Determinación de vitamina B2 (Riboflamina) por Cromatografía líquida de alta eficacia (HPLC) en el Laboratorio ECUACHEMLAB Cía. Ltda.

Abstract

The validation of analytical methods is an essential process that laboratories must implement to ensure that the results of analyzes are reliable and reproducible with a basis and statistical analysis. Similarly, the methodology used for validation shows a great performance in working conditions of Ecuachemlab. For the determination of Vitamin B2 (Riboflavin) using high-performance liquid chromatography (HPLC), the methodology described in the AOAC 970.65 Method modified and ISO / IEC 17025 were used. They describe the guidelines and parameters for validation of the method used to ensure the validity results. The analytical parameters determined for method validation are linearity, working range, detection and quantification limits, precision, accuracy and uncertainty. The results, for linearity show a coefficient of correlation higher than r = 0.999 for each calibration curve, LD is equal to 0.002 ug / ml and the LC is 0.08 ug / ml. The working range comprises a concentration of 0.008 mg / ml of riboflavin as a lower limit and 1,003 ug / ml of riboflavin as an upper limit. The coefficient of variation for repeatability and reproducibility is less than 3 and 5% respectively. To determine the accuracy, a Z-score within the limits for each MRC was considered. Finally it was identified a combined and expanded uncertainty for the first three levels set of +/- 0.04 and +/- 0.08 mg vitamin B2 per 100 g of sample respectively. For the last level value it was established an uncertainty value of 0.05 +/- combined and expanded uncertainty equal to +/- 0.09 mg vitamin B2 per 100 g of sample, working at a confidence interval of 95% with a coverage factor k = 2.