Artificial Intelligence In Microbiology by Dr. Prince C P
Phase modifications
1. CounterCurrent Separation
Theory and practice
§Minimal sample preparation
(direct chromatography of crude extracts)
§High mass – High resolution
§No sample loss (support-free
chromatography)
§Reproducibility
(scale-up or scale down)
§Flexibility
§Mild conditions for sensitive molecules
15. Influence of pH on K
Countercurrent purification of the tetrahydro iso-α acids derived from Humulus lupulus L. JOURNAL OF SEPARATION SCIENCE
Volume 33, Issue 17-18, September 2010, Pages: 2828–2832, Clinton J. Dahlberg, James S. Traub, Matthew L. Tripp, Jeffrey S. Bland and Brian J. Carroll
Figure 2. Overlay of the UV
chromatogram from the CS
experiment (entry 3, Table 1, 254 nm)
with a reconstructed CS
chromatogram (254 nm) from the
HPLC analysis of collected
28 mL fractions.
HEMWat (acetate pH 5.3)
7:3:5:5
18. Ion Pairs
Fig. 2. HSCCC-chromatogram of C18-enriched pigment extract (1677 mg) I. lindeniiVan Houtte leaves.
HSCCC conditions: flow rate: 3.0 mL/min; ‘head-to-tail mode’; biphasic solvent system TBME–1-BuOH–
ACN–H2O (0.7% HFBA) 1:3:1:5 (v/v/v/v).Below, HPLC retention times of the separated major
betacyanins detected in each HSCCC fraction.
Separation of amaranthine-type betacyanins by ion-pair high-speed countercurrent
chromatography
Journal of Chromatography A, Volume 1344, 30 May 2014, Pages 42-50
Gerold Jerz, Nadine Gebers, Dominika Szot, Maciej Szaleniec, Peter Winterhalter,
Slawomir Wybraniec
https://nl.wikipedia.org/wiki/Betanine#/media/File:Betanin.svg
19. Separation of amaranthine-type betacyanins by ion-pair high-speed countercurrent chromatography
Journal of Chromatography A, Volume 1344, 30 May 2014, Pages 42-50
Gerold Jerz, Nadine Gebers, Dominika Szot, Maciej Szaleniec, Peter Winterhalter, Slawomir Wybraniec
Target-guided separation of Bougainvillea glabra betacyanins by direct coupling of preparative ion-pair
high-speed countercurrent chromatography and electrospray ionization mass-spectrometry Journal of
Chromatography A, Volume 1217, Issue 27, 2 July 2010, Pages 4544-4554 Gerold Jerz, Sławomir
Wybraniec, Nadine Gebers, Peter Winterhalter
terBuAcWat (HFBA) 1:3:1:5
terBuAcWat (TFA) 2:2:1:5
Separation of betalains from berries of Phytolacca americana by ion-pair high-speed counter-current
chromatography Journal of Chromatography A, Volume 1190, Issues 1–2, 9 May 2008, Pages 63-73 Gerold
Jerz, Tanja Skotzki, Kathrin Fiege, Peter Winterhalter, Sławomir Wybraniec
BuAcWat (TFA) 5:1:6 (0.7%)
Separation of polar betalain pigments from cacti fruits of Hylocereus polyrhizus by ion-pair high-speed
countercurrent chromatography
Journal of Chromatography A, Volume 1216, Issue 41, 9 October 2009, Pages 6890-6899
Sławomir Wybraniec, Paweł Stalica, Gerold Jerz, Bettina Klose, Nadine Gebers, Peter Winterhalter, Aneta
Spórna, Maciej Szaleniec, Yosef Mizrahi
BuAcWat (TFA) 5:1:6 (0.7%)
terBuAcWat (TFA/HFBA) 2:2:1:5 (0.7%)
New solvent systems for gradient counter-current chromatography in separation of betanin and its
derivatives from processed Beta vulgaris L. juice.
Journal of Chromatography A, Volume 1380, 6 February 2015, Pages 29-37
Aneta Spórna-Kucab, Ian Garrard, Svetlana Ignatova, Sławomir Wybraniec
BuEtWat/Aq. NaCl/Aq. H3PO4
1300:200-1000:1300:700:2.5-10
Ion-pair high-speed countercurrent chromatography in fractionation of a high-molecular weight variation
of acyl-oligosaccharide linked betacyanins from purple bracts of Bougainvillea glabra
Journal of Chromatography B, Volume 878, Issues 5–6, 15 February 2010, Pages 538-550
Sławomir Wybraniec, Gerold Jerz, Nadine Gebers, Peter Winterhalter
BuAcWat (TFA) 5:1:6 (0.7%)
terBuAcWat (TFA) 2:2:1:5 (0.7%)
Fig. 3. IP–HSCCC chromatogram of directly recovered crude pigment extracts (755
mg) of violet B. glabra bracts in the solvent system terBuAcWat 2:2:1:5 using 0.7%
TFA. HSCCC conditions: flow rate: 3.0 mL/min; CCC-operation: head-to-tail mode;
velocity: 850 rpm and detection wavelength 540 nm.
Ion Pairs