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Fatty Acid Profiling and Desaturase Activity

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Total lipids were extracted from the plasma samples and converted to their methyl ester equivalents as described previously (12 (link), 22 (link)), and analyzed using an Agilent 7820A GC using flame ionization detection on a SP-2560 polar fused silica capillary column (100 m × 0.25 mm × 0.2 μm; Supelco Inc., PA, USA) with nitrogen as the carrier gas (12 (link)). The FA peaks were identified by comparing retention times with those of a standard mixture of GLC-68A, GLC-481B, GLC-532, GLC-744 (Nu-Chek Prep), 37 FAME, trans 16:1n-7, trans 18:1n-7, trans 18:1n-9, and cis/trans 18:2n-6 (all obtained from Supelco Inc., or Sigma). 13:0 free fatty acid (10 μg) was added as an internal standard. FA composition was expressed as the weight of a percentage of the total weight of carbon-12 to carbon-24 FAs (wt%). The estimated desaturase activity was calculated using the ratio of FA in plasma (11 (link), 17 (link), 23 (link)); SCD1(16) activity = (16:1n-7/16:0), SCD1(18) activity = (18:1n-9/18:0), D5D activity = (20:4n-6/20:3n-6), and D6D activity = (20:3n-6/18:2n-6).

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Hua M.C., Su H.M., Lai M.W., Yao T.C., Tsai M.H., Liao S.L., Lai S.H, & Huang J.L. (2021). Palmitoleic and Dihomo-γ-Linolenic Acids Are Positively Associated With Abdominal Obesity and Increased Metabolic Risk in Children. Frontiers in Pediatrics, 9, 628496.

Publication 2021

Agilent 7820A GC SP-2560 GLC-68A GLC-532 GLC-744 cis/trans 18:2n-6

Capillary Carbon Ester Flame ionization Free fatty acid Lipids Nitrogen Plasma Retention Silica

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Variable analysis

independent variables

Total lipids extracted from plasma samples
Lipids converted to methyl ester equivalents

dependent variables

Fatty acid (FA) composition (expressed as weight percentage of total carbon-12 to carbon-24 FAs)
Estimated desaturase activity (SCD1(16), SCD1(18), D5D, D6D)

control variables

Agilent 7820A GC using flame ionization detection on a SP-2560 polar fused silica capillary column (100 m × 0.25 mm × 0.2 μm) with nitrogen as the carrier gas
Comparison of FA peaks with retention times of standard mixtures (GLC-68A, GLC-481B, GLC-532, GLC-744, 37 FAME, trans 16:1n-7, trans 18:1n-7, trans 18:1n-9, and cis/trans 18:2n-6)
Addition of 13:0 free fatty acid (10 μg) as an internal standard

controls

Positive controls: Standard mixtures of fatty acid methyl esters (GLC-68A, GLC-481B, GLC-532, GLC-744, 37 FAME, trans 16:1n-7, trans 18:1n-7, trans 18:1n-9, and cis/trans 18:2n-6)
Negative control: Not explicitly mentioned

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