Standard Preparation:
100 mg of standard ascorbic acid was weighed precisely and transferred to a 100 ml volumetric flask, added 70 ml of 0.5% sodium metabisulphite and dissolved by shaking. The volume was made up to the mark with 0.5% sodium metabisulphite for getting a concentration of 1 mg/ml. 2 ml of this solution was taken into another 100 ml volumetric flask and made the volume up to the mark with 0.5% sodium metabisulphite which resulted in concentration of 0.02 mg/ml. The solution was filtered through 0.45 µ nylon syringe filter.
Sample Preparation:
2.5 g of sample was weighed accurately and transferred to a 100 ml volumetric flask. 60 ml of 0.5% sodium metabisulphite was added and dissolved by sonication for 1 minute. The volume was
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The % RSD values were calculated.
Interday precision:
The Interday precision this method was checked by injecting six different preparations of proposed sample weight (100%) on next day of method precision. The % RSD values were calculated.
Accuracy:
Reliability and accuracy of the methods were studied based on recovery studies. Analysis was carried in three replicates with the spiking standard ascorbic acid (99.9% purity) concentration of 0.02 mg/ml at the levels of 50%, 100%, and 150%. The % recovery was calculated.
Range:
The data generated in precision and linearity was considered for establishment of Range. The sample weight of 1250 mg to 3750 mg was taken for analysis and it was found to be within the linearity and precise.
Robustness:
Reliability of the method was done about deliberate variations in method parameters like
a. Variation in the flow rate of mobile phase (±0.1 ml/min)
b. Variation in cooler temperature (±2 nm)
The entire sample, standard and diluent was analyzed for small changes in method parameters by injecting duplicate injections of single preparation.
RESULTS AND