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RESEARCH ARTICLE
Article Info
Article history
Received 25 Sep 2013
Revised 8 Oct 2013
Accepted 17 Oct 2013
Available online 30 Oct 2013
Keywords
Caffeine,
validation.
UV-spectroscopy,
Abstract
INTRODUCTION
Caffeine is a central nervous system and metabolic
stimulant, and is used both recreationally and
medically to reduce physical fatigue and to restore
alertness when drowsiness occurs [1-3]. It produces
increased wakefulness, faster and clearer flow of
thought, increased focus, and better general body
coordination. According to literature survey, it
revealed that caffeine was quantitatively assayed in
Tea, coffee and food beverages using liquid
chromatography, UV-Visible spectroscopy How-ever
no UV-Spectrophotometry method was proposed
for the estimation of caffeine by using of water as a
solvent in Tablet dosage forms [4-7]. In the present
study to develop a simple, accurate and precise UV
spectroscopic method for estimation of caffeine in
tablet dosage form. The validation carried out as per
ICH guidelines [8-10].
EXPERIMENTAL WORK
Apparatus
A Systronics double beam UV visible
spectrophotometer model 2202, band width of 2nm
wavelength accuracy 0.5nm and two matched
quartz cells with 1cm pathlength was used for all
spectral measurements.
Materials
S.Sethuraman
Email: su_sethuraman@yahoo.com
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S.Sethuraman et al., Analytical Method Development And Validation Of Caffeine In Tablet Dosage Form By Using
UV- Spectroscopy
UV-Vis Spectrophotometer in the range 200-400nm
against distilled water as blank and the wavelength
corresponding to maximum absorbance was noted
which is its -max i.e. at 270nm.
Fig 1. -max of Caffeine
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S.Sethuraman et al., Analytical Method Development And Validation Of Caffeine In Tablet Dosage Form By Using
UV- Spectroscopy
consecutive days and the absorbance result mean,
standard deviation and %RSD was calculated and
summarised results given in table 4.
Specificity
Got spectrum in the range of 200nm to
400nm for appropriate concentration of sample,
Blank, and placebo.
Fig 3. Spectrum for blank
Robustness
Robustness of the method was determined
by carrying out the analysis under different -max
i.e. at 268nm and at 272nm. The respective
absorbances of 50g/ml were noted and the result
was indicated as %RSD and given in table no.1
Ruggedness
Ruggedness of the method was determined
by carrying out the analysis by different analyst and
the respective absorbance of 50 g/ml was noted.
The result was indicated as %RSD and given in table
no.2
RESULTS AND DISCUSSION
The drug was analyzed at 270nm in distilled
water using UV-Visible spectrophotometer. Optical
characteristics such as Beers law limits, intercept and
slope has been calculated using regression equation,
which has been presented in Table 3.
Precision was determined by analyzing the
drug at particular concentration for five times on the
same day. Inter day precision was determined
similarly, analyzing the samples daily, for three
consecutive days. The results are summarised in
Table 4.
To ensure the accuracy method, recovery
studies were performed by standard addition
method at 80%,100% and 120% levels of drug
concentration, to the pre-analyzed samples and
percent recovery values were calculated. Recovery
experiment indicated the absence of interferences
from the commonly encountered pharmaceutical
additives and excipients.
The linearity studies were performed by
plotting different concentration of standard solution
against their respective absorbances. Caffeine were
found to be linear in the concentration range of 1050g/ml. Correlation co-efficient value were found
to be 0.999, calibration curve shows that it obeys
Beers law limit within the concentration range.
The proposed method was found to be
simple, accurate, precise, simple, sensitive, robust
and cost effective. The results of the validation tests
were found to be satisfactory and therefore this
method can be applied successfully for the
estimation of Caffeine in Tablet dosage form.
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S.Sethuraman et al., Analytical Method Development And Validation Of Caffeine In Tablet Dosage Form By Using
Uv- Spectroscopy
Table 1. Robustness
Concentration(g/ml)
At 268nm
Absorbance
50
50
50
50
50
50
Statistical
analysis
Mean=0.5353
0.535
0.535
0.536
0.537
0.535
0.534
Concentration(g/ml)
At 272nm
Absorbance
50
50
50
50
50
50
SD=0.00103
%RSD=0.19
Statistical
analysis
Mean=0.5348
0.535
0.536
0.535
0.534
0.533
0.536
SD=0.00116
%RSD=0.21
Table 2. Ruggedness
Concentration(g/ml)
Analyst 1
Absorbance
50
50
50
50
50
50
0.535
0.536
0.535
0.535
0.537
0.535
Statistical
analysis
Mean=0.5355
Concentration(g/ml)
50
50
50
50
50
50
SD=0.000837
%RSD=0.15
Analyst 2
Absorbance
0.534
0.535
0.535
0.534
0.536
0.535
Statistical
analysis
Mean=0.5348
SD=0.000753
%RSD=0.14
10-50g/ml
Correlation coefficient
0.999
Regression equation(Y*)
0.009x + 0.042
Slope
0.009
Intercept
0.042
Result
0.999
0.26%
99.9044%
No interferences of impurity
Range
10g-50g/ml
0.009x + 0.042
0.19%
99.89%
REFERENCE
1. Wanyika HN, Gatebe EG, Gitu LM, Ngumba EK
and Maritim CW. Determination of caffeine
VOLUME 3 | NUMBER 4 | OCT | 2013
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UV- Spectroscopy
2.
3.
4.
5.
6.
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