Biosynthesis of isotopically enriched selenomethionine: application to its accurate determination in selenium-enriched yeast by isotope dilution analysis-HPLC-ICP-MS
作者： L. Hinojosa Reyes , F. Moreno Sanz , P. Herrero Espílez , J. M. Marchante-Gayón , J. I. García Alonso
DOI: 10.1039/B403662J
关键词:
摘要: Isotope dilution analysis is proposed for the determination of selenomethionine (SeMet) in Se-enriched yeast material by high performance liquid chromatography inductively coupled plasma mass spectrometry (HPLC-ICP-MS) using a 77Se-enriched spike obtained growth on 77Se-rich culture medium. Different methods enrichment with selenium were evaluated to determine most effective one. The use two different media, amount added, carbon source (glucose) and harvest time order characterize total incorporated cells. proteins hydrolysed protease XIV, isotopically enriched SeMet isolated anion exchange chromatography. isotopic composition solution was determined on-line HPLC-ICP-MS reverse isotope used characterization means natural standard. In material, sample spiked SeMet, hydrolyzed non-specific enzyme (protease XIV) at 37 °C 20 hours or an ultrasonic probe 1 min. Both 78Se/77Se 80Se/77Se ratios measured ICP-MS collision reaction cell after separation HPLC. method applied candidate reference (SEAS project), results agreement those reported other laboratories.
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