药学学报

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
A0	13.14	[M+Na]⁺	C₁₇H₂₄O₁₀Na	411.126 1	411.124 8	-3.2	249.072 8 [M+H-C₆H₁₀O₅]⁺ 231.062 1 [M+Na-C₆H₁₀O₅-H₂O]⁺	Geniposide, parent	Plasma, bile, urine
A1	9.03	[M+Na]⁺	C₁₇H₂₂O₁₁Na	425.105 4	425.107 5	4.9	249.072 8 [M+H-C₆H₈O₆]⁺ 231.062 5 [M+Na-C₆H₈O₆-H₂O]⁺ 199.021 7 [GlucA-H₂O+Na]⁺	Glucuronidation of A0	Plasma, bile, urine
A2	13.97	[M+Na]⁺	C₁₇H₂₄O₁₁Na	427.121 1	427.120 7	-0.9	233.078 9 [M+Na-C₆H₈O₆-H₂O]⁺	Ring-opened of A1	Plasma
A3-1	13.71	[M+H]⁺	C₁₆H₂₅O₈	345.154 4	345.153 7	-2.0	169.121 8 [M+H-C₆H₈O₆]⁺ 151.111 3 [M+H-C₆H₈O₆-H₂O]⁺	Cleavage and dihydroxylation of A2	Bile, feces
A3-2	14.01	[M+H]⁺	C₁₆H₂₅O₈	345.154 4	345.153 6	-2.3	169.121 8 [M+H-C₆H₈O₆]⁺ 151.111 3 [M+H-C₆H₈O₆-H₂O]⁺	Cleavage and dihydroxylation of A2	Bile, feces
A4-1	9.57	[M+H]⁺	C₂₀H₃₀NO₁₁S	492.153 4	492.152 0	-2.8	330.100 4 [M+H-C₆H₁₀O₅]⁺ 312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺	Cys conjugation of A0	Plasma, bile, urine
A4-2	13.35	[M+H]⁺	C₂₀H₃₀NO₁₁S	492.153 4	492.152 3	-2.2	330.100 4 [M+H-C₆H₁₀O₅]⁺ 312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺	Cys conjugation of A0	Plasma, bile, urine
A5	13.50	[M+Na]⁺	C₁₆H₂₂O₉Na	381.115 6	381.114 4	-3.1	219.065 0 [M+Na-C₆H₁₀O₅]⁺ 201.052 1 [M+Na-C₆H₁₀O₅-H₂O]⁺	Hydroxymethylene loss of A0	Plasma, bile
A6	10.50	[M+H]⁺	C₁₁H₁₅O₅	227.091 4	227.090 9	-2.2	213.076 2 [M+H-CH₂]⁺ 209.080 5 [M+H-H₂O]⁺ 191.070 3 [M+H-2H₂O]⁺	Genipin	Plasma, urine
A7	9.13	[M+H]⁺	C₁₀H₁₃O₅	213.075 8	213.075 1	-3.3	195.065 3 [M+H-H₂O]⁺ 185.080 8 [M+H-CO]⁺ 151.075 2 [M+H-H₂O-CO₂]⁺	Hydrolysis of A6	Plasma, urine, feces
A8	13.95	[M+H]⁺	C₁₁H₁₃O₅	225.075 8	225.075 2	-2.7	207.065 1 [M+H-H₂O]⁺ 195.059 4 [M+H-OCH₂]⁺ 151.075 5 [M+H-OCH₂-CO₂]⁺	Oxidation of A6	Plasma, bile, feces
A9-1	12.35	[M+H]⁺	C₁₁H₁₃O₄	209.080 8	209.080 4	-1.9	191.070 5 [M+H-H₂O]⁺ 177.054 3 [M+H-CH₂-H₂O]⁺	Dehydration of A6	Plasma, bile, urine, feces
A9-2	13.09	[M+H]⁺	C₁₁H₁₃O₄	209.080 8	209.080 2	-2.9	181.085 8 [M+H-CO]⁺ 177.054 5 [M+H-CH₂-H₂O]⁺	Dehydration of A6	Plasma, bile, urine, feces
A10	11.94	[M+H]⁺	C₁₀H₁₃O₄	197.080 8	197.080 2	-3.0	179.070 3 [M+H-H₂O]⁺ 169.085 8 [M+H-CO]⁺ 151.075 3 [M+H-CO-H₂O]⁺	Hydroxymethylene loss of A6	Plasma, bile
A11	2.11	[M+H]⁺	C₁₀H₁₅O₅	215.091 4	215.090 7	-3.3	197.080 8 [M+H-H₂O]⁺ 169.085 9 [M+H-H₂O-CO]⁺	Ring-opened of A7	Plasma, urine

No.

t_R /min

Ion form

Formula

m/z

Error /ppm

MS/MS fragment ion

Identification/ reaction

Source

Theoretical

Experimental

13.14

[M+Na]⁺

C₁₇H₂₄O₁₀Na

411.126 1

411.124 8

-3.2

249.072 8 [M+H-C₆H₁₀O₅]⁺
231.062 1 [M+Na-C₆H₁₀O₅-H₂O]⁺

Geniposide, parent

Plasma, bile, urine

9.03

[M+Na]⁺

C₁₇H₂₂O₁₁Na

425.105 4

425.107 5

4.9

249.072 8 [M+H-C₆H₈O₆]⁺
231.062 5 [M+Na-C₆H₈O₆-H₂O]⁺
199.021 7 [GlucA-H₂O+Na]⁺

Glucuronidation of A0

Plasma, bile, urine

13.97

[M+Na]⁺

C₁₇H₂₄O₁₁Na

427.121 1

427.120 7

-0.9

233.078 9 [M+Na-C₆H₈O₆-H₂O]⁺

Ring-opened of A1

Plasma

A3-1

13.71

[M+H]⁺

C₁₆H₂₅O₈

345.154 4

345.153 7

-2.0

169.121 8 [M+H-C₆H₈O₆]⁺
151.111 3 [M+H-C₆H₈O₆-H₂O]⁺

Cleavage and dihydroxylation of A2

Bile, feces

A3-2

14.01

[M+H]⁺

C₁₆H₂₅O₈

345.154 4

345.153 6

-2.3

169.121 8 [M+H-C₆H₈O₆]⁺
151.111 3 [M+H-C₆H₈O₆-H₂O]⁺

Cleavage and dihydroxylation of A2

Bile, feces

A4-1

9.57

[M+H]⁺

C₂₀H₃₀NO₁₁S

492.153 4

492.152 0

-2.8

330.100 4 [M+H-C₆H₁₀O₅]⁺
312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺

Cys conjugation of A0

Plasma, bile, urine

A4-2

13.35

[M+H]⁺

C₂₀H₃₀NO₁₁S

492.153 4

492.152 3

-2.2

330.100 4 [M+H-C₆H₁₀O₅]⁺
312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺

Cys conjugation of A0

Plasma, bile, urine

13.50

[M+Na]⁺

C₁₆H₂₂O₉Na

381.115 6

381.114 4

-3.1

219.065 0 [M+Na-C₆H₁₀O₅]⁺
201.052 1 [M+Na-C₆H₁₀O₅-H₂O]⁺

Hydroxymethylene loss of A0

Plasma, bile

10.50

[M+H]⁺

C₁₁H₁₅O₅

227.091 4

227.090 9

-2.2

213.076 2 [M+H-CH₂]⁺
209.080 5 [M+H-H₂O]⁺
191.070 3 [M+H-2H₂O]⁺

Genipin

Plasma, urine

9.13

[M+H]⁺

C₁₀H₁₃O₅

213.075 8

213.075 1

-3.3

195.065 3 [M+H-H₂O]⁺
185.080 8 [M+H-CO]⁺
151.075 2 [M+H-H₂O-CO₂]⁺

Hydrolysis of A6

Plasma, urine, feces

13.95

[M+H]⁺

C₁₁H₁₃O₅

225.075 8

225.075 2

-2.7

207.065 1 [M+H-H₂O]⁺
195.059 4 [M+H-OCH₂]⁺
151.075 5 [M+H-OCH₂-CO₂]⁺

Oxidation of A6

Plasma, bile, feces

A9-1

12.35

[M+H]⁺

C₁₁H₁₃O₄

209.080 8

209.080 4

-1.9

191.070 5 [M+H-H₂O]⁺
177.054 3 [M+H-CH₂-H₂O]⁺

Dehydration of A6

Plasma, bile, urine, feces

A9-2

13.09

[M+H]⁺

C₁₁H₁₃O₄

209.080 8

209.080 2

-2.9

181.085 8 [M+H-CO]⁺
177.054 5 [M+H-CH₂-H₂O]⁺

Dehydration of A6

Plasma, bile, urine, feces

A10

11.94

[M+H]⁺

C₁₀H₁₃O₄

197.080 8

197.080 2

-3.0

179.070 3 [M+H-H₂O]⁺
169.085 8 [M+H-CO]⁺
151.075 3 [M+H-CO-H₂O]⁺

Hydroxymethylene loss of A6

Plasma, bile

A11

2.11

[M+H]⁺

C₁₀H₁₅O₅

215.091 4

215.090 7

-3.3

197.080 8 [M+H-H₂O]⁺
169.085 9 [M+H-H₂O-CO]⁺

Ring-opened of A7

Plasma, urine

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
A0	13.14	[M+Na]⁺	C₁₇H₂₄O₁₀Na	411.126 1	411.124 8	-3.2	249.072 8 [M+H-C₆H₁₀O₅]⁺ 231.062 1 [M+Na-C₆H₁₀O₅-H₂O]⁺	Geniposide, parent	Plasma, bile, urine
A1	9.03	[M+Na]⁺	C₁₇H₂₂O₁₁Na	425.105 4	425.107 5	4.9	249.072 8 [M+H-C₆H₈O₆]⁺ 231.062 5 [M+Na-C₆H₈O₆-H₂O]⁺ 199.021 7 [GlucA-H₂O+Na]⁺	Glucuronidation of A0	Plasma, bile, urine
A2	13.97	[M+Na]⁺	C₁₇H₂₄O₁₁Na	427.121 1	427.120 7	-0.9	233.078 9 [M+Na-C₆H₈O₆-H₂O]⁺	Ring-opened of A1	Plasma
A3-1	13.71	[M+H]⁺	C₁₆H₂₅O₈	345.154 4	345.153 7	-2.0	169.121 8 [M+H-C₆H₈O₆]⁺ 151.111 3 [M+H-C₆H₈O₆-H₂O]⁺	Cleavage and dihydroxylation of A2	Bile, feces
A3-2	14.01	[M+H]⁺	C₁₆H₂₅O₈	345.154 4	345.153 6	-2.3	169.121 8 [M+H-C₆H₈O₆]⁺ 151.111 3 [M+H-C₆H₈O₆-H₂O]⁺	Cleavage and dihydroxylation of A2	Bile, feces
A4-1	9.57	[M+H]⁺	C₂₀H₃₀NO₁₁S	492.153 4	492.152 0	-2.8	330.100 4 [M+H-C₆H₁₀O₅]⁺ 312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺	Cys conjugation of A0	Plasma, bile, urine
A4-2	13.35	[M+H]⁺	C₂₀H₃₀NO₁₁S	492.153 4	492.152 3	-2.2	330.100 4 [M+H-C₆H₁₀O₅]⁺ 312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺	Cys conjugation of A0	Plasma, bile, urine
A5	13.50	[M+Na]⁺	C₁₆H₂₂O₉Na	381.115 6	381.114 4	-3.1	219.065 0 [M+Na-C₆H₁₀O₅]⁺ 201.052 1 [M+Na-C₆H₁₀O₅-H₂O]⁺	Hydroxymethylene loss of A0	Plasma, bile
A6	10.50	[M+H]⁺	C₁₁H₁₅O₅	227.091 4	227.090 9	-2.2	213.076 2 [M+H-CH₂]⁺ 209.080 5 [M+H-H₂O]⁺ 191.070 3 [M+H-2H₂O]⁺	Genipin	Plasma, urine
A7	9.13	[M+H]⁺	C₁₀H₁₃O₅	213.075 8	213.075 1	-3.3	195.065 3 [M+H-H₂O]⁺ 185.080 8 [M+H-CO]⁺ 151.075 2 [M+H-H₂O-CO₂]⁺	Hydrolysis of A6	Plasma, urine, feces
A8	13.95	[M+H]⁺	C₁₁H₁₃O₅	225.075 8	225.075 2	-2.7	207.065 1 [M+H-H₂O]⁺ 195.059 4 [M+H-OCH₂]⁺ 151.075 5 [M+H-OCH₂-CO₂]⁺	Oxidation of A6	Plasma, bile, feces
A9-1	12.35	[M+H]⁺	C₁₁H₁₃O₄	209.080 8	209.080 4	-1.9	191.070 5 [M+H-H₂O]⁺ 177.054 3 [M+H-CH₂-H₂O]⁺	Dehydration of A6	Plasma, bile, urine, feces
A9-2	13.09	[M+H]⁺	C₁₁H₁₃O₄	209.080 8	209.080 2	-2.9	181.085 8 [M+H-CO]⁺ 177.054 5 [M+H-CH₂-H₂O]⁺	Dehydration of A6	Plasma, bile, urine, feces
A10	11.94	[M+H]⁺	C₁₀H₁₃O₄	197.080 8	197.080 2	-3.0	179.070 3 [M+H-H₂O]⁺ 169.085 8 [M+H-CO]⁺ 151.075 3 [M+H-CO-H₂O]⁺	Hydroxymethylene loss of A6	Plasma, bile
A11	2.11	[M+H]⁺	C₁₀H₁₅O₅	215.091 4	215.090 7	-3.3	197.080 8 [M+H-H₂O]⁺ 169.085 9 [M+H-H₂O-CO]⁺	Ring-opened of A7	Plasma, urine

No.

t_R /min

Ion form

Formula

m/z

Error /ppm

MS/MS fragment ion

Identification/ reaction

Source

Theoretical

Experimental

13.14

[M+Na]⁺

C₁₇H₂₄O₁₀Na

411.126 1

411.124 8

-3.2

249.072 8 [M+H-C₆H₁₀O₅]⁺
231.062 1 [M+Na-C₆H₁₀O₅-H₂O]⁺

Geniposide, parent

Plasma, bile, urine

9.03

[M+Na]⁺

C₁₇H₂₂O₁₁Na

425.105 4

425.107 5

4.9

249.072 8 [M+H-C₆H₈O₆]⁺
231.062 5 [M+Na-C₆H₈O₆-H₂O]⁺
199.021 7 [GlucA-H₂O+Na]⁺

Glucuronidation of A0

Plasma, bile, urine

13.97

[M+Na]⁺

C₁₇H₂₄O₁₁Na

427.121 1

427.120 7

-0.9

233.078 9 [M+Na-C₆H₈O₆-H₂O]⁺

Ring-opened of A1

Plasma

A3-1

13.71

[M+H]⁺

C₁₆H₂₅O₈

345.154 4

345.153 7

-2.0

169.121 8 [M+H-C₆H₈O₆]⁺
151.111 3 [M+H-C₆H₈O₆-H₂O]⁺

Cleavage and dihydroxylation of A2

Bile, feces

A3-2

14.01

[M+H]⁺

C₁₆H₂₅O₈

345.154 4

345.153 6

-2.3

169.121 8 [M+H-C₆H₈O₆]⁺
151.111 3 [M+H-C₆H₈O₆-H₂O]⁺

Cleavage and dihydroxylation of A2

Bile, feces

A4-1

9.57

[M+H]⁺

C₂₀H₃₀NO₁₁S

492.153 4

492.152 0

-2.8

330.100 4 [M+H-C₆H₁₀O₅]⁺
312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺

Cys conjugation of A0

Plasma, bile, urine

A4-2

13.35

[M+H]⁺

C₂₀H₃₀NO₁₁S

492.153 4

492.152 3

-2.2

330.100 4 [M+H-C₆H₁₀O₅]⁺
312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺

Cys conjugation of A0

Plasma, bile, urine

13.50

[M+Na]⁺

C₁₆H₂₂O₉Na

381.115 6

381.114 4

-3.1

219.065 0 [M+Na-C₆H₁₀O₅]⁺
201.052 1 [M+Na-C₆H₁₀O₅-H₂O]⁺

Hydroxymethylene loss of A0

Plasma, bile

10.50

[M+H]⁺

C₁₁H₁₅O₅

227.091 4

227.090 9

-2.2

213.076 2 [M+H-CH₂]⁺
209.080 5 [M+H-H₂O]⁺
191.070 3 [M+H-2H₂O]⁺

Genipin

Plasma, urine

9.13

[M+H]⁺

C₁₀H₁₃O₅

213.075 8

213.075 1

-3.3

195.065 3 [M+H-H₂O]⁺
185.080 8 [M+H-CO]⁺
151.075 2 [M+H-H₂O-CO₂]⁺

Hydrolysis of A6

Plasma, urine, feces

13.95

[M+H]⁺

C₁₁H₁₃O₅

225.075 8

225.075 2

-2.7

207.065 1 [M+H-H₂O]⁺
195.059 4 [M+H-OCH₂]⁺
151.075 5 [M+H-OCH₂-CO₂]⁺

Oxidation of A6

Plasma, bile, feces

A9-1

12.35

[M+H]⁺

C₁₁H₁₃O₄

209.080 8

209.080 4

-1.9

191.070 5 [M+H-H₂O]⁺
177.054 3 [M+H-CH₂-H₂O]⁺

Dehydration of A6

Plasma, bile, urine, feces

A9-2

13.09

[M+H]⁺

C₁₁H₁₃O₄

209.080 8

209.080 2

-2.9

181.085 8 [M+H-CO]⁺
177.054 5 [M+H-CH₂-H₂O]⁺

Dehydration of A6

Plasma, bile, urine, feces

A10

11.94

[M+H]⁺

C₁₀H₁₃O₄

197.080 8

197.080 2

-3.0

179.070 3 [M+H-H₂O]⁺
169.085 8 [M+H-CO]⁺
151.075 3 [M+H-CO-H₂O]⁺

Hydroxymethylene loss of A6

Plasma, bile

A11

2.11

[M+H]⁺

C₁₀H₁₅O₅

215.091 4

215.090 7

-3.3

197.080 8 [M+H-H₂O]⁺
169.085 9 [M+H-H₂O-CO]⁺

Ring-opened of A7

Plasma, urine

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
B0	9.38	[M+Na]⁺	C₁₆H₂₂O₁₀Na	397.110 5	397.109 0	-3.8	235.052 6 [M+Na-C₆H₁₀O₅]⁺ 217.047 1 [M+Na-C₆H₁₀O₅-H₂O]⁺	Geniposidic acid, parent	Plasma, bile, urine
B1	10.53	[M+H]⁺	C₁₉H₂₈NO₁₁S	478.137 8	478.136 1	-3.6	316.076 4 [M+H-C₆H₁₀O₅]⁺ 298.074 4 [M+H-C₆H₁₀O₅-H₂O]⁺ 280.063 5 [M+H-C₆H₁₀O₅-2H₂O]⁺	Cys conjugation of B0	Plasma, urine
B2	15.48	[M+H]⁺	C₁₈H₂₉O₁₁	421.170 4	421.168 8	-3.8	375.125 6 [M+H-C₂H₄-H₂O]⁺ 213.075 7 [M+H-C₆H₁₀O₅-C₂H₄-H₂O]⁺	Hydroxylation and ethylation of B0	Plasma
B3	10.36	[M+H]⁺	C₁₆H₂₅O₁₀	377.144 2	377.144 0	-0.5	331.138 5 [M+H-CO₂]⁺ 215.091 4 [M+H-C₆H₁₀O₅]⁺ 197.080 8 [M+H-C₆H₁₀O₅-H₂O]⁺	Double-bond reduction of B0	Plasma, bile, urine, feces
B4	13.84	[M+H]⁺	C₁₆H₂₇O₇	331.175 1	331.174 1	-3.0	313.152 6 [M+H-H₂O]⁺ 169.122 3 [M+H-C₆H₁₀O₅]⁺	Ring-opened and cleavage of B0	Plasma

No.

t_R
/min

Ion form

Formula

m/z

Error
/ppm

MS/MS fragment ion

Identification/
reaction

Source

Theoretical

Experimental

9.38

[M+Na]⁺

C₁₆H₂₂O₁₀Na

397.110 5

397.109 0

-3.8

235.052 6 [M+Na-C₆H₁₀O₅]⁺
217.047 1 [M+Na-C₆H₁₀O₅-H₂O]⁺

Geniposidic acid, parent

Plasma, bile, urine

10.53

[M+H]⁺

C₁₉H₂₈NO₁₁S

478.137 8

478.136 1

-3.6

316.076 4 [M+H-C₆H₁₀O₅]⁺
298.074 4 [M+H-C₆H₁₀O₅-H₂O]⁺
280.063 5 [M+H-C₆H₁₀O₅-2H₂O]⁺

Cys conjugation of B0

Plasma, urine

15.48

[M+H]⁺

C₁₈H₂₉O₁₁

421.170 4

421.168 8

-3.8

375.125 6 [M+H-C₂H₄-H₂O]⁺
213.075 7 [M+H-C₆H₁₀O₅-C₂H₄-H₂O]⁺

Hydroxylation and ethylation of B0

Plasma

10.36

[M+H]⁺

C₁₆H₂₅O₁₀

377.144 2

377.144 0

-0.5

331.138 5 [M+H-CO₂]⁺
215.091 4 [M+H-C₆H₁₀O₅]⁺
197.080 8 [M+H-C₆H₁₀O₅-H₂O]⁺

Double-bond reduction of B0

Plasma, bile, urine, feces

13.84

[M+H]⁺

C₁₆H₂₇O₇

331.175 1

331.174 1

-3.0

313.152 6 [M+H-H₂O]⁺
169.122 3 [M+H-C₆H₁₀O₅]⁺

Ring-opened and cleavage of B0

Plasma

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
B0	9.38	[M+Na]⁺	C₁₆H₂₂O₁₀Na	397.110 5	397.109 0	-3.8	235.052 6 [M+Na-C₆H₁₀O₅]⁺ 217.047 1 [M+Na-C₆H₁₀O₅-H₂O]⁺	Geniposidic acid, parent	Plasma, bile, urine
B1	10.53	[M+H]⁺	C₁₉H₂₈NO₁₁S	478.137 8	478.136 1	-3.6	316.076 4 [M+H-C₆H₁₀O₅]⁺ 298.074 4 [M+H-C₆H₁₀O₅-H₂O]⁺ 280.063 5 [M+H-C₆H₁₀O₅-2H₂O]⁺	Cys conjugation of B0	Plasma, urine
B2	15.48	[M+H]⁺	C₁₈H₂₉O₁₁	421.170 4	421.168 8	-3.8	375.125 6 [M+H-C₂H₄-H₂O]⁺ 213.075 7 [M+H-C₆H₁₀O₅-C₂H₄-H₂O]⁺	Hydroxylation and ethylation of B0	Plasma
B3	10.36	[M+H]⁺	C₁₆H₂₅O₁₀	377.144 2	377.144 0	-0.5	331.138 5 [M+H-CO₂]⁺ 215.091 4 [M+H-C₆H₁₀O₅]⁺ 197.080 8 [M+H-C₆H₁₀O₅-H₂O]⁺	Double-bond reduction of B0	Plasma, bile, urine, feces
B4	13.84	[M+H]⁺	C₁₆H₂₇O₇	331.175 1	331.174 1	-3.0	313.152 6 [M+H-H₂O]⁺ 169.122 3 [M+H-C₆H₁₀O₅]⁺	Ring-opened and cleavage of B0	Plasma

No.

t_R
/min

Ion form

Formula

m/z

Error
/ppm

MS/MS fragment ion

Identification/
reaction

Source

Theoretical

Experimental

9.38

[M+Na]⁺

C₁₆H₂₂O₁₀Na

397.110 5

397.109 0

-3.8

235.052 6 [M+Na-C₆H₁₀O₅]⁺
217.047 1 [M+Na-C₆H₁₀O₅-H₂O]⁺

Geniposidic acid, parent

Plasma, bile, urine

10.53

[M+H]⁺

C₁₉H₂₈NO₁₁S

478.137 8

478.136 1

-3.6

316.076 4 [M+H-C₆H₁₀O₅]⁺
298.074 4 [M+H-C₆H₁₀O₅-H₂O]⁺
280.063 5 [M+H-C₆H₁₀O₅-2H₂O]⁺

Cys conjugation of B0

Plasma, urine

15.48

[M+H]⁺

C₁₈H₂₉O₁₁

421.170 4

421.168 8

-3.8

375.125 6 [M+H-C₂H₄-H₂O]⁺
213.075 7 [M+H-C₆H₁₀O₅-C₂H₄-H₂O]⁺

Hydroxylation and ethylation of B0

Plasma

10.36

[M+H]⁺

C₁₆H₂₅O₁₀

377.144 2

377.144 0

-0.5

331.138 5 [M+H-CO₂]⁺
215.091 4 [M+H-C₆H₁₀O₅]⁺
197.080 8 [M+H-C₆H₁₀O₅-H₂O]⁺

Double-bond reduction of B0

Plasma, bile, urine, feces

13.84

[M+H]⁺

C₁₆H₂₇O₇

331.175 1

331.174 1

-3.0

313.152 6 [M+H-H₂O]⁺
169.122 3 [M+H-C₆H₁₀O₅]⁺

Ring-opened and cleavage of B0

Plasma

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
C0	13.98	[M+H]⁺	C₁₇H₂₅O₁₁	405.139 1	405.138 6	-1.2	391.119 2 [M+H-CH₂]⁺ 373.121 6 [M+H-CH₂-H₂O]⁺ 243.085 4 [M+H-C₆H₁₀O₅]⁺	Secoxylogain, parent	Plasma, bile, urine
C1	10.34	[M+H]⁺	C₁₁H₁₅O₆	243.086 3	243.085 5	-3.3	225.076 7 [M+H-H₂O]⁺ 207.065 2 [M+H-2H₂O]⁺	Glucoside removal of C0	Plasma, urine
C2	12.26	[M+H]⁺	C₁₀H₁₃O₆	229.070 7	229.069 9	-3.5	211.060 1 [M+H-H₂O]⁺ 201.074 8 [M+H-CO]⁺	Hydrolysis of C1	Plasma, bile
C3-1	11.51	[M+H]⁺	C₁₉H₃₁O₁₂	451.181 0	451.182 3	2.9	243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺ 225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺	Hydroxylation and ethylation of C0	Bile
C3-2	14.11	[M+H]⁺	C₁₉H₃₁O₁₂	451.181 0	451.181 3	0.7	243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺ 225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺	Hydroxylation and ethylation of C0	Bile
C4	12.18	[M+H]⁺	C₁₆H₂₃O₁₁	391.123 5	391.124 2	1.8	373.112 6 [M+H-H₂O]⁺	Hydrolysis of C0	Plasma, bile, urine
C5	12.50	[M+H]⁺	C₁₆H₂₃O₁₀	375.128 6	375.127 5	-2.9	357.113 8 [M+H-H₂O]⁺ 347.133 1 [M+H-CO]⁺ 195.065 2 [M+H-C₆H₁₀O₅-H₂O]⁺	Hydroxymethylene loss of C0	Plasma, bile, urine

No.

t_R
/min

Ion form

Formula

m/z

Error
/ppm

MS/MS fragment ion

Identification/
reaction

Source

Theoretical

Experimental

13.98

[M+H]⁺

C₁₇H₂₅O₁₁

405.139 1

405.138 6

-1.2

391.119 2 [M+H-CH₂]⁺
373.121 6 [M+H-CH₂-H₂O]⁺
243.085 4 [M+H-C₆H₁₀O₅]⁺

Secoxylogain, parent

Plasma, bile, urine

10.34

[M+H]⁺

C₁₁H₁₅O₆

243.086 3

243.085 5

-3.3

225.076 7 [M+H-H₂O]⁺
207.065 2 [M+H-2H₂O]⁺

Glucoside removal of C0

Plasma, urine

12.26

[M+H]⁺

C₁₀H₁₃O₆

229.070 7

229.069 9

-3.5

211.060 1 [M+H-H₂O]⁺
201.074 8 [M+H-CO]⁺

Hydrolysis of C1

Plasma, bile

C3-1

11.51

[M+H]⁺

C₁₉H₃₁O₁₂

451.181 0

451.182 3

2.9

243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺
225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺

Hydroxylation and ethylation of C0

Bile

C3-2

14.11

[M+H]⁺

C₁₉H₃₁O₁₂

451.181 0

451.181 3

0.7

243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺
225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺

Hydroxylation and ethylation of C0

Bile

12.18

[M+H]⁺

C₁₆H₂₃O₁₁

391.123 5

391.124 2

1.8

373.112 6 [M+H-H₂O]⁺

Hydrolysis of C0

Plasma, bile, urine

12.50

[M+H]⁺

C₁₆H₂₃O₁₀

375.128 6

375.127 5

-2.9

357.113 8 [M+H-H₂O]⁺
347.133 1 [M+H-CO]⁺
195.065 2 [M+H-C₆H₁₀O₅-H₂O]⁺

Hydroxymethylene loss of C0

Plasma, bile, urine

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
C0	13.98	[M+H]⁺	C₁₇H₂₅O₁₁	405.139 1	405.138 6	-1.2	391.119 2 [M+H-CH₂]⁺ 373.121 6 [M+H-CH₂-H₂O]⁺ 243.085 4 [M+H-C₆H₁₀O₅]⁺	Secoxylogain, parent	Plasma, bile, urine
C1	10.34	[M+H]⁺	C₁₁H₁₅O₆	243.086 3	243.085 5	-3.3	225.076 7 [M+H-H₂O]⁺ 207.065 2 [M+H-2H₂O]⁺	Glucoside removal of C0	Plasma, urine
C2	12.26	[M+H]⁺	C₁₀H₁₃O₆	229.070 7	229.069 9	-3.5	211.060 1 [M+H-H₂O]⁺ 201.074 8 [M+H-CO]⁺	Hydrolysis of C1	Plasma, bile
C3-1	11.51	[M+H]⁺	C₁₉H₃₁O₁₂	451.181 0	451.182 3	2.9	243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺ 225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺	Hydroxylation and ethylation of C0	Bile
C3-2	14.11	[M+H]⁺	C₁₉H₃₁O₁₂	451.181 0	451.181 3	0.7	243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺ 225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺	Hydroxylation and ethylation of C0	Bile
C4	12.18	[M+H]⁺	C₁₆H₂₃O₁₁	391.123 5	391.124 2	1.8	373.112 6 [M+H-H₂O]⁺	Hydrolysis of C0	Plasma, bile, urine
C5	12.50	[M+H]⁺	C₁₆H₂₃O₁₀	375.128 6	375.127 5	-2.9	357.113 8 [M+H-H₂O]⁺ 347.133 1 [M+H-CO]⁺ 195.065 2 [M+H-C₆H₁₀O₅-H₂O]⁺	Hydroxymethylene loss of C0	Plasma, bile, urine

No.

t_R
/min

Ion form

Formula

m/z

Error
/ppm

MS/MS fragment ion

Identification/
reaction

Source

Theoretical

Experimental

13.98

[M+H]⁺

C₁₇H₂₅O₁₁

405.139 1

405.138 6

-1.2

391.119 2 [M+H-CH₂]⁺
373.121 6 [M+H-CH₂-H₂O]⁺
243.085 4 [M+H-C₆H₁₀O₅]⁺

Secoxylogain, parent

Plasma, bile, urine

10.34

[M+H]⁺

C₁₁H₁₅O₆

243.086 3

243.085 5

-3.3

225.076 7 [M+H-H₂O]⁺
207.065 2 [M+H-2H₂O]⁺

Glucoside removal of C0

Plasma, urine

12.26

[M+H]⁺

C₁₀H₁₃O₆

229.070 7

229.069 9

-3.5

211.060 1 [M+H-H₂O]⁺
201.074 8 [M+H-CO]⁺

Hydrolysis of C1

Plasma, bile

C3-1

11.51

[M+H]⁺

C₁₉H₃₁O₁₂

451.181 0

451.182 3

2.9

243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺
225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺

Hydroxylation and ethylation of C0

Bile

C3-2

14.11

[M+H]⁺

C₁₉H₃₁O₁₂

451.181 0

451.181 3

0.7

243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺
225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺

Hydroxylation and ethylation of C0

Bile

12.18

[M+H]⁺

C₁₆H₂₃O₁₁

391.123 5

391.124 2

1.8

373.112 6 [M+H-H₂O]⁺

Hydrolysis of C0

Plasma, bile, urine

12.50

[M+H]⁺

C₁₆H₂₃O₁₀

375.128 6

375.127 5

-2.9

357.113 8 [M+H-H₂O]⁺
347.133 1 [M+H-CO]⁺
195.065 2 [M+H-C₆H₁₀O₅-H₂O]⁺

Hydroxymethylene loss of C0

Plasma, bile, urine

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
D0	12.28	[M+H]⁺	C₁₆H₁₉O₉	355.102 4	355.101 6	-2.3	181.049 2 [M+H-C₇H₁₀O₅]⁺ 163.038 4 [M+H-C₇H₁₀O₅-H₂O]⁺ 149.022 8 [M+H-C₇H₁₀O₅-CH₂-H₂O]⁺	Chlorogenic acid	Plasma, urine
D1-1	12.54	[M+H]⁺	C₁₇H₂₁O₉	369.118 0	369.116 9	-3.0	351.107 0 [M+H-H₂O]⁺ 195.065 1 [M+H-C₇H₁₀O₅]⁺ 177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺	Methylation of D0	Plasma, urine, feces
D1-2	14.00	[M+H]⁺	C₁₇H₂₁O₉	369.118 0	369.117 0	-2.7	351.107 0 [M+H-H₂O]⁺ 305.095 8 [M+H-CO-2H₂O]⁺ 177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺	Methylation of D0	Plasma, urine, feces
D1-3	14.33	[M+H]⁺	C₁₇H₂₁O₉	369.118 0	369.116 8	-3.3	351.107 0 [M+H-H₂O]⁺ 195.065 1 [M+H-C₇H₁₀O₅]⁺ 177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺	Methylation of D0	Plasma, urine, feces
D2	13.12	[M+H]⁺	C₁₀H₁₁O₄	195.064 5	195.063 9	-3.1	163.039 0 [M+H-CH₂-H₂O]⁺ 153.054 6 [M+H-CH₂-CO]⁺	Hydrolysis and methylation of D0	Plasma, urine, feces
D3	3.34	[M+H]⁺	C₉H₁₁O₄	183.065 2	183.064 8	-2.2	165.054 6 [M+H-H₂O]⁺ 137.059 2 [M+H-H₂O-CO]⁺	Hydrolysis and double-bond reduction of D0	Plasma, bile
D4	8.53	[M+H]⁺	C₁₅H₁₉O₇	311.112 5	311.112 0	-1.6	183.065 0 [M+H-C₆H₈O₃]⁺ 165.054 4 [M+H-C₆H₈O₃-H₂O]⁺	Decarboxylation of D0	Plasma, urine
D5	12.50	[M+H]⁺	C₁₈H₂₁O₁₀	397.112 9	397.111 9	-2.5	379.102 4 [M+H-H₂O]⁺ 353.086 7 [M+H-HCOCH₃]⁺	Acetylation of D0	Plasma
D6-1	8.30	[M+H]⁺	C₂₂H₃₁N₂O₁₂S	547.159 2	547.157 5	-3.1	369.118 0 [M+H-Cys-Gly]⁺ 351.106 9 [M+H-Cys-Gly-H₂O]⁺ 179.047 9 [M+H-C₁₇H₂₀O₉]⁺	Methylation and Cys-Gly conjugation of D0	Bile
D6-2	9.43	[M+H]⁺	C₂₂H₃₁N₂O₁₂S	547.159 2	547.157 7	-2.7	369.118 0 [M+H-Cys-Gly]⁺ 177.054 6 [M+H-Cys-Gly-C₇H₁₀O₅-H₂O]⁺	Methylation and Cys-Gly conjugation of D0	Bile
D7-1	9.36	[M+H]⁺	C₂₇H₃₈N₃O₁₅S	676.201 8	676.200 3	-2.2	369.117 8 [M+H-GSH]⁺ 308.091 0 [M+H-C₁₇H₂₀O₉]⁺ 233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺	Methylation and GSH conjugation of D0	Bile
D7-2	10.47	[M+H]⁺	C₂₇H₃₈N₃O₁₅S	676.201 8	676.200 4	-2.1	369.117 8 [M+H-GSH]⁺ 308.091 0 [M+H-C₁₇H₂₀O₉]⁺ 233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺	Methylation and GSH conjugation of D0	Bile
D8	10.85	[M+H]⁺	C₂₀H₂₈N₃O₁₀S	502.149 0	502.147 8	-2.4	308.091 1 [M+H-C₁₀H₁₀O₄]⁺ 233.058 5 [M+H-C₁₀H₁₀O₄-C₂H₅NO₂]⁺ 195.064 0 [M+H-GSH]⁺	GSH conjugation of D2	Bile

No.

t_R
/min

Ion form

Formula

m/z

Error
/ppm

MS/MS fragment ion

Identification/
reaction

Source

Theoretical

Experimental

12.28

[M+H]⁺

C₁₆H₁₉O₉

355.102 4

355.101 6

-2.3

181.049 2 [M+H-C₇H₁₀O₅]⁺
163.038 4 [M+H-C₇H₁₀O₅-H₂O]⁺
149.022 8 [M+H-C₇H₁₀O₅-CH₂-H₂O]⁺

Chlorogenic acid

Plasma, urine

D1-1

12.54

[M+H]⁺

C₁₇H₂₁O₉

369.118 0

369.116 9

-3.0

351.107 0 [M+H-H₂O]⁺
195.065 1 [M+H-C₇H₁₀O₅]⁺
177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺

Methylation of D0

Plasma, urine, feces

D1-2

14.00

[M+H]⁺

C₁₇H₂₁O₉

369.118 0

369.117 0

-2.7

351.107 0 [M+H-H₂O]⁺
305.095 8 [M+H-CO-2H₂O]⁺
177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺

Methylation of D0

Plasma, urine, feces

D1-3

14.33

[M+H]⁺

C₁₇H₂₁O₉

369.118 0

369.116 8

-3.3

351.107 0 [M+H-H₂O]⁺
195.065 1 [M+H-C₇H₁₀O₅]⁺
177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺

Methylation of D0

Plasma, urine, feces

13.12

[M+H]⁺

C₁₀H₁₁O₄

195.064 5

195.063 9

-3.1

163.039 0 [M+H-CH₂-H₂O]⁺
153.054 6 [M+H-CH₂-CO]⁺

Hydrolysis and methylation of D0

Plasma, urine, feces

3.34

[M+H]⁺

C₉H₁₁O₄

183.065 2

183.064 8

-2.2

165.054 6 [M+H-H₂O]⁺
137.059 2 [M+H-H₂O-CO]⁺

Hydrolysis and double-bond reduction of D0

Plasma, bile

8.53

[M+H]⁺

C₁₅H₁₉O₇

311.112 5

311.112 0

-1.6

183.065 0 [M+H-C₆H₈O₃]⁺
165.054 4 [M+H-C₆H₈O₃-H₂O]⁺

Decarboxylation of D0

Plasma, urine

12.50

[M+H]⁺

C₁₈H₂₁O₁₀

397.112 9

397.111 9

-2.5

379.102 4 [M+H-H₂O]⁺
353.086 7 [M+H-HCOCH₃]⁺

Acetylation of D0

Plasma

D6-1

8.30

[M+H]⁺

C₂₂H₃₁N₂O₁₂S

547.159 2

547.157 5

-3.1

369.118 0 [M+H-Cys-Gly]⁺
351.106 9 [M+H-Cys-Gly-H₂O]⁺
179.047 9 [M+H-C₁₇H₂₀O₉]⁺

Methylation and Cys-Gly conjugation of D0

Bile

D6-2

9.43

[M+H]⁺

C₂₂H₃₁N₂O₁₂S

547.159 2

547.157 7

-2.7

369.118 0 [M+H-Cys-Gly]⁺
177.054 6 [M+H-Cys-Gly-C₇H₁₀O₅-H₂O]⁺

Methylation and Cys-Gly conjugation of D0

Bile

D7-1

9.36

[M+H]⁺

C₂₇H₃₈N₃O₁₅S

676.201 8

676.200 3

-2.2

369.117 8 [M+H-GSH]⁺
308.091 0 [M+H-C₁₇H₂₀O₉]⁺
233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺

Methylation and GSH conjugation of D0

Bile

D7-2

10.47

[M+H]⁺

C₂₇H₃₈N₃O₁₅S

676.201 8

676.200 4

-2.1

369.117 8 [M+H-GSH]⁺
308.091 0 [M+H-C₁₇H₂₀O₉]⁺
233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺

Methylation and GSH conjugation of D0

Bile

10.85

[M+H]⁺

C₂₀H₂₈N₃O₁₀S

502.149 0

502.147 8

-2.4

308.091 1 [M+H-C₁₀H₁₀O₄]⁺
233.058 5 [M+H-C₁₀H₁₀O₄-C₂H₅NO₂]⁺
195.064 0 [M+H-GSH]⁺

GSH conjugation of D2

Bile

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
D0	12.28	[M+H]⁺	C₁₆H₁₉O₉	355.102 4	355.101 6	-2.3	181.049 2 [M+H-C₇H₁₀O₅]⁺ 163.038 4 [M+H-C₇H₁₀O₅-H₂O]⁺ 149.022 8 [M+H-C₇H₁₀O₅-CH₂-H₂O]⁺	Chlorogenic acid	Plasma, urine
D1-1	12.54	[M+H]⁺	C₁₇H₂₁O₉	369.118 0	369.116 9	-3.0	351.107 0 [M+H-H₂O]⁺ 195.065 1 [M+H-C₇H₁₀O₅]⁺ 177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺	Methylation of D0	Plasma, urine, feces
D1-2	14.00	[M+H]⁺	C₁₇H₂₁O₉	369.118 0	369.117 0	-2.7	351.107 0 [M+H-H₂O]⁺ 305.095 8 [M+H-CO-2H₂O]⁺ 177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺	Methylation of D0	Plasma, urine, feces
D1-3	14.33	[M+H]⁺	C₁₇H₂₁O₉	369.118 0	369.116 8	-3.3	351.107 0 [M+H-H₂O]⁺ 195.065 1 [M+H-C₇H₁₀O₅]⁺ 177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺	Methylation of D0	Plasma, urine, feces
D2	13.12	[M+H]⁺	C₁₀H₁₁O₄	195.064 5	195.063 9	-3.1	163.039 0 [M+H-CH₂-H₂O]⁺ 153.054 6 [M+H-CH₂-CO]⁺	Hydrolysis and methylation of D0	Plasma, urine, feces
D3	3.34	[M+H]⁺	C₉H₁₁O₄	183.065 2	183.064 8	-2.2	165.054 6 [M+H-H₂O]⁺ 137.059 2 [M+H-H₂O-CO]⁺	Hydrolysis and double-bond reduction of D0	Plasma, bile
D4	8.53	[M+H]⁺	C₁₅H₁₉O₇	311.112 5	311.112 0	-1.6	183.065 0 [M+H-C₆H₈O₃]⁺ 165.054 4 [M+H-C₆H₈O₃-H₂O]⁺	Decarboxylation of D0	Plasma, urine
D5	12.50	[M+H]⁺	C₁₈H₂₁O₁₀	397.112 9	397.111 9	-2.5	379.102 4 [M+H-H₂O]⁺ 353.086 7 [M+H-HCOCH₃]⁺	Acetylation of D0	Plasma
D6-1	8.30	[M+H]⁺	C₂₂H₃₁N₂O₁₂S	547.159 2	547.157 5	-3.1	369.118 0 [M+H-Cys-Gly]⁺ 351.106 9 [M+H-Cys-Gly-H₂O]⁺ 179.047 9 [M+H-C₁₇H₂₀O₉]⁺	Methylation and Cys-Gly conjugation of D0	Bile
D6-2	9.43	[M+H]⁺	C₂₂H₃₁N₂O₁₂S	547.159 2	547.157 7	-2.7	369.118 0 [M+H-Cys-Gly]⁺ 177.054 6 [M+H-Cys-Gly-C₇H₁₀O₅-H₂O]⁺	Methylation and Cys-Gly conjugation of D0	Bile
D7-1	9.36	[M+H]⁺	C₂₇H₃₈N₃O₁₅S	676.201 8	676.200 3	-2.2	369.117 8 [M+H-GSH]⁺ 308.091 0 [M+H-C₁₇H₂₀O₉]⁺ 233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺	Methylation and GSH conjugation of D0	Bile
D7-2	10.47	[M+H]⁺	C₂₇H₃₈N₃O₁₅S	676.201 8	676.200 4	-2.1	369.117 8 [M+H-GSH]⁺ 308.091 0 [M+H-C₁₇H₂₀O₉]⁺ 233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺	Methylation and GSH conjugation of D0	Bile
D8	10.85	[M+H]⁺	C₂₀H₂₈N₃O₁₀S	502.149 0	502.147 8	-2.4	308.091 1 [M+H-C₁₀H₁₀O₄]⁺ 233.058 5 [M+H-C₁₀H₁₀O₄-C₂H₅NO₂]⁺ 195.064 0 [M+H-GSH]⁺	GSH conjugation of D2	Bile

No.

t_R
/min

Ion form

Formula

m/z

Error
/ppm

MS/MS fragment ion

Identification/
reaction

Source

Theoretical

Experimental

12.28

[M+H]⁺

C₁₆H₁₉O₉

355.102 4

355.101 6

-2.3

181.049 2 [M+H-C₇H₁₀O₅]⁺
163.038 4 [M+H-C₇H₁₀O₅-H₂O]⁺
149.022 8 [M+H-C₇H₁₀O₅-CH₂-H₂O]⁺

Chlorogenic acid

Plasma, urine

D1-1

12.54

[M+H]⁺

C₁₇H₂₁O₉

369.118 0

369.116 9

-3.0

351.107 0 [M+H-H₂O]⁺
195.065 1 [M+H-C₇H₁₀O₅]⁺
177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺

Methylation of D0

Plasma, urine, feces

D1-2

14.00

[M+H]⁺

C₁₇H₂₁O₉

369.118 0

369.117 0

-2.7

351.107 0 [M+H-H₂O]⁺
305.095 8 [M+H-CO-2H₂O]⁺
177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺

Methylation of D0

Plasma, urine, feces

D1-3

14.33

[M+H]⁺

C₁₇H₂₁O₉

369.118 0

369.116 8

-3.3

351.107 0 [M+H-H₂O]⁺
195.065 1 [M+H-C₇H₁₀O₅]⁺
177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺

Methylation of D0

Plasma, urine, feces

13.12

[M+H]⁺

C₁₀H₁₁O₄

195.064 5

195.063 9

-3.1

163.039 0 [M+H-CH₂-H₂O]⁺
153.054 6 [M+H-CH₂-CO]⁺

Hydrolysis and methylation of D0

Plasma, urine, feces

3.34

[M+H]⁺

C₉H₁₁O₄

183.065 2

183.064 8

-2.2

165.054 6 [M+H-H₂O]⁺
137.059 2 [M+H-H₂O-CO]⁺

Hydrolysis and double-bond reduction of D0

Plasma, bile

8.53

[M+H]⁺

C₁₅H₁₉O₇

311.112 5

311.112 0

-1.6

183.065 0 [M+H-C₆H₈O₃]⁺
165.054 4 [M+H-C₆H₈O₃-H₂O]⁺

Decarboxylation of D0

Plasma, urine

12.50

[M+H]⁺

C₁₈H₂₁O₁₀

397.112 9

397.111 9

-2.5

379.102 4 [M+H-H₂O]⁺
353.086 7 [M+H-HCOCH₃]⁺

Acetylation of D0

Plasma

D6-1

8.30

[M+H]⁺

C₂₂H₃₁N₂O₁₂S

547.159 2

547.157 5

-3.1

369.118 0 [M+H-Cys-Gly]⁺
351.106 9 [M+H-Cys-Gly-H₂O]⁺
179.047 9 [M+H-C₁₇H₂₀O₉]⁺

Methylation and Cys-Gly conjugation of D0

Bile

D6-2

9.43

[M+H]⁺

C₂₂H₃₁N₂O₁₂S

547.159 2

547.157 7

-2.7

369.118 0 [M+H-Cys-Gly]⁺
177.054 6 [M+H-Cys-Gly-C₇H₁₀O₅-H₂O]⁺

Methylation and Cys-Gly conjugation of D0

Bile

D7-1

9.36

[M+H]⁺

C₂₇H₃₈N₃O₁₅S

676.201 8

676.200 3

-2.2

369.117 8 [M+H-GSH]⁺
308.091 0 [M+H-C₁₇H₂₀O₉]⁺
233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺

Methylation and GSH conjugation of D0

Bile

D7-2

10.47

[M+H]⁺

C₂₇H₃₈N₃O₁₅S

676.201 8

676.200 4

-2.1

369.117 8 [M+H-GSH]⁺
308.091 0 [M+H-C₁₇H₂₀O₉]⁺
233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺

Methylation and GSH conjugation of D0

Bile

10.85

[M+H]⁺

C₂₀H₂₈N₃O₁₀S

502.149 0

502.147 8

-2.4

308.091 1 [M+H-C₁₀H₁₀O₄]⁺
233.058 5 [M+H-C₁₀H₁₀O₄-C₂H₅NO₂]⁺
195.064 0 [M+H-GSH]⁺

GSH conjugation of D2

Bile

静脉给药热毒宁注射液大鼠血浆、胆汁、尿液和粪便中代谢产物的鉴定

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王春雪 ¹ , 王玲 ² , 钱大玮 ¹^,^* , 张黄琴 ³ , 黄文哲 ⁴ , 王团结 ⁴ , 朱振华 ² , 宿树兰 ³ , 郭盛 ³ , 段金廒 ²

药学学报 | 研究论文·药物分析 2018,53(7): 1148-1155

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药学学报 | 研究论文·药物分析 2018, 53(7): 1148-1155

静脉给药热毒宁注射液大鼠血浆、胆汁、尿液和粪便中代谢产物的鉴定

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王春雪¹, 王玲², 钱大玮¹^,^*, 张黄琴³, 黄文哲⁴, 王团结⁴, 朱振华², 宿树兰³, 郭盛³, 段金廒²

作者信息

1.南京中医药大学江苏省方剂高技术研究重点实验室, 江苏南京 210023

2.南京中医药大学江苏省中药资源产业化过程协同创新中心, 江苏南京 210023

3.南京中医药大学中药资源产业化与方剂创新药物国家地方联合工程研究中心, 江苏南京 210023

4.江苏康缘药业股份有限公司, 江苏连云港 222001

通讯作者:

* 钱大玮, Tel/Fax:86-25-85811916, E-mail:qiandwnj@126.com

Identification of metabolites of Reduning injection in rat plasma, bile, urine and feces after intravenous administration

Chun-xue WANG¹, Ling WANG², Da-wei QIAN¹^,^*, Huang-qin ZHANG³, Wen-zhe HUANG⁴, Tuan-jie WANG⁴, Zhen-hua ZHU², Shu-lan SU³, Sheng GUO³, Jin-ao DUAN²

Affiliations

1. Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China

2. Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing 210023, China

3. National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China

4. Kanion Pharmaceutical Co., Ltd., Lianyungang 222001, China

出版时间: 2018-07-12 doi: 10.16438/j.0513-4870.2018-0052

文章导航

摘要

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研究热毒宁注射液在大鼠体内的代谢产物。采用超高效液相色谱-线性离子阱-静电轨道阱串联质谱联用方法（UHPLC-LTQ-Orbitrap-MS）对大鼠尾静脉注射热毒宁后血浆、胆汁、尿液和粪便进行分析，利用质量亏损过滤（MDF）技术进行京尼平苷、京尼平苷酸、断氧化马钱子苷、绿原酸代谢产物分析鉴定，推测出大鼠体内京尼平苷氧化、脱水、去甲氧基、水解、开环、半胱氨酸结合和苷元葡萄糖醛酸结合等14个代谢产物；京尼平苷酸脱水、开环、双键还原和半胱氨酸结合等9个代谢产物；断氧化马钱子苷水解、去甲氧基、羟基化和乙基化等6个代谢产物；绿原酸脱羧、水解、甲基化、乙酰化、半胱氨酰甘氨酸结合和谷胱甘肽结合等12个代谢产物。为进一步探讨热毒宁注射液体内过程，阐明其功效物质基础提供依据。

关键词

热毒宁注射液 / 京尼平苷 / 京尼平苷酸 / 断氧化马钱子苷 / 绿原酸 / 代谢产物

Abstract

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This study was designed to determine the metabolites of Renduining injection in rats. The ultra-high performance liquid chromatography-LTQ Orbitrap mass spectrometric (UHPLC-LTQ-Orbitrap-MS) and mass defect filter techniques were applied to analyze the metabolites of Reduning injection in rat plasma, bile, urine and feces. As a result, we determined 14 metabolites of geniposide, including oxidation, dehydration, hydroxymethylene loss, hydrolysis, ring-opened, cysteine conjugation and glucuronidation conjugation of aglycone; 9 metabolites of geniposidic acid, consisting of dehydration, ring-opened, double-bond reduction and cysteine conjugation; 6 metabolites of secoxylogain including hydrolysis, hydroxymethylene loss, hydroxylation and ethylation; 12 metabolites of chlorogenic acid, containing decarboxylation, hydrolysis, methylation, acetylation, cysteinylglycine conjugation and glutathione conjugation. It provided information for the therapeutic effect of Reduning in vivo.

Key words

Reduning injection / geniposide / geniposidic acid / secoxylogain / chlorogenic acid / metabolite

引用本文

王春雪, 王玲, 钱大玮, 张黄琴, 黄文哲, 王团结, 朱振华, 宿树兰, 郭盛, 段金廒. 静脉给药热毒宁注射液大鼠血浆、胆汁、尿液和粪便中代谢产物的鉴定. 药学学报, 2018 , 53 (7) : 1148 -1155 . DOI: 10.16438/j.0513-4870.2018-0052

Chun-xue WANG, Ling WANG, Da-wei QIAN, Huang-qin ZHANG, Wen-zhe HUANG, Tuan-jie WANG, Zhen-hua ZHU, Shu-lan SU, Sheng GUO, Jin-ao DUAN. Identification of metabolites of Reduning injection in rat plasma, bile, urine and feces after intravenous administration[J]. Acta Pharmaceutica Sinica, 2018 , 53 (7) : 1148 -1155 . DOI: 10.16438/j.0513-4870.2018-0052

正文

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热毒宁注射液由青蒿、栀子和金银花3味中药组合而成, 主要用于外感风热所致流感、高热不退、咳嗽及上呼吸道感染等症, 临床上退热作用迅速、抗病毒效果显著^[1]。研究表明, 其主要有效成分为京尼平苷、京尼平苷酸、断氧化马钱子苷和绿原酸等^{[2, 3]}。虽有京尼平苷、京尼平苷酸及绿原酸体内代谢的文献报道^{[4, 5]}, 但这些报道仅限于对单味药提取物或单体成分的代谢研究, 而热毒宁注射液在体内代谢产物研究尚未见报道。

药物化学结构的多样性和体内生物转化途径的复杂性使得药物代谢产物鉴定研究具有挑战性, 且由于生物样品中代谢产物含量低, 将样品收集再经色谱等方法分离得纯品后鉴定结构的方法, 既费时又费力。LC-MS联用技术由于兼有优秀的色谱分离能力和高灵敏度、高专属的质谱检测能力, 近年来在药物体内过程研究中的应用日益广泛, 其中高分辨质谱的应用使得代谢产物结构推测的可靠性得到极大的提高, 已成为药物代谢产物鉴定的主要方法之一^{[6, 7]}。而高分辨率的线性离子阱-静电轨道阱串联质谱(LTQ-Orbitrap-MS)在线性离子阱灵敏、快速的基础上增加了轨道阱技术的高分辨率、高质量准确度的特点, 可短时间内同时实现母、子离子的高分辨采集和多级质谱碎裂, 更适用于中药复杂物质体系中代谢产物的快速鉴定^[8]。本文采用UHPLC串联LTQ-Orbitrap-MS技术研究大鼠注射热毒宁注射液后主要化学成分在其体内代谢产物, 共鉴定出36个代谢物, 为进一步探讨热毒宁注射液的体内过程, 阐明其功效物质基础提供依据。

材料与方法

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仪器与试药 UltiMate 3000型超高效液相色谱、LTQ-Orbitrap Velos Pro质谱仪(美国Thermo公司); Xcalibur 3.0质谱工作站软件、Metworks 1.3及Mass Frontier 7.0分析软件(美国Thermo公司); DZ 30-32高速离心机(上海安亭科学仪器厂); WH-微型涡旋混合仪(上海沪西分析仪器厂有限公司); 电子天平(BT125, 赛多利斯科学仪器有限公司)。

京尼平苷(批号160729)、京尼平(批号160918)、京尼平苷酸(批号160916)、断氧化马钱子苷(批号160720)、绿原酸(批号160903)均购自上海融禾医药科技有限公司, 纯度均大于98%。热毒宁注射液(批号161010, 10 mL/支)由江苏康缘药业股份有限公司提供, 其中京尼平苷、京尼平苷酸、断氧化马钱子苷、绿原酸含量分别为9.95、0.830、1.13和6.33 mg·mL^-1。乙腈、甲酸(色谱级, 德国Merck公司)。

实验动物 清洁级雄性SD大鼠, 约8周龄, 体重(250 ± 10) g, 由南京市江宁区青龙山动物繁殖场提供, 合格证号SCXK (苏) 2017-0001。

UHPLC检测条件 色谱柱: Acquity UPLC BEH T₃ (100 mm × 2.1 mm, 1.7 μm)柱; 流动相: 0.1%甲酸(A)-乙腈(B), 洗脱梯度(0~3 min, 1% B; 3~9 min, 1%~10% B; 9~12 min, 10%~20% B; 12~13 min, 20%~25% B; 13~15 min, 25% B; 15~18 min, 25%~40% B; 18~20 min, 40%~99% B); 柱温30 ℃; 流速0.4 mL·min^-1, 进样量2 μL。

MS检测条件 电喷雾离子源(ESI), 采用正离子检测模式, m/z采集范围为110~1 000, 采集时间为0.8~20.8 min。主要质谱参数:毛细管温度350 ℃, 蒸发器温度350 ℃, 鞘气流40 kPa, 辅助气流15 kPa, 喷雾电压3.5 V, 源电流100 μA; MS²质谱的触发方式为动态数据依赖分析模式, 选取上一级最高峰进行碰撞诱导碎裂解离碎片扫描(CID), 碰撞能量设定为30%, 以离子阱打拿极检测。

对照品溶液的制备 分别精密称取京尼平苷、京尼平、京尼平苷酸、断氧化马钱子苷、绿原酸10 mg, 甲醇溶解并定容至10 mL得1 mg·mL^-1的储备液。

动物分组及给药 SD大鼠15只, 随机分为空白组、血浆组、胆汁组、尿液/粪便组, 空白组6只, 其余每组3只。根据临床剂量, 热毒宁注射液给药剂量为2 mL·kg^-1。分别连续3天给予热毒宁注射液及空白组的等体积生理盐水。

生物样品的采集与供试液的制备 大鼠第3天给药前禁食不禁水12 h, 取血浆组于30 min眼眶取血置于预先涂有1%肝素钠生理盐水的EP管中, 离心10 min (4 000 r·min^-1), 取上清得血浆样品; 取胆汁组10%水合氯醛麻醉, 胆管插入并固定后即刻给大鼠注射给药, 注射给药后开始收集0~30 min胆汁样品; 取尿液/粪便组分别置于代谢笼, 收集0~12 h的尿液和粪便。以上样品置于-20 ℃保存。

大鼠血浆样品供试液的制备 取血浆样品200 μL混合均匀后, 加入3倍乙腈沉淀蛋白, 涡旋混匀30 s, 离心10 min (12 000 r·min^-1), 取上清液于37 ℃氮气吹干, 以5%乙腈复溶, 离心5 min (12 000 r·min^-1), 取上清液为血浆样品供试液。

大鼠胆汁、尿液样品供试液的制备 取尿液或胆汁1 mL, 加等体积乙腈, 涡旋混匀30 s, 离心10 min (12 000 r·min^-1), 取上清液作为胆汁、尿液样品供试液。

大鼠粪便样品供试液的制备 取碾碎混匀的粪便1 g, 加蒸馏水3 mL溶解, 超声10 min, 取出后离心10 min (12 000 r·min^-1), 取上清液, 加入等体积乙腈, 涡旋混匀30 s, 离心5 min (12 000 r·min^-1), 取上清液作为粪便样品供试液。

数据处理 利用Metworks 1.3分析软件对空白和给药组的一级质谱进行自动比对, 结合待测化合物元素组成, 在误差小于5 ppm的条件下, 寻找符合代谢修饰的代谢产物准分子离子峰, 结合二级质谱分析代谢物结构特征, 并通过Mass Frontier 7.0谱图解析软件作进一步验证。

结果

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1 血浆、胆汁、尿液、粪便UHPLC-LTQ-Orbitrap-MS分析

给药血浆、胆汁、尿液、粪便样品正离子条件下原形化合物及其代谢产物总提取离子流图如图 1。

2 代谢产物分析

2.1 京尼平苷(A)代谢产物

大鼠体内共鉴定出14个京尼平苷代谢产物, 主要鉴定信息见表 1。

A0为京尼平苷: m/z 411.124 8为其加钠分子离子峰, m/z 249.072 8为糖苷键断裂的碎片, m/z 231.062 1由其进一步脱水形成, 与对照品相符。A1为京尼平苷苷元与葡萄糖醛酸结合产物: m/z 425.107 5为其加钠分子离子峰, m/z 199.021 7为葡萄糖醛酸正离子, 其余碎片与京尼平苷相同。A2为A1开环产物: m/z 427.120 7为其加钠分子离子峰, m/z 233.078 9为脱葡萄糖和水的碎片。A3-1和A3-2皆为A2去甲酸甲酯和羟基产物: m/z 345.153 7为其加氢分子离子峰, m/z 169.121 8为糖苷键断裂的碎片, m/z 151.111 3由其进一步脱水形成。A4-1和A4-2为半胱氨酸与京尼平苷不同碳位结合的同分异构产物^[9]: m/z 492.152 0为其加氢分子离子峰, m/z 330.100 4为糖苷键断裂的碎片, m/z 312.090 0由其进一步脱水形成。A5为京尼平苷去甲氧基产物: m/z 381.114 4为其加钠分子离子峰, m/z 219.065 0为糖苷键断裂的碎片, m/z 201.052 1由其进一步脱水生成。

A6为京尼平: m/z 227.090 9为其加氢分子离子峰, m/z 213.076 2为酯键断裂碎片, m/z 209.080 5为其脱水的碎片, m/z 191.070 3由其进一步脱水产生, 与对照品相符。A7为A6酯键水解产物: m/z 213.075 1为其加氢分子离子峰, m/z 195.065 3为其脱水的碎片, m/z 185.080 8为脱羰基碎片, m/z 151.075 2为其脱水并脱羧碎片。A8为A6氧化脱氢产物: m/z 225.075 2为其加氢分子离子峰, m/z 207.065 1为其脱水的碎片, m/z 195.059 4为脱甲氧基的碎片, m/z 151.075 5由进一步脱羧产生。A9-1和A9-2皆为A6不同位羟基脱水的同分异构产物: m/z 209.080 4为其加氢分子离子峰, m/z 191.070 5为其脱水的碎片, m/z 177.054 3由其去甲基后脱水产生。A10为A6脱甲氧基产物: m/z 197.080 2为其加氢分子离子峰, m/z 179.070 3为脱水的碎片, m/z 169.085 8为其脱羰基的碎片, m/z 151.075 3由其进一步脱水产生。A11为A7开环产物, m/z 215.090 7为其加氢分子离子峰, m/z 197.080 8为脱水的碎片, m/z 169.085 9为脱羰基的碎片。综上, 京尼平苷体内代谢途径见图 2a。

2.2 京尼平苷酸(B)代谢产物

大鼠体内共鉴定出9个京尼平苷酸代谢产物, 主要鉴定信息见表 2。

B0为京尼平苷酸: m/z 397.109 0为其加钠分子离子峰, m/z 235.052 6为糖苷键断裂的碎片, m/z 217.047 1由进一步脱水产生, 与对照品相符。B1为半胱氨酸结合产物: m/z 478.136 1为其加氢分子离子峰, m/z 316.076 4为糖苷键断裂的碎片, m/z 298.074 4和m/z 280.063 5由进一步连续脱水产生。B2为羟基化再乙基化产物: m/z 421.168 8为其加氢分子离子峰, m/z 375.125 6为脱乙基和水的碎片, m/z 213.075 7由其进一步脱葡萄糖产生。B3为双键还原产物: m/z 377.144 0为其加氢分子离子峰, m/z 331.138 5为脱羧基的碎片, m/z 215.091 4为糖苷键断裂的碎片, m/z 197.080 8由进一步脱水产生。B4为京尼平苷酸开环后去甲酸甲酯产物: m/z 331.174 1为其加氢分子离子峰, m/z 313.152 6为脱水的碎片, m/z 169.122 3为糖苷键断裂的碎片。B5为B0糖苷键水解产物, B6为B4脱糖与葡萄糖醛酸结合产物, B7-1和B7-2为B0脱羟基产物, B8则为B5开环产物, 产物B5、B6、B7-1、B7-2、B8与京尼平苷产物A7、A5、A3-1、A3-2、A11相同, 仅从结构无法判断基源, 其二级碎片详见表 1。综上, 京尼平苷酸体内代谢途径如图 2b。

2.3 断氧化马钱子苷(C)代谢产物

大鼠体内共鉴定出6个断氧化马钱子苷代谢产物, 主要鉴定信息见表 3。

C0为断氧化马钱子苷: m/z 405.138 6为其加氢分子离子峰, m/z 391.119 2为酯键断裂的碎片, m/z 373.121 6由其进一步脱水产生, m/z 243.085 4为糖苷键断裂的碎片, 与对照品相符。C1为断氧化马钱子苷苷元: m/z 243.085 5为其加氢分子离子峰, m/z 225.076 7为脱水的碎片, m/z 207.065 2由其进一步脱水产生。C2为C1酯键水解产物: m/z 229.069 9为其加氢分子离子峰, m/z 211.060 1为脱水的碎片, m/z 201.074 8为脱羰基的碎片。C3-1和C3-2同为不同碳位羟基化再乙基化的同分异构产物: m/z 451.182 3为加氢分子离子峰, m/z 243.086 0为其糖苷断裂后脱水脱羰基的碎片, m/z 225.112 3由其进一步脱水产生。C4为断氧化马钱子苷酯键水解产物: m/z 391.124 2为其加氢分子离子峰, m/z 373.112 6为其脱水的碎片。C5为去甲氧基产物: m/z 375.127 5为其加氢分子离子峰, m/z 357.113 8为脱水的碎片, m/z 195.065 2由进一步脱葡萄糖产生, m/z 347.133 1为脱羰基的碎片。综上, 断氧化马钱子苷的体内代谢途径如图 2c。

2.4 绿原酸(D)代谢产物

大鼠体内共鉴定出12个绿原酸代谢产物, 主要鉴定信息见表 4。

D0为绿原酸: m/z 355.101 6为加氢分子离子峰, m/z 181.049 2为酯键断裂的咖啡酸正离子, m/z 163.038 4由其进一步脱水产生, m/z 149.022 8为再去甲基产生, 与对照品相符。D1-1、D1-2和D1-3同为绿原酸不同位羟基甲基化的同分异构产物: m/z 369.116 9为其加氢分子离子峰, m/z 351.107 0为脱水的碎片, m/z 195.065 1为甲基化咖啡酸正离子, m/z 177.054 2由其进一步脱水产生。D2为甲基化咖啡酸: m/z 195.063 9为其加氢分子离子峰, m/z 163.039 0为其脱甲基和水的碎片, m/z 153.054 6为其脱甲基和脱羰基的碎片。D3为二氢咖啡酸: m/z 183.064 8为其加氢分子离子峰, m/z 165.054 6为脱水的碎片, m/z 137.059 2由进一步脱羰基产生。D4为绿原酸脱羧产物: m/z 311.112 0为其加氢分子离子峰, m/z 183.065 0为二氢咖啡酸正离子, m/z 165.054 4由进一步脱水产生。D5为绿原酸乙酰化产物: m/z 397.111 9为其加氢离子峰, m/z 379.102 4为其脱水碎片, m/z 353.086 7为丢失乙酰基的碎片。

D6-1和D6-2皆为绿原酸不同位羟基甲基化的同分异构体与半胱氨酰甘氨酸结合的产物: m/z 547.157 5为其加氢分子离子峰, m/z 369.118 0为甲基化绿原酸正离子, m/z 351.106 9由其进一步脱水产生, m/z 179.047 9为半胱氨酰甘氨酸正离子。D7-1和D7-2同为绿原酸不同位羟基甲基化的同分异构体与谷胱甘肽结合的产物^[10]: m/z676.200 3为其加氢分子离子峰, m/z 369.117 8为甲基化绿原酸正离子, m/z 308.091 0为谷胱甘肽正离子, m/z 233.058 9由其进一步脱甘氨酸产生。D8为甲基化咖啡酸与谷胱甘肽结合产物: m/z 502.147 8为其加氢分子离子峰, m/z 195.064 0为甲基化咖啡酸正离子, 其余碎片与D7相同。综上, 绿原酸体内代谢途径如图 2d。

讨论

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栀子药材中主要活性成分为环烯醚萜类, 代表性成分为含量达3.06%~4.12%的京尼平苷, 而其苷元京尼平的含量却只有0.005%~0.01%^[11]。本文的研究结果显示, 热毒宁注射液中京尼平的含量低于检测限, 但在大鼠注射给药热毒宁注射液后的血浆与尿液中却检测到京尼平, 由此可确定此注射液中京尼平苷代谢为其苷元京尼平的这一代谢途径, 并可进行共存成分对这一代谢的影响研究。有文献分别研究京尼平苷在人体和大鼠中的代谢途径, 发现羟基化是人类特有的代谢途径^{[12, 13]}, 在本研究同样未在大鼠体内检测到京尼平苷羟基化产物。

大鼠静脉注射绿原酸后, 胆汁中存在大量其与氨基酸、谷胱甘肽结合的产物, 这是由于绿原酸的烯酮双键具有很强的亲电性, 可与谷胱甘肽结合^{[14, 15]}。热毒宁注射液中的绿原酸在静脉注射给药后亦有如此的代谢途径。但口服给药却未检测到其与氨基酸、谷胱甘肽结合产物的报道, 推测其原因可能是绿原酸口服给药后, 由于肠道菌的代谢作用^[16], 使其原形进入体内的实际量减少, 以致代谢产物量偏少而无法检测出。

本文研究结果显示, 热毒宁注射液经静脉给药后, 其主要成分在大鼠体内发生以下代谢反应: Ⅰ相代谢主要包括氧化、水解和脱羟基等; Ⅱ相代谢可直接共价结合; 亦可在Ⅰ相代谢基础上进一步生成甲基化、乙酰化、葡萄糖醛酸化、氨基酸或谷胱甘肽结合物等。

基金

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基于药物相互作用研究热毒宁注射液联合用药(FJGJS-2015-14)
热毒宁注射液标准化建设(ZYBZH-C-JS-31)
江苏高校优势学科建设工程资助项目(PAPD)

参考文献

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文献

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参考文献引证文献

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2018年第53卷第7期

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文章信息

doi: 10.16438/j.0513-4870.2018-0052

接收时间：2018-01-13
首发时间：2026-01-15
出版时间：2018-07-12

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出版历史

收稿日期：2018-01-13
修回日期：2018-03-16

基金

基于药物相互作用研究热毒宁注射液联合用药(FJGJS-2015-14)

热毒宁注射液标准化建设(ZYBZH-C-JS-31)

江苏高校优势学科建设工程资助项目(PAPD)

作者信息

1.南京中医药大学江苏省方剂高技术研究重点实验室, 江苏南京 210023

2.南京中医药大学江苏省中药资源产业化过程协同创新中心, 江苏南京 210023

3.南京中医药大学中药资源产业化与方剂创新药物国家地方联合工程研究中心, 江苏南京 210023

4.江苏康缘药业股份有限公司, 江苏连云港 222001

通讯作者:

* 钱大玮, Tel/Fax:86-25-85811916, E-mail:qiandwnj@126.com

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
A0	13.14	[M+Na]⁺	C₁₇H₂₄O₁₀Na	411.126 1	411.124 8	-3.2	249.072 8 [M+H-C₆H₁₀O₅]⁺ 231.062 1 [M+Na-C₆H₁₀O₅-H₂O]⁺	Geniposide, parent	Plasma, bile, urine
A1	9.03	[M+Na]⁺	C₁₇H₂₂O₁₁Na	425.105 4	425.107 5	4.9	249.072 8 [M+H-C₆H₈O₆]⁺ 231.062 5 [M+Na-C₆H₈O₆-H₂O]⁺ 199.021 7 [GlucA-H₂O+Na]⁺	Glucuronidation of A0	Plasma, bile, urine
A2	13.97	[M+Na]⁺	C₁₇H₂₄O₁₁Na	427.121 1	427.120 7	-0.9	233.078 9 [M+Na-C₆H₈O₆-H₂O]⁺	Ring-opened of A1	Plasma
A3-1	13.71	[M+H]⁺	C₁₆H₂₅O₈	345.154 4	345.153 7	-2.0	169.121 8 [M+H-C₆H₈O₆]⁺ 151.111 3 [M+H-C₆H₈O₆-H₂O]⁺	Cleavage and dihydroxylation of A2	Bile, feces
A3-2	14.01	[M+H]⁺	C₁₆H₂₅O₈	345.154 4	345.153 6	-2.3	169.121 8 [M+H-C₆H₈O₆]⁺ 151.111 3 [M+H-C₆H₈O₆-H₂O]⁺	Cleavage and dihydroxylation of A2	Bile, feces
A4-1	9.57	[M+H]⁺	C₂₀H₃₀NO₁₁S	492.153 4	492.152 0	-2.8	330.100 4 [M+H-C₆H₁₀O₅]⁺ 312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺	Cys conjugation of A0	Plasma, bile, urine
A4-2	13.35	[M+H]⁺	C₂₀H₃₀NO₁₁S	492.153 4	492.152 3	-2.2	330.100 4 [M+H-C₆H₁₀O₅]⁺ 312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺	Cys conjugation of A0	Plasma, bile, urine
A5	13.50	[M+Na]⁺	C₁₆H₂₂O₉Na	381.115 6	381.114 4	-3.1	219.065 0 [M+Na-C₆H₁₀O₅]⁺ 201.052 1 [M+Na-C₆H₁₀O₅-H₂O]⁺	Hydroxymethylene loss of A0	Plasma, bile
A6	10.50	[M+H]⁺	C₁₁H₁₅O₅	227.091 4	227.090 9	-2.2	213.076 2 [M+H-CH₂]⁺ 209.080 5 [M+H-H₂O]⁺ 191.070 3 [M+H-2H₂O]⁺	Genipin	Plasma, urine
A7	9.13	[M+H]⁺	C₁₀H₁₃O₅	213.075 8	213.075 1	-3.3	195.065 3 [M+H-H₂O]⁺ 185.080 8 [M+H-CO]⁺ 151.075 2 [M+H-H₂O-CO₂]⁺	Hydrolysis of A6	Plasma, urine, feces
A8	13.95	[M+H]⁺	C₁₁H₁₃O₅	225.075 8	225.075 2	-2.7	207.065 1 [M+H-H₂O]⁺ 195.059 4 [M+H-OCH₂]⁺ 151.075 5 [M+H-OCH₂-CO₂]⁺	Oxidation of A6	Plasma, bile, feces
A9-1	12.35	[M+H]⁺	C₁₁H₁₃O₄	209.080 8	209.080 4	-1.9	191.070 5 [M+H-H₂O]⁺ 177.054 3 [M+H-CH₂-H₂O]⁺	Dehydration of A6	Plasma, bile, urine, feces
A9-2	13.09	[M+H]⁺	C₁₁H₁₃O₄	209.080 8	209.080 2	-2.9	181.085 8 [M+H-CO]⁺ 177.054 5 [M+H-CH₂-H₂O]⁺	Dehydration of A6	Plasma, bile, urine, feces
A10	11.94	[M+H]⁺	C₁₀H₁₃O₄	197.080 8	197.080 2	-3.0	179.070 3 [M+H-H₂O]⁺ 169.085 8 [M+H-CO]⁺ 151.075 3 [M+H-CO-H₂O]⁺	Hydroxymethylene loss of A6	Plasma, bile
A11	2.11	[M+H]⁺	C₁₀H₁₅O₅	215.091 4	215.090 7	-3.3	197.080 8 [M+H-H₂O]⁺ 169.085 9 [M+H-H₂O-CO]⁺	Ring-opened of A7	Plasma, urine

No.

t_R /min

Ion form

Formula

m/z

Error /ppm

MS/MS fragment ion

Identification/ reaction

Source

Theoretical

Experimental

13.14

[M+Na]⁺

C₁₇H₂₄O₁₀Na

411.126 1

411.124 8

-3.2

249.072 8 [M+H-C₆H₁₀O₅]⁺
231.062 1 [M+Na-C₆H₁₀O₅-H₂O]⁺

Geniposide, parent

Plasma, bile, urine

9.03

[M+Na]⁺

C₁₇H₂₂O₁₁Na

425.105 4

425.107 5

4.9

249.072 8 [M+H-C₆H₈O₆]⁺
231.062 5 [M+Na-C₆H₈O₆-H₂O]⁺
199.021 7 [GlucA-H₂O+Na]⁺

Glucuronidation of A0

Plasma, bile, urine

13.97

[M+Na]⁺

C₁₇H₂₄O₁₁Na

427.121 1

427.120 7

-0.9

233.078 9 [M+Na-C₆H₈O₆-H₂O]⁺

Ring-opened of A1

Plasma

A3-1

13.71

[M+H]⁺

C₁₆H₂₅O₈

345.154 4

345.153 7

-2.0

169.121 8 [M+H-C₆H₈O₆]⁺
151.111 3 [M+H-C₆H₈O₆-H₂O]⁺

Cleavage and dihydroxylation of A2

Bile, feces

A3-2

14.01

[M+H]⁺

C₁₆H₂₅O₈

345.154 4

345.153 6

-2.3

169.121 8 [M+H-C₆H₈O₆]⁺
151.111 3 [M+H-C₆H₈O₆-H₂O]⁺

Cleavage and dihydroxylation of A2

Bile, feces

A4-1

9.57

[M+H]⁺

C₂₀H₃₀NO₁₁S

492.153 4

492.152 0

-2.8

330.100 4 [M+H-C₆H₁₀O₅]⁺
312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺

Cys conjugation of A0

Plasma, bile, urine

A4-2

13.35

[M+H]⁺

C₂₀H₃₀NO₁₁S

492.153 4

492.152 3

-2.2

330.100 4 [M+H-C₆H₁₀O₅]⁺
312.090 0 [M+H-C₆H₁₀O₅-H₂O]⁺

Cys conjugation of A0

Plasma, bile, urine

13.50

[M+Na]⁺

C₁₆H₂₂O₉Na

381.115 6

381.114 4

-3.1

219.065 0 [M+Na-C₆H₁₀O₅]⁺
201.052 1 [M+Na-C₆H₁₀O₅-H₂O]⁺

Hydroxymethylene loss of A0

Plasma, bile

10.50

[M+H]⁺

C₁₁H₁₅O₅

227.091 4

227.090 9

-2.2

213.076 2 [M+H-CH₂]⁺
209.080 5 [M+H-H₂O]⁺
191.070 3 [M+H-2H₂O]⁺

Genipin

Plasma, urine

9.13

[M+H]⁺

C₁₀H₁₃O₅

213.075 8

213.075 1

-3.3

195.065 3 [M+H-H₂O]⁺
185.080 8 [M+H-CO]⁺
151.075 2 [M+H-H₂O-CO₂]⁺

Hydrolysis of A6

Plasma, urine, feces

13.95

[M+H]⁺

C₁₁H₁₃O₅

225.075 8

225.075 2

-2.7

207.065 1 [M+H-H₂O]⁺
195.059 4 [M+H-OCH₂]⁺
151.075 5 [M+H-OCH₂-CO₂]⁺

Oxidation of A6

Plasma, bile, feces

A9-1

12.35

[M+H]⁺

C₁₁H₁₃O₄

209.080 8

209.080 4

-1.9

191.070 5 [M+H-H₂O]⁺
177.054 3 [M+H-CH₂-H₂O]⁺

Dehydration of A6

Plasma, bile, urine, feces

A9-2

13.09

[M+H]⁺

C₁₁H₁₃O₄

209.080 8

209.080 2

-2.9

181.085 8 [M+H-CO]⁺
177.054 5 [M+H-CH₂-H₂O]⁺

Dehydration of A6

Plasma, bile, urine, feces

A10

11.94

[M+H]⁺

C₁₀H₁₃O₄

197.080 8

197.080 2

-3.0

179.070 3 [M+H-H₂O]⁺
169.085 8 [M+H-CO]⁺
151.075 3 [M+H-CO-H₂O]⁺

Hydroxymethylene loss of A6

Plasma, bile

A11

2.11

[M+H]⁺

C₁₀H₁₅O₅

215.091 4

215.090 7

-3.3

197.080 8 [M+H-H₂O]⁺
169.085 9 [M+H-H₂O-CO]⁺

Ring-opened of A7

Plasma, urine

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
B0	9.38	[M+Na]⁺	C₁₆H₂₂O₁₀Na	397.110 5	397.109 0	-3.8	235.052 6 [M+Na-C₆H₁₀O₅]⁺ 217.047 1 [M+Na-C₆H₁₀O₅-H₂O]⁺	Geniposidic acid, parent	Plasma, bile, urine
B1	10.53	[M+H]⁺	C₁₉H₂₈NO₁₁S	478.137 8	478.136 1	-3.6	316.076 4 [M+H-C₆H₁₀O₅]⁺ 298.074 4 [M+H-C₆H₁₀O₅-H₂O]⁺ 280.063 5 [M+H-C₆H₁₀O₅-2H₂O]⁺	Cys conjugation of B0	Plasma, urine
B2	15.48	[M+H]⁺	C₁₈H₂₉O₁₁	421.170 4	421.168 8	-3.8	375.125 6 [M+H-C₂H₄-H₂O]⁺ 213.075 7 [M+H-C₆H₁₀O₅-C₂H₄-H₂O]⁺	Hydroxylation and ethylation of B0	Plasma
B3	10.36	[M+H]⁺	C₁₆H₂₅O₁₀	377.144 2	377.144 0	-0.5	331.138 5 [M+H-CO₂]⁺ 215.091 4 [M+H-C₆H₁₀O₅]⁺ 197.080 8 [M+H-C₆H₁₀O₅-H₂O]⁺	Double-bond reduction of B0	Plasma, bile, urine, feces
B4	13.84	[M+H]⁺	C₁₆H₂₇O₇	331.175 1	331.174 1	-3.0	313.152 6 [M+H-H₂O]⁺ 169.122 3 [M+H-C₆H₁₀O₅]⁺	Ring-opened and cleavage of B0	Plasma

No.

t_R
/min

Ion form

Formula

m/z

Error
/ppm

MS/MS fragment ion

Identification/
reaction

Source

Theoretical

Experimental

9.38

[M+Na]⁺

C₁₆H₂₂O₁₀Na

397.110 5

397.109 0

-3.8

235.052 6 [M+Na-C₆H₁₀O₅]⁺
217.047 1 [M+Na-C₆H₁₀O₅-H₂O]⁺

Geniposidic acid, parent

Plasma, bile, urine

10.53

[M+H]⁺

C₁₉H₂₈NO₁₁S

478.137 8

478.136 1

-3.6

316.076 4 [M+H-C₆H₁₀O₅]⁺
298.074 4 [M+H-C₆H₁₀O₅-H₂O]⁺
280.063 5 [M+H-C₆H₁₀O₅-2H₂O]⁺

Cys conjugation of B0

Plasma, urine

15.48

[M+H]⁺

C₁₈H₂₉O₁₁

421.170 4

421.168 8

-3.8

375.125 6 [M+H-C₂H₄-H₂O]⁺
213.075 7 [M+H-C₆H₁₀O₅-C₂H₄-H₂O]⁺

Hydroxylation and ethylation of B0

Plasma

10.36

[M+H]⁺

C₁₆H₂₅O₁₀

377.144 2

377.144 0

-0.5

331.138 5 [M+H-CO₂]⁺
215.091 4 [M+H-C₆H₁₀O₅]⁺
197.080 8 [M+H-C₆H₁₀O₅-H₂O]⁺

Double-bond reduction of B0

Plasma, bile, urine, feces

13.84

[M+H]⁺

C₁₆H₂₇O₇

331.175 1

331.174 1

-3.0

313.152 6 [M+H-H₂O]⁺
169.122 3 [M+H-C₆H₁₀O₅]⁺

Ring-opened and cleavage of B0

Plasma

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
C0	13.98	[M+H]⁺	C₁₇H₂₅O₁₁	405.139 1	405.138 6	-1.2	391.119 2 [M+H-CH₂]⁺ 373.121 6 [M+H-CH₂-H₂O]⁺ 243.085 4 [M+H-C₆H₁₀O₅]⁺	Secoxylogain, parent	Plasma, bile, urine
C1	10.34	[M+H]⁺	C₁₁H₁₅O₆	243.086 3	243.085 5	-3.3	225.076 7 [M+H-H₂O]⁺ 207.065 2 [M+H-2H₂O]⁺	Glucoside removal of C0	Plasma, urine
C2	12.26	[M+H]⁺	C₁₀H₁₃O₆	229.070 7	229.069 9	-3.5	211.060 1 [M+H-H₂O]⁺ 201.074 8 [M+H-CO]⁺	Hydrolysis of C1	Plasma, bile
C3-1	11.51	[M+H]⁺	C₁₉H₃₁O₁₂	451.181 0	451.182 3	2.9	243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺ 225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺	Hydroxylation and ethylation of C0	Bile
C3-2	14.11	[M+H]⁺	C₁₉H₃₁O₁₂	451.181 0	451.181 3	0.7	243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺ 225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺	Hydroxylation and ethylation of C0	Bile
C4	12.18	[M+H]⁺	C₁₆H₂₃O₁₁	391.123 5	391.124 2	1.8	373.112 6 [M+H-H₂O]⁺	Hydrolysis of C0	Plasma, bile, urine
C5	12.50	[M+H]⁺	C₁₆H₂₃O₁₀	375.128 6	375.127 5	-2.9	357.113 8 [M+H-H₂O]⁺ 347.133 1 [M+H-CO]⁺ 195.065 2 [M+H-C₆H₁₀O₅-H₂O]⁺	Hydroxymethylene loss of C0	Plasma, bile, urine

No.

t_R
/min

Ion form

Formula

m/z

Error
/ppm

MS/MS fragment ion

Identification/
reaction

Source

Theoretical

Experimental

13.98

[M+H]⁺

C₁₇H₂₅O₁₁

405.139 1

405.138 6

-1.2

391.119 2 [M+H-CH₂]⁺
373.121 6 [M+H-CH₂-H₂O]⁺
243.085 4 [M+H-C₆H₁₀O₅]⁺

Secoxylogain, parent

Plasma, bile, urine

10.34

[M+H]⁺

C₁₁H₁₅O₆

243.086 3

243.085 5

-3.3

225.076 7 [M+H-H₂O]⁺
207.065 2 [M+H-2H₂O]⁺

Glucoside removal of C0

Plasma, urine

12.26

[M+H]⁺

C₁₀H₁₃O₆

229.070 7

229.069 9

-3.5

211.060 1 [M+H-H₂O]⁺
201.074 8 [M+H-CO]⁺

Hydrolysis of C1

Plasma, bile

C3-1

11.51

[M+H]⁺

C₁₉H₃₁O₁₂

451.181 0

451.182 3

2.9

243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺
225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺

Hydroxylation and ethylation of C0

Bile

C3-2

14.11

[M+H]⁺

C₁₉H₃₁O₁₂

451.181 0

451.181 3

0.7

243.086 0 [M+H-C₆H₁₀O₅-H₂O-CO]⁺
225.112 3 [M+H-C₆H₁₀O₅-2H₂O-CO]⁺

Hydroxylation and ethylation of C0

Bile

12.18

[M+H]⁺

C₁₆H₂₃O₁₁

391.123 5

391.124 2

1.8

373.112 6 [M+H-H₂O]⁺

Hydrolysis of C0

Plasma, bile, urine

12.50

[M+H]⁺

C₁₆H₂₃O₁₀

375.128 6

375.127 5

-2.9

357.113 8 [M+H-H₂O]⁺
347.133 1 [M+H-CO]⁺
195.065 2 [M+H-C₆H₁₀O₅-H₂O]⁺

Hydroxymethylene loss of C0

Plasma, bile, urine

No.	t_R /min	Ion form	Formula	m/z	Error /ppm	MS/MS fragment ion	Identification/ reaction	Source
D0	12.28	[M+H]⁺	C₁₆H₁₉O₉	355.102 4	355.101 6	-2.3	181.049 2 [M+H-C₇H₁₀O₅]⁺ 163.038 4 [M+H-C₇H₁₀O₅-H₂O]⁺ 149.022 8 [M+H-C₇H₁₀O₅-CH₂-H₂O]⁺	Chlorogenic acid	Plasma, urine
D1-1	12.54	[M+H]⁺	C₁₇H₂₁O₉	369.118 0	369.116 9	-3.0	351.107 0 [M+H-H₂O]⁺ 195.065 1 [M+H-C₇H₁₀O₅]⁺ 177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺	Methylation of D0	Plasma, urine, feces
D1-2	14.00	[M+H]⁺	C₁₇H₂₁O₉	369.118 0	369.117 0	-2.7	351.107 0 [M+H-H₂O]⁺ 305.095 8 [M+H-CO-2H₂O]⁺ 177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺	Methylation of D0	Plasma, urine, feces
D1-3	14.33	[M+H]⁺	C₁₇H₂₁O₉	369.118 0	369.116 8	-3.3	351.107 0 [M+H-H₂O]⁺ 195.065 1 [M+H-C₇H₁₀O₅]⁺ 177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺	Methylation of D0	Plasma, urine, feces
D2	13.12	[M+H]⁺	C₁₀H₁₁O₄	195.064 5	195.063 9	-3.1	163.039 0 [M+H-CH₂-H₂O]⁺ 153.054 6 [M+H-CH₂-CO]⁺	Hydrolysis and methylation of D0	Plasma, urine, feces
D3	3.34	[M+H]⁺	C₉H₁₁O₄	183.065 2	183.064 8	-2.2	165.054 6 [M+H-H₂O]⁺ 137.059 2 [M+H-H₂O-CO]⁺	Hydrolysis and double-bond reduction of D0	Plasma, bile
D4	8.53	[M+H]⁺	C₁₅H₁₉O₇	311.112 5	311.112 0	-1.6	183.065 0 [M+H-C₆H₈O₃]⁺ 165.054 4 [M+H-C₆H₈O₃-H₂O]⁺	Decarboxylation of D0	Plasma, urine
D5	12.50	[M+H]⁺	C₁₈H₂₁O₁₀	397.112 9	397.111 9	-2.5	379.102 4 [M+H-H₂O]⁺ 353.086 7 [M+H-HCOCH₃]⁺	Acetylation of D0	Plasma
D6-1	8.30	[M+H]⁺	C₂₂H₃₁N₂O₁₂S	547.159 2	547.157 5	-3.1	369.118 0 [M+H-Cys-Gly]⁺ 351.106 9 [M+H-Cys-Gly-H₂O]⁺ 179.047 9 [M+H-C₁₇H₂₀O₉]⁺	Methylation and Cys-Gly conjugation of D0	Bile
D6-2	9.43	[M+H]⁺	C₂₂H₃₁N₂O₁₂S	547.159 2	547.157 7	-2.7	369.118 0 [M+H-Cys-Gly]⁺ 177.054 6 [M+H-Cys-Gly-C₇H₁₀O₅-H₂O]⁺	Methylation and Cys-Gly conjugation of D0	Bile
D7-1	9.36	[M+H]⁺	C₂₇H₃₈N₃O₁₅S	676.201 8	676.200 3	-2.2	369.117 8 [M+H-GSH]⁺ 308.091 0 [M+H-C₁₇H₂₀O₉]⁺ 233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺	Methylation and GSH conjugation of D0	Bile
D7-2	10.47	[M+H]⁺	C₂₇H₃₈N₃O₁₅S	676.201 8	676.200 4	-2.1	369.117 8 [M+H-GSH]⁺ 308.091 0 [M+H-C₁₇H₂₀O₉]⁺ 233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺	Methylation and GSH conjugation of D0	Bile
D8	10.85	[M+H]⁺	C₂₀H₂₈N₃O₁₀S	502.149 0	502.147 8	-2.4	308.091 1 [M+H-C₁₀H₁₀O₄]⁺ 233.058 5 [M+H-C₁₀H₁₀O₄-C₂H₅NO₂]⁺ 195.064 0 [M+H-GSH]⁺	GSH conjugation of D2	Bile

No.

t_R
/min

Ion form

Formula

m/z

Error
/ppm

MS/MS fragment ion

Identification/
reaction

Source

Theoretical

Experimental

12.28

[M+H]⁺

C₁₆H₁₉O₉

355.102 4

355.101 6

-2.3

181.049 2 [M+H-C₇H₁₀O₅]⁺
163.038 4 [M+H-C₇H₁₀O₅-H₂O]⁺
149.022 8 [M+H-C₇H₁₀O₅-CH₂-H₂O]⁺

Chlorogenic acid

Plasma, urine

D1-1

12.54

[M+H]⁺

C₁₇H₂₁O₉

369.118 0

369.116 9

-3.0

351.107 0 [M+H-H₂O]⁺
195.065 1 [M+H-C₇H₁₀O₅]⁺
177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺

Methylation of D0

Plasma, urine, feces

D1-2

14.00

[M+H]⁺

C₁₇H₂₁O₉

369.118 0

369.117 0

-2.7

351.107 0 [M+H-H₂O]⁺
305.095 8 [M+H-CO-2H₂O]⁺
177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺

Methylation of D0

Plasma, urine, feces

D1-3

14.33

[M+H]⁺

C₁₇H₂₁O₉

369.118 0

369.116 8

-3.3

351.107 0 [M+H-H₂O]⁺
195.065 1 [M+H-C₇H₁₀O₅]⁺
177.054 2 [M+H-C₇H₁₀O₅-H₂O]⁺

Methylation of D0

Plasma, urine, feces

13.12

[M+H]⁺

C₁₀H₁₁O₄

195.064 5

195.063 9

-3.1

163.039 0 [M+H-CH₂-H₂O]⁺
153.054 6 [M+H-CH₂-CO]⁺

Hydrolysis and methylation of D0

Plasma, urine, feces

3.34

[M+H]⁺

C₉H₁₁O₄

183.065 2

183.064 8

-2.2

165.054 6 [M+H-H₂O]⁺
137.059 2 [M+H-H₂O-CO]⁺

Hydrolysis and double-bond reduction of D0

Plasma, bile

8.53

[M+H]⁺

C₁₅H₁₉O₇

311.112 5

311.112 0

-1.6

183.065 0 [M+H-C₆H₈O₃]⁺
165.054 4 [M+H-C₆H₈O₃-H₂O]⁺

Decarboxylation of D0

Plasma, urine

12.50

[M+H]⁺

C₁₈H₂₁O₁₀

397.112 9

397.111 9

-2.5

379.102 4 [M+H-H₂O]⁺
353.086 7 [M+H-HCOCH₃]⁺

Acetylation of D0

Plasma

D6-1

8.30

[M+H]⁺

C₂₂H₃₁N₂O₁₂S

547.159 2

547.157 5

-3.1

369.118 0 [M+H-Cys-Gly]⁺
351.106 9 [M+H-Cys-Gly-H₂O]⁺
179.047 9 [M+H-C₁₇H₂₀O₉]⁺

Methylation and Cys-Gly conjugation of D0

Bile

D6-2

9.43

[M+H]⁺

C₂₂H₃₁N₂O₁₂S

547.159 2

547.157 7

-2.7

369.118 0 [M+H-Cys-Gly]⁺
177.054 6 [M+H-Cys-Gly-C₇H₁₀O₅-H₂O]⁺

Methylation and Cys-Gly conjugation of D0

Bile

D7-1

9.36

[M+H]⁺

C₂₇H₃₈N₃O₁₅S

676.201 8

676.200 3

-2.2

369.117 8 [M+H-GSH]⁺
308.091 0 [M+H-C₁₇H₂₀O₉]⁺
233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺

Methylation and GSH conjugation of D0

Bile

D7-2

10.47

[M+H]⁺

C₂₇H₃₈N₃O₁₅S

676.201 8

676.200 4

-2.1

369.117 8 [M+H-GSH]⁺
308.091 0 [M+H-C₁₇H₂₀O₉]⁺
233.059 1 [M+H-C₁₇H₂₀O₉-C₂H₅NO₂]⁺

Methylation and GSH conjugation of D0

Bile

10.85

[M+H]⁺

C₂₀H₂₈N₃O₁₀S

502.149 0

502.147 8

-2.4

308.091 1 [M+H-C₁₀H₁₀O₄]⁺
233.058 5 [M+H-C₁₀H₁₀O₄-C₂H₅NO₂]⁺
195.064 0 [M+H-GSH]⁺

GSH conjugation of D2

Bile