Desarrollo y validación de un método analítico para la determinación de vitaminas hidrosolubles en una forma farmacéutica líquida (jarabe) por HPLC en la empresa Neofarmaco Cia. Ltda.

Abstract

This work describes the development, optimization and validation of a reversed phase high-performance liquid chromatography (HPLC) method coupled with an ultraviolet visible detector (UV-VIS) by means of the conjugated ion pair technique (CIP), for the quantification of ascorbic acid, thiamine, riboflavin, nicotinamide, and pyridoxine in combined standard and loaded placebo. A LiChrospher® 100 RP-18 endcapped column (125 x 4.6 mm, 5 um) was used. UV detection was performed with a gradient of 0.0-6.0 min at 267 nm, 6.1-12.0 min at 290 nm, 12.1-18 min at 248 nm, and 18.1-24 min at 340 nm. The mobile phase consisted of a CIP 7 buffer pH 2.0, methanol and acetonitrile using the gradient: 0-16 min (80: 17.5: 2.5), 16-21 min (17.5: 80: 2.5), and 21-24 min (80: 17.5: 2.5). The flow was 1 ml / min at 35 ° C. The injection volume of the sample was 10 ul. The method was validated after the development and optimization of it, according to category I of analytical procedures for the quantification of the main components of bulk drugs or active ingredients (including preservatives) in finished pharmaceutical products, described in the USP for the validation, proving to be selective with a percentage of discrepancy less than 3.465%, linear (r> 0.998), precise (RSD <1.8247), accurate (recovery of 97.2630% for ascorbic acid, 98.3540% for thiamine, 99.8853% for riboflavin, 97.8945% for nicotinamide and 98.9522% for pyridoxine), suitable and robust; furthermore, in the robustness test chromatographic parameters were compared with octylsilane C8, showing a discrepancy coefficient of less than 5%. According to these results, it was demonstrated that the analytical method developed, optimized and validated is appropriate for the identification and quantification of water-soluble vitamins in a liquid pharmaceutical form and can be applied for routine control in the laboratory.