Hydroxy-capric acidMetabolite 2

Hydroxy-capric acidMetabolite 3

Hydroxy-capric acidMetabolite 1

OH

O

OH

OH

O OH

OH

O

OH

Supplemental Figure 2: GC-MS analyses of metabolites produced by CYP703A2 on capric, lauric and myristic acids.

Hydroxy-lauric acidMetabolite 1

Hydroxy-lauric acidMetabolite 2

Hydroxy-lauric acidMetabolite 3

OH

O

OH

OH

O OH

OH

O

OH

Hydroxy-lauric acidMetabolite 4

OH

O OH

Spectrum not shown for hydroxy-myristic acidLauric acid derived-metabolite 1 showed after derivatisation ions at m/z (relative intensity %) 73 (54%) ((CH3)3Si+), 159 (base peak 100%) and 245 (57%) representing cleavage on both sides of the carbon carrying the hydroxyl silylated, 255 (8%) (M-47) (loss of methanol from the (M-15) fragment), 271 (4%) (M-31) (loss of OCH3 from the methyl ester), 287 (2%) (M-15) (loss of a methyl from the TMSi group). This fragmentation pattern is as expected characteristic from derivatised 8-hydroxylauric acid (M=302 g/mol). Mass spectrum of the metabolite 2 of lauric acid after derivatisation showed ions at m/z (relative intensity %) 73 (62%) ((CH3)3Si+), 173 (base peak 100%) and 231 (70%) come from cleavage on both sides of the carbon carrying the hydroxyl silylated, 159 (9%) (CH3-O+=C+(OSi(CH3)3)CH=CH2) 255 (9%) (M-47) (loss of methanol from the (M-15) fragment), 271 (5%) (M-31) (loss of OCH3 from the methyl ester), 287 (3%) (M-15) (loss of a methyl from the TMSi group). This fragmentation pattern is as expected from the derivatized 7-hydroxylauric acid (M=302 g/mol). Mass spectrum of metabolite 3 of lauric acid after derivatisation showed ions at m/z (relative intensity %) 73 (85%) ((CH3)3Si+), 187 (base peak 100%) and 217 (62%) come from cleavage on both sides of the carbon carrying the hydroxyl silylated, 159 (10%) (CH3-O+=C+(OSi(CH3)3)CH=CH2), 255 (10%) (M-47) (loss of methanol from the (M-15) fragment), 271 (8%) (M-31) (loss of OCH3 from the methyl ester), 287 (8%) (M-15) (loss of a methyl from the TMSi group).This fragmentation pattern is as expected from the derivatized 6-hydroxylauric acid (M=302 g/mol). Lauric acid derived-metabolite 4 showed after derivatisation ions at m/z (relative intensity %) 73 (59%) ((CH3)3Si+), 145 (base peak 100%) and 259 (44%) representing cleavage on both sides of the carbon carrying the hydroxyl silylated, 255 (7%) (M-47) (loss of methanol from the (M-15) fragment), 287 (2%) (M-15) (loss of a methyl from the TMSi group) This fragmentation pattern is as expected from the derivatized 9-hydroxylauric acid (M=302 g/mol).

Mass spectrum of metabolite 1 of capric acid after derivatisation showed ions at m/z (relative intensity %) 73 (65%) ((CH3)3Si+), 131 (base peak 100%) and 245 (50%) come from cleavage on both sides of the carbon carrying the hydroxyl silylated, 159 (10%) (CH3-O+=C+(OSi(CH3)3)CH=CH2), 227 (11%) (M-47) (loss of methanol from the (M-15) fragment), 243 (4%) (M-31) (loss of OCH3 from the methyl ester), 259 (2%) (M-15) (loss of a methyl from the TMSi group). This fragmentation pattern is characteristic of the derivative of 8-hydroxycapric acid (M=274 g/mol). Mass spectrum of the metabolite 2 of capric acid after derivatisation showed ions at m/z (relative intensity %) 73 (69%) ((CH3)3Si+), 145 (base peak 100%) and 231 (46%) come from cleavage on both sides of the carbon carrying the hydroxyl silylated, 159 (10%) (CH3-O+=C+(OSi(CH3)3)CH=CH2), 227 (10%) (M-47) (loss of methanol from the (M-15) fragment), 243 (4%) (M-31) (loss of OCH3 from the methyl ester), 259 (3%) (M-15) (loss of a methyl from the TMSi group). This fragmentation pattern is characteristic of the derivative of 7-hydroxycapric acid (M=274 g/mol). Mass spectrum of the metabolite 3 of capric acid after derivatisation showed ions at m/z (relative intensity %) 73 (62%) ((CH3)3Si+), 159 (24%) and 217 (10%) come from cleavage on both sides of the carbon carrying the hydroxyl silylated, 159 (24%) also come from (CH3-O+=C+(OSi(CH3)3)CH=CH2), 227 (3%) (M-47) (loss of methanol from the (M-15) fragment), 243 (base peak 100%) (M-31) (loss of OCH3 from the methyl ester), 259 (2%) (M-15) (loss of a methyl from the TMSi group). This fragmentation pattern is characteristic of the derivative of 6-hydroxycapric acid (M=274 g/mol).

Mass spectrum of metabolite 1 of myristic acid after derivatisation showed ions at m/z (relative intensity %) 73 (base peak 100%) ((CH3)3Si+), 187 (88%) and 245 (83%) come from cleavage on both sides of the carbon carrying the hydroxyl silylated, 283 (12%) (M-47) (loss of methanol from the (M-15) fragment), 299 (21%) (M-31) (loss of OCH3 from the methyl ester), and 315 (5%) (M-15) (loss of a methyl from the TMSi group). This fragmentation pattern is characteristic of the derivative of 7-hydroxymyristic acid (M=330 g/mol). Mass spectrum of metabolite 2 of myristic acid after derivatisation showed ions at m/z (relative intensity %) 73 (base peak 100%) ((CH3)3Si+), 201 (50%) and 231 (44%) come from cleavage on both sides of the carbon carrying the hydroxyl silylated, 283 (10%) (M-47) (loss of methanol from the (M-15) fragment), 299 (8%) (M-31) (loss of OCH3 from the methyl ester), and 315 (3%) (M-15) (loss of a methyl from the TMSi group). This fragmentation pattern is characteristic of the derivative of 6-hydroxymyristic acid (M=330 g/mol). Mass spectrum of metabolite 3 of myristic acid after derivatisation showed ions at m/z (relative intensity %) 73 (base peak 100%) ((CH3)3Si+), 215 (60%) and 217 (58%) come from cleavage on both sides of the carbon carrying the hydroxyl silylated, 283 (14%) (M-47) (loss of methanol from the (M-15) fragment), 299 (14%) (M-31) (loss of OCH3 from the methyl ester), and 315 (8%) (M-15) (loss of a methyl from the TMSi group). This fragmentation pattern is characteristic of the derivative of 5-hydroxymyristic acid (M=330 g/mol).