“This new generation system provides improved hardware and software features that answer the pharmaceutical industry’s demand for an efficient and robust prep SFC system.” “Working with the Enabling Technologies Consortium, and hearing the voice of the pharmaceutical industry, has been an incredibly valuable learning experience for us at Shimadzu,” said Patrick Fromal, VP Sales, Shimadzu Scientific Instruments, based in Columbia, Maryland, USA. “The Nexera UC Prep system is an improved purification instrumentation that meets the specifications and requirements of SFC users across the pharmaceutical industry” said Mirlinda Biba, PhD, Principal Scientist, Merck & Co. It simplifies configuration of parameter settings and greatly increases efficiency of the preparative workflow. The newly developed, easy-to-use “Prep Solution” software streamlines operations. Its elegant design reduces the total size of the recovery system and allows for easy rinsing compared to earlier cyclone-style or centrifugal type gas-liquid separating systems. The novel and patented “LotusStream” gas-liquid separator technology ensures higher recovery and lower carryover than earlier prep SFC systems.
Innovative technologies include a flexible format, combination injector/fraction collector, and CO2 pump with integrated chiller requiring less lab space, thereby enabling benchtop use.
This complete SFC solution reduces the need for costly and hazardous solvents used in normal phase prep LC, while shortening purification run time and dry down time. A Nexera UC Prep system can be configured according to user specifications for optimum performance of the desired purification function, including chiral or achiral purifications, single injections, stacked injections and fraction collections from several microliters to liters. The Nexera UC Prep is part of Shimadzu’s Nexera UC platform, which accommodates a wide variety of analyses and purifications. It provides flexible system configurations in a compact design, requiring low installation space and enabling maximum use of lab resources. Nexera UC Prep resolves a number of issues in preparative tasks, reducing labor and improving efficiency while fitting into pre-existing workflows. Finally, compared with established fractionation/pre-concentration approaches, semi-preparative SFC is superior in terms of versatility, as it involved only volatile modifiers and salt additives facilitating any follow-up use such as qualitative or quantitate analysis or further purification down to the single lipid species level.Unparalleled performance, unparalleled flexibility Thus, the novel workflow is ideally suited for lipid class-specific purification/isolation from milligram amounts of sample while not compromising on omics type of analysis (identification and quantification). Indeed, for 70 lipids, the consensus values available for this sample could be met. Proof-of-principle experiments using a standard reference material (SRM 1950, NIST) for human plasma showed that the proposed strategy enabled quantitative lipidomics. A significantly increased coverage of low abundant lipids improving lipid identification by numbers and degree (species and molecular level) was obtained in Pichia pastoris when comparing high-resolution mass spectrometry based lipidomics with and without prior fractionation. The powerful SFC approach offered separation of a wide polarity range for lipids, enabled enrichment (up to 3 orders of magnitude) of lipids, selective fractionation of 14 lipid classes/subclasses, and increased dynamic range enabling in-depth characterization.
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Abstract : In this work, a lipidomics workflow based on offline semi-preparative lipid class-specific fractionation by supercritical fluid chromatography (SFC) followed by high-resolution mass spectrometry was introduced.
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