Ultrasound-assisted extraction of Ca, K and Mg from in vitro citrus culture

Abstract

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 ...

Ultrasound-assisted Extraction of Polyphenols from Red-grape (Vitis Vinifera) Residues

“…Wang and Weller (2006) cited that scanning electron micrographs (SEM) have provided evidence of the mechanical effects of ultrasound, mainly shown by the destruction of cell walls and release of cell contents. Our results also agree with some other authors (Minami, 1996;Simal et al, 1998;Nascentes et al,2001;Yebra and Moreno, 2002;Arruda et al, 2003;Banjoo and Nelson, 2005;Gu et al, 2005;and Herrera and Luque de Castro, 2005), who have stated that ultrasound increases the extraction levels not only of polyphenols but ions (Cu, Fe, K, Mg Mn, Pb, Se, Zn) and other compounds such as polycyclic aromatic hydrocarbons and water that are present in different biological materials such as citric cultures, seafood, tobacco, strawberries, lettuce, cabbage and apples. …”

“…Studies of ultrasound application during osmotic dehydration of apples have demonstrated that there is an increase of mass transfer after applying ultrasound waves on samples (Simal et al, 1998). It has also been demonstrated that ultrasonic waves improve the extraction level of calcium (Ca), potassium (K) and magnesium (Mg) from in vitro citrus culture (Arruda et al, 2003), the extraction level of polyphenols from tobacco (Gu et al, 2005) and the extraction level of polyphenols from strawberries (Herrera and Luque de Castro, 2005). Wang and Weller (2006) cited several studies that provide evidences about benefits and potential applications of ultrasound-assisted extraction of bioactive compounds from plants.…”

Optimized extraction of polysaccharides from Taxus chinensis var. mairei fruits and its antitumor activity

“…The main advantage of the microwave approach is speed [13,14], but a disadvantage is inhomogeneous heating. The possible benefits of ultrasonic extraction are mass transfer intensification, cell disruption, and improved penetration effects [15]. Consequently, the combination of ultrasound and microwave radiation is a complementary technique that may exhibit the advantages of both methods.…”

Use of Ultrasound Bath in the Extraction and Quantification of Ester-Linked Phenolic Acids in Tropical Forages

“…There have been many studies applying ultrasound bath in the sample treatment of soils and sediments [28][29][30][31], biological [32][33][34], nutritious [35,36]. However, in the case of forage samples, no study has yet been reported in the literature describing this technique for the extraction and quantification of phenolic acids.…”