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Updated: Jul 30, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
Advantages and drawbacks of popular supercritical fluid chromatography/mass interfacing approaches--a user's
1Procter & Gamble Pharmaceuticals, Health Care Research Center, 8700 Mason-Montgomery Rd, PO Box 8006, Mason, OH 45040, USA. pinkston.jd@pg.com
Abstract:
Mobile phases in supercritical fluid chromatography (SFC) have low viscosities and high diffusion coefficients with respect to those of traditional high performance liquid chromatography (HPLC). These properties allow higher mobile phase flow rates and/or longer columns in SFC, resulting in rapid analyses and high efficiency separations. In addition, chiral SFC is becoming especially popular. Mass spectrometry (MS) is arguably the most popular "informative" detector for chromatographic separations. Most SFC/MS is performed with atmospheric pressure ionization (API) sources. Unlike LC/MS, the interface between the SFC column and the API source must allow control of the downstream (post-column) pressure while also providing good chromatographic fidelity. Here we compare and contrast the popular interfacing approaches. Some are simple, such as direct effluent introduction with no active back-pressure-regulator (BPR) in high speed bioanalytical applications. The pressure-regulating-fluid interface is more versatile and provides excellent chromatographic fidelity, but is less user friendly. The pre-BPR- split interface and an interface which provides total-flow-introduction with a mechanical BPR are good compromises between user friendliness and performance, and have become the most popular among practitioners. Applications of SFC/MS using these various interfaces are also discussed.
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