title: Automated semipreparative purification with mass spectrometric fraction collection trigger: modeling and experimental evaluation of a setup employing passive splitting.
abstract: 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.
authors.affiliation: Department of Chemistry, Instrumental Analysis and Bioanalysis, Saarland University, Saarbrücken, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Steiner
authors.forename: Anis
authors.initials: A
authors.lastname: Mahsunah
authors.forename: Frank
authors.initials: F
authors.lastname: Arnold
authors.forename: Thomas
authors.initials: T
authors.lastname: Piecha
authors.forename: Christian G
authors.initials: CG
authors.lastname: Huber
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D008422:Materials Testing; D008433:Mathematics; D008962:Models, Theoretical; D010226:Parabens / Q000032:analysis; D011310:Preservatives, Pharmaceutical / Q000032:analysis; D013056:Spectrophotometry, Ultraviolet
chemical_list: D010226:Parabens; D011310:Preservatives, Pharmaceutical