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O escopo da presente pesquisa foi desenvolver e validar uma metodologia para extracao simultânea de colesterol e oxidos de colesterol utilizando saponificacao direta em amostras de leite fluido para posterior analise por cromatografia liquida de alta eficiencia em fase reversa com deteccao por arranjo de diodos (RP-HPLC-DAD). Foram avaliados os parâmetros analiticos de seletividade, linearidade e faixa de aplicacao, precisao inter e intradia, recuperacao, limites de deteccao e quantificacao do metodo. Os resultados demonstraram que a metodologia pode ser empregada com seguranca e confiabilidade para analise do colesterol e oxidos do leite. The aim of the present work was to develop and validate a reversed-phase highperformance liquid chromatography with diode array detection (RP-HPLC-DAD) method for determination of cholesterol and cholesterol oxides extracted from milk samples. The method consisted of direct samples saponification and analytes extraction, followed by their determination using RP-HPLC-DAD. Suitable analytical selectivity, linearity and range of application, intra-assay and intermediary precision, recovery, detection and quantification limits (LOD and LOQ) were achieved and, the results showed that the developed RP-HPLC-DAD method can be applied to routine determination of cholesterol and cholesterol oxides in milk samples.

Method Validation for Simultaneous Determination of Cholesterol and Cholesterol Oxides in Milk by RP-HPLC-DAD