99.81 99.81
System suitability
System suitability test is an integral part of analytical method development. System suitabil-
ity parameters are established by injecting the diluent and freshly prepared standard solution Linearity:
(each active 50ppm) in to the system for five replicate injections and calculated the percentage Linearity was performed by analyzing eight different concentrations of the standard solution of
relative standard deviation for area and retention time, results found to be satisfactory. Five all three active ingredients mixture. Calibration curve was constructed by plotting area against
replicate standard solution chromatograms represented in figure-5 and tabulated the results in concentration and area. The results were shown in table-6. The correlation coefficient value was
within the limit 0.999 for all three compounds. All linearity solutions overlaid and represented
table-2, 3 and 4.
in figure-6.
Table-2: System suitability (Area %RSD)
Table-6: Linearity Results.
Active Ingredient Standard solution Area
Name Injection-1 Injection-2 Injection-3 Injection-4 Injection-5 Average %RSD
Linearity solutions area Active Ingredient Name
Psuedophedrine 1293288 1283931 1289692 1289377 1278835 1287025 0.44 Psuedophedrine Fexofenadine Loratadine
Fexofenadine 3220316 3197781 3178660 3159160 3154911 3182166 0.86
Loratadine 5011359 4909016 4932364 4926712 4952375 4946365 0.80 Level-1 (5ppm) 126200 321000 490013
Level-2 12.5ppm 317255 800945 1241908
Table-3: System suitability (Retention time %RSD) Level-3 (25ppm) 627796 1602371 2489320
Level-4 (37.5ppm) 944042 2365772 3696274
Active Ingredient Standard solution Retention time (min) Level-5 (50ppm) 1290914 3188314 4953353
Name Injection-1 Injection-2 Injection-3 Injection-4 Injection-5 Average %RSD Level-6 (62.5ppm) 1608032 3937688 6096792
Level-6 (75ppm) 1929033 4740215 7361086
Psuedophedrine 4.355 4.335 4.359 4.342 4.337 4.35 0.25 Level-7 (87.5ppm) 2241482 5579386 8728110
Fexofenadine 10.424 10.405 10.411 10.393 10.394 10.41 0.12 Level-8 (100ppm) 2569051 6386648 9863271
Loratadine 17.093 17.049 17.001 16.984 16.960 17.02 0.31 Co-relation Coefficient 0.9999 0.9999 0.9999

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 Kullai Reddy Ulavapalli et al. / Journal of Pharmacy Research 2011,4(4),1219-1221
for all ingredients (Pseudoephedrine, Fexofenadine and Loratadine) and applicable for routine
quality control analysis.

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Source of support: Nil, Conflict of interest: None Declared

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