Supplementary Table S2 . List of Internal Standards Used in LC-MS/MS

Supplementary Table S1. Fatty acid composition of experimental diets. Chow diet and high fat diet contain 4.46 g and 32.94 g of fatty acids per 100 g, respectively. Calorie values for chow diet are 53 % carbohydrate, 36 % protein, 11 % fat; for high fat diet 22 % carbohydrate, 24 % protein and 54 % fat. Numbers in bold italics highlight fatty acids with highest wt % gains in high fat diet.

Fatty acid / Chow diet (FED) control / High fat diet (HFD)
wt % of diet / wt % of total FA / wt % of diet / wt % of total FA
10:0 / 0.04 / 0.1
12:0 / 0.07 / 0.2
14:0 / 0.01 / 0.2 / 0.44 / 1.3
16:0 / 0.56 / 12.5 / 7.22 / 23.4
16:1 / 0.02 / 0.5 / 0.94 / 2.9
18:0 / 0.14 / 3.1 / 4.34 / 13.2
18:1 / 0.96 / 21.5 / 13.57 / 41.2
18:2 / 2.42 / 54.3 / 4.75 / 14.4
18:3 / 0.31 / 6.7 / 0.51 / 1.6
20:0 / 0.02 / 0.5 / 0.02 / 0.06
20:1 / 0.02 / 0.5 / 0.01 / 0.03
20:4 / 0.53 / 1.6

Supplementary Table S2. List of internal standards used in LC-MS/MS

Lipid class / Internal standards used / Concentration of internal standards in autosampler vial
PS / 12:0/13:0 14:1/17:0 17:0/20:4 21:0/22:6 / 2 µM
SM / LM 6002 SP mix (10 compounds) / 1 µM
PI / 12:0/13:0 14:1/17:0 17:0/20:4 21:0/22:6 / 3 µM
LPC / 17:0 / 1 µM
PE / 12:0/13:0 14:1/17:0 17:0/20:4 21:0/22:6 / 2 µM
PC / 12:0/13:0 14:1/17:0 17:0/20:4 21:0/22:6 / 1 µM
DG / LM 6001 DG mix (9 compounds) / 4 µM
TG / LM 6000 TG mix (9 compounds) / 1.7 µM

Supplementary Table S3. Summary of TG species determined by LC-MS/MS in murine hepatocyte LD from FED, FAS and HFD samples. Absolute amounts (n=3 animals) are calculated in µmol, and are normalized to gram total acylgycerols (InfinityTM Triglycerides Reagent kit). Corresponding ‰ values (n=6 animals) are relative to total amount of TGspecies. Molecular species are derived from possible fatty acyl combinations as determined by fatty acid fragment ions (neutral losses) detected in the corresponding product ion spectra. Fatty acyls in bold are major compositions.

Species / µmol / g / ‰ / Molecular species
TG / 28:0 / FED / 0.04 / ± / 0.01 / 0.01 / ± / 0.00
FAS / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.01 / ± / 0.00 / 0.00 / ± / 0.00
TG / 30:0 / FED / 0.05 / ± / 0.00 / 0.01 / ± / 0.00
FAS / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.01 / ± / 0.00 / 0.00 / ± / 0.00
TG / 32:0 / FED / 0.56 / ± / 0.06 / 0.25 / ± / 0.14 / 8:0_8:0_16:0
FAS / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.22 / ± / 0.07 / 0.07 / ± / 0.03
TG / 32:1 / FED / 0.19 / ± / 0.01 / 0.05 / ± / 0.03
FAS / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.06 / ± / 0.02 / 0.02 / ± / 0.01
TG / 34:0 / FED / 0.28 / ± / 0.03 / 0.09 / ± / 0.05 / 8:0_8:0_18:0 / 8:0_10:0_16:0
FAS / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.10 / ± / 0.03 / 0.03 / ± / 0.02
TG / 34:1 / FED / 2.01 / ± / 0.16 / 0.58 / ± / 0.13
FAS / 0.03 / ± / 0.01 / 0.02 / ± / 0.01
HFD / 1.19 / ± / 0.36 / 0.45 / ± / 0.16
TG / 34:2 / FED / 1.75 / ± / 0.13 / 0.47 / ± / 0.12
FAS / 0.05 / ± / 0.02 / 0.03 / ± / 0.02
HFD / 0.19 / ± / 0.07 / 0.08 / ± / 0.02 / 8:0_8:0_18:2
TG / 36:0 / FED / 0.04 / ± / 0.02 / 0.01 / ± / 0.00
FAS / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.02 / ± / 0.00 / 0.01 / ± / 0.00
TG / 36:1 / FED / 0.23 / ± / 0.03 / 0.06 / ± / 0.02
FAS / 0.01 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.12 / ± / 0.04 / 0.04 / ± / 0.02 / 8:0_10:0_18:1
TG / 36:2 / FED / 0.19 / ± / 0.01 / 0.06 / ± / 0.02
FAS / 0.01 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.02 / ± / 0.01 / 0.01 / ± / 0.00
TG / 36:3 / FED / 0.06 / ± / 0.01 / 0.02 / ± / 0.00
FAS / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.02 / ± / 0.01 / 0.01 / ± / 0.00
TG / 36:4 / FED / 0.46 / ± / 0.04 / 0.19 / ± / 0.11
FAS / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.17 / ± / 0.06 / 0.05 / ± / 0.04 / 8:0_8:0_20:4
TG / 38:0 / FED / 0.03 / ± / 0.01 / 0.01 / ± / 0.00
TG continued
FAS / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.01 / ± / 0.00 / 0.01 / ± / 0.00
TG / 38:1 / FED / 0.09 / ± / 0.00 / 0.02 / ± / 0.00
FAS / 0.01 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.03 / ± / 0.01 / 0.01 / ± / 0.00
TG / 38:2 / FED / 0.08 / ± / 0.01 / 0.02 / ± / 0.00 / 8:0_12:0_18:2 / 10:0_10:0_18:2
FAS / 0.01 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.01 / ± / 0.00 / 0.00 / ± / 0.00
TG / 38:4 / FED / 0.04 / ± / 0.01 / 0.01 / ± / 0.00
FAS / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
HFD / 0.01 / ± / 0.01 / 0.01 / ± / 0.00
TG / 40:0 / FED / 0.30 / ± / 0.05 / 0.22 / ± / 0.19
FAS / 0.02 / ± / 0.00 / 0.01 / ± / 0.00
HFD / 0.17 / ± / 0.02 / 0.10 / ± / 0.03
TG / 40:1 / FED / 0.57 / ± / 0.08 / 0.30 / ± / 0.19
FAS / 0.07 / ± / 0.01 / 0.05 / ± / 0.03
HFD / 0.24 / ± / 0.06 / 0.15 / ± / 0.07
TG / 40:2 / FED / 0.29 / ± / 0.02 / 0.09 / ± / 0.04
FAS / 0.07 / ± / 0.02 / 0.04 / ± / 0.02
HFD / 0.05 / ± / 0.01 / 0.03 / ± / 0.01
TG / 42:1 / FED / 6.14 / ± / 1.08 / 2.21 / ± / 0.87
FAS / 0.89 / ± / 0.28 / 0.29 / ± / 0.07
HFD / 4.34 / ± / 0.40 / 2.33 / ± / 0.66
TG / 42:2 / FED / 10.69 / ± / 1.32 / 4.38 / ± / 1.97
FAS / 1.87 / ± / 0.80 / 1.09 / ± / 0.53
HFD / 2.06 / ± / 0.42 / 1.14 / ± / 0.33
TG / 42:3 / FED / 1.50 / ± / 0.08 / 0.51 / ± / 0.17
FAS / 0.29 / ± / 0.10 / 0.18 / ± / 0.09
HFD / 0.22 / ± / 0.03 / 0.11 / ± / 0.03
TG / 42:4 / FED / 0.12 / ± / 0.01 / 0.03 / ± / 0.01
FAS / 0.05 / ± / 0.02 / 0.02 / ± / 0.01
HFD / 0.03 / ± / 0.01 / 0.01 / ± / 0.00
TG / 44:1 / FED / 1.01 / ± / 0.26 / 0.31 / ± / 0.07
FAS / 0.41 / ± / 0.09 / 0.11 / ± / 0.06
HFD / 0.64 / ± / 0.10 / 0.29 / ± / 0.04
TG / 44:2 / FED / 3.99 / ± / 0.48 / 1.23 / ± / 0.30
FAS / 2.73 / ± / 0.83 / 1.02 / ± / 0.17
HFD / 1.65 / ± / 0.17 / 0.72 / ± / 0.16
TG / 44:3 / FED / 3.87 / ± / 0.58 / 1.22 / ± / 0.42
FAS / 1.78 / ± / 0.86 / 1.08 / ± / 0.55
HFD / 0.99 / ± / 0.23 / 0.51 / ± / 0.15
TG / 44:4 / FED / 5.56 / ± / 0.66 / 2.28 / ± / 1.00
FAS / 0.68 / ± / 0.31 / 0.42 / ± / 0.21
HFD / 2.67 / ± / 0.56 / 1.16 / ± / 0.19
TG / 44:5 / FED / 0.56 / ± / 0.01 / 0.17 / ± / 0.04
FAS / 0.10 / ± / 0.04 / 0.05 / ± / 0.02
HFD / 0.13 / ± / 0.02 / 0.06 / ± / 0.01
TG / 46:1 / FED / 0.66 / ± / 0.25 / 0.18 / ± / 0.04
FAS / 0.94 / ± / 0.26 / 0.24 / ± / 0.15
TG continued
HFD / 0.63 / ± / 0.21 / 0.33 / ± / 0.07
TG / 46:2 / FED / 1.28 / ± / 0.27 / 0.35 / ± / 0.06
FAS / 6.24 / ± / 0.42 / 1.89 / ± / 0.54
HFD / 0.68 / ± / 0.10 / 0.36 / ± / 0.07
TG / 46:3 / FED / 0.89 / ± / 0.12 / 0.50 / ± / 0.32 / 8:0_20:3_18:0
FAS / 6.68 / ± / 1.97 / 2.71 / ± / 0.59 / 8:0_20:3_18:0
HFD / 0.37 / ± / 0.04 / 0.21 / ± / 0.06 / 10:0_18:1_18:2 / 10:1_18:1_18:1 / 12:0_16:1_18:2
TG / 46:4 / FED / 2.61 / ± / 0.35 / 1.15 / ± / 0.54
FAS / 2.29 / ± / 1.06 / 1.38 / ± / 0.70
HFD / 1.48 / ± / 0.17 / 0.67 / ± / 0.11
TG / 46:5 / FED / 1.52 / ± / 0.28 / 0.83 / ± / 0.49 / 8:0_22:5_16:0
FAS / 0.53 / ± / 0.25 / 0.35 / ± / 0.20 / 8:0_22:5_16:0
HFD / 1.04 / ± / 0.17 / 0.52 / ± / 0.11
TG / 46:6 / FED / 4.86 / ± / 0.40 / 2.40 / ± / 1.34 / 8:0_22:6_16:0
FAS / 0.45 / ± / 0.17 / 0.24 / ± / 0.11 / 8:0_22:6_16:0
HFD / 1.75 / ± / 0.25 / 1.05 / ± / 0.29 / 8:0_22:6_16:0
TG / 48:0 / FED / 1.35 / ± / 0.19 / 0.46 / ± / 0.14
FAS / 2.14 / ± / 0.25 / 0.75 / ± / 0.09
HFD / 0.68 / ± / 0.12 / 0.75 / ± / 0.51
TG / 48:1 / FED / 6.69 / ± / 0.95 / 1.87 / ± / 0.31
FAS / 5.74 / ± / 0.53 / 2.02 / ± / 0.16
HFD / 6.59 / ± / 0.91 / 3.74 / ± / 0.77
TG / 48:2 / FED / 9.77 / ± / 0.81 / 2.74 / ± / 0.43
FAS / 13.15 / ± / 0.09 / 4.62 / ± / 0.45
HFD / 8.48 / ± / 1.77 / 4.46 / ± / 0.71
TG / 48:3 / FED / 3.71 / ± / 0.20 / 1.15 / ± / 0.30
FAS / 16.33 / ± / 1.01 / 6.00 / ± / 0.42
HFD / 2.15 / ± / 0.35 / 1.30 / ± / 0.41
TG / 48:4 / FED / 0.62 / ± / 0.07 / 0.57 / ± / 0.46
FAS / 16.20 / ± / 4.17 / 6.15 / ± / 1.03
HFD / 0.46 / ± / 0.07 / 0.22 / ± / 0.03
TG / 48:5 / FED / 0.31 / ± / 0.09 / 0.10 / ± / 0.03
FAS / 4.27 / ± / 2.01 / 2.43 / ± / 1.16
HFD / 0.17 / ± / 0.02 / 0.10 / ± / 0.07
TG / 48:6 / FED / 0.88 / ± / 0.13 / 0.42 / ± / 0.22
FAS / 1.00 / ± / 0.55 / 0.65 / ± / 0.36
HFD / 0.32 / ± / 0.05 / 0.16 / ± / 0.02
TG / 48:7 / FED / 0.86 / ± / 0.06 / 0.37 / ± / 0.17
FAS / 0.19 / ± / 0.09 / 0.12 / ± / 0.07
HFD / 0.30 / ± / 0.03 / 0.15 / ± / 0.03
TG / 48:8 / FED / 0.24 / ± / 0.04 / 0.08 / ± / 0.03
FAS / 0.03 / ± / 0.01 / 0.02 / ± / 0.01
HFD / 0.05 / ± / 0.01 / 0.02 / ± / 0.00
TG / 50:0 / FED / 0.31 / ± / 0.04 / 0.21 / ± / 0.14
FAS / 1.09 / ± / 0.30 / 0.38 / ± / 0.06
HFD / 2.31 / ± / 0.32 / 1.36 / ± / 0.45
TG / 50:1 / FED / 40.87 / ± / 3.47 / 12.61 / ± / 2.52 / 16:0_16:0_18:1 / 14:0_16:0_20:1 / 14:0_18:0_18:1
FAS / 51.87 / ± / 8.00 / 17.12 / ± / 1.89 / 16:0_16:0_18:1 / 14:0_18:0_18:1
TG continued
TG / 50:2 / FED / 117.39 / ± / 12.72 / 29.49 / ± / 1.75 / 16:0_16:1_18:1 / 16:0_16:0_18:2 / 14:0_18:1_18:1
FAS / 86.33 / ± / 9.49 / 31.44 / ± / 2.51 / 16:0_16:1_18:1 / 16:0_16:0_18:2 / 14:0_18:1_18:1
HFD / 98.79 / ± / 19.08 / 46.73 / ± / 2.93 / 16:0_16:1_18:1 / 16:0_16:0_18:2 / 14:0_18:1_18:1
TG / 50:3 / FED / 88.65 / ± / 7.41 / 22.21 / ± / 2.01 / 16:0_16:1_18:2 / 16:1_16:1_18:1 / 14:0_18:1_18:2
FAS / 65.62 / ± / 1.13 / 25.03 / ± / 2.62 / 16:0_16:1_18:2 / 16:1_16:1_18:1 / 14:0_18:1_18:2
HFD / 51.38 / ± / 9.30 / 24.55 / ± / 2.25 / 16:0_16:1_18:2 / 14:0_18:1_18:2 / 16:0_16:2_18:1
16:0_16:0_18:3
TG / 50:4 / FED / 19.97 / ± / 0.66 / 5.45 / ± / 0.73
FAS / 50.56 / ± / 4.99 / 18.16 / ± / 1.23
HFD / 10.99 / ± / 1.73 / 5.66 / ± / 0.83
TG / 50:5 / FED / 2.19 / ± / 0.21 / 0.81 / ± / 0.30
FAS / 27.79 / ± / 4.39 / 10.42 / ± / 1.29
HFD / 1.78 / ± / 0.26 / 0.98 / ± / 0.22
TG / 50:6 / FED / 0.23 / ± / 0.08 / 0.23 / ± / 0.20
FAS / 8.86 / ± / 3.82 / 4.51 / ± / 1.84
HFD / 0.22 / ± / 0.04 / 0.15 / ± / 0.05
TG / 50:7 / FED / 0.10 / ± / 0.02 / 0.11 / ± / 0.09
FAS / 1.50 / ± / 0.75 / 0.99 / ± / 0.55
HFD / 0.04 / ± / 0.01 / 0.03 / ± / 0.02
TG / 50:8 / FED / 0.03 / ± / 0.01 / 0.01 / ± / 0.00
FAS / 0.15 / ± / 0.07 / 0.11 / ± / 0.07
HFD / 0.01 / ± / 0.00 / 0.00 / ± / 0.00
TG / 52:0 / FED / 0.09 / ± / 0.01 / 0.12 / ± / 0.11 / 16:0_18:0_18:0 / 16:0_16:0_20:0
FAS / 0.10 / ± / 0.04 / 0.07 / ± / 0.04
HFD / 0.92 / ± / 0.10 / 0.83 / ± / 0.48 / 16:0_18:0_18:0 / 16:0_16:0_20:0
TG / 52:1 / FED / 5.90 / ± / 0.42 / 3.93 / ± / 2.70 / 16:0_18:0_18:1
FAS / 16.78 / ± / 5.20 / 5.49 / ± / 0.99 / 16:0_18:0_18:1
HFD / 55.08 / ± / 7.72 / 26.08 / ± / 3.79 / 16:0_18:0_18:1
TG / 52:2 / FED / 361.72 / ± / 31.84 / 101.83 / ± / 11.35 / 16:0_18:1_18:1
FAS / 204.62 / ± / 29.22 / 62.45 / ± / 10.19 / 16:0_18:1_18:1
HFD / 401.46 / ± / 93.32 / 204.54 / ± / 20.19 / 16:0_18:1_18:1 / 16:0_18:0_18:2
TG / 52:3 / FED / 795.65 / ± / 78.97 / 182.90 / ± / 24.09 / 16:0_18:1_18:2 / 16:1_18:1_18:1
FAS / 345.33 / ± / 26.33 / 118.40 / ± / 3.30 / 16:0_18:1_18:2
HFD / 331.69 / ± / 69.57 / 153.44 / ± / 8.52 / 16:0_18:1_18:2 / 16:1_18:1_18:1
TG / 52:4 / FED / 507.88 / ± / 44.24 / 115.27 / ± / 16.93 / 16:0_18:2_18:2 / 16:0_18:1_18:3 / 16:1_18:1_18:2
16:2_16:2_20:0
FAS / 305.09 / ± / 7.68 / 107.08 / ± / 3.78 / 16:0_18:2_18:2 / 16:1_18:1_18:2 / 16:0_18:1_18:3
16:2_18:1_18:1
HFD / 133.35 / ± / 29.26 / 57.29 / ± / 6.21 / 16:0_18:2_18:2 / 16:1_18:1_18:2 / 16:0_18:1_18:3
16:2_18:1_18:1
TG / 52:5 / FED / 101.57 / ± / 3.29 / 26.76 / ± / 2.21 / 16:1_18:2_18:2 / 16:0_18:2_18:3 / 16:0_16:1_20:4
16:0_18:1_18:4
FAS / 145.33 / ± / 5.70 / 50.21 / ± / 1.81 / 16:1_18:2_18:2 / 16:0_18:2_18:3 / 16:0_16:1_20:4
16:0_18:1_18:4
HFD / 31.32 / ± / 4.32 / 15.34 / ± / 1.49 / 16:1_18:2_18:2 / 16:0_18:2_18:3 / 16:0_16:1_20:4
16:0_18:1_18:4
TG / 52:6 / FED / 10.26 / ± / 0.62 / 3.47 / ± / 1.08 / 16:1_18:2_18:3 / 16:0_18:2_18:4 / 16:0_16:2_20:4
16:3_18:1_18:2
TG continued
FAS / 62.86 / ± / 6.62 / 22.96 / ± / 1.87 / 16:1_18:2_18:3 / 16:0_18:2_18:4 / 16:0_16:2_20:4
16:3_18:1_18:2
HFD / 4.18 / ± / 0.61 / 2.44 / ± / 0.67 / 16:1_18:2_18:3 / 16:0_18:2_18:4 / 16:0_16:2_20:4
16:3_18:1_18:2
TG / 52:7 / FED / 1.03 / ± / 0.13 / 0.34 / ± / 0.09
FAS / 15.80 / ± / 5.52 / 7.58 / ± / 2.58
HFD / 0.44 / ± / 0.06 / 0.26 / ± / 0.08 / 16:1_18:2_18:4 / 16:2_18:2_18:3 / 16:3_18:2_18:2
16:4_18:1_18:2
TG / 52:8 / FED / 0.11 / ± / 0.03 / 0.03 / ± / 0.01
FAS / 2.66 / ± / 1.25 / 1.64 / ± / 0.85
HFD / 0.03 / ± / 0.01 / 0.01 / ± / 0.01 / 16:2_18:3_18:3 / 16:3_18:2_18:3 / 16:4_18:2_18:2
TG / 52:9 / FED / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
FAS / 0.29 / ± / 0.16 / 0.20 / ± / 0.11
HFD / 0.00 / ± / 0.00 / 0.00 / ± / 0.00 / 16:3_18:2_18:4 / 16:4_18:2_18:3 / 14:2_16:3_22:6
TG / 52:10 / FED / 0.02 / ± / 0.00 / 0.00 / ± / 0.00
FAS / 0.02 / ± / 0.01 / 0.02 / ± / 0.01
HFD / 0.01 / ± / 0.00 / 0.00 / ± / 0.00
TG / 54:0 / FED / 0.00 / ± / 0.00 / 0.07 / ± / 0.02
FAS / 0.02 / ± / 0.00 / 0.01 / ± / 0.01
HFD / 0.10 / ± / 0.01 / 0.10 / ± / 0.08
TG / 54:1 / FED / 1.34 / ± / 0.07 / 0.35 / ± / 0.04 / 16:0_18:1_20:0 / 16:0_18:0_20:1
FAS / 1.08 / ± / 0.47 / 0.76 / ± / 0.45 / 16:0_18:1_20:0 / 16:0_18:0_20:1 / 14:0_18:1_22:0
HFD / 18.99 / ± / 3.11 / 12.25 / ± / 3.97 / 16:0_18:1_20:0 / 16:0_18:0_20:1 / 14:0_18:1_22:0
TG / 54:2 / FED / 23.38 / ± / 1.23 / 14.07 / ± / 9.56 / 18:0_18:1_18:1 / 16:0_18:1_20:1 / 16:0_20:0_18:2
FAS / 37.76 / ± / 13.87 / 12.56 / ± / 2.54 / 18:0_18:1_18:1 / 16:0_18:1_20:1 / 16:0_20:0_18:2
14:0_18:1_22:1
HFD / 131.12 / ± / 20.74 / 54.56 / ± / 9.50 / 18:0_18:1_18:1 / 16:0_18:1_20:1 / 16:0_20:0_18:2
TG / 54:3 / FED / 190.24 / ± / 16.72 / 49.47 / ± / 2.00 / 18:1_18:1_18:1 / 16:0_18:2_20:1 / 16:0_18:1_20:2
FAS / 123.77 / ± / 15.89 / 38.73 / ± / 5.06 / 18:1_18:1_18:1 / 16:0_18:2_20:1 / 16:0_18:1_20:2
HFD / 105.36 / ± / 25.37 / 48.09 / ± / 4.11 / 18:1_18:1_18:1 / 16:0_18:2_20:1 / 16:0_18:1_20:2
TG / 54:4 / FED / 333.86 / ± / 33.90 / 74.69 / ± / 13.12 / 18:1_18:1_18:2 / 16:0_18:2_20:2 / 16:0_18:1_20:3
16:1_18:1_20:2
FAS / 174.17 / ± / 11.84 / 62.88 / ± / 2.94 / 18:1_18:1_18:2 / 16:0_18:2_20:2 / 16:0_18:1_20:3
16:1_18:1_20:2
HFD / 106.94 / ± / 17.07 / 45.01 / ± / 7.17 / 18:1_18:1_18:2
TG / 54:5 / FED / 305.18 / ± / 30.52 / 73.52 / ± / 6.62 / 16:0_18:1_20:4 / 16:1_18:1_20:3 / 18:1_18:1_18:3
18:1_18:2_18:2
FAS / 236.13 / ± / 5.14 / 86.55 / ± / 7.49 / 16:0_18:1_20:4 / 16:1_18:1_20:3 / 18:1_18:1_18:3
18:1_18:2_18:2
HFD / 106.64 / ± / 29.63 / 46.64 / ± / 4.91 / 18:1_18:2_18:2 / 16:0_18:1_20:4 / 16:0_18:2_20:3
18:1_18:1_18:3
TG / 54:6 / FED / 162.99 / ± / 11.82 / 45.00 / ± / 3.93 / 16:0_18:2_20:4 / 16:1_18:1_20:4 / 18:1_18:2_18:3
16:0_18:1_20:5
FAS / 243.81 / ± / 13.89 / 80.97 / ± / 7.96 / 16:0_18:2_20:4 / 16:1_18:1_20:4 / 18:1_18:2_18:3
16:0_18:1_20:5
HFD / 64.30 / ± / 12.13 / 30.93 / ± / 2.58 / 18:2_18:2_18:2 / 16:0_18:2_20:4 / 16:1_18:1_20:4
16:0_18:1_20:5
TG / 54:7 / FED / 30.51 / ± / 0.43 / 9.68 / ± / 2.27 / 16:0_18:3_20:4 / 16:0_18:2_20:5 / 16:0_16:1_22:6
TG continued
14:0_18:1_22:6
FAS / 105.73 / ± / 3.09 / 37.12 / ± / 1.53 / 16:0_18:3_20:4 / 16:0_18:2_20:5 / 16:0_16:1_22:6
HFD / 12.54 / ± / 1.86 / 7.92 / ± / 2.40 / 18:2_18:2_18:3 / 16:0_18:3_20:4 / 16:0_18:2_20:5
16:0_16:1_22:6
TG / 54:8 / FED / 3.00 / ± / 0.33 / 0.99 / ± / 0.29 / 16:1_16:1_22:6 / 16:1_18:2_20:5 / 16:1_18:3_20:4
18:2_18:3_18:3
FAS / 26.08 / ± / 7.89 / 11.23 / ± / 2.81
HFD / 0.90 / ± / 0.13 / 0.64 / ± / 0.27 / 18:2_18:3_18:3 / 18:2_18:2_18:4 / 16:1_16:1_22:6
16:1_18:2_20:5
TG / 54:9 / FED / 0.19 / ± / 0.07 / 0.05 / ± / 0.02
FAS / 3.53 / ± / 1.49 / 1.81 / ± / 0.89
HFD / 0.10 / ± / 0.03 / 0.06 / ± / 0.02
TG / 54:10 / FED / 0.00 / ± / 0.00 / 0.01 / ± / 0.00
FAS / 0.37 / ± / 0.19 / 0.24 / ± / 0.14
HFD / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
TG / 56:1 / FED / 0.22 / ± / 0.05 / 0.27 / ± / 0.25
FAS / 0.12 / ± / 0.04 / 0.10 / ± / 0.07
HFD / 0.89 / ± / 0.12 / 0.55 / ± / 0.19
TG / 56:2 / FED / 2.70 / ± / 0.13 / 2.50 / ± / 2.16 / 18:1_22:1_16:0 / 18:1_18:1_20:0 / 18:0_20:1_18:1
FAS / 1.90 / ± / 0.84 / 1.46 / ± / 0.89 / 18:1_22:1_16:0 / 18:1_18:1_20:0 / 18:2_18:0_20:0
18:0_20:1_18:1
HFD / 25.78 / ± / 4.07 / 12.25 / ± / 0.85 / 18:1_22:1_16:0 / 18:1_18:1_20:0 / 18:2_18:0_20:0
18:0_20:1_18:1
TG / 56:3 / FED / 17.43 / ± / 1.78 / 8.24 / ± / 4.27 / 18:1_18:1_20:1 / 18:1_18:2_20:0 / 18:1_22:2_16:0
18:1_16:1_22:1
FAS / 21.85 / ± / 7.05 / 6.76 / ± / 1.49 / 18:1_18:1_20:1 / 18:1_18:2_20:0 / 18:1_22:2_16:0
18:1_16:1_22:1
HFD / 43.37 / ± / 8.42 / 15.40 / ± / 5.68 / 18:1_18:1_20:1 / 18:1_18:2_20:0 / 18:1_22:2_16:0
18:1_16:1_22:1
TG / 56:4 / FED / 37.48 / ± / 4.35 / 11.16 / ± / 1.89 / 18:1_20:3_18:0 / 18:1_22:3_16:0 / 18:0_18:0_20:4
16:0_20:2_20:2
FAS / 23.50 / ± / 3.58 / 7.22 / ± / 1.17 / 18:1_18:2_20:1 / 18:1_20:3_18:0 / 18:1_22:3_16:0
18:0_18:0_20:4
HFD / 12.71 / ± / 3.32 / 5.28 / ± / 1.32 / 18:1_18:2_20:1 / 18:1_20:3_18:0 / 18:1_22:3_16:0
18:0_18:0_20:4
TG / 56:5 / FED / 54.07 / ± / 6.33 / 14.33 / ± / 3.21 / 18:1_16:0_22:4 / 18:0_18:1_20:4 / 18:0_20:3_18:2
18:0_16:0_22:5
FAS / 18.52 / ± / 2.25 / 6.19 / ± / 1.06 / 18:1_18:2_20:2 / 18:1_16:0_22:4 / 18:0_20:3_18:2
16:0_20:1_20:4
HFD / 43.25 / ± / 15.47 / 16.18 / ± / 5.45 / 18:1_18:2_20:2 / 18:1_16:0_22:4 / 18:0_20:3_18:2
16:0_20:1_20:4
TG / 56:6 / FED / 107.62 / ± / 9.92 / 26.92 / ± / 2.93 / 18:1_18:1_20:4 / 16:0_18:1_22:5 / 16:0_18:2_22:4
18:0_18:2_20:4
FAS / 35.17 / ± / 6.11 / 10.81 / ± / 2.44 / 18:1_18:1_20:4 / 16:0_18:1_22:5 / 16:0_18:2_22:4
18:0_18:2_20:4
HFD / 59.90 / ± / 18.78 / 25.73 / ± / 4.58 / 18:1_18:1_20:4 / 16:0_18:1_22:5 / 16:0_18:2_22:4
18:0_18:2_20:4
TG continued
TG / 56:7 / FED / 153.35 / ± / 17.75 / 39.82 / ± / 1.87 / 16:0_18:1_22:6 / 16:1_18:1_22:5 / 18:2_18:1_20:4
18:0_18:3_20:4
FAS / 66.58 / ± / 9.85 / 24.98 / ± / 3.51 / 16:0_18:1_22:6 / 16:1_18:1_22:5 / 18:2_18:1_20:4
18:0_18:3_20:4
HFD / 71.57 / ± / 14.83 / 36.68 / ± / 4.41 / 16:0_18:1_22:6 / 18:2_18:1_20:4 / 16:1_18:1_22:5
18:0_18:3_20:4
TG / 56:8 / FED / 118.01 / ± / 6.15 / 31.75 / ± / 2.59 / 16:0_18:2_22:6 / 16:1_18:2_22:5 / 18:2_18:1_20:5
18:1_18:3_20:4
FAS / 100.17 / ± / 3.48 / 32.69 / ± / 3.02 / 16:0_18:2_22:6 / 16:1_18:2_22:5 / 18:2_18:1_20:5
18:1_18:3_20:4
HFD / 26.13 / ± / 5.12 / 15.78 / ± / 3.83 / 16:0_18:2_22:6 / 16:0_20:4_20:4
18:2_18:1_20:5
TG / 56:9 / FED / 9.38 / ± / 0.65 / 2.92 / ± / 0.66 / 16:1_18:2_22:6 / 18:2_18:2_20:5 / 18:2_18:3_20:4
FAS / 35.40 / ± / 6.99 / 13.34 / ± / 1.64 / 16:1_18:2_22:6 / 18:2_18:2_20:5 / 18:2_18:3_20:4
14:0_18:3_24:6
HFD / 2.16 / ± / 0.36 / 1.48 / ± / 0.54 / 16:1_18:2_22:6 / 18:2_18:2_20:5 / 18:2_18:3_20:4
TG / 56:10 / FED / 0.61 / ± / 0.08 / 0.18 / ± / 0.05
FAS / 5.61 / ± / 2.24 / 2.84 / ± / 1.08
HFD / 0.13 / ± / 0.02 / 0.10 / ± / 0.04
TG / 56:11 / FED / 0.00 / ± / 0.00 / 0.01 / ± / 0.00
FAS / 0.47 / ± / 0.25 / 0.29 / ± / 0.15
HFD / 0.01 / ± / 0.00 / 0.01 / ± / 0.00
TG / 58:2 / FED / 0.43 / ± / 0.11 / 0.34 / ± / 0.26
FAS / 0.25 / ± / 0.09 / 0.23 / ± / 0.17
HFD / 1.00 / ± / 0.14 / 0.50 / ± / 0.08
TG / 58:3 / FED / 1.61 / ± / 0.07 / 1.26 / ± / 1.01
FAS / 1.33 / ± / 0.75 / 0.94 / ± / 0.56
HFD / 7.16 / ± / 1.01 / 2.86 / ± / 0.68
TG / 58:4 / FED / 3.84 / ± / 0.11 / 2.01 / ± / 1.19
FAS / 3.29 / ± / 1.13 / 1.03 / ± / 0.23
HFD / 3.90 / ± / 0.74 / 1.41 / ± / 0.50
TG / 58:5 / FED / 5.03 / ± / 0.98 / 2.32 / ± / 1.16
FAS / 1.60 / ± / 0.30 / 0.50 / ± / 0.12
HFD / 4.15 / ± / 1.47 / 1.41 / ± / 0.60
TG / 58:6 / FED / 13.04 / ± / 1.18 / 3.31 / ± / 0.65
FAS / 4.34 / ± / 1.05 / 1.11 / ± / 0.54
HFD / 9.47 / ± / 3.24 / 3.22 / ± / 1.45
TG / 58:7 / FED / 34.11 / ± / 2.45 / 8.32 / ± / 1.16
FAS / 9.82 / ± / 1.60 / 2.56 / ± / 1.08
HFD / 11.41 / ± / 3.05 / 5.54 / ± / 0.74
TG / 58:8 / FED / 58.41 / ± / 9.84 / 14.44 / ± / 1.73
FAS / 17.23 / ± / 1.58 / 5.13 / ± / 1.15
HFD / 13.32 / ± / 2.99 / 6.48 / ± / 0.82
TG / 58:9 / FED / 51.41 / ± / 3.53 / 12.70 / ± / 1.61 / 18:1_20:4_20:4
FAS / 27.54 / ± / 3.40 / 8.56 / ± / 1.87 / 18:1_18:2_22:6 / 18:1_20:4_20:4
HFD / 8.30 / ± / 1.16 / 4.40 / ± / 0.60 / 18:1_18:2_22:6 / 18:1_20:4_20:4
TG / 58:10 / FED / 14.87 / ± / 1.43 / 4.45 / ± / 0.88
FAS / 37.90 / ± / 3.30 / 12.28 / ± / 1.51
TG continued
HFD / 3.23 / ± / 0.48 / 2.05 / ± / 0.59
TG / 58:11 / FED / 1.37 / ± / 0.24 / 0.39 / ± / 0.07
FAS / 8.24 / ± / 2.46 / 3.37 / ± / 0.73
HFD / 0.23 / ± / 0.03 / 0.20 / ± / 0.11
TG / 58:12 / FED / 0.09 / ± / 0.04 / 0.02 / ± / 0.01
FAS / 0.67 / ± / 0.29 / 0.39 / ± / 0.18
HFD / 0.02 / ± / 0.00 / 0.01 / ± / 0.01
TG / 60:3 / FED / 0.26 / ± / 0.07 / 0.16 / ± / 0.10
FAS / 0.17 / ± / 0.06 / 0.16 / ± / 0.12
HFD / 0.27 / ± / 0.03 / 0.12 / ± / 0.02
TG / 60:4 / FED / 0.37 / ± / 0.06 / 0.27 / ± / 0.20
FAS / 0.49 / ± / 0.28 / 0.34 / ± / 0.19
HFD / 0.50 / ± / 0.06 / 0.18 / ± / 0.07
TG / 60:5 / FED / 0.53 / ± / 0.08 / 0.36 / ± / 0.25
FAS / 0.61 / ± / 0.19 / 0.19 / ± / 0.04
HFD / 0.70 / ± / 0.11 / 0.26 / ± / 0.11
TG / 60:6 / FED / 0.93 / ± / 0.12 / 0.46 / ± / 0.27
FAS / 0.43 / ± / 0.11 / 0.10 / ± / 0.06
HFD / 0.82 / ± / 0.31 / 0.24 / ± / 0.16
TG / 60:7 / FED / 3.02 / ± / 0.22 / 0.77 / ± / 0.15
FAS / 0.66 / ± / 0.26 / 0.15 / ± / 0.14
HFD / 1.19 / ± / 0.49 / 0.38 / ± / 0.22
TG / 60:8 / FED / 7.80 / ± / 1.04 / 1.81 / ± / 0.30
FAS / 0.75 / ± / 0.24 / 0.20 / ± / 0.12
HFD / 1.14 / ± / 0.18 / 0.48 / ± / 0.09
TG / 60:9 / FED / 6.90 / ± / 0.88 / 2.04 / ± / 0.39
FAS / 0.71 / ± / 0.58 / 0.20 / ± / 0.15
HFD / 1.75 / ± / 0.42 / 0.87 / ± / 0.11
TG / 60:10 / FED / 6.11 / ± / 0.30 / 1.84 / ± / 0.31
FAS / 1.17 / ± / 0.17 / 0.31 / ± / 0.13
HFD / 2.18 / ± / 0.44 / 1.28 / ± / 0.29
TG / 60:11 / FED / 3.84 / ± / 0.13 / 1.25 / ± / 0.31
FAS / 4.13 / ± / 0.25 / 1.38 / ± / 0.13
HFD / 1.55 / ± / 0.22 / 1.04 / ± / 0.34
TG / 60:12 / FED / 2.42 / ± / 0.17 / 0.72 / ± / 0.13
FAS / 5.93 / ± / 1.16 / 2.00 / ± / 0.24
HFD / 0.61 / ± / 0.12 / 0.60 / ± / 0.35
TG / 60:13 / FED / 0.20 / ± / 0.04 / 0.05 / ± / 0.01
FAS / 0.73 / ± / 0.33 / 0.36 / ± / 0.14
HFD / 0.03 / ± / 0.00 / 0.02 / ± / 0.02
TG / 62:11 / FED / 1.30 / ± / 0.16 / 0.27 / ± / 0.07
FAS / 0.05 / ± / 0.01 / 0.02 / ± / 0.01
HFD / 0.31 / ± / 0.02 / 0.16 / ± / 0.04
TG / 62:12 / FED / 1.51 / ± / 0.14 / 0.41 / ± / 0.05
FAS / 0.48 / ± / 0.10 / 0.11 / ± / 0.07
HFD / 0.19 / ± / 0.01 / 0.19 / ± / 0.11
TG / 62:13 / FED / 0.93 / ± / 0.03 / 0.25 / ± / 0.05
FAS / 0.93 / ± / 0.08 / 0.30 / ± / 0.07
HFD / 0.08 / ± / 0.01 / 0.05 / ± / 0.01
TG continued
TG / 62:14 / FED / 0.38 / ± / 0.12 / 0.10 / ± / 0.03
FAS / 0.79 / ± / 0.32 / 0.29 / ± / 0.08
HFD / 0.01 / ± / 0.01 / 0.01 / ± / 0.00
TG / 62:15 / FED / 0.00 / ± / 0.00 / 0.00 / ± / 0.00
FAS / 0.05 / ± / 0.03 / 0.03 / ± / 0.02
HFD / 0.00 / ± / 0.00 / 0.00 / ± / 0.00

Supplementary Table S4. Summary of DG species determined by LC-MS/MS in murine hepatocyte LD from FED, FAS and HFD samples. Absolute amounts (n=3 animals) are calculated in nmol, and are normalized to gram total acylgycerols (InfinityTM Triglycerides Reagent kit). Corresponding % values (n=6) are relative to totalamount of DG species.Molecular species are derived from possible fatty acyl combinations as determined by fatty acid fragment ions (neutral losses) detected in the corresponding product ion spectra. Fatty acyls in bold are major compositions.

Species / nmol / g / % / Molecular species
DG / 32:0 / FED / 37.0 / ± / 3.5 / 4.6 / ± / 3.7
FAS / 146.0 / ± / 0.5 / 3.0 / ± / 0.8
HFD / 93.2 / ± / 10.1 / 4.3 / ± / 1.9
DG / 32:1 / FED / 129.5 / ± / 15.9 / 4.9 / ± / 1.0
FAS / 312.3 / ± / 61.5 / 4.5 / ± / 0.8
HFD / 308.7 / ± / 41.0 / 9.2 / ± / 1.5
DG / 32:2 / FED / 49.5 / ± / 6.2 / 1.5 / ± / 1.0
FAS / 300.1 / ± / 52.0 / 3.3 / ± / 1.7
HFD / 29.4 / ± / 2.9 / 0.6 / ± / 0.2
DG / 34:0 / FED / 30.7 / ± / 3.7 / 11.8 / ± / 12.6 / 16:0_18:0
FAS / 52.0 / ± / 6.6 / 1.2 / ± / 0.5 / 16:0_18:0
HFD / 97.5 / ± / 26.8 / 3.4 / ± / 0.9 / 16:0_18:0
DG / 34:1 / FED / 1856.7 / ± / 87.9 / 121.7 / ± / 52.7 / 16:0_18:1
FAS / 4148.5 / ± / 222.8 / 81.9 / ± / 17.7 / 16:0_18:1
HFD / 7614.1 / ± / 831.9 / 283.2 / ± / 82.7 / 16:0_18:1
DG / 34:2 / FED / 4024.7 / ± / 237.3 / 158.0 / ± / 21.3 / 16:0_18:2 / 16:1_18:1
FAS / 10952.0 / ± / 1963.2 / 201.6 / ± / 30.9 / 16:0_18:2 / 16:1_18:1
HFD / 5050.2 / ± / 573.4 / 141.0 / ± / 16.1 / 16:0_18:2 / 16:1_18:1
DG / 34:3 / FED / 614.4 / ± / 16.1 / 20.7 / ± / 5.5
FAS / 2049.4 / ± / 423.3 / 29.2 / ± / 5.8 / 16:0_18:3
HFD / 395.1 / ± / 62.2 / 9.6 / ± / 1.9 / 16:0_18:3 / 16:1_18:2
DG / 34:4 / FED / 41.2 / ± / 8.5 / 1.3 / ± / 0.7
FAS / 273.0 / ± / 5.0 / 4.5 / ± / 0.7
HFD / 30.8 / ± / 10.5 / 0.5 / ± / 0.4
DG / 36:1 / FED / 208.8 / ± / 6.6 / 13.4 / ± / 5.8 / 18:0_18:1
FAS / 424.7 / ± / 40.8 / 7.3 / ± / 1.2 / 18:0_18:1
HFD / 793.3 / ± / 27.1 / 18.3 / ± / 5.1 / 18:0_18:1
DG / 36:2 / FED / 2792.9 / ± / 220.9 / 117.6 / ± / 15.6 / 18:1_18:1 / 18:0_18:2
FAS / 5741.6 / ± / 511.8 / 98.2 / ± / 8.8 / 18:1_18:1 / 18:0_18:2
HFD / 7160.1 / ± / 859.4 / 175.1 / ± / 36.2 / 18:1_18:1
DG / 36:3 / FED / 5005.2 / ± / 226.3 / 170.7 / ± / 40.3 / 18:1_18:2 / 18:0_18:3
FAS / 13983.0 / ± / 2055.5 / 244.8 / ± / 27.0 / 18:1_18:2 / 18:0_18:3
HFD / 3338.2 / ± / 423.8 / 80.2 / ± / 16.8 / 18:1_18:2
DG / 36:4 / FED / 3349.7 / ± / 159.0 / 105.7 / ± / 36.2 / 16:0_20:4 / 18:2_18:2
FAS / 10812.4 / ± / 1783.5 / 135.8 / ± / 44.0 / 18:2_18:2 / 18:1_18:3
HFD / 2524.0 / ± / 249.6 / 51.4 / ± / 22.0 / 18:2_18:2 / 16:0_20:4 / 18:1_18:3
DG / 36:5 / FED / 321.2 / ± / 25.4 / 9.3 / ± / 4.6
DG continued
FAS / 1236.2 / ± / 104.4 / 17.9 / ± / 3.8
HFD / 82.8 / ± / 27.1 / 1.5 / ± / 1.0
DG / 38:1 / FED / 22.5 / ± / 7.9 / 0.9 / ± / 0.3
FAS / 33.1 / ± / 3.1 / 0.3 / ± / 0.3
HFD / 107.4 / ± / 1.8 / 3.3 / ± / 1.5
DG / 38:2 / FED / 231.8 / ± / 48.3 / 17.2 / ± / 13.6
FAS / 293.6 / ± / 22.4 / 6.9 / ± / 2.4
HFD / 675.6 / ± / 18.1 / 18.2 / ± / 2.6
DG / 38:3 / FED / 521.9 / ± / 102.5 / 28.6 / ± / 15.5
FAS / 767.7 / ± / 82.6 / 13.2 / ± / 2.5
HFD / 690.9 / ± / 68.4 / 15.6 / ± / 4.7
DG / 38:4 / FED / 1230.2 / ± / 104.8 / 58.9 / ± / 13.4 / 18:0_20:4
FAS / 1737.3 / ± / 130.5 / 27.3 / ± / 3.6 / 18:0_20:4
HFD / 1829.3 / ± / 335.8 / 48.7 / ± / 6.6 / 18:0_20:4 / 18:1_20:3
DG / 38:5 / FED / 966.1 / ± / 43.8 / 34.6 / ± / 6.5 / 18:1_20:4
FAS / 1550.8 / ± / 105.0 / 22.2 / ± / 4.0
HFD / 1519.4 / ± / 263.2 / 37.5 / ± / 7.2 / 18:1_20:4 / 16:0_22:5
DG / 38:6 / FED / 1265.1 / ± / 178.5 / 47.2 / ± / 7.7 / 16:0_22:6
FAS / 3212.2 / ± / 581.9 / 45.7 / ± / 8.8
HFD / 1570.5 / ± / 346.5 / 54.1 / ± / 13.0 / 16:0_22:6 / 18:2_20:4
DG / 38:7 / FED / 98.9 / ± / 8.6 / 2.6 / ± / 1.7
FAS / 236.4 / ± / 14.6 / 3.7 / ± / 0.5
HFD / 71.9 / ± / 7.7 / 1.8 / ± / 0.3
DG / 40:3 / FED / 40.7 / ± / 3.2 / 1.7 / ± / 0.2
FAS / 27.9 / ± / 7.6 / 0.4 / ± / 0.1
HFD / 30.7 / ± / 6.0 / 0.9 / ± / 0.2
DG / 40:4 / FED / 45.9 / ± / 3.4 / 4.4 / ± / 3.4
FAS / 25.8 / ± / 9.6 / 0.3 / ± / 0.1
HFD / 148.4 / ± / 10.6 / 3.5 / ± / 1.1
DG / 40:5 / FED / 89.2 / ± / 23.3 / 5.3 / ± / 2.9
FAS / 242.3 / ± / 10.1 / 3.2 / ± / 1.2
HFD / 289.6 / ± / 55.4 / 6.4 / ± / 2.5
DG / 40:6 / FED / 180.3 / ± / 24.5 / 10.9 / ± / 4.4
FAS / 557.6 / ± / 124.9 / 11.4 / ± / 3.5
HFD / 565.9 / ± / 39.2 / 15.4 / ± / 2.7 / 18:0_22:6
DG / 40:7 / FED / 711.1 / ± / 89.2 / 26.1 / ± / 4.7
FAS / 1352.0 / ± / 145.7 / 17.7 / ± / 5.3
HFD / 540.4 / ± / 49.3 / 13.5 / ± / 2.4
DG / 40:8 / FED / 386.3 / ± / 30.8 / 16.0 / ± / 3.0 / 18:2_22:6
FAS / 966.8 / ± / 101.6 / 12.0 / ± / 4.3
HFD / 65.4 / ± / 12.1 / 1.5 / ± / 0.4
DG / 42:7 / FED / 47.4 / ± / 16.9 / 1.9 / ± / 1.0
FAS / 71.8 / ± / 82.0 / 1.3 / ± / 1.6
HFD / 37.7 / ± / 33.0 / 1.0 / ± / 0.8
DG / 42:8 / FED / 43.0 / ± / 7.7 / 2.7 / ± / 1.6
FAS / 109.8 / ± / 95.9 / 1.1 / ± / 1.3
HFD / 13.4 / ± / 1.6 / 0.3 / ± / 0.2

Supplementary Table S5. Summary of PC species determined by LC-MS/MS in murine hepatocyte LD from FED, FAS and HFD samples. Absolute amounts (n=3 animals) are calculated in nmol, and are normalized to gram total acylgycerols (InfinityTM Triglycerides Reagent kit). Corresponding ‰ values (n=6 animals) are relative to total amount of PC species. Molecular species are derived from possible fatty acyl combinations as determined by fatty acid fragment ions (neutral losses) detected in the corresponding product ion spectra. Fatty acyls in bold are major compositions.

Species / nmol / g / ‰ / Molecular species
PC / 32:0 / FED / 341.5 / ± / 23.3 / 11.9 / ± / 8.0
FAS / 103.8 / ± / 15.9 / 1.8 / ± / 0.3
HFD / 294.7 / ± / 43.0 / 4.2 / ± / 0.9
PC / 32:1 / FED / 343.8 / ± / 45.6 / 5.4 / ± / 0.9
FAS / 250.7 / ± / 20.3 / 3.5 / ± / 0.3
HFD / 310.5 / ± / 33.8 / 3.6 / ± / 0.5
PC / 32:2 / FED / 79.5 / ± / 9.3 / 1.1 / ± / 0.1
FAS / 106.8 / ± / 2.4 / 1.8 / ± / 0.4
HFD / 34.9 / ± / 12.6 / 0.4 / ± / 0.1
PC / 34:1 / FED / 5723.0 / ± / 360.9 / 97.9 / ± / 19.1 / 16:0_18:1
FAS / 6009.1 / ± / 752.9 / 78.9 / ± / 7.7 / 16:0_18:1
HFD / 9531.5 / ± / 1513.5 / 104.0 / ± / 13.5 / 16:0_18:1
PC / 34:2 / FED / 18018.7 / ± / 2496.2 / 228.4 / ± / 36.4 / 16:0_18:2 / 16:1_18:1
FAS / 22571.9 / ± / 1270.3 / 318.6 / ± / 10.3 / 16:0_18:2 / 16:1_18:1
HFD / 8362.2 / ± / 1193.1 / 99.6 / ± / 6.4 / 16:0_18:2
PC / 34:3 / FED / 1212.8 / ± / 126.5 / 15.0 / ± / 3.0 / 16:0_18:3 / 16:1_18:2
FAS / 1588.1 / ± / 51.8 / 24.1 / ± / 2.5 / 16:1_18:2 / 16:0_18:3
HFD / 547.9 / ± / 59.9 / 5.4 / ± / 1.4 / 16:1_18:2 / 16:0_18:3
PC / 34:4 / FED / 49.8 / ± / 19.0 / 0.9 / ± / 0.5 / 16:0_18:3 / 16:1_18:2
FAS / 105.4 / ± / 10.1 / 1.5 / ± / 0.3 / 16:1_18:2 / 16:0_18:3
HFD / 74.9 / ± / 11.7 / 0.7 / ± / 0.2 / 16:1_18:2 / 16:0_18:3
PC / 36:1 / FED / 637.4 / ± / 47.1 / 11.0 / ± / 3.5 / 18:0_18:1
FAS / 422.7 / ± / 64.2 / 4.5 / ± / 1.7 / 18:0_18:1
HFD / 1362.4 / ± / 249.9 / 13.9 / ± / 2.9 / 18:0_18:1
PC / 36:2 / FED / 6307.7 / ± / 313.2 / 105.4 / ± / 18.2 / 18:1_18:1 / 18:0_18:2
FAS / 6544.0 / ± / 650.5 / 95.6 / ± / 7.8 / 18:1_18:1 / 18:0_18:2
HFD / 6089.6 / ± / 1045.5 / 76.3 / ± / 11.0 / 18:1_18:1
PC / 36:3 / FED / 3975.6 / ± / 553.6 / 45.4 / ± / 14.7 / 18:1_18:2 / 16:0_20:3
FAS / 3238.4 / ± / 375.2 / 45.4 / ± / 4.1 / 18:1_18:2 / 16:0_20:3
HFD / 4237.7 / ± / 539.4 / 42.8 / ± / 12.1 / 18:1_18:2 / 16:0_20:3
PC / 36:4 / FED / 10752.2 / ± / 575.8 / 149.9 / ± / 11.5 / 16:0_20:4
FAS / 8973.4 / ± / 1385.0 / 132.8 / ± / 9.6 / 16:0_20:4 / 18:2_18:2
HFD / 17241.8 / ± / 2564.9 / 199.5 / ± / 16.9 / 16:0_20:4 / 18:2_18:2
PC / 36:5 / FED / 314.3 / ± / 64.3 / 4.9 / ± / 0.8 / 18:1_20:4 / 16:0_22:5
FAS / 210.7 / ± / 10.5 / 5.2 / ± / 2.5 / 18:1_20:4 / 16:0_22:5
PC continued
HFD / 528.0 / ± / 83.4 / 5.3 / ± / 1.3 / 18:1_20:4 / 16:0_22:5
PC / 36:6 / FED / 26.0 / ± / 9.3 / 0.8 / ± / 0.6
FAS / 44.3 / ± / 5.5 / 1.4 / ± / 1.1
HFD / 59.9 / ± / 8.5 / 1.0 / ± / 0.3
PC / 38:2 / FED / 157.2 / ± / 23.6 / 1.9 / ± / 0.7
FAS / 29.0 / ± / 6.5 / 0.4 / ± / 0.1
HFD / 153.4 / ± / 25.3 / 1.1 / ± / 0.8
PC / 38:3 / FED / 403.0 / ± / 48.4 / 9.8 / ± / 5.3
FAS / 102.4 / ± / 11.1 / 2.1 / ± / 0.8
HFD / 1718.4 / ± / 213.7 / 18.8 / ± / 3.8
PC / 38:4 / FED / 7013.4 / ± / 347.7 / 112.5 / ± / 14.9 / 18:0_20:4
FAS / 5817.9 / ± / 853.8 / 76.3 / ± / 8.5 / 18:0_20:4
HFD / 14095.6 / ± / 1925.0 / 190.3 / ± / 28.7 / 18:0_20:4
PC / 38:5 / FED / 1708.1 / ± / 366.7 / 24.4 / ± / 3.5
FAS / 1285.2 / ± / 58.2 / 19.2 / ± / 1.9
HFD / 4205.8 / ± / 853.0 / 50.4 / ± / 4.5
PC / 38:6 / FED / 9176.0 / ± / 1361.6 / 120.6 / ± / 15.1 / 16:0_22:6
FAS / 9310.4 / ± / 847.1 / 136.3 / ± / 7.5 / 16:0_22:6 / 18:2_20:4
HFD / 9562.3 / ± / 1336.8 / 126.4 / ± / 13.3 / 16:0_22:6 / 18:2_20:4
PC / 38:7 / FED / 260.6 / ± / 34.2 / 3.0 / ± / 0.8
FAS / 278.9 / ± / 3.6 / 3.1 / ± / 1.5
HFD / 291.9 / ± / 68.7 / 3.1 / ± / 0.6
PC / 40:4 / FED / 91.7 / ± / 5.6 / 1.7 / ± / 0.5
FAS / 28.2 / ± / 25.5 / 0.4 / ± / 0.2
HFD / 182.4 / ± / 44.3 / 2.0 / ± / 0.3
PC / 40:5 / FED / 117.3 / ± / 8.2 / 2.7 / ± / 1.2
FAS / 77.0 / ± / 23.4 / 1.8 / ± / 0.9
HFD / 439.4 / ± / 51.5 / 5.5 / ± / 0.6
PC / 40:6 / FED / 1656.0 / ± / 225.3 / 30.7 / ± / 7.9 / 18:0_22:6
FAS / 2529.4 / ± / 179.4 / 29.9 / ± / 6.9 / 18:0_22:6
HFD / 2400.6 / ± / 300.6 / 31.4 / ± / 4.5 / 18:0_22:6 / 18:1_22:5
PC / 40:7 / FED / 768.4 / ± / 75.1 / 11.0 / ± / 1.3 / 18:1_22:6
FAS / 714.3 / ± / 57.3 / 10.9 / ± / 1.6 / 18:1_22:6
HFD / 812.3 / ± / 115.4 / 10.8 / ± / 1.6 / 18:1_22:6
PC / 40:8 / FED / 298.1 / ± / 21.3 / 3.8 / ± / 0.6 / 18:2_22:6 / 20:4_20:4
FAS / 311.2 / ± / 9.9 / 4.6 / ± / 0.5 / 18:2_22:6 / 20:4_20:4
HFD / 324.8 / ± / 47.7 / 3.7 / ± / 0.4 / 18:2_22:6 / 20:4_20:4

Supplementary Table S6. Summary of LPC molecular species determined by LC-MS/MS in murine hepatocyte LD from FED, FAS and HFD samples. Absolute amounts (n=3 animals) are calculated in nmol, and are normalized to gram total acylgycerols (InfinityTM Triglycerides Reagent kit). Corresponding % values (n=6 animals) are relatative to total amount of LPC species.

Molecular species / nmol / g / %
LPC / 16:0 / FED / 1752.3 / ± / 498.3 / 31.5 / ± / 5.1
FAS / 2595.3 / ± / 253.2 / 32.3 / ± / 2.7
HFD / 2844.7 / ± / 246.4 / 22.5 / ± / 2.4
LPC / 18:0 / FED / 1142.6 / ± / 433.7 / 18.6 / ± / 5.8
FAS / 864.6 / ± / 60.9 / 12.0 / ± / 1.1
HFD / 2048.7 / ± / 116.2 / 16.4 / ± / 1.9
LPC / 18:1 / FED / 511.8 / ± / 175.9 / 8.4 / ± / 2.1
FAS / 578.2 / ± / 58.9 / 7.6 / ± / 0.8
HFD / 1105.7 / ± / 58.7 / 8.7 / ± / 1.0
LPC / 18:2 / FED / 564.4 / ± / 141.7 / 15.1 / ± / 4.9
FAS / 1744.0 / ± / 198.6 / 21.4 / ± / 2.4
HFD / 1105.8 / ± / 91.3 / 8.9 / ± / 0.8
LPC / 20:3 / FED / 76.0 / ± / 26.5 / 2.1 / ± / 0.7
FAS / 29.5 / ± / 6.1 / 1.2 / ± / 0.9
HFD / 407.9 / ± / 50.8 / 4.0 / ± / 0.6
LPC / 20:4 / FED / 472.8 / ± / 72.5 / 12.3 / ± / 3.5
FAS / 771.7 / ± / 43.2 / 10.7 / ± / 1.0
HFD / 2515.3 / ± / 66.1 / 23.1 / ± / 2.2
LPC / 22:5 / FED / 32.8 / ± / 29.0 / 0.7 / ± / 0.3
FAS / 36.0 / ± / 9.9 / 0.7 / ± / 0.3
HFD / 200.7 / ± / 55.2 / 1.6 / ± / 0.4
LPC / 22:6 / FED / 489.5 / ± / 79.3 / 11.3 / ± / 2.1
FAS / 1079.0 / ± / 9.8 / 14.1 / ± / 1.1
HFD / 1408.9 / ± / 60.8 / 14.9 / ± / 3.1

Supplementary Table S7. Summary of PE species determined by LC-MS/MS in murine hepatocyte LD from FED, FAS and HFD samples. Absolute amounts (n=3 animals) are calculated in µmol, and are normalized to gram total acylgycerols (InfinityTM Triglycerides Reagent kit). Corresponding % values (n=6 animals) are relative to total amounts of PE species. Molecular species are derived from possible fatty acyl combinations as determined by fatty acid fragment ions (neutral losses) detected in the corresponding product ion spectra. Fatty acyls in bold are major compositions.

Species / µmol / g / % / Molecular species
PE / 34:2 / FED / 1.04 / ± / 0.11 / 3.96 / ± / 0.52
FAS / 1.34 / ± / 0.07 / 6.81 / ± / 1.55
HFD / 0.24 / ± / 0.03 / 0.85 / ± / 0.33 / 16:0_18:2
PE / 34:3 / FED / 0.06 / ± / 0.02 / 0.19 / ± / 0.08
FAS / 0.09 / ± / 0.01 / 0.49 / ± / 0.11
HFD / 0.02 / ± / 0.00 / 0.08 / ± / 0.02
PE / 36:2 / FED / 1.04 / ± / 0.25 / 5.13 / ± / 1.71
FAS / 1.64 / ± / 0.14 / 6.67 / ± / 0.68
HFD / 0.71 / ± / 0.08 / 3.43 / ± / 0.67
PE / 36:3 / FED / 0.73 / ± / 0.16 / 2.70 / ± / 0.44
FAS / 0.96 / ± / 0.09 / 3.58 / ± / 0.74
HFD / 0.38 / ± / 0.02 / 1.23 / ± / 0.67 / 18:1_18:2
PE / 36:4 / FED / 2.26 / ± / 0.12 / 8.56 / ± / 0.67 / 16:0_20:4
FAS / 1.87 / ± / 0.08 / 9.39 / ± / 2.08 / 16:0_20:4 / 18:2_18:2
HFD / 1.94 / ± / 0.27 / 9.02 / ± / 1.35 / 16:0_20:4
PE / 36:5 / FED / 0.17 / ± / 0.04 / 0.54 / ± / 0.16
FAS / 0.11 / ± / 0.00 / 0.63 / ± / 0.21
HFD / 0.11 / ± / 0.01 / 0.44 / ± / 0.17
PE / 38:4 / FED / 9.45 / ± / 0.98 / 36.61 / ± / 2.31 / 18:0_20:4
FAS / 7.32 / ± / 0.86 / 28.23 / ± / 4.05 / 18:0_20:4
HFD / 10.27 / ± / 0.90 / 52.46 / ± / 7.30 / 18:0_20:4
PE / 38:5 / FED / 1.98 / ± / 0.30 / 6.81 / ± / 2.02 / 18:1_20:4
FAS / 1.37 / ± / 0.05 / 4.57 / ± / 1.41 / 18:1_20:4
HFD / 2.12 / ± / 0.16 / 8.46 / ± / 3.21 / 18:1_20:4 / 16:0_22:5
PE / 38:6 / FED / 5.43 / ± / 0.24 / 21.23 / ± / 1.29 / 16:0_22:6
FAS / 5.17 / ± / 0.27 / 24.41 / ± / 3.57 / 16:0_22:6 / 18:2_20:4
HFD / 3.30 / ± / 0.35 / 15.63 / ± / 2.27 / 16:0_22:6 / 18:2_20:4
PE / 38:7 / FED / 0.14 / ± / 0.08 / 0.55 / ± / 0.23
FAS / 0.23 / ± / 0.02 / 1.02 / ± / 0.18
HFD / 0.13 / ± / 0.02 / 0.58 / ± / 0.19
PE / 40:6 / FED / 1.60 / ± / 0.13 / 9.34 / ± / 3.33 / 18:0_22:6
FAS / 2.54 / ± / 0.25 / 9.01 / ± / 2.03 / 18:0_22:6
HFD / 0.96 / ± / 0.08 / 4.87 / ± / 0.98 / 18:0_22:6 / 18:1_22:5
PE / 40:7 / FED / 1.16 / ± / 0.05 / 4.12 / ± / 0.61
FAS / 0.91 / ± / 0.04 / 4.40 / ± / 0.75
HFD / 0.53 / ± / 0.08 / 2.58 / ± / 0.43
PE / 40:8 / FED / 0.09 / ± / 0.01 / 0.26 / ± / 0.11
FAS / 0.17 / ± / 0.01 / 0.79 / ± / 0.17
HFD / 0.07 / ± / 0.01 / 0.37 / ± / 0.13

Supplementary Table S8. Summary of PI species determined by LC-MS/MS in murine hepatocyte LD from FED, FAS and HFD samples. Absolute amounts (n=3 animals) are calculated in nmol, and are normalized to gram total acylgycerols (InfinityTM Triglycerides Reagent kit). Corresponding % values (n=6 animals) are relative to total amounts of PI species. Molecular species are derived from possible fatty acyl combinations as determined by fatty acid fragment ions (carboxylates) detected in the corresponding product ion spectra. Fatty acyls in bold are major compositions.

Species / nmol / g / % / Molecular species
PI / 34:2 / FED / 48.5 / ± / 17.4 / 0.5 / ± / 0.3
FAS / 28.9 / ± / 3.5 / 0.5 / ± / 0.1
HFD / 8.2 / ± / 2.0 / 0.2 / ± / 0.1
PI / 36:2 / FED / 139.4 / ± / 71.8 / 1.4 / ± / 0.5
FAS / 96.5 / ± / 5.0 / 1.9 / ± / 0.4
HFD / 36.2 / ± / 2.9 / 5.7 / ± / 11.9
PI / 36:3 / FED / 79.5 / ± / 31.3 / 0.9 / ± / 0.4
FAS / 34.8 / ± / 3.8 / 0.7 / ± / 0.1
HFD / 20.4 / ± / 5.3 / 0.4 / ± / 0.1
PI / 36:4 / FED / 247.6 / ± / 137.3 / 3.3 / ± / 1.3 / 16:0_20:4
FAS / 195.5 / ± / 12.3 / 3.5 / ± / 1.2 / 16:0_20:4
HFD / 168.4 / ± / 10.0 / 2.6 / ± / 1.3 / 16:0_20:4
PI / 38:3 / FED / 201.3 / ± / 60.2 / 8.7 / ± / 7.0
FAS / 16.0 / ± / 3.8 / 7.3 / ± / 7.9 / 18:0_20:3
HFD / 149.2 / ± / 32.6 / 7.0 / ± / 6.0 / 18:0_20:3
PI / 38:4 / FED / 7228.6 / ± / 1117.6 / 80.9 / ± / 8.8 / 18:0_20:4
FAS / 4641.2 / ± / 246.7 / 83.0 / ± / 9.7 / 18:0_20:4
HFD / 5318.3 / ± / 399.8 / 80.6 / ± / 17.0 / 18:0_20:4
PI / 38:5 / FED / 440.3 / ± / 68.5 / 3.4 / ± / 2.0
FAS / 137.8 / ± / 5.6 / 2.3 / ± / 0.7
HFD / 184.6 / ± / 28.9 / 2.7 / ± / 0.9
PI / 38:6 / FED / 24.0 / ± / 9.5 / 0.2 / ± / 0.1
FAS / 22.0 / ± / 2.4 / 0.4 / ± / 0.1
HFD / 15.8 / ± / 1.2 / 0.2 / ± / 0.1
PI / 40:6 / FED / 78.5 / ± / 12.8 / 0.8 / ± / 0.2
FAS / 30.1 / ± / 2.8 / 0.5 / ± / 0.2
HFD / 43.0 / ± / 4.0 / 0.7 / ± / 0.2

Supplementary Table S9. Summary of PS species determined by LC-MS/MS in murine hepatocyte LD from FED, FAS and HFDsamples. Absolute amounts (n=3 animals) are calculated in nmol, and are normalized to gramtotal acylgycerols(InfinityTM Triglycerides Reagent kit). Corresponding % values (n=6 animals) are relative to total amounts of PS species.Molecular species are derived from possible fatty acyl combinations as determined by fatty acid fragment ions (neutral losses) detected in the corresponding product ion spectra. Fatty acyls in bold are major compositions.

Species / nmol / g / % / Molecular species
PS / 36:4 / FED / 35.7 / ± / 9.5 / 4.5 / ± / 4.2
FAS / 42.5 / ± / 6.1 / 6.5 / ± / 2.3
HFD / 62.1 / ± / 5.3 / 6.2 / ± / 4.2
PS / 38:4 / FED / 236.2 / ± / 27.6 / 33.7 / ± / 23.2 / 18:0_20:4
FAS / 176.9 / ± / 18.4 / 28.1 / ± / 8.7
HFD / 439.2 / ± / 38.1 / 63.1 / ± / 14.5
PS / 38:6 / FED / 48.1 / ± / 4.7 / 6.0 / ± / 5.4 / 16:0_22:6
FAS / 97.7 / ± / 7.6 / 14.5 / ± / 5.4 / 16:0_22:6
HFD / 46.2 / ± / 6.4 / 4.9 / ± / 2.9 / 16:0_22:6
PS / 40:6 / FED / 138.3 / ± / 6.3 / 55.8 / ± / 34.5 / 18:0_22:6
FAS / 203.5 / ± / 13.4 / 50.9 / ± / 16.3 / 18:0_22:6
HFD / 79.4 / ± / 7.9 / 25.8 / ± / 16.0 / 18:0_22:6

Supplementary Table S10. Summary of SM molecular species determined by LC-MS/MS in murine hepatocyte LD from FED, FAS and HFD samples.Absolute amounts (n=3 animals) are calculated in nmol, and are normalized to gram total acylgycerols (InfinityTM Triglycerides Reagent kit). Corresponding % values (n=6 animals) are relatative to total amounts og SM species. Sphingosine base assumed to be d18:1 (26).

Molecular species / nmol / g / %
SM / 16:0 / FED / 156.6 / ± / 4.6 / 3.3 / ± / 3.3
FAS / 190.1 / ± / 21.5 / 4.9 / ± / 1.2
HFD / 233.2 / ± / 7.7 / 5.0 / ± / 2.9
SM / 16:1 / FED / 1.1 / ± / 0.4 / 0.0 / ± / 0.0
FAS / 22.5 / ± / 3.7 / 0.4 / ± / 0.2
HFD / 18.3 / ± / 2.7 / 0.3 / ± / 0.3
SM / 22:0 / FED / 687.2 / ± / 183.3 / 24.2 / ± / 9.9
FAS / 1354.7 / ± / 148.6 / 27.2 / ± / 13.2
HFD / 1732.3 / ± / 85.3 / 37.4 / ± / 21.4
SM / 22:1 / FED / 109.5 / ± / 15.3 / 31.6 / ± / 32.2
FAS / 43.0 / ± / 13.3 / 8.4 / ± / 8.4
HFD / 154.9 / ± / 33.7 / 31.7 / ± / 31.6
SM / 24:0 / FED / 640.5 / ± / 55.2 / 14.6 / ± / 12.7
FAS / 851.6 / ± / 88.5 / 32.9 / ± / 11.5
HFD / 461.7 / ± / 46.7 / 9.6 / ± / 6.1
SM / 24:1 / FED / 967.8 / ± / 58.6 / 26.2 / ± / 16.4
FAS / 1073.5 / ± / 191.4 / 26.1 / ± / 6.7
HFD / 682.2 / ± / 85.5 / 16.0 / ± / 7.9