LC/MS Analysis of Triacylglycerols (TAG) in Human Plasma on Ascentis® Express C18, NARP-HPLC-APCI-qMS Method

LC/MS Analysis of Triacylglycerols (TAG) in Human Plasma on Ascentis® Express C18, NARP-HPLC-APCI-qMS Method

Conditions

column Ascentis Express C18, 15 cm x 4.6 mm I.D., 2.7 µm particles (53829-U)
mobile phase [A] acetonitrile; [B] isopropanol
gradient 0 to 70% B in 50 min; held for 5 min; to 0% B in 6 min
flow rate 1.0 mL/min
sample 10 mg/mL
injection 5.0 µL
detector MS, APCI(+), m/z 250-1100
detector MS parameters: 250-1100 m/z mass range; scan speed: 4000 amu/s; nebulizing gas (nitrogen) flow rate: 2.5 L/min; event time: 0.2 sec; (detector voltage: 1.5 kV; interface voltage: 4.5 kV; interface temperature: 400 °C; CDL temperature: 250 °C; heat block temperature: 200 °C)

Description

Analysis Note Individual TAGs were identified on the basis of their positive-ion APCI mass spectra exploiting protonated ions ([M+H]+), and diglyceride fragment ions ([DG]+) for the identification of individual FAs. Additionally, the predicted elution order [increasing partition number (PN), the latter equal to the carbon number (CN) minus two times the double bond number (DB)], under NARP (non-aqueous reversed-phase) conditions, was used to support TAG identification.
Categories Analytical Chromatography, Clinical, Lipids, lipids
Featured Industry Clinical
Life Science and Biopharma
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC
Other Notes The plasma lipid fraction of each sample was extracted according to the Folch method. Specifically, 1 mL of serum, for each sample, was placed in a centrifuge tube with 9 mL of a chloroform/methanol (2:1) mixture, extracted twice and centrifuged. The lower lipid-containing organic phase was dried with anhydrous Na2SO4, filtered, then distilled with a rotating evaporator.
suitability application for LC-MS

Materials

     
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