药学学报

No.	Identified compound	t_R _/min	[M+HCOO]^- (m/z)	ESI-MS/MS (-) (m/z)	ESI-MS/MS (+) (m/z)	Elemental composition	Substituent Group
1^*	Sibiricose A3	2.64	507.136 5	507.135 0	3.0	507 [M+HCOO]^-; 323 [M-H-HB]^-; 137 [HB-H]^-	-	C₁₉H₂₆O₁₃	R₁=H, R₂=HB
2^*	Arillanin C	3.30	563.159 0	563.161 2	-3.9	563 [M+HCOO]^-; 323 [M-H-MHC]^-	519 [M+H]⁺; 325 [M+H-MHC]⁺	C₂₂H₃₀O₁₄	R₁=HDC, R₂=H
3^*	Sibiricose A6	3.43	593.173 8	593.171 8	3.4	593 [M+HCOO]^-; 323 [M-H-TMC]^-; 223 [TMC-H]^-	549 [M+H]⁺; 325 [M+H-TMC]⁺	C₂₃H₃₂O₁₅	R₁=HDC, R₂=H
4^*	Sibiricose A1	3.76	593.171 6	593.171 8	-0.3	593 [M+HCOO]^-; 323 [M-H-TMC]^-; 223 [TMC-H]^-	549 [M+H]⁺; 325 [M+H-TMC]⁺	C₂₃H₃₂O₁₅	R₁=H, R₂=HDC
5^*	Sibiricose A2	4.53	607.187 0	607.187 4	-0.7	607[M+HCOO]^-; 223 [TMC-H]^-; 323 [M-H-TMC]^-	563 [M+H]⁺; 339 [M+H-TMC]⁺	C₂₄H₃₄O₁₅	R₁=TMC, R₂=H
6	Tenuifoliside B	4.73	713.192 4	713.192 9	-0.7	713 [M+HCOO]^-; 323 [M-H-HB-HDC]^-	669 [M+H]⁺; 549 [M+H-HB]⁺; 325 [M+H-HB-HDC]⁺	C₃₀H₃₆O₁₇	R₁=HDC, R₂=HB
7	3, 6- Disinapoylsucrose & Sibiricose A4	4.80	799.225 7	799.229 7	-5.0	799 [M+HCOO]^-; 323 [M-H-2HDC]^-	755 [M+H]⁺; 549 [M+H-HDC]⁺; 325 [M+H-2HDC]⁺	C₃₄H₄₂O₁₉	R₁=R₂=HDC
8^*	Sibiricose A4 & 3, 6- Disinapoylsucrose	5.36	799.233 1	799.229 7	4.3	799 [M+HCOO]^-; 547 [M-H-HDC]^-; 323 [M-H-2HDC]^-	755 [M+H]⁺; 549 [M+H-HDC]⁺; 325 [M+H-2HDC]⁺	C₃₄H₄₂O₁₉	R₁=R₂=HDC
9	Arillanin A	5.47	769.220 1	769.219 1	1.3	769 [M+HCOO]^-; 499 [M-H-HDC]^-; 529 [M-H-MHC]^-; 323 [M-H-HDC-MHC]^-	725 [M+H]⁺; 325 [M+H-HDC-MHC]⁺	C₃₃H₄₀O₁₈	R₁=R₂=HDC
10^*	Tenuifoliside A	5.89	727.209 0	727.208 6	0.6	727 [M+HCOO]^-; 543 [M-H-HB]^-; 443 [M-H-TMC]^-	683 [M+H]⁺; 545 [M+H-HB]⁺	C₃₁H₃₈O₁₇	R₁=TMC, R₂=HB
11^*	Tenuifoliose K	6.20	1 311.382 1	1 311.382 7	-0.4	1 311 [M+HCOO]^-; 1 119 [M-H-HC]^-; 1 077 [M-H-HC-C₂H₂O]^-	1 267 [M+H]⁺; 1 121 [M+H-HC]⁺; 1 079 [M+H-HC-C₂H₂O]⁺	C₅₇H₇₀O₃₂	R₁=R₃=R₄=H, R₂=Ac
12^*	Tenuifoliose C & Tenuifoliose E	6.37	1 341.392 4	1 341.393 2	-0.5	1 341 [M+HCOO]^-; 1 119 [M-H-MHC]^-	1 297 [M+H]⁺; 1 121 [M+H-MHC]⁺; 1 079 [M+H-MHC-C₂H₂O]⁺	C₅₈H₇₂O₃₃	R₁=Ac, R₂=OCH₃, R₃=R₄=H
13^*	Tenuifoliose I or Tenuifoliose J	6.52	1 353.389 1	1 353.393 2	-3.0	1 353 [M+HCOO]^-; 1 119 [M-H-HC-C₂H₂O]^-; 1 101 [M-H-HC-2C₂H₂O]^-	1 309 [M+H]⁺; 1 163 [M+H-HC]⁺; 1 121 [M+H-HC-C₂H₂O]⁺; 1 103 [M+H-HC-2C₂H₂O]⁺	C₆₁H₉₆O₃₀	R₁=R₃=Ac, R₂=R₄=H
14^*	TenuifolioseB & Tenuifoliose D	6.59	1 383.404 0	1 383.403 8	0.2	1 383 [M+HCOO]^-; 1 337 [M-H]^-; 1 161 [M-H-MHC]^-; 1 119 [M-H-MHC-C₂H₂O]^-; 1 077 [M-H-MHC-2C₂H₂O]^-; 973 [M-H-MHC-2C₂H₂O- (HB-OH)]^-	1 339 [M+H]⁺; 1 163 [M+H-MHC]⁺; 1 121 [M+H-MHC-C₂H₂O]⁺; 1 079 [M+H-MHC-2C₂H₂O]⁺	C₆₀H₇₄O₃₄	R₁=H, R₂=OCH₃, R₃=R₄=Ac
15	Tenuifoliose C & Tenuifoliose E	6.75	1 341.392 8	1 341.393 2	-0.2	1 341 [M+HCOO]^-; 1 119 [M-H-MHC]^-	1 297 [M+H]⁺; 1 121 [M+H-MHC]⁺; 1 079 [M+H-MHC-C₂H₂O]⁺	C₅₈H₇₂O₃₃	R₁=H, R₂=OCH₃, R₃=Ac, R₄=Glu
16^*	Tenuifoliose B & Tenuifoliose D	7.13	1 383.404 5	1 383.403 8	-0.3	1 383 [M+HCOO]^-; 1 161 [M-H-MHC]^-; 973 [M-H-MHC-2C₂H₂O- (HB-OH)]^-	1 339 [M+H]⁺; 1 163 [M+H-MHC]⁺; 1 079 [M+H-MHC-2C₂H₂O]⁺	C₆₀H₇₄O₃₄	R₁=R₃=Ac, R₂=OCH₃, R₄=H
17^*	Tenuifoliose A	7.43	1 425.414 0	1 425.414 4	-0.3	1 425 [M+HCOO]^-; 1 379 [M-H]^-; 1 203 [M-H-HHC]^-	1 381 [M+H]⁺; 1 205 [M+H-HHC]⁺; 1 163 [M+H-HHC-C₂H₂O]⁺; 1 179 [M+H-HHC-3C₂H₂O]⁺	C₆₂H₇₆O₃₅	R₁=R₃=R₄=Ac, R₂=CH₂OH
18^*	Reiniose J	7.59	1 383.401 6	1 338.403 8	-1.8	1 383 [M+HCOO]^-; 1 161 [M-H-MHC]^-	1 339 [M+H]⁺; 1 163 [M+H-MHC]⁺; 1 297 [M+H-C₂H₂O]⁺; 1 079 [M+H-MHC-2C₂H₂O]⁺	C₆₀H₇₄O₃₄	R₁=R₃=Ac, R₂=OCH₃, R₄=H
19	Tenuifoliose I & Tenuifoliose J	7.80	1 353.394 0	1 353.393 2	0.3	1 353 [M+HCOO]^-; 1 161 [M-H-HC]^-; 1 119 [M-H-HC-C₂H₂O]^-; 1 101 [M-H-HC-2C₂H₂O]^-	-	C₆₁H₉₆O₃₀	R₁=R₄=Ac, R₂=R₃=H,
20	Tenuifoliose N	11.02	1 455.420 5	1 455.424 9	-3.8	1 455 [M+HCOO]^-; 1 233 [M-H-MHC]^-; 1 191 [M-H-MHC-C₂H₂O]^-; 1 149 [M-H-MHC-2C₂H₂O]^-	-	C₆₃H₇₈O₃₆	R₁=R₃=R₄=Ac, R₂=O

No.

Identified
compound

t_R
_/min

[M+HCOO]^- (m/z)

ESI-MS/MS (-)
(m/z)

ESI-MS/MS (+)
(m/z)

Elemental composition

Substituent Group

Measured

Calculated

Δ
(ppm)

1^*

Sibiricose A3

2.64

507.136 5

507.135 0

3.0

507 [M+HCOO]^-;
323 [M-H-HB]^-;
137 [HB-H]^-

C₁₉H₂₆O₁₃

R₁=H,
R₂=HB

2^*

Arillanin C

3.30

563.159 0

563.161 2

-3.9

563 [M+HCOO]^-;
323 [M-H-MHC]^-

519 [M+H]⁺;
325 [M+H-MHC]⁺

C₂₂H₃₀O₁₄

R₁=HDC,
R₂=H

3^*

Sibiricose A6

3.43

593.173 8

593.171 8

3.4

593 [M+HCOO]^-;
323 [M-H-TMC]^-;
223 [TMC-H]^-

549 [M+H]⁺;
325 [M+H-TMC]⁺

C₂₃H₃₂O₁₅

R₁=HDC,
R₂=H

4^*

Sibiricose A1

3.76

593.171 6

593.171 8

-0.3

593 [M+HCOO]^-;
323 [M-H-TMC]^-;
223 [TMC-H]^-

549 [M+H]⁺;
325 [M+H-TMC]⁺

C₂₃H₃₂O₁₅

R₁=H,
R₂=HDC

5^*

Sibiricose A2

4.53

607.187 0

607.187 4

-0.7

607[M+HCOO]^-;
223 [TMC-H]^-;
323 [M-H-TMC]^-

563 [M+H]⁺;
339 [M+H-TMC]⁺

C₂₄H₃₄O₁₅

R₁=TMC,
R₂=H

Tenuifoliside B

4.73

713.192 4

713.192 9

-0.7

713 [M+HCOO]^-;
323 [M-H-HB-HDC]^-

669 [M+H]⁺;
549 [M+H-HB]⁺;
325 [M+H-HB-HDC]⁺

C₃₀H₃₆O₁₇

R₁=HDC,
R₂=HB

3, 6-
Disinapoylsucrose
& Sibiricose A4

4.80

799.225 7

799.229 7

-5.0

799 [M+HCOO]^-;
323 [M-H-2HDC]^-

755 [M+H]⁺;
549 [M+H-HDC]⁺;
325 [M+H-2HDC]⁺

C₃₄H₄₂O₁₉

R₁=R₂=HDC

8^*

Sibiricose A4 &
3, 6-
Disinapoylsucrose

5.36

799.233 1

799.229 7

4.3

799 [M+HCOO]^-;
547 [M-H-HDC]^-;
323 [M-H-2HDC]^-

755 [M+H]⁺;
549 [M+H-HDC]⁺;
325 [M+H-2HDC]⁺

C₃₄H₄₂O₁₉

R₁=R₂=HDC

Arillanin A

5.47

769.220 1

769.219 1

1.3

769 [M+HCOO]^-;
499 [M-H-HDC]^-;
529 [M-H-MHC]^-;
323 [M-H-HDC-MHC]^-

725 [M+H]⁺;
325 [M+H-HDC-MHC]⁺

C₃₃H₄₀O₁₈

R₁=R₂=HDC

10^*

Tenuifoliside A

5.89

727.209 0

727.208 6

0.6

727 [M+HCOO]^-;
543 [M-H-HB]^-;
443 [M-H-TMC]^-

683 [M+H]⁺;
545 [M+H-HB]⁺

C₃₁H₃₈O₁₇

R₁=TMC,
R₂=HB

11^*

Tenuifoliose K

6.20

1 311.382 1

1 311.382 7

-0.4

1 311 [M+HCOO]^-;
1 119 [M-H-HC]^-;
1 077 [M-H-HC-C₂H₂O]^-

1 267 [M+H]⁺;
1 121 [M+H-HC]⁺;
1 079 [M+H-HC-C₂H₂O]⁺

C₅₇H₇₀O₃₂

R₁=R₃=R₄=H,
R₂=Ac

12^*

Tenuifoliose C & Tenuifoliose E

6.37

1 341.392 4

1 341.393 2

-0.5

1 341 [M+HCOO]^-;
1 119 [M-H-MHC]^-

1 297 [M+H]⁺;
1 121 [M+H-MHC]⁺;
1 079 [M+H-MHC-C₂H₂O]⁺

C₅₈H₇₂O₃₃

R₁=Ac, R₂=OCH₃, R₃=R₄=H

13^*

Tenuifoliose I or Tenuifoliose J

6.52

1 353.389 1

1 353.393 2

-3.0

1 353 [M+HCOO]^-;
1 119 [M-H-HC-C₂H₂O]^-;
1 101 [M-H-HC-2C₂H₂O]^-

1 309 [M+H]⁺;
1 163 [M+H-HC]⁺;
1 121 [M+H-HC-C₂H₂O]⁺;
1 103 [M+H-HC-2C₂H₂O]⁺

C₆₁H₉₆O₃₀

R₁=R₃=Ac, R₂=R₄=H

14^*

TenuifolioseB & Tenuifoliose D

6.59

1 383.404 0

1 383.403 8

0.2

1 383 [M+HCOO]^-;
1 337 [M-H]^-;
1 161 [M-H-MHC]^-;
1 119 [M-H-MHC-C₂H₂O]^-;
1 077 [M-H-MHC-2C₂H₂O]^-;
973 [M-H-MHC-2C₂H₂O-
(HB-OH)]^-

1 339 [M+H]⁺;
1 163 [M+H-MHC]⁺;
1 121 [M+H-MHC-C₂H₂O]⁺;
1 079 [M+H-MHC-2C₂H₂O]⁺

C₆₀H₇₄O₃₄

R₁=H, R₂=OCH₃, R₃=R₄=Ac

Tenuifoliose C & Tenuifoliose E

6.75

1 341.392 8

1 341.393 2

-0.2

1 341 [M+HCOO]^-;
1 119 [M-H-MHC]^-

1 297 [M+H]⁺;
1 121 [M+H-MHC]⁺;
1 079 [M+H-MHC-C₂H₂O]⁺

C₅₈H₇₂O₃₃

R₁=H, R₂=OCH₃, R₃=Ac,
R₄=Glu

16^*

Tenuifoliose B & Tenuifoliose D

7.13

1 383.404 5

1 383.403 8

-0.3

1 383 [M+HCOO]^-;
1 161 [M-H-MHC]^-;
973 [M-H-MHC-2C₂H₂O-
(HB-OH)]^-

1 339 [M+H]⁺;
1 163 [M+H-MHC]⁺;
1 079 [M+H-MHC-2C₂H₂O]⁺

C₆₀H₇₄O₃₄

R₁=R₃=Ac, R₂=OCH₃,
R₄=H

17^*

Tenuifoliose A

7.43

1 425.414 0

1 425.414 4

-0.3

1 425 [M+HCOO]^-;
1 379 [M-H]^-;
1 203 [M-H-HHC]^-

1 381 [M+H]⁺;
1 205 [M+H-HHC]⁺;
1 163 [M+H-HHC-C₂H₂O]⁺;
1 179 [M+H-HHC-3C₂H₂O]⁺

C₆₂H₇₆O₃₅

R₁=R₃=R₄=Ac,
R₂=CH₂OH

18^*

Reiniose J

7.59

1 383.401 6

1 338.403 8

-1.8

1 383 [M+HCOO]^-;
1 161 [M-H-MHC]^-

1 339 [M+H]⁺;
1 163 [M+H-MHC]⁺;
1 297 [M+H-C₂H₂O]⁺;
1 079 [M+H-MHC-2C₂H₂O]⁺

C₆₀H₇₄O₃₄

R₁=R₃=Ac, R₂=OCH₃,
R₄=H

Tenuifoliose I & Tenuifoliose J

7.80

1 353.394 0

1 353.393 2

0.3

1 353 [M+HCOO]^-;
1 161 [M-H-HC]^-;
1 119 [M-H-HC-C₂H₂O]^-;
1 101 [M-H-HC-2C₂H₂O]^-

C₆₁H₉₆O₃₀

R₁=R₄=Ac, R₂=R₃=H,

Tenuifoliose N

11.02

1 455.420 5

1 455.424 9

-3.8

1 455 [M+HCOO]^-;
1 233 [M-H-MHC]^-;
1 191 [M-H-MHC-C₂H₂O]^-;
1 149 [M-H-MHC-2C₂H₂O]^-

C₆₃H₇₈O₃₆

R₁=R₃=R₄=Ac,
R₂=O

No.	Identified compound	t_R _/min	[M+HCOO]^- (m/z)	ESI-MS/MS (-) (m/z)	ESI-MS/MS (+) (m/z)	Elemental composition	Substituent Group
1^*	Sibiricose A3	2.64	507.136 5	507.135 0	3.0	507 [M+HCOO]^-; 323 [M-H-HB]^-; 137 [HB-H]^-	-	C₁₉H₂₆O₁₃	R₁=H, R₂=HB
2^*	Arillanin C	3.30	563.159 0	563.161 2	-3.9	563 [M+HCOO]^-; 323 [M-H-MHC]^-	519 [M+H]⁺; 325 [M+H-MHC]⁺	C₂₂H₃₀O₁₄	R₁=HDC, R₂=H
3^*	Sibiricose A6	3.43	593.173 8	593.171 8	3.4	593 [M+HCOO]^-; 323 [M-H-TMC]^-; 223 [TMC-H]^-	549 [M+H]⁺; 325 [M+H-TMC]⁺	C₂₃H₃₂O₁₅	R₁=HDC, R₂=H
4^*	Sibiricose A1	3.76	593.171 6	593.171 8	-0.3	593 [M+HCOO]^-; 323 [M-H-TMC]^-; 223 [TMC-H]^-	549 [M+H]⁺; 325 [M+H-TMC]⁺	C₂₃H₃₂O₁₅	R₁=H, R₂=HDC
5^*	Sibiricose A2	4.53	607.187 0	607.187 4	-0.7	607[M+HCOO]^-; 223 [TMC-H]^-; 323 [M-H-TMC]^-	563 [M+H]⁺; 339 [M+H-TMC]⁺	C₂₄H₃₄O₁₅	R₁=TMC, R₂=H
6	Tenuifoliside B	4.73	713.192 4	713.192 9	-0.7	713 [M+HCOO]^-; 323 [M-H-HB-HDC]^-	669 [M+H]⁺; 549 [M+H-HB]⁺; 325 [M+H-HB-HDC]⁺	C₃₀H₃₆O₁₇	R₁=HDC, R₂=HB
7	3, 6- Disinapoylsucrose & Sibiricose A4	4.80	799.225 7	799.229 7	-5.0	799 [M+HCOO]^-; 323 [M-H-2HDC]^-	755 [M+H]⁺; 549 [M+H-HDC]⁺; 325 [M+H-2HDC]⁺	C₃₄H₄₂O₁₉	R₁=R₂=HDC
8^*	Sibiricose A4 & 3, 6- Disinapoylsucrose	5.36	799.233 1	799.229 7	4.3	799 [M+HCOO]^-; 547 [M-H-HDC]^-; 323 [M-H-2HDC]^-	755 [M+H]⁺; 549 [M+H-HDC]⁺; 325 [M+H-2HDC]⁺	C₃₄H₄₂O₁₉	R₁=R₂=HDC
9	Arillanin A	5.47	769.220 1	769.219 1	1.3	769 [M+HCOO]^-; 499 [M-H-HDC]^-; 529 [M-H-MHC]^-; 323 [M-H-HDC-MHC]^-	725 [M+H]⁺; 325 [M+H-HDC-MHC]⁺	C₃₃H₄₀O₁₈	R₁=R₂=HDC
10^*	Tenuifoliside A	5.89	727.209 0	727.208 6	0.6	727 [M+HCOO]^-; 543 [M-H-HB]^-; 443 [M-H-TMC]^-	683 [M+H]⁺; 545 [M+H-HB]⁺	C₃₁H₃₈O₁₇	R₁=TMC, R₂=HB
11^*	Tenuifoliose K	6.20	1 311.382 1	1 311.382 7	-0.4	1 311 [M+HCOO]^-; 1 119 [M-H-HC]^-; 1 077 [M-H-HC-C₂H₂O]^-	1 267 [M+H]⁺; 1 121 [M+H-HC]⁺; 1 079 [M+H-HC-C₂H₂O]⁺	C₅₇H₇₀O₃₂	R₁=R₃=R₄=H, R₂=Ac
12^*	Tenuifoliose C & Tenuifoliose E	6.37	1 341.392 4	1 341.393 2	-0.5	1 341 [M+HCOO]^-; 1 119 [M-H-MHC]^-	1 297 [M+H]⁺; 1 121 [M+H-MHC]⁺; 1 079 [M+H-MHC-C₂H₂O]⁺	C₅₈H₇₂O₃₃	R₁=Ac, R₂=OCH₃, R₃=R₄=H
13^*	Tenuifoliose I or Tenuifoliose J	6.52	1 353.389 1	1 353.393 2	-3.0	1 353 [M+HCOO]^-; 1 119 [M-H-HC-C₂H₂O]^-; 1 101 [M-H-HC-2C₂H₂O]^-	1 309 [M+H]⁺; 1 163 [M+H-HC]⁺; 1 121 [M+H-HC-C₂H₂O]⁺; 1 103 [M+H-HC-2C₂H₂O]⁺	C₆₁H₉₆O₃₀	R₁=R₃=Ac, R₂=R₄=H
14^*	TenuifolioseB & Tenuifoliose D	6.59	1 383.404 0	1 383.403 8	0.2	1 383 [M+HCOO]^-; 1 337 [M-H]^-; 1 161 [M-H-MHC]^-; 1 119 [M-H-MHC-C₂H₂O]^-; 1 077 [M-H-MHC-2C₂H₂O]^-; 973 [M-H-MHC-2C₂H₂O- (HB-OH)]^-	1 339 [M+H]⁺; 1 163 [M+H-MHC]⁺; 1 121 [M+H-MHC-C₂H₂O]⁺; 1 079 [M+H-MHC-2C₂H₂O]⁺	C₆₀H₇₄O₃₄	R₁=H, R₂=OCH₃, R₃=R₄=Ac
15	Tenuifoliose C & Tenuifoliose E	6.75	1 341.392 8	1 341.393 2	-0.2	1 341 [M+HCOO]^-; 1 119 [M-H-MHC]^-	1 297 [M+H]⁺; 1 121 [M+H-MHC]⁺; 1 079 [M+H-MHC-C₂H₂O]⁺	C₅₈H₇₂O₃₃	R₁=H, R₂=OCH₃, R₃=Ac, R₄=Glu
16^*	Tenuifoliose B & Tenuifoliose D	7.13	1 383.404 5	1 383.403 8	-0.3	1 383 [M+HCOO]^-; 1 161 [M-H-MHC]^-; 973 [M-H-MHC-2C₂H₂O- (HB-OH)]^-	1 339 [M+H]⁺; 1 163 [M+H-MHC]⁺; 1 079 [M+H-MHC-2C₂H₂O]⁺	C₆₀H₇₄O₃₄	R₁=R₃=Ac, R₂=OCH₃, R₄=H
17^*	Tenuifoliose A	7.43	1 425.414 0	1 425.414 4	-0.3	1 425 [M+HCOO]^-; 1 379 [M-H]^-; 1 203 [M-H-HHC]^-	1 381 [M+H]⁺; 1 205 [M+H-HHC]⁺; 1 163 [M+H-HHC-C₂H₂O]⁺; 1 179 [M+H-HHC-3C₂H₂O]⁺	C₆₂H₇₆O₃₅	R₁=R₃=R₄=Ac, R₂=CH₂OH
18^*	Reiniose J	7.59	1 383.401 6	1 338.403 8	-1.8	1 383 [M+HCOO]^-; 1 161 [M-H-MHC]^-	1 339 [M+H]⁺; 1 163 [M+H-MHC]⁺; 1 297 [M+H-C₂H₂O]⁺; 1 079 [M+H-MHC-2C₂H₂O]⁺	C₆₀H₇₄O₃₄	R₁=R₃=Ac, R₂=OCH₃, R₄=H
19	Tenuifoliose I & Tenuifoliose J	7.80	1 353.394 0	1 353.393 2	0.3	1 353 [M+HCOO]^-; 1 161 [M-H-HC]^-; 1 119 [M-H-HC-C₂H₂O]^-; 1 101 [M-H-HC-2C₂H₂O]^-	-	C₆₁H₉₆O₃₀	R₁=R₄=Ac, R₂=R₃=H,
20	Tenuifoliose N	11.02	1 455.420 5	1 455.424 9	-3.8	1 455 [M+HCOO]^-; 1 233 [M-H-MHC]^-; 1 191 [M-H-MHC-C₂H₂O]^-; 1 149 [M-H-MHC-2C₂H₂O]^-	-	C₆₃H₇₈O₃₆	R₁=R₃=R₄=Ac, R₂=O

No.

Identified
compound

t_R
_/min

[M+HCOO]^- (m/z)

ESI-MS/MS (-)
(m/z)

ESI-MS/MS (+)
(m/z)

Elemental composition

Substituent Group

Measured

Calculated

Δ
(ppm)

1^*

Sibiricose A3

2.64

507.136 5

507.135 0

3.0

507 [M+HCOO]^-;
323 [M-H-HB]^-;
137 [HB-H]^-

C₁₉H₂₆O₁₃

R₁=H,
R₂=HB

2^*

Arillanin C

3.30

563.159 0

563.161 2

-3.9

563 [M+HCOO]^-;
323 [M-H-MHC]^-

519 [M+H]⁺;
325 [M+H-MHC]⁺

C₂₂H₃₀O₁₄

R₁=HDC,
R₂=H

3^*

Sibiricose A6

3.43

593.173 8

593.171 8

3.4

593 [M+HCOO]^-;
323 [M-H-TMC]^-;
223 [TMC-H]^-

549 [M+H]⁺;
325 [M+H-TMC]⁺

C₂₃H₃₂O₁₅

R₁=HDC,
R₂=H

4^*

Sibiricose A1

3.76

593.171 6

593.171 8

-0.3

593 [M+HCOO]^-;
323 [M-H-TMC]^-;
223 [TMC-H]^-

549 [M+H]⁺;
325 [M+H-TMC]⁺

C₂₃H₃₂O₁₅

R₁=H,
R₂=HDC

5^*

Sibiricose A2

4.53

607.187 0

607.187 4

-0.7

607[M+HCOO]^-;
223 [TMC-H]^-;
323 [M-H-TMC]^-

563 [M+H]⁺;
339 [M+H-TMC]⁺

C₂₄H₃₄O₁₅

R₁=TMC,
R₂=H

Tenuifoliside B

4.73

713.192 4

713.192 9

-0.7

713 [M+HCOO]^-;
323 [M-H-HB-HDC]^-

669 [M+H]⁺;
549 [M+H-HB]⁺;
325 [M+H-HB-HDC]⁺

C₃₀H₃₆O₁₇

R₁=HDC,
R₂=HB

3, 6-
Disinapoylsucrose
& Sibiricose A4

4.80

799.225 7

799.229 7

-5.0

799 [M+HCOO]^-;
323 [M-H-2HDC]^-

755 [M+H]⁺;
549 [M+H-HDC]⁺;
325 [M+H-2HDC]⁺

C₃₄H₄₂O₁₉

R₁=R₂=HDC

8^*

Sibiricose A4 &
3, 6-
Disinapoylsucrose

5.36

799.233 1

799.229 7

4.3

799 [M+HCOO]^-;
547 [M-H-HDC]^-;
323 [M-H-2HDC]^-

755 [M+H]⁺;
549 [M+H-HDC]⁺;
325 [M+H-2HDC]⁺

C₃₄H₄₂O₁₉

R₁=R₂=HDC

Arillanin A

5.47

769.220 1

769.219 1

1.3

769 [M+HCOO]^-;
499 [M-H-HDC]^-;
529 [M-H-MHC]^-;
323 [M-H-HDC-MHC]^-

725 [M+H]⁺;
325 [M+H-HDC-MHC]⁺

C₃₃H₄₀O₁₈

R₁=R₂=HDC

10^*

Tenuifoliside A

5.89

727.209 0

727.208 6

0.6

727 [M+HCOO]^-;
543 [M-H-HB]^-;
443 [M-H-TMC]^-

683 [M+H]⁺;
545 [M+H-HB]⁺

C₃₁H₃₈O₁₇

R₁=TMC,
R₂=HB

11^*

Tenuifoliose K

6.20

1 311.382 1

1 311.382 7

-0.4

1 311 [M+HCOO]^-;
1 119 [M-H-HC]^-;
1 077 [M-H-HC-C₂H₂O]^-

1 267 [M+H]⁺;
1 121 [M+H-HC]⁺;
1 079 [M+H-HC-C₂H₂O]⁺

C₅₇H₇₀O₃₂

R₁=R₃=R₄=H,
R₂=Ac

12^*

Tenuifoliose C & Tenuifoliose E

6.37

1 341.392 4

1 341.393 2

-0.5

1 341 [M+HCOO]^-;
1 119 [M-H-MHC]^-

1 297 [M+H]⁺;
1 121 [M+H-MHC]⁺;
1 079 [M+H-MHC-C₂H₂O]⁺

C₅₈H₇₂O₃₃

R₁=Ac, R₂=OCH₃, R₃=R₄=H

13^*

Tenuifoliose I or Tenuifoliose J

6.52

1 353.389 1

1 353.393 2

-3.0

1 353 [M+HCOO]^-;
1 119 [M-H-HC-C₂H₂O]^-;
1 101 [M-H-HC-2C₂H₂O]^-

1 309 [M+H]⁺;
1 163 [M+H-HC]⁺;
1 121 [M+H-HC-C₂H₂O]⁺;
1 103 [M+H-HC-2C₂H₂O]⁺

C₆₁H₉₆O₃₀

R₁=R₃=Ac, R₂=R₄=H

14^*

TenuifolioseB & Tenuifoliose D

6.59

1 383.404 0

1 383.403 8

0.2

1 383 [M+HCOO]^-;
1 337 [M-H]^-;
1 161 [M-H-MHC]^-;
1 119 [M-H-MHC-C₂H₂O]^-;
1 077 [M-H-MHC-2C₂H₂O]^-;
973 [M-H-MHC-2C₂H₂O-
(HB-OH)]^-

1 339 [M+H]⁺;
1 163 [M+H-MHC]⁺;
1 121 [M+H-MHC-C₂H₂O]⁺;
1 079 [M+H-MHC-2C₂H₂O]⁺

C₆₀H₇₄O₃₄

R₁=H, R₂=OCH₃, R₃=R₄=Ac

Tenuifoliose C & Tenuifoliose E

6.75

1 341.392 8

1 341.393 2

-0.2

1 341 [M+HCOO]^-;
1 119 [M-H-MHC]^-

1 297 [M+H]⁺;
1 121 [M+H-MHC]⁺;
1 079 [M+H-MHC-C₂H₂O]⁺

C₅₈H₇₂O₃₃

R₁=H, R₂=OCH₃, R₃=Ac,
R₄=Glu

16^*

Tenuifoliose B & Tenuifoliose D

7.13

1 383.404 5

1 383.403 8

-0.3

1 383 [M+HCOO]^-;
1 161 [M-H-MHC]^-;
973 [M-H-MHC-2C₂H₂O-
(HB-OH)]^-

1 339 [M+H]⁺;
1 163 [M+H-MHC]⁺;
1 079 [M+H-MHC-2C₂H₂O]⁺

C₆₀H₇₄O₃₄

R₁=R₃=Ac, R₂=OCH₃,
R₄=H

17^*

Tenuifoliose A

7.43

1 425.414 0

1 425.414 4

-0.3

1 425 [M+HCOO]^-;
1 379 [M-H]^-;
1 203 [M-H-HHC]^-

1 381 [M+H]⁺;
1 205 [M+H-HHC]⁺;
1 163 [M+H-HHC-C₂H₂O]⁺;
1 179 [M+H-HHC-3C₂H₂O]⁺

C₆₂H₇₆O₃₅

R₁=R₃=R₄=Ac,
R₂=CH₂OH

18^*

Reiniose J

7.59

1 383.401 6

1 338.403 8

-1.8

1 383 [M+HCOO]^-;
1 161 [M-H-MHC]^-

1 339 [M+H]⁺;
1 163 [M+H-MHC]⁺;
1 297 [M+H-C₂H₂O]⁺;
1 079 [M+H-MHC-2C₂H₂O]⁺

C₆₀H₇₄O₃₄

R₁=R₃=Ac, R₂=OCH₃,
R₄=H

Tenuifoliose I & Tenuifoliose J

7.80

1 353.394 0

1 353.393 2

0.3

1 353 [M+HCOO]^-;
1 161 [M-H-HC]^-;
1 119 [M-H-HC-C₂H₂O]^-;
1 101 [M-H-HC-2C₂H₂O]^-

C₆₁H₉₆O₃₀

R₁=R₄=Ac, R₂=R₃=H,

Tenuifoliose N

11.02

1 455.420 5

1 455.424 9

-3.8

1 455 [M+HCOO]^-;
1 233 [M-H-MHC]^-;
1 191 [M-H-MHC-C₂H₂O]^-;
1 149 [M-H-MHC-2C₂H₂O]^-

C₆₃H₇₈O₃₆

R₁=R₃=R₄=Ac,
R₂=O

No.	Identified compound	t_R _/min	[M+H]⁺ (m/z)	ESI-MS/MS (-) (m/z)	ESI-MS/MS (+) (m/z)	Elemental composition
M1	Tenuifoliside B-(HDC)	4.29	463.144 8	463.145 2	-1.8	507 [M+HCOO]^-; 137 [HB-H]^-	463 [M+H]⁺; 325 [M-HB+H]⁺; 139 [HB+H]⁺	C₁₉H₂₆O₁₃
M2	Tenuifoliose K-(CH₂O)	4.90	1 239.396 0	1 239.397 9	-3.4	1 237 [M-H]^-; 1 136 [M-H-C₈H₇O+H₂O]^-; 1 034 [M-H-2C₈H₇O+H₂O]^-	1 239 [M+H]⁺; 1 138 [M+H-C₈H₇O+H₂O]⁺; 1 036 [M+H-2C₈H₇O+H₂O]⁺	C₅₆H₇₀O₃₁
M3	3, 4, 5-Trimethoxycinnamylic acid	6.92	239.091 0	239.091 9	-3.8	283 [M+HCOO]^-; 237 [M-H]^-; 221 [M-O-H]^-	239 [M+H]⁺; 223 [M-O+H]⁺; 209 [M-O-CH₂+H]⁺	C₁₂H₁₄O₅
M4	3, 4, 5-Trimethoxycinnamylic acid-(CH₂)	7.35	225.074 1	225.076 3	-4.1	223 [M-H]^-; 209 [M-O+H]⁺; 269 [M+HCOO]^-	225 [M+H]⁺; 209 [M-O+H]⁺	C₁₁H₁₂O₅

No.

Identified compound

t_R
_/min

[M+H]⁺ (m/z)

ESI-MS/MS (-)
(m/z)

ESI-MS/MS (+)
(m/z)

Elemental
composition

Measured

Calculated

Δ(ppm)

Tenuifoliside B-(HDC)

4.29

463.144 8

463.145 2

-1.8

507 [M+HCOO]^-; 137 [HB-H]^-

463 [M+H]⁺; 325 [M-HB+H]⁺; 139 [HB+H]⁺

C₁₉H₂₆O₁₃

Tenuifoliose K-(CH₂O)

4.90

1 239.396 0

1 239.397 9

-3.4

1 237 [M-H]^-; 1 136 [M-H-C₈H₇O+H₂O]^-; 1 034 [M-H-2C₈H₇O+H₂O]^-

1 239 [M+H]⁺; 1 138 [M+H-C₈H₇O+H₂O]⁺; 1 036 [M+H-2C₈H₇O+H₂O]⁺

C₅₆H₇₀O₃₁

3, 4, 5-Trimethoxycinnamylic acid

6.92

239.091 0

239.091 9

-3.8

283 [M+HCOO]^-; 237 [M-H]^-; 221 [M-O-H]^-

239 [M+H]⁺; 223 [M-O+H]⁺; 209 [M-O-CH₂+H]⁺

C₁₂H₁₄O₅

3, 4, 5-Trimethoxycinnamylic acid-(CH₂)

7.35

225.074 1

225.076 3

-4.1

223 [M-H]^-; 209 [M-O+H]⁺; 269 [M+HCOO]^-

225 [M+H]⁺; 209 [M-O+H]⁺

C₁₁H₁₂O₅

No.	Identified compound	t_R _/min	[M+H]⁺ (m/z)	ESI-MS/MS (-) (m/z)	ESI-MS/MS (+) (m/z)	Elemental composition
M1	Tenuifoliside B-(HDC)	4.29	463.144 8	463.145 2	-1.8	507 [M+HCOO]^-; 137 [HB-H]^-	463 [M+H]⁺; 325 [M-HB+H]⁺; 139 [HB+H]⁺	C₁₉H₂₆O₁₃
M2	Tenuifoliose K-(CH₂O)	4.90	1 239.396 0	1 239.397 9	-3.4	1 237 [M-H]^-; 1 136 [M-H-C₈H₇O+H₂O]^-; 1 034 [M-H-2C₈H₇O+H₂O]^-	1 239 [M+H]⁺; 1 138 [M+H-C₈H₇O+H₂O]⁺; 1 036 [M+H-2C₈H₇O+H₂O]⁺	C₅₆H₇₀O₃₁
M3	3, 4, 5-Trimethoxycinnamylic acid	6.92	239.091 0	239.091 9	-3.8	283 [M+HCOO]^-; 237 [M-H]^-; 221 [M-O-H]^-	239 [M+H]⁺; 223 [M-O+H]⁺; 209 [M-O-CH₂+H]⁺	C₁₂H₁₄O₅
M4	3, 4, 5-Trimethoxycinnamylic acid-(CH₂)	7.35	225.074 1	225.076 3	-4.1	223 [M-H]^-; 209 [M-O+H]⁺; 269 [M+HCOO]^-	225 [M+H]⁺; 209 [M-O+H]⁺	C₁₁H₁₂O₅

No.

Identified compound

t_R
_/min

[M+H]⁺ (m/z)

ESI-MS/MS (-)
(m/z)

ESI-MS/MS (+)
(m/z)

Elemental
composition

Measured

Calculated

Δ(ppm)

Tenuifoliside B-(HDC)

4.29

463.144 8

463.145 2

-1.8

507 [M+HCOO]^-; 137 [HB-H]^-

463 [M+H]⁺; 325 [M-HB+H]⁺; 139 [HB+H]⁺

C₁₉H₂₆O₁₃

Tenuifoliose K-(CH₂O)

4.90

1 239.396 0

1 239.397 9

-3.4

1 237 [M-H]^-; 1 136 [M-H-C₈H₇O+H₂O]^-; 1 034 [M-H-2C₈H₇O+H₂O]^-

1 239 [M+H]⁺; 1 138 [M+H-C₈H₇O+H₂O]⁺; 1 036 [M+H-2C₈H₇O+H₂O]⁺

C₅₆H₇₀O₃₁

3, 4, 5-Trimethoxycinnamylic acid

6.92

239.091 0

239.091 9

-3.8

283 [M+HCOO]^-; 237 [M-H]^-; 221 [M-O-H]^-

239 [M+H]⁺; 223 [M-O+H]⁺; 209 [M-O-CH₂+H]⁺

C₁₂H₁₄O₅

3, 4, 5-Trimethoxycinnamylic acid-(CH₂)

7.35

225.074 1

225.076 3

-4.1

223 [M-H]^-; 209 [M-O+H]⁺; 269 [M+HCOO]^-

225 [M+H]⁺; 209 [M-O+H]⁺

C₁₁H₁₂O₅

UHPLC-MS法鉴定大鼠灌服开心散后血中远志糖酯类化合物及其代谢产物

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汪娜 ¹ , AHMAD Hassan ¹ , 贾永明 ¹ , 薛迪 ¹ , CHAULAGAIN Ram ¹ , 黄树明 ² , 刘学伟 ¹^,^*

药学学报 | 研究论文 2017,52(10): 1592-1598

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药学学报 | 研究论文 2017, 52(10): 1592-1598

UHPLC-MS法鉴定大鼠灌服开心散后血中远志糖酯类化合物及其代谢产物

全屏

汪娜¹, AHMAD Hassan¹, 贾永明¹, 薛迪¹, CHAULAGAIN Ram¹, 黄树明², 刘学伟¹^,^*

作者信息

1.齐齐哈尔医学院, 黑龙江齐齐哈尔 161006

2.黑龙江中医药大学, 黑龙江哈尔滨 150040

通讯作者:

* 刘学伟, Tel/Fax:86-452-2663804, E-mail:liuxuewei2001@163.com

Identification of polygala oligosaccharide esters and their metabolites in rat plasma after oral administration of ethanol extract of Kai Xin San by UHPLC-MS

Na WANG¹, AHMAD Hassan¹, Yong-ming JIA¹, Di XUE¹, CHAULAGAIN Ram¹, Shu-ming HUANG², Xue-wei LIU¹^,^*

Affiliations

1. Qiqihar Medical University, Qiqihar 161006, China

2. Heilongjiang University of Chinese Medicine, Harbin 150040, China

出版时间: 2017-10-12 doi: 10.16438/j.0513-4870.2017-0373

文章导航

摘要

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本研究利用超高效液相色谱-飞行时间质谱联用技术（UHPLC-TOF/MS），借助液质联用工作站Masslynx V4.1分析软件，结合现有文献及相关数据库，鉴定了开心散60%乙醇提取物（KXS-60% E）中20个远志糖酯类成分；在此基础上，进一步指认并鉴定了灌服KXS-60% E后大鼠血中的14个远志糖酯类化合物及4个来源于远志糖酯类成分的代谢产物。以上研究结果表明，远志糖酯类化合物及其代谢产物是KXS-60% E的主要血中移行成分，原形及代谢产物可能是开心散防治AD的潜在药效物质基础。

关键词

开心散 / 血中移行成分 / 远志糖酯 / 代谢产物

Abstract

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Using a UHPLC-TOF/MS method combined with software of Masslynx V4.1 and database in the literatures, a total of 20 polygala oligosaccharide esters (POEs) were identified in 60% ethanol extract of Kai Xin San (KXS-60%E). Furthermore, 14 POEs as well as 4 POE-metabolites were identified in rat plasma. The results revealed that POEs and POE-metabolites which were identified in rat plasma, were key components in KXS-60%E and the potential bioactive compounds for KXS action in the treatment of AD.

Key words

Kai Xin San / blood transitional component / polygala oligosaccharide ester / metabolite

引用本文

汪娜, AHMAD Hassan, 贾永明, 薛迪, CHAULAGAIN Ram, 黄树明, 刘学伟. UHPLC-MS法鉴定大鼠灌服开心散后血中远志糖酯类化合物及其代谢产物. 药学学报, 2017 , 52 (10) : 1592 -1598 . DOI: 10.16438/j.0513-4870.2017-0373

Na WANG, AHMAD Hassan, Yong-ming JIA, Di XUE, CHAULAGAIN Ram, Shu-ming HUANG, Xue-wei LIU. Identification of polygala oligosaccharide esters and their metabolites in rat plasma after oral administration of ethanol extract of Kai Xin San by UHPLC-MS[J]. Acta Pharmaceutica Sinica, 2017 , 52 (10) : 1592 -1598 . DOI: 10.16438/j.0513-4870.2017-0373

正文

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开心散由“人参、远志、茯苓、石菖蒲”组成, 首见于唐代孙思邈的《备急千金要方》, 是治疗以“好忘”为主要症状的中医情志疾病的经典方剂。近年来研究证实, 开心散在治疗痴呆、抑郁及神经衰弱等疾病中具有明显的药理作用^[1-4], 且开心散的组成药物远志、石菖蒲、人参及茯苓在治疗痴呆类疾病中的使用频率明显高于其他药物^[5], 进一步提示该方具有防治痴呆的药效学特征。

课题组近年来发现开心散60%乙醇提取物(KXS-60% E, 人参-茯苓-远志-石菖蒲比例为3:3:2:2) 能明显缓解由β-淀粉样蛋白(β-amyloid, Aβ)引起的动物认知功能障碍、离体及在体细胞损伤、神经元凋亡及LTP抑制等^[6-8], 提示KXS-60%E中防治AD的药效活性成分客观存在。前期, 课题组建立了KXS-60% E体外成分的UHPLC-MS分析方法, 并对体外成分及大鼠灌服药物后含药血清样品进行了初步研究, 发现在KXS-60% E中远志对全方的贡献度最大^{[9, 10]}, 但具体的化合物结构鉴定工作尚未进行。

糖酯类成分是远志中特有的一类成分, 该类成分以蔗糖为母核, 并以不同的糖苷键连接单糖。近年来对开心散中糖酯成分的活性研究逐渐引起人们的重视。研究表明, 糖酯类化合物可作为抗老化和脑保护新药开发的先导化合物^[11]。Sibiricose A5、A6, 远志酮Ⅲ和3, 6-二芥子酰基蔗糖酯等相继被证实为活性成分^[12]。目前, 3, 6-二芥子酰基蔗糖已收录于2010年《中国药典》, 作为评价远志药材质量标准的标志性成分, 提示糖酯类化合物可能是远志中重要的生物活性物质。

鉴于前期研究中KXS-60% E防治AD的药理作用及目前糖酯类化合物的药理活性, 有必要对KXS-60% E中远志糖酯类成分进行深入的研究。因此, 本研究在前期研究基础上, 进一步对大鼠口服KXS-60% E后的血清移行成分—远志糖酯类成分及代谢产物进行鉴定, 为糖酯类化合物的药理活性及开心散防治AD的直接药效物质基础提供实验依据。

材料与方法

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仪器美国Waters Acquity^TM UPLC液相色谱仪(四元梯度泵-在线真空脱气机-自动进样器-二极管阵列检测器-柱温箱); 美国Waters LCT Premier XE飞行时间质谱仪(电喷雾离子源-正负离子扫描方法-Lockspray); MassLynx V4.1工作站等。

药品与试剂 乙腈(色谱级, 德国, Merck公司); 甲醇(色谱级, 美国, Fisher公司); 蒸馏水(中国, 广州屈臣氏食品饮料有限公司); 甲酸(色谱级, 中国, 天津科密欧化学试剂有限公司); KXS-60% E由课题组前期制备^[9]。

实验动物 Wistar大鼠, 雄性, 体重220~240 g, 由黑龙江中医药大学实验动物中心提供(清洁级, 许可证号为: SCXK沪2006-003), 室温饲养, 自由摄食饮水。于实验前12 h禁食不禁水。

空白及含药血清样品的制备 雄性Wistar大鼠10只, 随机分为空白组及给药组。给药组按5.4 g·kg^-1体重灌服KXS-60% E药粉(用蒸馏水按1.08 g·mL^-1浓度配制, 超声混匀), 给药30 min后采用乙醚吸入麻醉法, 待动物麻醉后, 自眼底留取静脉血1.5 mL, 4 ℃、13 000 r·min^-1离心10 min, 分离获得含药血清备用。空白组给予同体积蒸馏水, 采用相同方法制备空白血清。取大鼠空白血清及给药血清各0.5 mL上样到预先以甲醇2 mL、水2 mL活化平衡好的SPE柱上, 分别以水4 mL和甲醇2 mL淋洗, 淋洗液弃去, 再以甲醇3 mL洗脱, 收集洗脱液, 35 ℃下空气流吹干, 残渣用流动相100 μL复溶, 超声1 min, 再混悬振荡30 s后于4 ℃、13 000 r·min^-1离心10 min, 取上层清液, 进样2 μL供UHPLC分析。

色谱条件^[9] 色谱柱: ACQUITY UPLC^TM BEH C18 column (50 mm × 2.1 mm ID, 1.7 μm); 流速0.4 mL·min^-1; 流动相: A: 0.1%甲酸-水溶液, B: 0.1%甲酸-乙腈溶液; 流动相梯度洗脱: 0~5 min, 5%~30% A; 5~12 min, 30%~50% A; 12~15 min, 50%~80% A; 15~16 min, 80%~100% A; 柱温: 40 ℃; 进样体积: 2 μL。

质谱条件^[9] 电喷雾离子源(ESI), 采用正负离子扫描检测; 毛细管电压为1 000 V; 样本锥孔电压为40 V; 离子源温度为100 ℃; 脱溶剂温度为350 ℃; 脱溶剂气流量扫描为700 L·h^-1, 锥孔气流量为10 L·h^-1; 微通道板电压为2 300 V, 扫描时间为0.3 s; 准确质量校正采用亮氨酸-脑啡肽(leucine-enkephalin, [M+H]⁺ m/z 556.277 1), 浓度为1.0 ng·mL^-1, 校正溶液进样速度为30 μL·min^-1, 校正频率为15 s; 扫描方式为全扫描, 质量扫描范围m/z 100~2 000。

远志糖酯类成分的结构鉴定 UHPLC-TOF-MS检出的信息可以确定化合物的分子质量及色谱峰的来源归属, 能够界定化合物的检索范围。对单个化合物进行Q-TOF-MS/MS扫描可以获得碎片离子信息。同时, 利用Masslynx V 4.1的数据平台, 可以确定其分子式的可能组成。在鉴定时, 利用化合物的精确分子质量或分子式检索现有的化合物, 综合对比化合物的归属、母离子、子离子的信息, 并参考相关研究的文献, 确定化合物的结构。

结果

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1 开心散中远志糖酯类成分的鉴定

在KXS-60% E色谱峰中, 鉴定了全部20个远志糖酯类成分(图 1a, b)。根据结构不同分为两类:母核中含有2个单糖的蔗糖酯(图 1c)和3个以上的寡糖酯(图 1d)。其主要化合物鉴定信息及分子组成等见表 1。

2 开心散中远志糖酯类成分及其代谢物入血成分鉴定

通过对比分析含药血清、空白血清中色谱峰的保留时间及质谱数据, 结合体外成分分析, 发现Peak 1~5、8、10~14、16~18均以原形成分吸收入血(图 2、表 1), 另发现4个代谢产物(图 2、表 2)。

代谢产物的结构鉴定与原形成分鉴定略有不同。以M3为例, 在负正模式下分别以m/z = 283 [M+HCOO]^-及m/z = 239 [M+H]⁺被检出, 其分子质量可以确定为238。利用Masslynx V 4.1的数据平台中“Elemental composition”功能, 可以推测其分子组成是C₁₂H₁₄O₅ (i-Fit = 0) 和C₁₉H₁₁ (i-Fit = 3.3)。二级质谱数据中, M3在正离子模式下给出的数据是m/z = 223和m/z = 209。分析这些离子碎片, 发现与3, 4, 5-三甲氧基肉桂酸(3, 4, 5-trimethoxycinnamylic acid)离子裂解后碎片所产生的离子峰相符, 离子碎片223是该化合物羧基被还原成醛基, 209是进一步脱去了甲基而形成的羟基, 因此, M3被确定为3, 4, 5-三甲氧基肉桂酸。通过这种方式, 鉴定了其余的3个代谢产物, 化合物的相应信息见表 2。

讨论

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本研究鉴定了开心散醇提取物中的远志糖酯类成分, 并对该类成分的大鼠血中移行情况进行分析。研究结果表明, 鉴定的20个糖酯类化合物主要分为两类:母核中含有2个单糖的蔗糖酯和3个单糖以上的寡糖酯。体外成分分析中两类成分的色谱峰集中分布, 提示能够通过改变不同的溶液浓度, 实现对组分的进一步细化与富集。体内分析结果显示, 有14个原形成分是大鼠血中移行成分, 占体外该类成分的70%, 提示该类成分是开心散醇提取物中的主要入血成分, Liu^[13]的研究结果也显示, 在灌服了开心散水提取物的大鼠血清中, 有8个寡糖酯类成分被吸收入血, 与本研究结果相似。而对开心散的水提取物研究结果显示, 在开心散中有26个原形成分被吸收入血, 主要以人参皂苷为主, 而血中检测到的3个代谢物也主要来源于人参皂苷, 这与本研究的分析结果存在着一定差异^{[14, 15]}, 可能是由于提取物不同及检测条件与方法上的差异而引起的, 提示在药效物质基础判定应充分建立在药理学基础之上。

寡糖酯类成分是远志中特有的一类糖酯类成分, 该类成分以蔗糖为母核, 并以不同的糖苷键连接单糖(以葡萄糖为主, 少数为鼠李糖)。Yang等^[11]的研究结果表明, 寡糖酯类化合物可能是远志中主要的生物活性物质, 并可作为抗老化和脑保护新药开发的先导化合物。近年来对开心散中寡糖酯的活性研究逐渐引起人们的重视, 石任兵教授所领导的课题组^{[12, 16]}研究发现, 开心散中sibiricose A5、A6, 远志酮Ⅲ和3, 6-二芥子酰基蔗糖等均为有效成分, 并建立了开心散类药有效组分的远志特征图谱质量表征方法, 并分别对3, 6-二芥子酰基蔗糖等进行了药动学研究, 为该类研究提供了可供借鉴的方法, 也反映出远志中寡糖酯类成分的潜在的活性作用。目前, 3, 6-二芥子酰基蔗糖已收录于2010年《中国药典》, 作为评价远志药材质量标准的标志性成分。

值得注意的是, 糖酯类成分存在于多种植物中, 但母核结构中含有三糖及以上的寡糖多酯, 只在远志属植物中被发现, 因此有研究认为, 该类成分可能具有某些特殊的药理作用^[17]。本研究结果表明, tenuifoliside A、tenuifoliside C (E)、tenuifoliose B (D)、tenuifoliside K及tenuifoliside I (J)是药物的吸收入血成分, 有些化合物已被证实具有明显的生物活性, 如tenuifoliside A可以促进原代培养大鼠海马神经元的存活, 具有神经营养作用, 其机制可能与激活Trek/BDNF/ERK循环通路有关^[18]。Tenuifoliside B能够通过增强胆碱能系统而改善东莨菪碱诱导的小鼠/大鼠被动回避行为的损伤, 提示tenuifoliside B是增强认知功能和脑保护作用的活性成分^[19]。

值得一提的是, 代谢产物3, 4, 5-trimethoxycinnamylic acid及3, 4, 5-trimethoxycinnamylic acid -(CH₂)既是远志皂苷类如onjisaponin G、E的代谢产物, 又是远志糖酯类成分如sibiricose A1、sibiricose A6、tenuifoliside B、3, 6'-disinapoyl sucrose和arillanin A的代谢产物, 研究表明, 二者均易透过胃肠单层细胞膜, 在大鼠体内吸收良好, 生物利用度高, 且活性作用已有部分阐明, 因此, 应进一步对其作用机制进行深入分析, 以全面阐明开心散及远志的直接药效物质基础。

基金

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国家自然科学基金青年基金资助项目(81303248)
黑龙江省普通本科高等学校青年创新人才培养计划(UNPYSCT-2016116)
齐齐哈尔医学院校内博士创新基金资助项目(QY2016B-09)
黑龙江省自然科学基金资助项目(H2015028)
黑龙江省教育厅基本科研业务费基础研究项目(2016-KYYWF-0865)

参考文献

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文献

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参考文献引证文献

排序方式：

[1]

Chu H, Zhang A, Han Y, et al. Metabolomics approach to explore the effects of Kai-Xin-San on Alzheimer's disease using UPLC/ESI-Q-TOF mass spectrometry[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2016, 1015-1016:50-61.

[2]

Lin Z, Gu J, Xiu J, et al. Traditional Chinese medicine for senile dementia[J]. Evid Based Complement Altern Med, 2012, 2012:692621-692633.

[3]

Sun ZK, Yang HQ, Chen SD. Traditional Chinese medicine:a promising candidate for the treatment of Alzheimer's disease[J]. Transl Neurodegener, 2013, 2:6.

[4]

Yan L, Hu Q, Mak MS, et al. A Chinese herbal decoction, reformulated from Kai-Xin-San, relieves the depression-like symptoms in stressed rats and induces neurogenesis in cultured neurons[J]. Sci Rep, 2016, 6:30014.

[5]

May BH, Lu C, Lu Y, et al. Chinese herbs for memory disorders:a review and systematic analysis of classical herbal literature[J]. J Acupunct Meridian Stud, 2013, 6:2-11.

[6]

Li Y, Liu B, Liu XW, et al. Mechanism research of Kai-xin granule on Aβ-induced cognitive impairment in rats[J]. Inf Tradit Chin Med (中医药信息), 2016, 33:34-37.

[7]

Wen W, Zhang C, Liu M, et al. The protective effect of the serum containing drugs of Kaixin san against Aβ-induced toxicity in PC12 cells[J]. Inf Tradit Chin Med (中医药信息), 2012, 29:80-81.

[8]

Huang SM, Ren LM, Li JY, et al. Kaixin san improving Aβ1-42 inhibition in vivo hippocampal LTP[J]. Inf Tradit Chin Med (中医药信息), 2013, 30:61-63.

[9]

Liu XW, Liu S, Huang SM, et al. Plasma pharmacochemis-try study of effective extractive from Kai-xinsan on Alzhemer's disease[J]. Chin J Exp Tradit Med Form (中国实验方剂学杂志), 2014, 20:179-183.

[10]

Liu XW, Zhang B, Wang N, et al. Research overview of Kai xinsan in preventing and treating AD based on serum pharmacochemistry of TCM[J]. Acta Chin Med and Pharm (中医药学报), 2016, 44:101-104.

[11]

Yang XD, Zhang L, Liang B, et al. Oligosaccharide esters isolated from plants of Polygalaceae[J]. Chin Tradit Herb Drugs (中草药), 2002, 33:954-958.

[12]

Jiang YY, Dai Y, Ba YY, et al. Quality representation of Yuanzhi (Radix Polygalae) based on characteristic chroma-tograms of drug-like effective components of Kai Xin San-Yuanzhi[J]. J Beijing Univ Tradit Chin Med (北京中医药大学学报), 2011, 34:544-547.

[13]

Liu CF, Yang WZ, Liu KD, et al. Characterization of chemical constituents and in vivo metabolites of Kai-Xin-San prescription by HPLC/DAD/ESI-MSn[J]. J Chin Pharm Sci, 2012, 21:569-576.

[14]

Lü CX, He BS, Sui ZY, et al. Identification and determina-tion of the major constituents in Kai-Xin-San by UPLC-Q/TOF MS and UFLC-MS/MS method[J]. J Mass Spectrom, 2016, 51:479-490.

[15]

Zhang XW, Li Q, Lü CX, et al. Characterization of multiple constituents in Kai-Xin-San prescription and rat plasma after oral administration by liquid chromatography with quadrupole time-of-flight tandem mass spectrometry[J]. J Sep Sci, 2015, 38:2068-2075.

[16]

Ba YY, Jiang YY, Liu Y, et al. Comparative pharmacokinetics of 3, 6'-disinapoyl sucrose after oral administration of pure 3, 6'-disinapoyl Sucrose, Radix Polygalae extract and Kaixinsan in acquired dysmnesia model rats[J]. Chin J Exp Tradit Med Form (中国实验方剂学杂志), 2012, 18:138-142.

[17]

Li CJ. The Bioactivity of Chemical Constituents in Polygala tenuifolia and Polygala glomerata Lour (远志和华南远志的化学成分及其生物活性研究)[D]. Beijing: Chinese Academy of Medical Sciences & Peking Union Medical College, 2008.

[18]

Yu BY. The Mechanism of Antidepressive Effects by Kai-Xin-San on CMS rats and Studies on Neuroprotective Effects of Tenuifoliside A via ERK and PI3K Pathway (开心散治疗CMS抑郁模型大鼠的作用机制及活性成分Tenuifoliside A基于ERK和PI3K通路介导的神经保护机制研究)[D]. Zhangjiakou: Hebei North University, 2013.

[19]

Ikeya Y, Takeda S, Tunakawa M, et al. Cognitive improving and cerebral protective effects of acylated oligosaccharides in Polygala tenuifolia[J]. Biol Pharm Bull, 2004, 27:1081-1085.

2017年第52卷第10期

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文章信息

doi: 10.16438/j.0513-4870.2017-0373

接收时间：2017-04-17
首发时间：2026-01-14
出版时间：2017-10-12

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收稿日期：2017-04-17
修回日期：2017-06-30

基金

国家自然科学基金青年基金资助项目(81303248)

黑龙江省普通本科高等学校青年创新人才培养计划(UNPYSCT-2016116)

齐齐哈尔医学院校内博士创新基金资助项目(QY2016B-09)

黑龙江省自然科学基金资助项目(H2015028)

黑龙江省教育厅基本科研业务费基础研究项目(2016-KYYWF-0865)

作者信息

1.齐齐哈尔医学院, 黑龙江齐齐哈尔 161006

2.黑龙江中医药大学, 黑龙江哈尔滨 150040

通讯作者:

* 刘学伟, Tel/Fax:86-452-2663804, E-mail:liuxuewei2001@163.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.	Identified compound	t_R _/min	[M+HCOO]^- (m/z)	ESI-MS/MS (-) (m/z)	ESI-MS/MS (+) (m/z)	Elemental composition	Substituent Group
1^*	Sibiricose A3	2.64	507.136 5	507.135 0	3.0	507 [M+HCOO]^-; 323 [M-H-HB]^-; 137 [HB-H]^-	-	C₁₉H₂₆O₁₃	R₁=H, R₂=HB
2^*	Arillanin C	3.30	563.159 0	563.161 2	-3.9	563 [M+HCOO]^-; 323 [M-H-MHC]^-	519 [M+H]⁺; 325 [M+H-MHC]⁺	C₂₂H₃₀O₁₄	R₁=HDC, R₂=H
3^*	Sibiricose A6	3.43	593.173 8	593.171 8	3.4	593 [M+HCOO]^-; 323 [M-H-TMC]^-; 223 [TMC-H]^-	549 [M+H]⁺; 325 [M+H-TMC]⁺	C₂₃H₃₂O₁₅	R₁=HDC, R₂=H
4^*	Sibiricose A1	3.76	593.171 6	593.171 8	-0.3	593 [M+HCOO]^-; 323 [M-H-TMC]^-; 223 [TMC-H]^-	549 [M+H]⁺; 325 [M+H-TMC]⁺	C₂₃H₃₂O₁₅	R₁=H, R₂=HDC
5^*	Sibiricose A2	4.53	607.187 0	607.187 4	-0.7	607[M+HCOO]^-; 223 [TMC-H]^-; 323 [M-H-TMC]^-	563 [M+H]⁺; 339 [M+H-TMC]⁺	C₂₄H₃₄O₁₅	R₁=TMC, R₂=H
6	Tenuifoliside B	4.73	713.192 4	713.192 9	-0.7	713 [M+HCOO]^-; 323 [M-H-HB-HDC]^-	669 [M+H]⁺; 549 [M+H-HB]⁺; 325 [M+H-HB-HDC]⁺	C₃₀H₃₆O₁₇	R₁=HDC, R₂=HB
7	3, 6- Disinapoylsucrose & Sibiricose A4	4.80	799.225 7	799.229 7	-5.0	799 [M+HCOO]^-; 323 [M-H-2HDC]^-	755 [M+H]⁺; 549 [M+H-HDC]⁺; 325 [M+H-2HDC]⁺	C₃₄H₄₂O₁₉	R₁=R₂=HDC
8^*	Sibiricose A4 & 3, 6- Disinapoylsucrose	5.36	799.233 1	799.229 7	4.3	799 [M+HCOO]^-; 547 [M-H-HDC]^-; 323 [M-H-2HDC]^-	755 [M+H]⁺; 549 [M+H-HDC]⁺; 325 [M+H-2HDC]⁺	C₃₄H₄₂O₁₉	R₁=R₂=HDC
9	Arillanin A	5.47	769.220 1	769.219 1	1.3	769 [M+HCOO]^-; 499 [M-H-HDC]^-; 529 [M-H-MHC]^-; 323 [M-H-HDC-MHC]^-	725 [M+H]⁺; 325 [M+H-HDC-MHC]⁺	C₃₃H₄₀O₁₈	R₁=R₂=HDC
10^*	Tenuifoliside A	5.89	727.209 0	727.208 6	0.6	727 [M+HCOO]^-; 543 [M-H-HB]^-; 443 [M-H-TMC]^-	683 [M+H]⁺; 545 [M+H-HB]⁺	C₃₁H₃₈O₁₇	R₁=TMC, R₂=HB
11^*	Tenuifoliose K	6.20	1 311.382 1	1 311.382 7	-0.4	1 311 [M+HCOO]^-; 1 119 [M-H-HC]^-; 1 077 [M-H-HC-C₂H₂O]^-	1 267 [M+H]⁺; 1 121 [M+H-HC]⁺; 1 079 [M+H-HC-C₂H₂O]⁺	C₅₇H₇₀O₃₂	R₁=R₃=R₄=H, R₂=Ac
12^*	Tenuifoliose C & Tenuifoliose E	6.37	1 341.392 4	1 341.393 2	-0.5	1 341 [M+HCOO]^-; 1 119 [M-H-MHC]^-	1 297 [M+H]⁺; 1 121 [M+H-MHC]⁺; 1 079 [M+H-MHC-C₂H₂O]⁺	C₅₈H₇₂O₃₃	R₁=Ac, R₂=OCH₃, R₃=R₄=H
13^*	Tenuifoliose I or Tenuifoliose J	6.52	1 353.389 1	1 353.393 2	-3.0	1 353 [M+HCOO]^-; 1 119 [M-H-HC-C₂H₂O]^-; 1 101 [M-H-HC-2C₂H₂O]^-	1 309 [M+H]⁺; 1 163 [M+H-HC]⁺; 1 121 [M+H-HC-C₂H₂O]⁺; 1 103 [M+H-HC-2C₂H₂O]⁺	C₆₁H₉₆O₃₀	R₁=R₃=Ac, R₂=R₄=H
14^*	TenuifolioseB & Tenuifoliose D	6.59	1 383.404 0	1 383.403 8	0.2	1 383 [M+HCOO]^-; 1 337 [M-H]^-; 1 161 [M-H-MHC]^-; 1 119 [M-H-MHC-C₂H₂O]^-; 1 077 [M-H-MHC-2C₂H₂O]^-; 973 [M-H-MHC-2C₂H₂O- (HB-OH)]^-	1 339 [M+H]⁺; 1 163 [M+H-MHC]⁺; 1 121 [M+H-MHC-C₂H₂O]⁺; 1 079 [M+H-MHC-2C₂H₂O]⁺	C₆₀H₇₄O₃₄	R₁=H, R₂=OCH₃, R₃=R₄=Ac
15	Tenuifoliose C & Tenuifoliose E	6.75	1 341.392 8	1 341.393 2	-0.2	1 341 [M+HCOO]^-; 1 119 [M-H-MHC]^-	1 297 [M+H]⁺; 1 121 [M+H-MHC]⁺; 1 079 [M+H-MHC-C₂H₂O]⁺	C₅₈H₇₂O₃₃	R₁=H, R₂=OCH₃, R₃=Ac, R₄=Glu
16^*	Tenuifoliose B & Tenuifoliose D	7.13	1 383.404 5	1 383.403 8	-0.3	1 383 [M+HCOO]^-; 1 161 [M-H-MHC]^-; 973 [M-H-MHC-2C₂H₂O- (HB-OH)]^-	1 339 [M+H]⁺; 1 163 [M+H-MHC]⁺; 1 079 [M+H-MHC-2C₂H₂O]⁺	C₆₀H₇₄O₃₄	R₁=R₃=Ac, R₂=OCH₃, R₄=H
17^*	Tenuifoliose A	7.43	1 425.414 0	1 425.414 4	-0.3	1 425 [M+HCOO]^-; 1 379 [M-H]^-; 1 203 [M-H-HHC]^-	1 381 [M+H]⁺; 1 205 [M+H-HHC]⁺; 1 163 [M+H-HHC-C₂H₂O]⁺; 1 179 [M+H-HHC-3C₂H₂O]⁺	C₆₂H₇₆O₃₅	R₁=R₃=R₄=Ac, R₂=CH₂OH
18^*	Reiniose J	7.59	1 383.401 6	1 338.403 8	-1.8	1 383 [M+HCOO]^-; 1 161 [M-H-MHC]^-	1 339 [M+H]⁺; 1 163 [M+H-MHC]⁺; 1 297 [M+H-C₂H₂O]⁺; 1 079 [M+H-MHC-2C₂H₂O]⁺	C₆₀H₇₄O₃₄	R₁=R₃=Ac, R₂=OCH₃, R₄=H
19	Tenuifoliose I & Tenuifoliose J	7.80	1 353.394 0	1 353.393 2	0.3	1 353 [M+HCOO]^-; 1 161 [M-H-HC]^-; 1 119 [M-H-HC-C₂H₂O]^-; 1 101 [M-H-HC-2C₂H₂O]^-	-	C₆₁H₉₆O₃₀	R₁=R₄=Ac, R₂=R₃=H,
20	Tenuifoliose N	11.02	1 455.420 5	1 455.424 9	-3.8	1 455 [M+HCOO]^-; 1 233 [M-H-MHC]^-; 1 191 [M-H-MHC-C₂H₂O]^-; 1 149 [M-H-MHC-2C₂H₂O]^-	-	C₆₃H₇₈O₃₆	R₁=R₃=R₄=Ac, R₂=O

No.

Identified
compound

t_R
_/min

[M+HCOO]^- (m/z)

ESI-MS/MS (-)
(m/z)

ESI-MS/MS (+)
(m/z)

Elemental composition

Substituent Group

Measured

Calculated

Δ
(ppm)

1^*

Sibiricose A3

2.64

507.136 5

507.135 0

3.0

507 [M+HCOO]^-;
323 [M-H-HB]^-;
137 [HB-H]^-

C₁₉H₂₆O₁₃

R₁=H,
R₂=HB

2^*

Arillanin C

3.30

563.159 0

563.161 2

-3.9

563 [M+HCOO]^-;
323 [M-H-MHC]^-

519 [M+H]⁺;
325 [M+H-MHC]⁺

C₂₂H₃₀O₁₄

R₁=HDC,
R₂=H

3^*

Sibiricose A6

3.43

593.173 8

593.171 8

3.4

593 [M+HCOO]^-;
323 [M-H-TMC]^-;
223 [TMC-H]^-

549 [M+H]⁺;
325 [M+H-TMC]⁺

C₂₃H₃₂O₁₅

R₁=HDC,
R₂=H

4^*

Sibiricose A1

3.76

593.171 6

593.171 8

-0.3

593 [M+HCOO]^-;
323 [M-H-TMC]^-;
223 [TMC-H]^-

549 [M+H]⁺;
325 [M+H-TMC]⁺

C₂₃H₃₂O₁₅

R₁=H,
R₂=HDC

5^*

Sibiricose A2

4.53

607.187 0

607.187 4

-0.7

607[M+HCOO]^-;
223 [TMC-H]^-;
323 [M-H-TMC]^-

563 [M+H]⁺;
339 [M+H-TMC]⁺

C₂₄H₃₄O₁₅

R₁=TMC,
R₂=H

Tenuifoliside B

4.73

713.192 4

713.192 9

-0.7

713 [M+HCOO]^-;
323 [M-H-HB-HDC]^-

669 [M+H]⁺;
549 [M+H-HB]⁺;
325 [M+H-HB-HDC]⁺

C₃₀H₃₆O₁₇

R₁=HDC,
R₂=HB

3, 6-
Disinapoylsucrose
& Sibiricose A4

4.80

799.225 7

799.229 7

-5.0

799 [M+HCOO]^-;
323 [M-H-2HDC]^-

755 [M+H]⁺;
549 [M+H-HDC]⁺;
325 [M+H-2HDC]⁺

C₃₄H₄₂O₁₉

R₁=R₂=HDC

8^*

Sibiricose A4 &
3, 6-
Disinapoylsucrose

5.36

799.233 1

799.229 7

4.3

799 [M+HCOO]^-;
547 [M-H-HDC]^-;
323 [M-H-2HDC]^-

755 [M+H]⁺;
549 [M+H-HDC]⁺;
325 [M+H-2HDC]⁺

C₃₄H₄₂O₁₉

R₁=R₂=HDC

Arillanin A

5.47

769.220 1

769.219 1

1.3

769 [M+HCOO]^-;
499 [M-H-HDC]^-;
529 [M-H-MHC]^-;
323 [M-H-HDC-MHC]^-

725 [M+H]⁺;
325 [M+H-HDC-MHC]⁺

C₃₃H₄₀O₁₈

R₁=R₂=HDC

10^*

Tenuifoliside A

5.89

727.209 0

727.208 6

0.6

727 [M+HCOO]^-;
543 [M-H-HB]^-;
443 [M-H-TMC]^-

683 [M+H]⁺;
545 [M+H-HB]⁺

C₃₁H₃₈O₁₇

R₁=TMC,
R₂=HB

11^*

Tenuifoliose K

6.20

1 311.382 1

1 311.382 7

-0.4

1 311 [M+HCOO]^-;
1 119 [M-H-HC]^-;
1 077 [M-H-HC-C₂H₂O]^-

1 267 [M+H]⁺;
1 121 [M+H-HC]⁺;
1 079 [M+H-HC-C₂H₂O]⁺

C₅₇H₇₀O₃₂

R₁=R₃=R₄=H,
R₂=Ac

12^*

Tenuifoliose C & Tenuifoliose E

6.37

1 341.392 4

1 341.393 2

-0.5

1 341 [M+HCOO]^-;
1 119 [M-H-MHC]^-

1 297 [M+H]⁺;
1 121 [M+H-MHC]⁺;
1 079 [M+H-MHC-C₂H₂O]⁺

C₅₈H₇₂O₃₃

R₁=Ac, R₂=OCH₃, R₃=R₄=H

13^*

Tenuifoliose I or Tenuifoliose J

6.52

1 353.389 1

1 353.393 2

-3.0

1 353 [M+HCOO]^-;
1 119 [M-H-HC-C₂H₂O]^-;
1 101 [M-H-HC-2C₂H₂O]^-

1 309 [M+H]⁺;
1 163 [M+H-HC]⁺;
1 121 [M+H-HC-C₂H₂O]⁺;
1 103 [M+H-HC-2C₂H₂O]⁺

C₆₁H₉₆O₃₀

R₁=R₃=Ac, R₂=R₄=H

14^*

TenuifolioseB & Tenuifoliose D

6.59

1 383.404 0

1 383.403 8

0.2

1 383 [M+HCOO]^-;
1 337 [M-H]^-;
1 161 [M-H-MHC]^-;
1 119 [M-H-MHC-C₂H₂O]^-;
1 077 [M-H-MHC-2C₂H₂O]^-;
973 [M-H-MHC-2C₂H₂O-
(HB-OH)]^-

1 339 [M+H]⁺;
1 163 [M+H-MHC]⁺;
1 121 [M+H-MHC-C₂H₂O]⁺;
1 079 [M+H-MHC-2C₂H₂O]⁺

C₆₀H₇₄O₃₄

R₁=H, R₂=OCH₃, R₃=R₄=Ac

Tenuifoliose C & Tenuifoliose E

6.75

1 341.392 8

1 341.393 2

-0.2

1 341 [M+HCOO]^-;
1 119 [M-H-MHC]^-

1 297 [M+H]⁺;
1 121 [M+H-MHC]⁺;
1 079 [M+H-MHC-C₂H₂O]⁺

C₅₈H₇₂O₃₃

R₁=H, R₂=OCH₃, R₃=Ac,
R₄=Glu

16^*

Tenuifoliose B & Tenuifoliose D

7.13

1 383.404 5

1 383.403 8

-0.3

1 383 [M+HCOO]^-;
1 161 [M-H-MHC]^-;
973 [M-H-MHC-2C₂H₂O-
(HB-OH)]^-

1 339 [M+H]⁺;
1 163 [M+H-MHC]⁺;
1 079 [M+H-MHC-2C₂H₂O]⁺

C₆₀H₇₄O₃₄

R₁=R₃=Ac, R₂=OCH₃,
R₄=H

17^*

Tenuifoliose A

7.43

1 425.414 0

1 425.414 4

-0.3

1 425 [M+HCOO]^-;
1 379 [M-H]^-;
1 203 [M-H-HHC]^-

1 381 [M+H]⁺;
1 205 [M+H-HHC]⁺;
1 163 [M+H-HHC-C₂H₂O]⁺;
1 179 [M+H-HHC-3C₂H₂O]⁺

C₆₂H₇₆O₃₅

R₁=R₃=R₄=Ac,
R₂=CH₂OH

18^*

Reiniose J

7.59

1 383.401 6

1 338.403 8

-1.8

1 383 [M+HCOO]^-;
1 161 [M-H-MHC]^-

1 339 [M+H]⁺;
1 163 [M+H-MHC]⁺;
1 297 [M+H-C₂H₂O]⁺;
1 079 [M+H-MHC-2C₂H₂O]⁺

C₆₀H₇₄O₃₄

R₁=R₃=Ac, R₂=OCH₃,
R₄=H

Tenuifoliose I & Tenuifoliose J

7.80

1 353.394 0

1 353.393 2

0.3

1 353 [M+HCOO]^-;
1 161 [M-H-HC]^-;
1 119 [M-H-HC-C₂H₂O]^-;
1 101 [M-H-HC-2C₂H₂O]^-

C₆₁H₉₆O₃₀

R₁=R₄=Ac, R₂=R₃=H,

Tenuifoliose N

11.02

1 455.420 5

1 455.424 9

-3.8

1 455 [M+HCOO]^-;
1 233 [M-H-MHC]^-;
1 191 [M-H-MHC-C₂H₂O]^-;
1 149 [M-H-MHC-2C₂H₂O]^-

C₆₃H₇₈O₃₆

R₁=R₃=R₄=Ac,
R₂=O

No.	Identified compound	t_R _/min	[M+H]⁺ (m/z)	ESI-MS/MS (-) (m/z)	ESI-MS/MS (+) (m/z)	Elemental composition
M1	Tenuifoliside B-(HDC)	4.29	463.144 8	463.145 2	-1.8	507 [M+HCOO]^-; 137 [HB-H]^-	463 [M+H]⁺; 325 [M-HB+H]⁺; 139 [HB+H]⁺	C₁₉H₂₆O₁₃
M2	Tenuifoliose K-(CH₂O)	4.90	1 239.396 0	1 239.397 9	-3.4	1 237 [M-H]^-; 1 136 [M-H-C₈H₇O+H₂O]^-; 1 034 [M-H-2C₈H₇O+H₂O]^-	1 239 [M+H]⁺; 1 138 [M+H-C₈H₇O+H₂O]⁺; 1 036 [M+H-2C₈H₇O+H₂O]⁺	C₅₆H₇₀O₃₁
M3	3, 4, 5-Trimethoxycinnamylic acid	6.92	239.091 0	239.091 9	-3.8	283 [M+HCOO]^-; 237 [M-H]^-; 221 [M-O-H]^-	239 [M+H]⁺; 223 [M-O+H]⁺; 209 [M-O-CH₂+H]⁺	C₁₂H₁₄O₅
M4	3, 4, 5-Trimethoxycinnamylic acid-(CH₂)	7.35	225.074 1	225.076 3	-4.1	223 [M-H]^-; 209 [M-O+H]⁺; 269 [M+HCOO]^-	225 [M+H]⁺; 209 [M-O+H]⁺	C₁₁H₁₂O₅

No.

Identified compound

t_R
_/min

[M+H]⁺ (m/z)

ESI-MS/MS (-)
(m/z)

ESI-MS/MS (+)
(m/z)

Elemental
composition

Measured

Calculated

Δ(ppm)

Tenuifoliside B-(HDC)

4.29

463.144 8

463.145 2

-1.8

507 [M+HCOO]^-; 137 [HB-H]^-

463 [M+H]⁺; 325 [M-HB+H]⁺; 139 [HB+H]⁺

C₁₉H₂₆O₁₃

Tenuifoliose K-(CH₂O)

4.90

1 239.396 0

1 239.397 9

-3.4

1 237 [M-H]^-; 1 136 [M-H-C₈H₇O+H₂O]^-; 1 034 [M-H-2C₈H₇O+H₂O]^-

1 239 [M+H]⁺; 1 138 [M+H-C₈H₇O+H₂O]⁺; 1 036 [M+H-2C₈H₇O+H₂O]⁺

C₅₆H₇₀O₃₁

3, 4, 5-Trimethoxycinnamylic acid

6.92

239.091 0

239.091 9

-3.8

283 [M+HCOO]^-; 237 [M-H]^-; 221 [M-O-H]^-

239 [M+H]⁺; 223 [M-O+H]⁺; 209 [M-O-CH₂+H]⁺

C₁₂H₁₄O₅

3, 4, 5-Trimethoxycinnamylic acid-(CH₂)

7.35

225.074 1

225.076 3

-4.1

223 [M-H]^-; 209 [M-O+H]⁺; 269 [M+HCOO]^-

225 [M+H]⁺; 209 [M-O+H]⁺

C₁₁H₁₂O₅