1. 1 2. 2 : GC-MS : : 1393 3. 3Determination of amino acids inselenium-enriched yeast bygas chromatography-mass spectrometryafter microwave assisted hydrolysisXu Zhang,Lu Yang, Zoltan MesterNational Research Council Canada, Ottawa, Ontario K1A 0R6, CanadaAnalytica Chimica Acta 744 (2012) 54 59 4. 4 : GC-MS 5. : 1500 : : : - 5- - 6. . B-Complex : 6Vitamin B1 ThiamineVitamin B2 RiboflavinVitamin B3 NiacinVitamin B5PantothenicacidVitamin B6 PyridoxineVitamin B7 BiotinVitamin B9 Folic AcidVitamin B12 Cobalamin 7. 7 : - - . (Selenite) Selenomethionine % 19 . Selenous acid (Selenite) Selenomethionine 8. 8 . . 9. : : HCl - 9 : : =110 Ct = 20 24 h HCl , 6M : =150 C: - MSA , 4M 10. : ion exchange chromatography (IEC) : capillary electrophoresis mass spectrometry (CE-MS) high-performance liquid chromatography mass spectrometry (HPLCMS) gas chromatography mass spectrometry (GCMS) : GC-MS 10 11. 11 GC-MS :: - HexamethldisilazaneorAAs Trimethylsilyl :: - : 60 = Methyl chloroformAAst = 60 Sec : 12. :CEM Focused Microwave TM48 position 2 / 45 : 12 13. :CEM Focused Microwave TM13 14. : GC-MS 14 Hewlett-Packard HP 6890 GC fitted with a HP 7683 Automatic Injector- injector temperature was 280 C Detection was achieved with an HP model 5973 Enhanced Chemstation software (Version B.01.00) column used for separation was DB-5MS Helium carrier gas- flow rate was 1.5 mL min1 at a pressure of 17.6 psi oven- temperature program :started at 120 C with a hold of2 min and was ramped at 15 C min1 to 290 C MS ion source temperature was 250 C transfer line temperature was 260 C 15. : GC-MS 15GC-MS GC-MS Injector 16. Mass GC 16 17. : HCl 1% (v/v) 1) mmol L ( 1(98% purity) methanesulfonic acid (MSA) DIW 17Methanesulfonic acid 18. :GC-MS Dithiothreitol (DTT) (50 mmol L ( 1Environmental grade ammonium hydroxide Methyl chlorofomate (MCF) (99% purity) Methanol (99.9% purity) Pyridine (99.9% purity) Chloroform 18 19. :19 4.5 mlDIW1.5 mlMSAMSA4M 20. :GC-MS 20 PH=3ammoniumhydroxide 12000RPM5 minDTT Cysteine 21. :DTT 21Cystine 2 DTT 22. :GC-MS 22300lMCF 900l / 1:31 mlCF2000 RPM10 min200 l 1 min 1 min GC-MS 23. 23:GC-MS 24. :24Standard Addition Calibration 25. 25:: standard addition calibration 1) VX CX = VS CS2) S = m Vs + b S = 0 Vs = b/m = . . . GC-MS 3 5 26. 26: . Arg , Gln , Ser - - . HPLC-MS . Rt Pro & Asn Ala & Gly 27. 27: 28. 28 : : 10-180 min : 300 150 C : 50 mg 29. 29 : : 120-180 C : 200 30 min : 0.050 g 6 ml , 4M MSA 30. 30 :: . 150-300 W 150 C : 30 min : 50 mg 200 : W - ) 3 (- 31. 31 ::MSA 1-10 M : 200 30 min : 0.050 g 150 C : 32. 32 : Standard Addition Calibration 150 C : - 30 min : - 200 W : - 4 M :MSA - MCF 33. 33 : 34. 34 : 17 GC-MS 35. 35