Chiralm selective chromatography

1. Chiralm selective chromatography D. Kozma ed. Handbook of Optical Resolutions via Diastereomeric Salt Formation. CRC Handbooks, 2001. Z Juvancz, G. Seres Appendix

2. Definition of chirality • Chiral molecules are asymmetric. • Enantiomers or optical isomers are mirror image of each others, but they are not same. • Diastereomeric molecules are not • mirror images, but they have same constitutions. Central chirality with for different substituents.

3. Enantiomers DNS molecules Kajtár M., Változatok négy elemre, Gondolat, 1984.

4. Application of chiral selective analysis Checking the chiral selective synthesis: Pharmaceutical, pesticide industry Metabolite research: (pl. Warfarin, Ibuprofen, pheromonos) Adulteration, bacterial infectiouns: ( pl. wine, essential oils) Racemisation: (pl. heat treatment, age determination)

5. Different effects of enantiomeric pharmaceuticals

6. Share of enantiomeric pure pharmceuticals

7. Different odor of enatiomers

8. Three point interaction is necessary for the chiral recognation I. V.R. Meyer, M. Rais, Chirality 1 (1989) 167.

9. Two point interactions are not enough II. V.R. Meyer, M. Rais, Chirality 1 (1989) 167.

10. Certain rigidity is necessary for chiral recognition V.R. Meyer, M. Rais, Chirality 1 (1989) 167.

11. Interaction points can be surface or axis π acid - π base, and h_bridge are the key interactions.

12. Working theory of chiral stationer phase, (CSP) C. Welch et al., Review of Stereochemistry

13. Working theory of chiral mobile phase addtitive (CMA) C. Welch et al., Review of Stereochemistry

14. Rigid structure offers high selectivity. Flexible structure offer broad selectivity range. Most of the separation bases on three point interaction. Functional groups close to asymmetric center improves the resolution. Repulsion interactions havea big role. Low analysis temperature improves the separations. General rules of chiral selective chromatography

15. Temeperature dependence of chiral chromatography ln α = S+- (S0sm)/R - H+- (H0sm)/RT  +- : difference between the two isomers (+and -) • sm: difference between two phases (stationary and mobile). Generally the low analysis temperature improve the separation. Using high efficiency stationaryphase, 0,1kJ/mol (α: 1,01) energy difference is enough between the enantiomers for Rs 1.5 resolution value.

16. Importance of minor first elution order

17. Most frequently used CSP are cyclodextrin based molecules .

18. Shape of derivatized a CD The direction and length of bonds are differents..

19. Reason of good chiral recognation properties of cyclodextrins • There are several chiral center in a CD • The surrounding of chiral centers are of uniform in a glucose unit. • The shape of glucose units are different • Substituted derivates are bunch of isomers • The chiral recognition depends on the type of substituents • The selectivity can depend on the ionized state • Cyclodextrins have flexible structure (induce fit)

20. Cyclodextrins can separate even the functionless hydrocarbons enantiomers

21. Selecticvity vs. Concetration of chiral selector The acharial silicone matrix offers a good efficiency.

22. Selecticvity vs. Concetration of chiral selector

23. Low analysis temperature results in high selectivity

24. Cyclic derivatives give high selectivity

25. Separation of enetiomers of essential aminoacids with Chirasil-Val Orginal CSP ImprovedCSP

26. Chiral selective HPLC

27. Most frequently used CSps in

28. Key interaction of polysaccharide CSps H-bridge ands π- π interactions

29. Selectivity depend on the substitution of cellulose

30. Cyclodextrin CSp in HPLC

31. Cyclodextris as CMAs (

32. Methylated cyclodexrin CMAs

33. Pirkle type CSP

34. Separation with π base CSP

35. Vancomycin, antibiotic CSP

36. Separation with Vancomycin CSP

37. Chiral separation in CE Neutral selector can not separate neutral molecules. The oppositely charged selector and sample offer high resolution. Separation has maximum value in the function of the selector concentration.

38. Separation vs. concentration of selector

39. High selectivity with oppositelly charged sample and selector

40. Used chiral selectors in CE

41. Separations of pyretroic acids

42. Chiral separation with β-HDMACD 0,5 mMol concentration of selectror is enough for baseline separation.

43. Chiral selective separation of γ-PhoCD reversed migration orders TRIMEB

44. 50 mM acetic acid / TRIS; pH 5.0 mAU 30 20 10 0 -10 -20 2 4 6 8 10 12 14 min 50 mM acetic acid / TRIS; pH 5.0 mAU + 1 mM MAbCD 30 20 10 Kass : 100 – 370 M-1 0 -10 -20 2 4 6 8 10 12 14 min 50 mM acetic acid / TRIS; pH 5.0 mAU + 10 mM PMMAbCD 30 20 10 Kass : 1.3 – 70 M-1 0 mAU -10 20 -20 4MePPA 15 + 1 mM MAbCD + 10 mM PMMAbCD 2 4 6 8 10 12 14 min 50 mM acetic acid / TRIS; pH 5.0 10 5 Fenoprop 0 PPA - 5 - 10 5 10 15 20 25 min mecoprop, 2,4-DP, 3ClPPA Dual CD seletor system R. Iványi et al.

45. The selectivity of ionizable CDs depend on their ionized states

46. Partially filling technique No EOF (acrylamide coated column). The sample and selector migrate in opposite directions. The selector nigrates away, when the sample reaches the detector windows.

47. Vancomicine selector

48. Chiral selector with chiral MKEC agent

49. Chips chiral separation EKC Designed for Martian space rockett

50. Chiral CEC Monolit CSP selector