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Automated semipreparative purification with mass spectrometric fraction collection trigger: Modeling and experimental evaluation of a setup employing passive splitting
Author(s) -
Steiner Frank,
Mahsunah Anis,
Arnold Frank,
Piecha Thomas,
Huber Christian G.
Publication year - 2007
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200700059
Subject(s) - fraction (chemistry) , chromatography , mass spectrometry , mass fraction , chemistry , analytical chemistry (journal) , organic chemistry
A semipreparative HPLC setup was evaluated for automated fractionation with both photometric‐ and mass spectrometric trigger. The goals of the work were to systematically study and optimize the flow‐splitting setup for mass‐directed purifications by mathematical modeling and experimental verification. The system comprised a passive splitting device with make‐up flow capability, which directed a small fraction of the column effluent to the mass spectrometer and the remainder to the fraction collector. Tubing lengths and diameters of the splitter as well as make‐up flow rates were varied in order to address and optimize peak dispersion, delay times between mass detector and fraction collector, and mass spectrometric signal quality. A paraben standard mixture was analyzed and purified on both microparticulate and monolithic columns with 10 mm inner diameter and at typical flow rates of 5–10 mL/min. Fraction purities and recoveries close to 100% were achieved. The system allowed mass‐triggered fractionations on a 1 mg scale at flow rates of 10 mL/min in combination with monolithic columns in less than 2 min. Finally, the system was successfully applied to the fully automated isolation of milligram quantities of degradation products in a pharmaceutical preparation to successfully allow for structure elucidation with NMR spectroscopy.