Synthesis of Dimethyl Selenone and Studies on the Biomethylation Pt 2

Chapter III Data 1. Elemental analysis of synthesized dimethyl selenone. Element Theoretical Determined Difference Hydrogen 4.25% 4.09% 0.16% Oxygen 22.75% 21.1% 1.65% Carbon Selenium 17% 56% 16.86% 57.96% 0.14% 1.96% 2. FTIR experimental and literature spectra of dimethyl selenone. (Figure 1) 3. FTIR spectrum of dimethyl selenone from 800 to 1100 wavenumbers. (Figure 2) 4. NMR spectrums of dimethyl selenone and dimethyl selenone as it decayed. (Figure 3) 5. Melting point of dimethyl selenone. The melting point of dimethyl selenone was 1470C-1480C. Before it was recrystallized, the melting point was 137-1380C. Ten days after synthesis, the melting point of dimethyl selenone dropped to 140-1410C. 6. The chromatograms of autoclaved medium and filter sterilized medium. (Figure 4) 7. The chromatogram of 1 mM sodium selenate dosed K27 culture. (Figure 5) 8. The chromatogram of 1 mM dimethyl selenone dosed K27 culture. (Figure 6) 9. The chromatogram of 10 mM dimethyl selenone in medium blank with DMS and DMDS. (Figure 7) 10. The chromatogram of 10 mM sodium selenate in medium blank with DMS and DMDS. (Figure 8) 11. The chromatogram of 10 mM sodium selenite in medium blank with DMS and DMDS. (Figure 9) Sorry! Graphic won't be "Distilled" by Adobe Acrobat! Figure 1. FTIR spectra of dimethyl selenone. Upper spectrum is experimental spectrum. Lower spectrum is from Paetzold and Bochmann, 1968. 1.5 886.8 cm-1 Absorbance 1.3 1.1 916.6 cm-1 0.9 0.7 0.5 0.3 800 850 900 950 1000 1050 1100 Wavenumbers Figure 2. The two characteristic absorptions of dimethyl selenone in our experimental FTIR spectrum. These absorptions are due to SeO2 stretching. water 6 5 Dimethyl selenone 4 3 2 water Figure 3. Nuclear magnetic resonance spectra of dimethyl sele- none in deuterium oxide. Spectrum A was performed soon after it was synthesized. Spectrum B was performed one month after the compound was synthesized. DMTS DMDS Autoclaved CL Intensity TSN Medium 0 5 10 15 Time (min) 20 Sterile-Filtered TSN Medium 0 5 10 15 20 Time (min) Figure 4. Headspace analysis of media blanks. Chromatogram A is from autoclaved medium. Chromatogram B is from filter sterilized medium. DMTS DMSeS ? DMDSe DMDS DMSe CH SH 3 CL Intensity 0 5 10 15 Time (min) Figure 5. Headspace analysis of 1 mM sodium selenate in K27 culture. Culture was sampled 24 hours after being poisoned. 20 DMDSe DMSeS ? DMS DMSe CH SH 3 CL Intensity 0 5 10 15 20 Time (min) Figure 6. Headspace analysis of 1 mM dimethyl selenone in K27 culture. Culture was sampled 24 hours after being poisoned. DMDSe DMSeS ? DMDS DMSe DMS CL Intensity 0 5 10 15 20 Time (min) Figure 7. Headspace analysis of 10 mM dimethyl selenone in me- dium blank with DMS and DMDS. Solution was sampled 24 hours after being prepared. DMDS DMS CL Intensity 0 5 10 15 20 Time (min) Figure 8. Headspace analysis of 10 mM sodium selenate in me- dium blank with DMS and DMDS. Solution was sampled 24 hours after being prepared. DMDS DMS CL Intensity 0 5 10 15 20 Time (min) Figure 9. Headspace analysis of 10 mM sodium selenite in medium blank with DMS and DMDS. Solution was sampled 24 hours after being prepared. The early eluting peak is not methanethiol. 12. Table of integrated peak area of samples in 10 mL media volumes. All peak areas are normalized to 1 mL headspace gas and splitless injection. (Table 1) Sample Name TSN blank K27 Blank K27 + 1 mM SeO4 2K27 + 10 mM SeO4 2K27 + 1 mM DMSeO2 K27 + 10 mM DMSeO2 TSN + 10 mM DMSeO2 DMS & DMDS TSN + 10 mM SeO4 2- + DMS & DMDS TSN + 10 mM SeO3 2- + DMS & DMDS CH3 OH DMS DMSe DMDS DMSeS DMDSe DMTS 0 2.50e + 06 3.10e + 08 0 1.67e + 06 3.52e + 06 0 0 2.17e + 07 0 6.17e + 06 6.43e + 07 0 0 5.75e + 07 0 0 1.13e + 08 0 3.02e + 06 1.30e + 07 1.39e + 07 1.44e + 06 9.87e + 06 6.50e + 07 9.55e + 07 9.81e + 07 1.38e + 07 2.99e + 10 2.82e + 09 1.03e + 11 5.54e + 08 1.64e + 10 1.42e + 11 0 0 5.51e + 08 5.65e + 09 0 2.51e + 09 5.12e + 10 0 0 6.48e + 07 1.43e + 07 1.89e + 07 4.83e + 07 1.02e + 07 0 1.29e + 06 5.42e + 07 0 2.08e + 07 0 0 1.58e + 06 0 7.38e + 07 0 2.37e + 07 0 0 0 13. Table of the retention times of volatile sulfur and selenium compounds using the chromatographic system described in this work. (Table 2) Compound Name Retention Time Methanethiol 2.05 min Dimethyl sulfide 4.34 min Dimethyl selenide 6.04 min Dimethyl disulfide 9.72 min Dimethyl selenenyl sulfide 10.93 min Dimethyl diselenide 11.989 min Dimethyl trisulfide 13.2 min