Otimização e validação de método analítico para determinação simultânea de citrato e fosfato em soluções anticoagulantes de bolsas de sangue por CLAE

Blood bags are products of high complexity, classified as risk III, which are designed to collect, store and transfer blood in an efficient and safe manner. The national reference for analyzes of this product is the National Institute of Quality Control in Health (INCQS / FIOCRUZ), responsible for the pre-registration and control of these products. Resolution RDC no 35 of 2014 establishes the conditions required for blood bags with anticoagulant and / or preservative solutions. Total citrate and diacid sodium phosphate monohydrate, present in the anticoagulant and / or preservative solutions, are of great importance in maintaining the viability of the collected blood. The proposed method uses ion chromatography and allows the simultaneous determination of the total citrate and sodium diacid phosphate monohydrate contents in the anticoagulant solutions, reducing the analysis time. In this context, the objective of this study was to perform the optimization and validation of the analytical method for the simultaneous determination of total citrate and sodium diacid phosphate monohydrate in anticoagulant solutions of blood bags by high performance liquid chromatography using ionic detection. This study is divided into four stages: the retrospective evaluation of the samples, the comparison of the analytical methods, the planning of the analytical validation, the adequacy of the system and the optimization of the methodology and the analytical validation. The selection of the work samples was performed considering on the retrospective evaluation of the samples submitted to the INCQS in the period from 2012 to 2016. The analytical conditions of the individual and concomitant analysis of these substances were evaluated. The optimization of the analytical methodology, second stage, was evaluated the flow of the mobile phase and the dilution of the samples and standards allowing the reduction of the time of analysis and the operating cost with efficiency of separation proved by the parameters of suitability of the chromatographic system. The analytical validation, the third stage of the study, was performed according to the parameters recommended by Anvisa, in RE no 899 of 2003 and Resolution no 166 of 2017, and by Inmetro, in the guideline DOQ-CGCRE008 of 2016. In the comparison of methods, the statistical evaluation showed that there is no significant difference between the two methods for the analyzes performed. In the analytical validation stage, the selectivity, linearity, limits of detection and quantification, precision, accuracy and robustness presented satisfactory results that proved the reliability of the method, indicating that the alternative method is reproducible. The study allowed to compare the analytical conditions and to present the relevant aspects of the implementation of the alternative method, mainly, to allow to simultaneously determine the analytes of interest. The validation of this alternative method meets the criteria of both Anvisa and Inmetro, the new method allows a reduction of the cost of exports and the measure of current legislation is not a reference in blood bags not in Brazil. In addition, it suggests new advances without product quality control and reiterates INCQS participation in sanitary quality regulations.