title: Blind column selection protocol for two-dimensional high performance liquid chromatography.
abstract: The selection of two orthogonal columns for two-dimensional high performance liquid chromatography (LC×LC) separation of natural product extracts can be a labour intensive and time consuming process and in many cases is an entirely trial-and-error approach. This paper introduces a blind optimisation method for column selection of a black box of constituent components. A data processing pipeline, created in the open source application OpenMS®, was developed to map the components within the mixture of equal mass across a library of HPLC columns; LC×LC separation space utilisation was compared by measuring the fractional surface coverage, fcoverage. It was found that for a test mixture from an opium poppy (Papaver somniferum) extract, the combination of diphenyl and C18 stationary phases provided a predicted fcoverage of 0.48 and was matched with an actual usage of 0.43. OpenMS®, in conjunction with algorithms designed in house, have allowed for a significantly quicker selection of two orthogonal columns, which have been optimised for a LC×LC separation of crude extractions of plant material.
authors.affiliation: Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, Waurn Ponds, Vic 3216, Australia.
authors.forename: Niki K
authors.initials: NK
authors.lastname: Burns
authors.affiliation: Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, Waurn Ponds, Vic 3216, Australia.
authors.forename: Luke M
authors.initials: LM
authors.lastname: Andrighetto
authors.affiliation: Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, Waurn Ponds, Vic 3216, Australia.
authors.forename: Xavier A
authors.initials: XA
authors.lastname: Conlan
authors.affiliation: Sun Pharmaceutical Industries Australia Pty, Port Fairy, Vic 3284, Australia.
authors.forename: Stuart D
authors.initials: SD
authors.lastname: Purcell
authors.affiliation: Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, Waurn Ponds, Vic 3216, Australia.
authors.forename: Neil W
authors.initials: NW
authors.lastname: Barnett
authors.affiliation: School of Information Technology, Deakin University, Waurn Ponds, Vic 3216, Australia.
authors.forename: Jacquie
authors.initials: J
authors.lastname: Denning
authors.affiliation: Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, Waurn Ponds, Vic 3216, Australia.
authors.forename: Paul S
authors.initials: PS
authors.lastname: Francis
authors.affiliation: Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, Waurn Ponds, Vic 3216, Australia. Electronic address: paul.stevenson@deakin.edu.au.
authors.forename: Paul G
authors.initials: PG
authors.lastname: Stevenson
mesh_terms: D002851:Chromatography, High Pressure Liquid
keywords: Electrospray mass spectrometry; Multidimensional high performance liquid chromatography; OpenMS®; Plant extracts