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The results of the validation are presented in Table 1. Table 1 Statistical analysis of nicotinamide content in its commercial drugs using the proposed RPHPTLC-densitometric method. 3.4. Precision (Repeatability) The precision of the method was calculated in terms [%] relative standard deviation (coefficient of variation, CV) of intraday and interday precision of the method. Intraday and interday precision of the assay were determined using three samples replicates of nicotinamide preparations (Product 1, Product 2, and Product 3) at different concentrations of nicotinamide (0.25, 0.30, and 0.35?mg/mL). Precision was estimated by densitometric measurements of the peak areas obtained for analyzed samples and it was expressed as coefficient of variation (CV, %). The validation data showed that coefficient of variation for [http://www.selleckchem.com/products/gw3965.html GW3965 research buy] the determination of peak [http://www.selleckchem.com/screening/selective-library.html Target Selective Inhibitor Library nmr] areas in the three examined pharmaceutical formulations was placed in the following range: from 1.02% to 1.52% (intraday precision) and from 1.75% to 2.42% (interday precision) for Product 1 (tablets), 0.78% do 1.22% (intraday precision) and from 1.32% to 1.78% (interday precision) for Product 2 (tablets), from 0.89% to 1.33% (intraday precision) and from 0.98% to 1.76% (interday precision) for Product 3. The results of CV less than 3% confirmed precision of proposed method. 3.5. Sensitivity (Limit of Detection (LOD) and Limit of Quantification (LOQ)) The LODs and LOQs of nicotinamide were calculated on the basis of standard deviation of the intercept of specific calibration curve according to the formulae 2. The limit of detection and the limit of quantification determined by the use of developed method were found to be 0.15?��g/spot and 0.45?��g/spot, respectively. Because the standard deviation depends on concentration (amount) of analyzed compound, there is a need to check the correctness of obtained LOD values with the use of the following relationships observed between LOD value and concentration of examined substance [44]: 10��LOD>CLOD[https://en.wikipedia.org/wiki/Adenine Adenine] reference samples. Our results of LOD fulfilled the conditions presented in 4. Thus it confirms that the amount of nicotinamide used in preparation of specific calibration plot was well selected. 3.6. Quantitative Determination of Nicotinamide in Commercial Samples (as Vitamin PP) The validity of the proposed RPHPTLC method in combination with densitometry was applied for the accurate determination of nicotinamide in its pharmaceutical preparations: Product 1 and Product 2 (tablets) and Product 3 (injection solution). In this study, sample solutions of the investigated preparations in quantity of 1.50?��g of nicotinamide/spot were chromatographed on silica gel 60 RP18WF254 and with the use of methanol-water (3?:?7, v/v).