Chapter 6. Conclusion 6.1 Determination of selenium content in biological samples by ICP -MS…………………………………………………………………………………..255 6.2 Selenium speciation of Se compounds/metabolites in urine and liver samples 6.2.1 Selenium speciation analysis of the four relevant human urinary selenium species: TMSe + , SeMet, SeGalNAc and SeGluNAc by HPLC -ICP -MS and HPLC -API -MS/MS……………………………………………………………………..255 6.2.2 Application of HPLC -ICP -MS and HPLC -APCI -MS/MS for selenium speciation in cytosolic samples of porcine liver…………………………………….256 6.2.3 The boundary of HPLC -ICP -MS and HPLC -API -MS/MS techniques...256 XIII 6.3 HPLC separation and mass spectrometric characterisations of Se-containing tocopherol and tocotrienol succinate derivative compounds…………………...257 6.4 Further considerations on the selenium speciation in biological samples 6.4.1 Further considerations on the selenium speciation in biological samples……258 6.4.2 Aspects for future studies……………………………………………………..259 Reference…………………………………………………………………………..261 XIV Appendix Appendix 1 1.1 Total selenium contents in food products in the local market (Crete, Greece)…263 1.2 General information on mammalian selenoproteins……………………………266 1.3 Selenium species of biological interests………………………………………..268 Reference…………………………………………………………………………...273 Appendix 2 2.1 Variety of biological specimens relevant to the diverse biochemical events in mammal body……………………………………………………………………….277 2.2 Reversed phase columns for highly aqueous mobile phase (or, for the separation of highly polar/hydrophilic compounds)……………………………………………278 2.3. Technical perspectives on capillary/nano liquid chromatography and nanoelectrospray ionization mass spectrometry…………………………………….279 Reference…………………………………………………………………………...286 Appendix 3 3.1 Supplemental HPLC parameters used in this study…………………….………289 3.2 Supplemental figures for chapter 3………………………………………….….292 3.3 ESI-MS and ESI-MS/MS of molecular ions of TMSe + and SeMet and their corresponding product ions of in this study………………………………………...300 3.4 Proposed fragmentation pathways for TMSe + , SeMet, SeGalNAc and SeGluNAc, and supplemental information obtained using multistage ion trap mass spectrometry………………………………………………………………………...303 3.5. Isobaric interferences in Se determination by ICP-MS………………………...307 3.6 Recovery of PerfectSil Target 100 C 8 -3 column (150 × 4.0 mm, 5 µm, Analysentechnik, Germany)………………………………………………………...308 3.7 Comparison of the signal and background levels in APCI -SRM and ESI -SRM determination of SeMet, SeGalNAc and SeGluNAc standards…………………….309 3.8 Supplemental data on determination of total selenium concentration in biological samples by ICP-MS…………………………………………………………………310 XV 3.9 Comparison of signal intensity of ESI-MS/MS detection of four selenium species separated respectively in two reversed phase columns……………………………..313 3.10 Quantification of TMSe + in crude human urine sample by HPLC-ESI-MS/MS……………………………………………………………………………...314 3.11 Quantification of SeGalNAc in crude human urine sample by HPLC-ESI-MS/MS………………………...