This manuscript exemplifies the prospective use of asymmetrical flow field flow fractionation (AF4) coupled to inductively coupled plasma mass spectrometry (ICP-MS) as a simple tool for chemical speciation of selenomethionine (SeMet) in selenized yeast. Several popular sample preparation methods were evaluated for their suitability to determine selenomethionine (SeMet) in selenized yeast by AF4-ICP-MS. These included water, methanesulfonic acid (MSA), formic acid (FA) and alkaline extractions. Alkaline extraction (using sodium dodecyl sulfate buffer) provided the best recovery/determination conditions for SeMet based on analysis of NRC certified reference material (CRM) SELM-1 since it minimized hydrolysis of the protein peptide bonds optimally required for the AF4 separation. The analytical performance of three different AF4 membranes (5, 10 and 500 kDa regenerated cellulose) was also evaluated. No significant difference in the recovery of SeMet was observed when using 5 and 10 kDa RC membranes, whereas the 500 kDa membrane resulted in a significant loss. The proposed method presents appropriate instrument and intra-assay precisions of 4.4-9.2% and 3.8% RSD, respectively, a detection limit of 0.49 μg L-1 SeMet as Se and good linearity with correlation coefficients (R) between 0.996 - 0.999. This is the first report of use of AF4-ICP-MS for species specific quantitation of SeMet in selenized yeast demonstrating its efficient use as an alternative method to other traditional chromatographic techniques.
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Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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Site | Modification | Modifier | Source | Reference |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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