On-line high speed lipid extraction for nanoflow liquid chromatography-tandem mass spectrometry

Ju Yong Lee, Joon Seon Yang, Se Mi Park, Seul Kee Byeon, Myeong Hee Moon

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)


An on-line lipid extraction method is introduced by utilizing a short capillary extraction column using HILIC and C4 particles prior to nanoflow liquid chromatography-tandem mass spectrometry (nLC–MS/MS). The on-line extraction using a urine sample spiked with PL standards showed similar or slightly higher recovery values (86%–96%) of phospholipids (PLs) compared to those obtained by the conventional off-line extraction based on the Folch method with or without using the air-exposed drying process. In this study, we demonstrated that PL oxidation can occur during the air-exposed drying process of lipid extracts in standard liquid–liquid extraction procedures, which was confirmed by the oxidized PL (OxPL) molecules that were generated from an off-line extraction using a few PL standards. Quantitative comparison of these OxPL species between on- and off-line extraction followed by nLC–MS/MS with multiple reaction monitoring (MRM) analysis showed a significant decrease (2–10 fold) in unwanted OxPL species when on-line extraction was employed. While the number of identified PLs from a urine sample was somewhat lower than those by off-line extraction, the number of OxPLs was significantly reduced (from 70 to 22) with on-line extraction. The new method offers high speed (∼ 5 min) automated extraction of PLs with nLC–MS/MS analysis and presents the possibility of handling a biological sample with a very limited amount of lipids.

Original languageEnglish
Pages (from-to)12-20
Number of pages9
JournalJournal of Chromatography A
Publication statusPublished - 2016 Sept 16

Bibliographical note

Funding Information:
This study was supported by a grant ( NRF-2015004677 ) from National Research Foundation (NRF) of Korea .

Publisher Copyright:
© 2016 Elsevier B.V.

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry


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