Absolute quantification of lipids in complex mixtures by HPLC without authentic standards

The identification and quantification of compounds in biological samples are commonly performed using a technique called reversed phase-HPLC- hyphenated to mass spectrometric detection (RP-HPLC-MS).

However, when authentic standards are not available, quantification becomes difficult due to the analyte-specific response of the detector, as the relative signal intensities in electrospray ionization-MS are dependent on ionization efficiency.

The choice of an appropriate standard is further limited by various parameters such as the chemical and physical properties of the analyte or the retention behavior.

Additionally, in lipid research, the analytes often lack chromophoric groups, which means that UV/vis detection is not feasible, or additional derivatization procedures become inevitable.

To overcome these challenges, a complementary quantification strategy called charged aerosol detection (CAD) is employed. CAD offers a consistent and uniform response for nearly all non-volatile substances, whilst ensuring both mass sensitivity and robustness.

In this webinar, quantification strategies without authentic standards based on HPLC-CAD will be showcased for determination of several lipid classes in complex sample matrices. Examples of these lipid classes include biosurfactants like rhamnolipids and liamocins, as well as membrane lipids such as phospholipids.

Learning points:

• What makes charged aerosol detection so powerful

• What benefits it offers

• What applications are ideal for charged aerosol detection?

Who should attend:

• Laboratory managers

• Technology leaders

• Principal scientists

• Laboratory analyst

Presenter: Prof. Dr. Heiko Hayen (Professor, Institute of Inorganic and Analytical Chemistry, University of Münster)

Heiko Hayen is professor for Analytical Chemistry at the University of Münster since 2014. He studied Food Chemistry and obtained his PhD in Analytical Chemistry at the University of Twente (Enschede, The Netherlands) in 2003.

After a postdoctoral research stay at the Institute for Marine Biosciences (National Research Council of Canada, Halifax, Nova Scotia, Canada), he moved to the Department of Bioanalytics, Leibniz-Institut für Analytische Wissenschaften – ISAS – e.V., Dortmund.

From 2010 to 2014 he held a professorship in Food Chemistry at the Faculty of Mathematics and Natural Sciences, University of Wuppertal.

His current research interests include the development of hyphenated mass spectrometric methods for the investigation of metabolites and lipids, like phospholipids and glycolipids, using HPLC-MS, SFC-MS and capillary IC-MS.

Presenter: Chris Tuczemskyi (Moderator, Product Marketing Manager, Thermo Fisher Scientific)

Chris Tuczemskyi is a chromatography specialist at Thermo Fisher Scientific, based in Cambridge, UK. After spending a number of years building up his experience with chromatography instruments in a water testing laboratory, he joined Thermo Fisher Scientific in 2014 as a chromatography service engineer for Unity Lab Services. Here he developed his passion for supporting customers using HPLC and in 2018 moved into the application specialist team. More recently, in 2022 he made the move to Product Marketing to help ensure we meet and exceed our customers expectation. In 2009, Chris graduated from the University of Essex in Biochemistry (Bsc Hons).