Analysis of neutral lipids

SC Shuqiang Chen
MW Ming Wang
LL Li Li
JW Jun Wang
XM Xuhui Ma
HZ Hengde Zhang
YC Yang Cai
BK Bin Kang
JH Jianlei Huang
BL Bo Li
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Neutral lipids (TAGs, DAGs and CEs) were analyzed using a modified version of reverse-phase HPLC/MRM as previously described [33, 34]. Briefly, the lipids were separated on a Phenomenex Kinetex 2.6 μm C18 column (i.d. 4.6 × 100 mm2) using an isocratic mobile phase of chloroform:methanol:0.1 M ammonium acetate (100:100:4) at a flow rate of 170 µl/min for 17 min. Levels of short, medium, and long-chain TAGs were calculated by referencing the spiked internal standards of TAG(14:0)3-d5, TAG(16:0)3-d5 and TAG(18:0)3-d5 obtained from CDN isotopes, respectively. DAGs were quantified using d5-DAG16:0/16:0 and d5-DAG18:1/18:1 (Avanti Polar Lipids) as internal standards. Free cholesterols and cholesteryl esters were analyzed as previously described with d6-cholesterol and d6-C18:0 cholesteryl ester (CE) (CDN isotopes) as internal standards.

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