Apresenta-se um procedimento para extração de Ca, K e Mg de amostras de cultivo in vitro de plantas, comparando-se culturas de calo com diferentes potenciais embriogênicos de Citrus sinensis e Citrus limonia, com o emprego de energia ultra-som. Foram investigados os parâmetros relacionados com a extração de metais, tais como tipo de amostragem do material, concentração do ácido e tempo de sonicação. Visando a comprovação da exatidão, o procedimento de extração ultra-sônica foi comparado com o de digestão ácida assistida por microondas, e não foram observadas diferenças estatísticas significativas em um nível de confiança de 95%. Com este procedimento de extração simples e preciso, foi possível estabelecer diferenças nas concentrações de Ca, K e Mg durante a formação/desenvolvimento dos embriões de Citrus, bem como entre as culturas (embriogênicas e não-embriogênicas). Finalmente, a extração ultra-sônica demonstrou-se uma excelente alternativa para menor manipulação das amostras e redução de custos operacionais.An ultrasound extraction procedure for Ca, K and Mg from in vitro plant cultures is proposed, comparing cultures of different embryogenic levels of Citrus sinensis and Citrus limonia, employing ultrasound energy. Parameters related to metals extraction, such as plant material sampling, acid concentration and sonication time were investigated. For accuracy check, the proposed ultrasound extraction procedure was compared with a microwave-assisted digestion procedure and no differences in the results were verified at 95% of the confidence level. With this simple and accurate extraction procedure, it was possible to determine differences in Ca, K and Mg concentrations during Citrus embryo formation/development and between cultures (embryogenic and non-embryogenic). Finally, the ultrasound extraction method demonstrated to be an excellent alternative for handless sampling and operational costs.
Keywords: ultrasound extraction, FAAS, FAES, Ca, K and Mg determination
IntroductionClassical sample preparation methods commonly used for biological studies are those based on dry or wet decomposition, 1,2 which, in general, require intensive sample handling and time/reagent consumption. In this context, the use of ultrasound can be an excellent alternative to minimize sample preparation [3][4][5] and to avoid solvent extraction, 6,7 because its energy facilitates and accelerates some analytical steps, permitting less time consumption and decreasing the cost per analysis. Besides these characteristics, accuracy, precision, and handless manipulation can be achieved when ultrasound-assisted extraction methods are used. 4 As an example, the application of ultrasound as an auxiliary energy source for solid sample treatment was made by Luque de Castro and Silva. 8 The authors emphasized different applications of ultrasound mainly related to sample treatment of agricultural, biological and environmental samples. Carvalho et al. 9 pointed out the sonochemical degradation that occurs inside extracted solutions when using the ...