ABSTRACT
Youden's test is a reliable method to evaluate the robustness of analytical methods, by means of an experiment design which involves seven analytical parameters combined in eight tests. In the present study, we assessed the robustness of a chromatographic method to quantify lumefantrine in raw material samples, using Youden's test. Hence, it was possible to determine the effect of each analytical parameter in the final analysis results. Youden's test showed to be a simple and feasible procedure to evaluate the robustness of chromatographic methods.
O teste de Youden constitui um método confiável para avaliação da robustez de métodos analíticos, por meio de um delineamento que envolve sete parâmetros analíticos combinados em oito experimentos. No presente estudo, a robustez de um método cromatográfico para quantificação de lumefantrina em matéria-prima farmacêutica foi avaliada utilizando-se o teste de Youden. Assim, foi possível determinar o efeito de cada parâmetro analítico nos resultados finais das análises. O teste de Youden se mostrou um procedimento simples e confiável para a avaliação da robustez de métodos cromatográficos.
Subject(s)
Antimalarials/analysis , /statistics & numerical data , /methods , Chromatography, Liquid/standards , Evaluation Studies as Topic , Analysis of Variance , Reference Standards/analysisABSTRACT
Background & objectives: β-thalassaemia is a genetic disorder and an important health problem around the world. Quantitative haemoglobin A2 (HbA2) levels are used for the diagnosis of β-thalassaemia. The conventional methods are high performance liquid chromatography (HPLC), electrophoresis, and microcolumn chromatography techniques. We established a fast protein liquid chromatography (FPLC) method, to measure quantitatively of HbA2 levels, and compared its efficacy with conventional methods. Methods: The FPLC method, using a DEAE Sepharose, Hi Trap anion-exchange column chromatography technique was set up for HbA2 measurement. In this study, 220 blood samples were screened for haemoglobin type by FPLC technique and also using HPLC, microcolumn chromatography and electrophoresis. Results: The FPLC results were highly correlated (r = 0.985, P<0.001) with those of HPLC for quantification of HbA2 as well as cellulose acetate electrophoresis (r = 0.977) and microcolumn chromatography (r = 0.980). The FPLC method showed 100 per cent sensitivity and specificity, positive and negative predictive value for β-thalassaemia diagnosis. In addition, the FPLC method was simple, rapid, low cost and reproducible. The HbA2/E range of FPLC for β-thalassaemia was 6-10 per cent, HbE trait was 10-40 per cent, β-thalassaemia/HbE was 40-60 per cent and homozygous HbE was more than 60 per cent. Interpretation & conclusions: Our findings suggested that FPLC method could be used as a cost-effective method for routine β-thalassaemia diagnosis.