Analytical approach to the metallomic of Nile tilapia (Oreochromis niloticus) liver tissue by SRXRF and FAAS after 2D-PAGE separation: Preliminary results |
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Authors: | Paula M. Lima Felipe A. dos Santos Marcelo O.A. da Silva Marco A.Z. Arruda Gustavo R. de Castro |
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Affiliation: | a Faculdade de Medicina Veterinária e Zootecnia/UNESP, Programa de Pós-Graduação em Zootecnia, C. Postal 31, 15385-000 Botucatu, SP, Brazil b Laboratório Nacional de Luz Síncronton, Campinas, SP, Brazil c Instituto de Química/UNICAMP, Grupo de Espectrometria, Preparo de Amostras e Mecanização-GEPAM, C. Postal 6154, 13084-971 Campinas, SP, Brazil d Instituto Nacional de Ciência e Tecnologia de Bioanalítica/UNICAMP, C. Postal 6154, 13084-971 Campinas, SP, Brazil e Instituto de Biociências/UNESP, Departamento de Química e Bioquímica, C. Postal 510, 18618-970 Botucatu, SP, Brazil |
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Abstract: | An investigation was made into calcium, iron and zinc in protein spots in samples of Nile tilapia (Oreochromis niloticus) liver tissue obtained after protein separation by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and subsequent qualitative and quantitative evaluation by synchrotron radiation X-ray fluorescence (SRXRF) and Flame Atomic Absorption Spectrometry (FAAS). An analysis of the fluorescence spectra indicated the presence of calcium, iron and zinc in 12, 6 and 8 liver protein spots, respectively. The metal ions found were distributed mainly in proteins with a molar mass of less than 40.00 kDa and more than 12.00 kDa, with pI in the range of 4.70-9.40. The only exception was a spot presenting protein with a molar mass of 10.10 kDa. In addition to calcium, iron and zinc, sulfur and phosphorus - which are non-metals that may be part of the protein structure, were also detected. After microwave-assisted acid mineralization of the proteins spots, a FAAS estimation of the concentration of calcium, iron and zinc ions bound to these proteins indicated a range of 1.08-5.80 mg g−1, 2.02-8.03 mg g−1 and 1.60-8.55 mg g−1, respectively. |
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Keywords: | Metallomics Protein spots 2D-PAGE SRXRF FAAS |
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