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Journal of Chromatography A
Volume 972, Issue 1, 27 September 2002, Pages 87-99
Believe it or not, the authors purified 200 mg of a crude peptide using three 4.6 x 50 mm columns strung together in a series.
Briefly, the sample is loaded in aqueous TFA, isocratically eluted with MeCN/TFA (MeCN amount should be 20% less than % required for elution of desired component in normal gradient HPLC). Next, % MeCN is increased (so that it is 15% lower than % required in gradient elution). This is held isocratically for 70 min, where eventually desired product is eluted in high purity and near quantitative recovery.
I know nothing about this technique. Can anyone give a little insight as far as pitfalls, etc?
Volume 972, Issue 1, 27 September 2002, Pages 87-99
Believe it or not, the authors purified 200 mg of a crude peptide using three 4.6 x 50 mm columns strung together in a series.
Briefly, the sample is loaded in aqueous TFA, isocratically eluted with MeCN/TFA (MeCN amount should be 20% less than % required for elution of desired component in normal gradient HPLC). Next, % MeCN is increased (so that it is 15% lower than % required in gradient elution). This is held isocratically for 70 min, where eventually desired product is eluted in high purity and near quantitative recovery.
I know nothing about this technique. Can anyone give a little insight as far as pitfalls, etc?
