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A Study of Validation and Uncertainty in Real Samples of Nicotine Polacrilex Gum by Reverse Phase HPLC
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A reverse phase HPLC method was developed for quantitative estimation of nicotine in nicotine polacrilex chewing gums excessively used in nicotine replacement therapy (NRT) adopted to replace the nicotine from cigarettes used by smokers. Further the method was validated to ensure the feasibility of the method for its application in routine analysis. Parameters such as specificity, linearity, quantitation limits, precision, and accuracy were determined. The minimum detection limit was 0.25 μg/ml and LOQ was 0.75 μg/ml. Studies were also carried out to estimate the uncertainty associated with the analytical method by applying a bottom-up approach. All data appearing in this study complies with NABL 17025 requirements. It was implemented in our laboratory as a routine method and our laboratory was accredited. The uncertainty in each step was estimated.. The expanded uncertainty in the estimation of nicotine was ±0.06
Keywords
Nicotine Gum, RP-HPLC, Validation, Uncertainty.
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