RP-HPLC Method Development and Validation for the Analyisis of ...

International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701
Table: Robustness Results
Sample condition (100µg/ml) Area % of Recovery
Flow rate change
0.7ml/min
0.9ml/min
Wavelength Change
256nm
252nm
Mobile phase ratio change
MeOH: ACN: 5%OP
78 17 5 (v/v/v)
82 13 5 (v/v/v)
548253
543251
543220
542545
544569
547140
100.64
99.72
99.72
99.53
99.97
100.44
Table: System suitability parameters
Parameter
Values
λ max (nm) 254
Beer’s law limit (µg/ml) 60-160
Correlation coefficient 0.999
Retention time
4.282min
Theoretical plates 9796
Tailing factor 1.36
Limit of detection (µg/ml) 5
Limit of quantification (µg/ml) 15
Ruggedness
Inter day variations were performed by using six
replicate injections of standard and sample
solutions of concentrations which were prepared
and analyzed by different analyst on three different
days over a period of one week. Ruggedness also
expressed in terms of percentage relative standard
deviation.
Ruggedness
Table: Ruggedness results
Sample
(100µg/ml)
Area
1 549585
2 546858
3 544990
4 546202
5 546623
6 550021
Result
R.S.D= 0.363
System suitability
The system suitability parameter like capacity
factor, asymmetry factor, tailing factor and number
of theoretical plates were also calculated. It was
observed that all the values are within the limits
(Table.8). The statistical evaluation of the proposed
method was revealed its good linearity,
reproducibility and its validation for different
parameters and let us to the conclusion that it could
be used for the rapid and reliable determination of
Telaprevir in tablet formulation. The results are
furnished in Table.
Assay of Telaprevir tablets
Weigh 20 Telaprevir (Gluconil-5mg) tablets and
calculate the average weight. Accurately weigh and
transfer the sample equivalent to 10mg of
Telaprevir in to a 10ml volumetric flask. Add
diluent and sonicate to dissolve it completely and
make volume up to the mark with diluents. Mix
well and filter through 0.45um filter. Further
pipette 1ml of the above stock solution into a 10ml
volumetric flask and dilute up to mark with diluent.
Mix well and filter through 0.45um filter. An
aliquot of this solution was injected into HPLC
system. Peak area of Telaprevir was measured for
the determination. The results are furnished in
Table.
Table: Formulation results of Telaprevir
Formulation Dosage Concentrati
Incovek
(Tablet)
on
Amount
found
375mg 100ppm 99.91pp
m
%
Estimated
99.91
CONCLUSION
A validated RP-HPLC method has been developed
for the determination of Telaprevir in tablet dosage
form. The proposed method is simple, rapid,
accurate, precise and specific. Its chromatographic
run time of 10 min allows the analysis of a large
number of samples in short period of time.
Therefore, it is suitable for the routine analysis of
Telaprevir in pharmaceutical dosage form.
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