RECYCLE CHROMATOGRAPHY

1. RECYCLECHROMATOGRAPHY Jiri SEVCIK Prague, the Czech Republic RECYCLE GC

2. ABOUT • CHARACTERISTICS • SEPARATION EFFICIENCY • RESOLUTION • RECYCLE CHROMATOGRAPHY HW RECYCLE GC

3. multidimensional chromatographySYSTEMS serial parallel KD1 <> KD2 <> KDd RECYCLE GC

4. SCHEMATICSof recycle chromatography INJECT R1 tM w0 INTERFACE tMex wew COLUMN tRa/ , tRb/ wa , wb R2 DETECT tMdet wdet LOOP COUNTER i RECYCLE GC

5. recycle chromatographyCOMPONENTS AND VARIATIONS RECYCLE GC

6. RECYCLE CHROMATOGRAPHYPARAMETERS • retention time • elution profile width • separation efficiency • resolution RECYCLE GC

7. RETENTION TIMEin recycle chromatography INJECT R1 tM INTERFACE tMex COLUMN tRa/ , tRb/ R2 DETECT tMdet LOOP COUNTER i RECYCLE GC

8. RECYCLE CHROMATOGRAPHYRETENTION TIME • from an injector to a detector • within a loop • after i loops RECYCLE GC

9. RECYCLE CHROMATOGRAPHYRETENTION TIME • retention time between consecutive cycles is constant • total retention time is additive RECYCLE GC

10. RECYCLE CHROMATOGRAPHYRETENTION TIME PROOF of ADDITIVITY RECYCLE GC

11. RECYCLE CHROMATOGRAPHYRETENTION TIME PROOF of ADDITIVITY RECYCLE GC

12. RETENTION TIMEIN RECYCLE CHROMATOGRAPHY RECYCLE GC

13. ELUTION PROFILE WIDTHof recycle chromatography INJECT R1 w0 INTERFACE wew COLUMN wa , wb R2 DETECT wdet LOOP COUNTER i RECYCLE GC

14. RECYCLE CHROMATOGRAPHYELUTION PROFILE WIDTH • from an injector to a detector • within a loop • after i loops RECYCLE GC

15. RECYCLE CHROMATOGRAPHYELUTION PROFILE WIDTH • elution profile width between consecutive cycles is constant • total peak width is additive in form of their variances RECYCLE GC

16. RECYCLE CHROMATOGRAPHYPEAK WIDTH PROOF of ADDITIVITY RECYCLE GC

17. RECYCLE CHROMATOGRAPHYPEAK WIDTH PROOF of ADDITIVITY RECYCLE GC

18. PEAK WIDTHIN RECYCLE CHROMATOGRAPHY RECYCLE GC

19. RECYCLE CHROMATOGRAPHYSEPARATION EFFICIENCY • number of effective plates • after a loop RECYCLE GC

20. RECYCLE CHROMATOGRAPHYSEPARATION EFFICIENCY • number of effective plates after i loops RECYCLE GC

21. RECYCLE SEPARATION EFFICIENCYPROOF of ADDITIVITY RECYCLE GC

22. RECYCLE CHROMATOGRAPHYEXTRA COLUMN EFFECTS • number of effective plates after i loops RECYCLE GC

23. RECYCLE CHROMATOGRAPHYEXTRA COLUMN EFFECTS • ratio ni / n1 RECYCLE GC

24. RECYCLE CHROMATOGRAPHYEXTRA COLUMN EFFECTS • ratio ni / n1 • aninlet part of a column is unefficient THUS • efficienty increases faster in recycle system than in nonrecycle system • it is not correct to express separation efficiency in terms of the theoretical plate number RECYCLE GC

25. RECYCLE CHROMATOGRAPHYEXTRA COLUMN EFFECTS RECYCLE GC

26. SEPARATION IN RECYCLE CHROMATOGRAPHY • retention interval of unseparated mixture • in one-dimensional system • in recycle system tD tb ta RECYCLE GC

27. SEPARATION IN RECYCLE CHROMATOGRAPHY • retention interval of unseparated mixture increases over the column lenght untill 1st peak leaves the column while the last one just enters the column • then peak will merge again THUS • maximum number of cycles tD tb ta RECYCLE GC

28. RECYCLE SEPARATION EFFICIENCYPROOF of ADDITIVITY RECYCLE GC

29. MAXIMUM NUMBER OF CYCLESIN RECYCLE CHROMATOGRAPHY tD tb ta RECYCLE GC

30. RESOLUTIONIN RECYCLE CHROMATOGRAPHY • retention order can not change because of the same st.phase (a remains constant) RECYCLE GC

31. MAXIMUM AVAILABLE RESOLUTIONIN RECYCLE CHROMATOGRAPHY RECYCLE GC

32. REQUIRED SEPARATION EFFICIENCYIN RECYCLE CHROMATOGRAPHY • for required resolution • a , k and n can be optimized • if Rab = 1 an optimum number of cycles results RECYCLE GC

33. RECYCLE SEPARATION EFFICIENCYNUMBER of CYCLES RECYCLE GC

34. THE OPTIMAL NUMBER OF CYCLESIN RECYCLE CHROMATOGRAPHY • for isomers a=1.01 , k=5and n=20,000 Rab1 = 0,29 and the optimal number of cycles iopt= 3.3 RECYCLE GC

35. RECYCLE CHROMATOGRAPHYMAXIMAL NUMBER of CYCLES merged at start maximal separation merged once again RECYCLE GC

36. MS-SIM separation of GC non separated isomers RECYCLE GC

37. recycle capillary GCPRINCIPLE • recycling analysed mixture through single capillary column • by means of peristaltic pump as mobile phase source • and controling actually achieved separation by means of end column splitter RECYCLE GC

38. recycle capillary GCCONSTRUCTION • peristaltic pump providing optimal column flow for columns from 5 to 50 meters • fixed end column splitter RECYCLE GC

39. INSTRUMENTATION - interface IN RECYCLE CHROMATOGRAPHY peristaltic pump R1 40 mm (injector) R2 40 mm (detector MS) flow column RECYCLE GC

40. recycle capillary GCAPPLICATION • suited for componds up to C10 • for limited number of compounds with similar physicochemical and chiral properties • first time separation of 3 stereoisomers of 3,4-dimethylhexane on β–cyclodextrine with 2,000,000 plates RECYCLE GC

41. nonselective separation • Carbowax 20M • 300 m, 250 µm • n = 500,000 • n-dodecene isomers RECYCLE GC

42. selectivity of separation • separation of isomers also with a functional group in middle of carbon chain • n-dodecenes RECYCLE GC

43. MS-SIM separation of GC non separated isomers RECYCLE GC

44. ABOUT • CHARACTERISTICS • SEPARATION EFFICIENCY • RESOLUTION • RECYCLE CHROMATOGRAPHY HW RECYCLE GC

45. sample preparation efficiency selectivity hyphenation sensitivity detection software ITP - GC recycle GC stationary phases comprehensive thermal modulation spectral detectors deconvolution NEW GC METHODS development toward reduced ultimate uncertainty RECYCLE GC