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Coffein Uv Vis
Coffein Uv Vis
1
Department of chemistry, SCSVMV University, Kanchipuram
2
Department of pharmaceutical analysis, Jagans college of pharmacy, Nellore.
INTRODUCTION
Caffeine is a central nervous system and metabolic All the chemicals used were of analytical
stimulant, and is used both recreationally and grade. A gift sample of caffeine obtained from
medically to reduce physical fatigue and to restore Microlabs, Hosur, was used as working standard. The
alertness when drowsiness occurs [1-3]. It produces formulation of caffeine tablet was purchased from
increased wakefulness, faster and clearer flow of online shopping.
thought, increased focus, and better general body
coordination. According to literature survey, it Solubility test
revealed that caffeine was quantitatively assayed in Solubility test for the drug Caffeine was
Tea, coffee and food beverages using liquid performed by using various solvents. The solvents
chromatography, UV-Visible spectroscopy How-ever include Water, Methanol, Ethanol, Acetonitrile,
no UV-Spectrophotometry method was proposed Hydrochloric Acid (HCl), Sodium Hydroxide, Nitric
for the estimation of caffeine by using of water as a acid, Sulphuric acid and Chloroform.
solvent in Tablet dosage forms [4-7]. In the present
study to develop a simple, accurate and precise UV DETERMINATION OF λ max
spectroscopic method for estimation of caffeine in Preparation of stock solution
tablet dosage form. The validation carried out as per Standard stock solution of Caffeine was
ICH guidelines [8-10]. prepared by dissolving 100mg of Caffeine in 100ml
of distilled water which gives 1000 µg/ml.
EXPERIMENTAL WORK
Apparatus Preparation of working solution
A Systronics double beam UV visible From the above stock solution 10ml was
spectrophotometer model 2202, band width of 2nm transferred into 100ml volumetric flask and volume
wavelength accuracy ±0.5nm and two matched was made up to the mark with distilled water to
quartz cells with 1cm pathlength was used for all make 100µg/ml. Then the sample was scanned with
spectral measurements.
To whom correspondence should be addressed:
Materials S.Sethuraman
Email: su_sethuraman@yahoo.com
Method validation
Validation is a process of establishing
documented evidence, which provides a high degree
of assurance that a specific activity will consistently
produce a desired result or product meeting its
predetermined specifications and quality
Preparation of calibration curve characteristics. The validation for UV method
One ml of this 100µg/ml solution was development was performed using parameters like
further diluted and the volume was made up to 10ml Linearity, Accuracy, Precision, Robustness, and
by using method to produce 10µg/ml solution. 2ml, Ruggedness.
3ml, 4ml and 5ml of 50µg/ml solution were diluted
and the volume was made up to 10ml using Linearity
methanol to produce 20µg/ml, 30µg/ml, 40µg/ml, Various aliquots were prepared from the
50µg/ml solutions respectively. Then the secondary stock solution (100 µg/ml) ranging from
construction of calibration curve was done by taking 10-50 µg/ml. The samples were scanned in UV-Vis
the above prepared solutions of different Spectrophotometer against distilled water as blank.
concentration ranging from 10-50µg/ml. Then taking It was found that the selected drug shows linearity
the absorbance calibration curve was plotted taking between the ranges of 10-50 µg/ml.(Table no.3)
concentration on x-axis and absorbance on y-axis
which showed a straight line. This straight line Accuracy
obeyed linearity in the concentration range of Solutions were prepared in triplicate at
10-50µg/ml. The correlation coefficient was found to levels 80%, 100% and 120% of test concentration
be 0.999 using Caffeine working Standard as per the test
method and taken absorbance of each solution in
Fig 2. Linearity curve of Caffeine triplicate. The recovery results showed that the
proposed method has an acceptable level of
accuracy for Caffeine which is from 80% - 120% of
test concentration is from 99.51 % - 100.01 %(Table
4).
Precision
Precision of the method was demonstrated
by intraday and interday variation studies. In
intraday variation study nine different solutions of
same concentration 50µg/ml were analyzed three
times in a day i.e. from morning, afternoon and
evening and the absorbance is note. In the interday
variation studies, solution of same concentration
50µg/ml were analyzed three times for the three
Table 1. Robustness
At 268nm At 272nm
Concentration(µg/ml) Absorbance Statistical Concentration(µg/ml) Absorbance Statistical
analysis analysis
50 0.535 Mean=0.5353 50 0.535 Mean=0.5348
50 0.535 50 0.536
50 0.536 SD=0.00103 50 0.535 SD=0.00116
50 0.537 50 0.534
50 0.535 %RSD=0.19 50 0.533 %RSD=0.21
50 0.534 50 0.536
Table 2. Ruggedness
Analyst 1 Analyst 2
Concentration(µg/ml) Absorbance Statistical Concentration(µg/ml) Absorbance Statistical
analysis analysis
50 0.535 Mean=0.5355 50 0.534 Mean=0.5348
50 0.536 50 0.535
50 0.535 SD=0.000837 50 0.535 SD=0.000753
50 0.535 50 0.534
50 0.537 %RSD=0.15 50 0.536 %RSD=0.14
50 0.535 50 0.535
Slope 0.009
Intercept 0.042
Parameter Result
Linearity indicated by correlation coefficient 0.999
Precision indicated by % RSD 0.26%
Accuracy indicated by % recovery 99.9044%
Specificity No interferences of impurity
Range 10µg-50µg/ml
Linear regression equation 0.009x + 0.042
Robustness indicated by %RSD 0.19%
Assay indicated by % recovery 99.89%
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