Elution-Extrusion Countercurrent Chromatography: Theory and Concepts in Metabolic Analysis
详细信息 查看全文
- 作者：Alain Berthod ; J. Brent Friesen ; Taichi Inui ; Guido F. Pauli
- 刊名：Analytical Chemistry
- 出版年：2007
- 出版时间：May 1, 2007
- 年：2007
- 卷：79
- 期：9
- 页码：3371 - 3382
- 全文大小：2986K
- 年卷期：v.79,no.9(May 1, 2007)
- ISSN：1520-6882
文摘
Elution-extrusion countercurrent chromatography(EECCC) takes full advantage of the liquid nature of thestationary phase in CCC by combining regular chromatographic elution with stationary-phase extrusion. EECCCis shown to be a three-stage process consisting of classicalelution (I), sweeping elution (II), and extrusion (III). Afteronly two column volumes of solvent, it rapidly yields ahigh-resolution chromatogram that covers an extendedpolarity range of solutes. As hydrophilicity/lipophilicitybalance is a crucial discriminatory property of analytesin highly complex mixtures such as metabolomic samples,the precise determination of CCC distribution constants(KD) is vital to the analysis of metabolomes and othercomplex biological matrixes. This work builds the EECCCconcept by performing a full theoretical treatment andproviding equations for retention volumes, peak widths,resolution factors, and distribution constants. Experimental validation utilizes natural products standards thatresemble the zero to infinity range of the polarity continuum. EECCC extends the "sweet spot" of high resolution in CCC and provides access to the otherwise practically unapproachable high-KD portion of the high-resolutionchromatograms in CCC. Its improved capabilities of KDtargeting make EECCC a promising tool for the specificanalysis of "small" molecules in complex samples suchas in metabolomic fingerprinting and footprinting.