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our consortium. It is clearly very well suited to larger scale separations and continuous operation.
The future of the project The project is just starting its second year and, having established a wide ranging applications portfolio for HPCCC, the next year of the project will focus on the following:
• Further simplify the method development protocols to make them even quicker and easier for a chromatographer to use.
• Extend the range of solvent systems available for use to enhance solubility/loading and enable the use of greener solvents.
Figure 5: Dynamic Extractions Maxi 18L prototype HPCCC instrument at Brunel University’s Advanced Bioprocessing Centre. ICcE – Target enriched in column
Crude Sample
NORMAL PHASE
Sample Loading Pumps
UP Pump V1 LP Detector Column 2 UP LP 4-Way REVERSED PHASE
Target in columns
UP Detector Column 1 V2 LP Pump
Non-polar impurities in upper phase (UP) fractions
• Further develop, understand and demonstrate the ICcE operating method.
All of these objectives will further enhance the capability of HPCCC instrumentation to easily integrate into existing workflows. Updates throughout the year and until the end of the project will be available at
www.dynamicextractions.com/TSB.
Conclusion Polar impurities in lower phase (LP) fractions
Figure 6: Diagrammatic representation of Intermittent Counter Current Extraction (ICcE) applied to the separation of a target material from a complex mixture. The photos give a pictorial indication of the separation of this crude mixture with the pure (clear) target fractions remaining in the columns while the impurities are washed away – the polar materials in one direction and the non polar materials in the other.
CCC holds considerable promise as a preparative technique to enhance current laboratory capability to rapidly react to separation problems. The challenge here is to integrate the instrumentation and control systems to allow CCC to take its place alongside existing preparative separation capability. This will give a greater overall chance of finding generic solutions to preparative separation problems quickly and efficiently. For larger scale separations CCC again offers the potential to lower overall costs, opening up the possibility of using preparative chromatography in new areas.
References 1 S. Ignatova , P. Wood, D. Hawes, L. Janaway, D. Keay and I. Sutherland, J. Chromatogr. A, 1151 (2007), 20
2 Y. Ito, M. Weinstein, I. Aoki, R. Harada, E. Kimura and K. Nunogaki, Nature, 212 (1966), 985
3 A. Marston, K. Hostettmann J. Chromatog. A, 1112 (2006), 181
4 J M. Roberts, S. R. Cole, J. Spadie, H. E. Weston, W. K. Young, American Pharmaceutical Review, Apr March 2010, 38 – 44
Figure 7 A: Separation of a target peak from a crude mixture - HPLC analytical separation of input material
5 P. Borman, J. Roberts, B. O'Reilly, R. Attrill, I. Barylski, K. Freebairn, Pharmaceutical Technology 2010, Volume 34, s6-s11
6 G. Harris, Presentation at CCC2010, Lyon, France, July 2010 7 D. Keay, Manufacturing Chemist, September 2008, 33-34 8 I.A. Sutherland, J. Chromatogr. A, 1151, (2007), 6 9 C. Thickitt, N. Douillet Presentation at SPICA2010, Stockholm, Sweden, September 2010
10 S. Ignatova, N. Douillet Presentation at SPICA2010, Stockholm, Sweden, September 2010
11 P. Hewitson, S. Ignatova, I. Sutherland, CCC2010 special issue, J. Chromatogr. A, 2010, doi: 10.1016/
j.chroma.2011.03.072
Figure 7 B: ICcE separation of a target peak from a crude mixture - HPLC analytical separation of pooled fractions isolated by CCC
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