药学学报

No.	Compound	Formula	t_R/min	Ion mode	Theoretical value	Measured value	Error (ppm)	Ion fragment	Classification	Source
1	5-Hydroxymethylfurfural^[21]	C₆H₆O₃	0.93	[M+H]⁺	127.038 9	127.037 2	-4.25	81.032 7, 109.026 9, 127.037 1	F	GZ
2^*	Quinic acid^[21]	C₇H₁₂O₆	1.09	[M-H]^-	191.056 1	191.054 6	-2.27	93.032 9, 128.038 3	A	GZ
3	Malic acid^[21]	C₄H₆O₅	1.28	[M-H]^-	133.014 2	133.012 5	4.81	71.012 1, 115.001 8	A	GZ
4	Trimethyl citrate^[21]	C₇H₁₀O₇	2.09	[M-H]^-	205.035 3	205.033 9	-1.99	87.068 8, 111.006 9, 143.033 6	A	GZ
5	Triethyl citrate^[21]	C₈H₁₂O₇	2.27	[M-H]^-	219.051 0	219.049 4	-2.59	87.001 0, 111.007 2, 157.075 3	A	GZ
6	2-Isopropylmalic acid^[21]	C₇H₁₂O₅	11.77	[M-H]^-	175.061 1	175.059 5	-3.19	85.064 1, 113.059 0, 115.038 3	A	GZ
7^*	Protocatechuic acid^[21]	C₇H₆O₄	13.83	[M-H]^-	153.019 3	153.017 6	-4.28	91.016 8, 108.051 4, 109.027 7	A	GZ
8^*	Ferulic acid^[21]	C₁₀H₁₀O₄	15.53	[M-H]^-	193.050 6	193.047 4	-7.11	134.035 6, 178.025 5, 193.047 3	A	GZ
9^*	Caffeic acid^[17]	C₉H₈O₄	18.32	[M-H]^-	179.034 9	179.033 4	-2.54	135.043 4, 179.033 4	A	DS
10	Danshensu^[17]	C₉H₁₀O₅	19.01	[M-H]^-	197.049 9	197.044 1	-1.72	178.996 9, 197.044 1	A	DS
11	Isorhamnetin^[23]	C₇H₆O₄	19.43	[M-H]^-	153.019 3	315.052 3	7.36	151.038 3, 315.052 2	D	SJ/SN
12	Scopolamine^[21]	C₁₀H₈O₄	19.63	[M+H]⁺	193.049 5	193.046 5	-5.77	122.058 2, 133.099 2, 149.958 7, 178.047 0, 193.046 4	C	GZ
13	Xenognoside B^[22]	C₁₆H₁₂O₅	20.14	[M-H]^-	283.061 1	283.060 1	-0.03	195.137 2, 224.046 2, 268.036 5, 283.060 0	D	JX
14	3-Hydroxyvestitone^[22]	C₁₆H₁₄O₆	20.17	[M-H]^-	301.071 7	301.070 5	-0.64	135.007 1, 301.070 4	D	JX
15	Homoeriodictyol^[22]	C₁₆H₁₄O₆	20.17	[M-H]^-	301.071 7	301.070 5	-0.64	121.027 2, 135.071 3, 177.017 6, 178.020 7, 268.036 5, 301.070 4	D	JX
16	2-2-(2, 4-Dimethoxy-phenyl)-2-oxoe-thoxy-4-hydrobenzoic acid^[22]	C₁₇H₁₆O₇	20.21	[M-H]^-	331.082 3	331.081 5	0.84	176.010 5, 255.030 3, 301.069 7, 331.081 5	A	JX
17	Myristicin^{[19, 20]}	C₁₁H₁₂O₃	20.7	[M+H]⁺	193.085 9	193.082 3	-8.54	105.068 7, 133.062 6, 165.087 8, 193.082 3	C	RDK
18	Ellagic acid^[23]	C₁₄H₆O₈	20.78	[M-H]^-	300.998 9	300.997 8	-0.41	185.059 4, 229.049 1, 257.116 3, 283.059 8	A	SN/SJ
19^*	Kaempferol^[23]	C₁₅H₁₀O₆	21.3	[M+H]⁺	287.055 0	287.054 1	-3.25	153.052 1, 165.052 1, 286.074 3, 287.054 0, 289.136 5	D	SN
20	α-Curcumene^[19]	C₁₅H₂₂	22.47	[M+H]⁺	203.179 4	203.176 0	-6.72	81.069 2, 95.084 6, 109.099 6, 147.114 6, 203.176 0	E	TX
21	(3R)-7, 3′-Dihydroxy-6, 2′, 4′-trime-thoxyisoflavanone^[22]	C₁₈H₁₈O₇	23.48	[M-H]^-	345.097 9	345.098 9	5.71	149.095 3, 271.135 6, 301.143 3, 315.049 9, 330.072 6, 345.098 8	D	JX
22	5, 7-Dihydroxy-2′, 3′, 4′-trimethoxyisofla-vanone^[22]	C₁₈H₁₈O₇	23.67	[M-H]^-	345.097 9	345.096 8	-0.23	135.007 6, 151.038 4, 177.017 9, 295.060 7, 315.049 3, 330.072 7, 345.096 8	D	JX
23	Fisetin^[22]	C₁₅H₁₀O₆	23.68	[M-H]^-	285.040 4	285.042 7	6.56	121.064 3, 149.022 4, 162.098 6, 239.164 1, 257.080 3, 285.042 6	D	JX
24	Fustin^[22]	C₁₅H₁₂O₆	24.18	[M-H]^-	287.056 1	287.058 2	4.09	109.027 5, 149.058 7, 225.111 3	D	JX
25	Genistein^[22]	C₁₅H₁₀O₅	24.34	[M-H]^-	269.045 5	269.044 5	0.14	133.064 3, 135.079 3, 159.100 6, 201.021 0, 269.044 4	D	JX
26	3′-O-Methylviolanone^[22]	C₁₈H₁₈O₆	24.38	[M-H]^-	329.103 0	329.101 6	-1.04	135.006 9, 148.016 4, 161.022 5, 284.099 4, 299.054 7, 314.005 4, 329.101 6	A	JX
27	Vestitone^[22]	C₁₆H₁₄O₅	25.77	[M-H]^-	285.076 8	285.076 3	1.89	135.007 4, 241.122 1, 270.052 0, 285.076 2	D	JX
28	Dehydrodiisoeugenol^[20]	C₂₀H₂₂O₄	26.16	[M+H]⁺	327.159 0	327.153 8	-6.12	137.057 6, 171.077 7, 203.103 3, 329.131 4	B	RDK
29	Machilin A^{[20, 21]}	C₂₀H₂₂O₄	26.16	[M+H]⁺	327.159 0	327.153 8	-6.12	137.057 6, 151.072 9, 171.077 7, 203.103 3	B	RDK
30	Ristiphenol (±)^[22]	C₁₆H₁₆O₄	26.58	[M-H]^-	271.097 5	271.096 7	0.82	109.027 9, 135.043 4, 256.109 1, 271.096 7	D	JX
31	Methyl eugenol^[22]	C₁₁H₁₄O₂	26.91	[M+H]⁺	179.106 6	179.103 6	-6.95	119.083 9, 151.072 8, 164.076 5, 179.103 6	C	RDK
32	Luteolin^[22]	C₁₅H₁₀O₆	27.13	[M-H]^-	285.040 4	285.039 4	0.01	241.122 7, 285.039 3	D	JX
33	2′-Methoxyisoliquiriti-genin^[22]	C₁₆H₁₄O₄	27.96	[M-H]^-	269.081 9	269.080 6	-0.76	226.094 4, 239.107 1, 254.057 2, 269.080 6	D	JX
34	Pterolinus B^[22]	C₁₆H₁₄O₄	27.96	[M-H]^-	269.081 9	269.080 6	-0.76	225.090 7, 239.107 1, 254.057 2, 269.080 6	F	JX
35^*	Cryptotanshinone^{[17, 18]}	C₁₉H₂₀O₃	28.31	[M+H]⁺	297.148 5	297.148 5	-5.98	279.133 4, 297.143 7	E	DS
36	Galbacin^{[18, 19]}	C₂₀H₂₀O₅	28.44	[M+H]⁺	341.138 3	341.130 6	-2.66	151.073 0, 163.052 8, 203.203 4, 341.130 6	B	RDK
37	Methylenedihydrotanshinquinone^[17]	C₁₈H₁₆O₃	28.45	[M-H]^-	279.102 6	279.101 4	-0.46	261.092 1, 279.101 4	E	DS
38	Dan shen spiroketal lactone^[17]	C₁₈H₂₂O₃	29	[M-H]^-	285.149 6	285.148 6	0.24	239.143 9, 285.148 5	E	DS
39	Tanshinone Ⅱ B^{[17, 18]}	C₁₉H₁₈O₄	29.12	[M+H]⁺	311.127 7	311.123 0	-5.31	247.107 0, 250.094 3, 275.103 1, 293.112 3, 311.123 0	E	DS
40	Danshenxinkun A^[17]	C₁₈H₁₆O₄	31.45	[M-H]^-	295.097 5	295.095 8	-2.22	277.180 7, 295.095 8	E	DS
41^*	Tanshinone I^{[17, 18]}	C₁₈H₁₂O₃	32.21	[M+H]⁺	277.085 9	277.081 5	-5.91	193.118 8, 203.103 5, 221.150 7, 249.087 0, 277.081 5	E	DS
42	Tanshinone IIA^{[17, 18]}	C₁₉H₁₈O₃	33.21	[M+H]⁺	295.132 8	295.128 3	-5.48	249.123 5, 262.095 0, 266.089 8, 277.118 1, 280.105 2, 295.128 3	E	DS

No.

Compound

Formula

t_R/min

Ion mode

Theoretical value

Measured value

Error (ppm)

Ion fragment

Classification

Source

5-Hydroxymethylfurfural^[21]

C₆H₆O₃

0.93

[M+H]⁺

127.038 9

127.037 2

-4.25

81.032 7, 109.026 9, 127.037 1

2^*

Quinic acid^[21]

C₇H₁₂O₆

1.09

[M-H]^-

191.056 1

191.054 6

-2.27

93.032 9, 128.038 3

Malic acid^[21]

C₄H₆O₅

1.28

[M-H]^-

133.014 2

133.012 5

4.81

71.012 1, 115.001 8

Trimethyl citrate^[21]

C₇H₁₀O₇

2.09

[M-H]^-

205.035 3

205.033 9

-1.99

87.068 8, 111.006 9, 143.033 6

Triethyl citrate^[21]

C₈H₁₂O₇

2.27

[M-H]^-

219.051 0

219.049 4

-2.59

87.001 0, 111.007 2, 157.075 3

2-Isopropylmalic acid^[21]

C₇H₁₂O₅

11.77

[M-H]^-

175.061 1

175.059 5

-3.19

85.064 1, 113.059 0, 115.038 3

7^*

Protocatechuic acid^[21]

C₇H₆O₄

13.83

[M-H]^-

153.019 3

153.017 6

-4.28

91.016 8, 108.051 4, 109.027 7

8^*

Ferulic acid^[21]

C₁₀H₁₀O₄

15.53

[M-H]^-

193.050 6

193.047 4

-7.11

134.035 6, 178.025 5, 193.047 3

9^*

Caffeic acid^[17]

C₉H₈O₄

18.32

[M-H]^-

179.034 9

179.033 4

-2.54

135.043 4, 179.033 4

Danshensu^[17]

C₉H₁₀O₅

19.01

[M-H]^-

197.049 9

197.044 1

-1.72

178.996 9, 197.044 1

Isorhamnetin^[23]

C₇H₆O₄

19.43

[M-H]^-

153.019 3

315.052 3

7.36

151.038 3, 315.052 2

SJ/SN

Scopolamine^[21]

C₁₀H₈O₄

19.63

[M+H]⁺

193.049 5

193.046 5

-5.77

122.058 2, 133.099 2, 149.958 7, 178.047 0, 193.046 4

Xenognoside B^[22]

C₁₆H₁₂O₅

20.14

[M-H]^-

283.061 1

283.060 1

-0.03

195.137 2, 224.046 2, 268.036 5, 283.060 0

3-Hydroxyvestitone^[22]

C₁₆H₁₄O₆

20.17

[M-H]^-

301.071 7

301.070 5

-0.64

135.007 1, 301.070 4

Homoeriodictyol^[22]

C₁₆H₁₄O₆

20.17

[M-H]^-

301.071 7

301.070 5

-0.64

121.027 2, 135.071 3, 177.017 6, 178.020 7, 268.036 5, 301.070 4

2-2-(2, 4-Dimethoxy-phenyl)-2-oxoe-thoxy-4-hydrobenzoic acid^[22]

C₁₇H₁₆O₇

20.21

[M-H]^-

331.082 3

331.081 5

0.84

176.010 5, 255.030 3, 301.069 7, 331.081 5

Myristicin^{[19, 20]}

C₁₁H₁₂O₃

20.7

[M+H]⁺

193.085 9

193.082 3

-8.54

105.068 7, 133.062 6, 165.087 8, 193.082 3

RDK

Ellagic acid^[23]

C₁₄H₆O₈

20.78

[M-H]^-

300.998 9

300.997 8

-0.41

185.059 4, 229.049 1, 257.116 3, 283.059 8

SN/SJ

19^*

Kaempferol^[23]

C₁₅H₁₀O₆

21.3

[M+H]⁺

287.055 0

287.054 1

-3.25

153.052 1, 165.052 1, 286.074 3, 287.054 0, 289.136 5

α-Curcumene^[19]

C₁₅H₂₂

22.47

[M+H]⁺

203.179 4

203.176 0

-6.72

81.069 2, 95.084 6, 109.099 6, 147.114 6, 203.176 0

(3R)-7, 3′-Dihydroxy-6, 2′, 4′-trime-thoxyisoflavanone^[22]

C₁₈H₁₈O₇

23.48

[M-H]^-

345.097 9

345.098 9

5.71

149.095 3, 271.135 6, 301.143 3, 315.049 9, 330.072 6, 345.098 8

5, 7-Dihydroxy-2′, 3′, 4′-trimethoxyisofla-vanone^[22]

C₁₈H₁₈O₇

23.67

[M-H]^-

345.097 9

345.096 8

-0.23

135.007 6, 151.038 4, 177.017 9, 295.060 7, 315.049 3, 330.072 7, 345.096 8

Fisetin^[22]

C₁₅H₁₀O₆

23.68

[M-H]^-

285.040 4

285.042 7

6.56

121.064 3, 149.022 4, 162.098 6, 239.164 1, 257.080 3, 285.042 6

Fustin^[22]

C₁₅H₁₂O₆

24.18

[M-H]^-

287.056 1

287.058 2

4.09

109.027 5, 149.058 7, 225.111 3

Genistein^[22]

C₁₅H₁₀O₅

24.34

[M-H]^-

269.045 5

269.044 5

0.14

133.064 3, 135.079 3, 159.100 6, 201.021 0, 269.044 4

3′-O-Methylviolanone^[22]

C₁₈H₁₈O₆

24.38

[M-H]^-

329.103 0

329.101 6

-1.04

135.006 9, 148.016 4, 161.022 5, 284.099 4, 299.054 7, 314.005 4, 329.101 6

Vestitone^[22]

C₁₆H₁₄O₅

25.77

[M-H]^-

285.076 8

285.076 3

1.89

135.007 4, 241.122 1, 270.052 0, 285.076 2

Dehydrodiisoeugenol^[20]

C₂₀H₂₂O₄

26.16

[M+H]⁺

327.159 0

327.153 8

-6.12

137.057 6, 171.077 7, 203.103 3, 329.131 4

RDK

Machilin A^{[20, 21]}

C₂₀H₂₂O₄

26.16

[M+H]⁺

327.159 0

327.153 8

-6.12

137.057 6, 151.072 9, 171.077 7, 203.103 3

RDK

Ristiphenol (±)^[22]

C₁₆H₁₆O₄

26.58

[M-H]^-

271.097 5

271.096 7

0.82

109.027 9, 135.043 4, 256.109 1, 271.096 7

Methyl eugenol^[22]

C₁₁H₁₄O₂

26.91

[M+H]⁺

179.106 6

179.103 6

-6.95

119.083 9, 151.072 8, 164.076 5, 179.103 6

RDK

Luteolin^[22]

C₁₅H₁₀O₆

27.13

[M-H]^-

285.040 4

285.039 4

0.01

241.122 7, 285.039 3

2′-Methoxyisoliquiriti-genin^[22]

C₁₆H₁₄O₄

27.96

[M-H]^-

269.081 9

269.080 6

-0.76

226.094 4, 239.107 1, 254.057 2, 269.080 6

Pterolinus B^[22]

C₁₆H₁₄O₄

27.96

[M-H]^-

269.081 9

269.080 6

-0.76

225.090 7, 239.107 1, 254.057 2, 269.080 6

35^*

Cryptotanshinone^{[17, 18]}

C₁₉H₂₀O₃

28.31

[M+H]⁺

297.148 5

-5.98

279.133 4, 297.143 7

Galbacin^{[18, 19]}

C₂₀H₂₀O₅

28.44

[M+H]⁺

341.138 3

341.130 6

-2.66

151.073 0, 163.052 8, 203.203 4, 341.130 6

RDK

Methylenedihydrotanshinquinone^[17]

C₁₈H₁₆O₃

28.45

[M-H]^-

279.102 6

279.101 4

-0.46

261.092 1, 279.101 4

Dan shen spiroketal lactone^[17]

C₁₈H₂₂O₃

[M-H]^-

285.149 6

285.148 6

0.24

239.143 9, 285.148 5

Tanshinone Ⅱ B^{[17, 18]}

C₁₉H₁₈O₄

29.12

[M+H]⁺

311.127 7

311.123 0

-5.31

247.107 0, 250.094 3, 275.103 1, 293.112 3, 311.123 0

Danshenxinkun A^[17]

C₁₈H₁₆O₄

31.45

[M-H]^-

295.097 5

295.095 8

-2.22

277.180 7, 295.095 8

41^*

Tanshinone I^{[17, 18]}

C₁₈H₁₂O₃

32.21

[M+H]⁺

277.085 9

277.081 5

-5.91

193.118 8, 203.103 5, 221.150 7, 249.087 0, 277.081 5

Tanshinone IIA^{[17, 18]}

C₁₉H₁₈O₃

33.21

[M+H]⁺

295.132 8

295.128 3

-5.48

249.123 5, 262.095 0, 266.089 8, 277.118 1, 280.105 2, 295.128 3

No.	Compound	Formula	t_R/min	Ion mode	Theoretical value	Measured value	Error (ppm)	Ion fragment	Classification	Source
1	5-Hydroxymethylfurfural^[21]	C₆H₆O₃	0.93	[M+H]⁺	127.038 9	127.037 2	-4.25	81.032 7, 109.026 9, 127.037 1	F	GZ
2^*	Quinic acid^[21]	C₇H₁₂O₆	1.09	[M-H]^-	191.056 1	191.054 6	-2.27	93.032 9, 128.038 3	A	GZ
3	Malic acid^[21]	C₄H₆O₅	1.28	[M-H]^-	133.014 2	133.012 5	4.81	71.012 1, 115.001 8	A	GZ
4	Trimethyl citrate^[21]	C₇H₁₀O₇	2.09	[M-H]^-	205.035 3	205.033 9	-1.99	87.068 8, 111.006 9, 143.033 6	A	GZ
5	Triethyl citrate^[21]	C₈H₁₂O₇	2.27	[M-H]^-	219.051 0	219.049 4	-2.59	87.001 0, 111.007 2, 157.075 3	A	GZ
6	2-Isopropylmalic acid^[21]	C₇H₁₂O₅	11.77	[M-H]^-	175.061 1	175.059 5	-3.19	85.064 1, 113.059 0, 115.038 3	A	GZ
7^*	Protocatechuic acid^[21]	C₇H₆O₄	13.83	[M-H]^-	153.019 3	153.017 6	-4.28	91.016 8, 108.051 4, 109.027 7	A	GZ
8^*	Ferulic acid^[21]	C₁₀H₁₀O₄	15.53	[M-H]^-	193.050 6	193.047 4	-7.11	134.035 6, 178.025 5, 193.047 3	A	GZ
9^*	Caffeic acid^[17]	C₉H₈O₄	18.32	[M-H]^-	179.034 9	179.033 4	-2.54	135.043 4, 179.033 4	A	DS
10	Danshensu^[17]	C₉H₁₀O₅	19.01	[M-H]^-	197.049 9	197.044 1	-1.72	178.996 9, 197.044 1	A	DS
11	Isorhamnetin^[23]	C₇H₆O₄	19.43	[M-H]^-	153.019 3	315.052 3	7.36	151.038 3, 315.052 2	D	SJ/SN
12	Scopolamine^[21]	C₁₀H₈O₄	19.63	[M+H]⁺	193.049 5	193.046 5	-5.77	122.058 2, 133.099 2, 149.958 7, 178.047 0, 193.046 4	C	GZ
13	Xenognoside B^[22]	C₁₆H₁₂O₅	20.14	[M-H]^-	283.061 1	283.060 1	-0.03	195.137 2, 224.046 2, 268.036 5, 283.060 0	D	JX
14	3-Hydroxyvestitone^[22]	C₁₆H₁₄O₆	20.17	[M-H]^-	301.071 7	301.070 5	-0.64	135.007 1, 301.070 4	D	JX
15	Homoeriodictyol^[22]	C₁₆H₁₄O₆	20.17	[M-H]^-	301.071 7	301.070 5	-0.64	121.027 2, 135.071 3, 177.017 6, 178.020 7, 268.036 5, 301.070 4	D	JX
16	2-2-(2, 4-Dimethoxy-phenyl)-2-oxoe-thoxy-4-hydrobenzoic acid^[22]	C₁₇H₁₆O₇	20.21	[M-H]^-	331.082 3	331.081 5	0.84	176.010 5, 255.030 3, 301.069 7, 331.081 5	A	JX
17	Myristicin^{[19, 20]}	C₁₁H₁₂O₃	20.7	[M+H]⁺	193.085 9	193.082 3	-8.54	105.068 7, 133.062 6, 165.087 8, 193.082 3	C	RDK
18	Ellagic acid^[23]	C₁₄H₆O₈	20.78	[M-H]^-	300.998 9	300.997 8	-0.41	185.059 4, 229.049 1, 257.116 3, 283.059 8	A	SN/SJ
19^*	Kaempferol^[23]	C₁₅H₁₀O₆	21.3	[M+H]⁺	287.055 0	287.054 1	-3.25	153.052 1, 165.052 1, 286.074 3, 287.054 0, 289.136 5	D	SN
20	α-Curcumene^[19]	C₁₅H₂₂	22.47	[M+H]⁺	203.179 4	203.176 0	-6.72	81.069 2, 95.084 6, 109.099 6, 147.114 6, 203.176 0	E	TX
21	(3R)-7, 3′-Dihydroxy-6, 2′, 4′-trime-thoxyisoflavanone^[22]	C₁₈H₁₈O₇	23.48	[M-H]^-	345.097 9	345.098 9	5.71	149.095 3, 271.135 6, 301.143 3, 315.049 9, 330.072 6, 345.098 8	D	JX
22	5, 7-Dihydroxy-2′, 3′, 4′-trimethoxyisofla-vanone^[22]	C₁₈H₁₈O₇	23.67	[M-H]^-	345.097 9	345.096 8	-0.23	135.007 6, 151.038 4, 177.017 9, 295.060 7, 315.049 3, 330.072 7, 345.096 8	D	JX
23	Fisetin^[22]	C₁₅H₁₀O₆	23.68	[M-H]^-	285.040 4	285.042 7	6.56	121.064 3, 149.022 4, 162.098 6, 239.164 1, 257.080 3, 285.042 6	D	JX
24	Fustin^[22]	C₁₅H₁₂O₆	24.18	[M-H]^-	287.056 1	287.058 2	4.09	109.027 5, 149.058 7, 225.111 3	D	JX
25	Genistein^[22]	C₁₅H₁₀O₅	24.34	[M-H]^-	269.045 5	269.044 5	0.14	133.064 3, 135.079 3, 159.100 6, 201.021 0, 269.044 4	D	JX
26	3′-O-Methylviolanone^[22]	C₁₈H₁₈O₆	24.38	[M-H]^-	329.103 0	329.101 6	-1.04	135.006 9, 148.016 4, 161.022 5, 284.099 4, 299.054 7, 314.005 4, 329.101 6	A	JX
27	Vestitone^[22]	C₁₆H₁₄O₅	25.77	[M-H]^-	285.076 8	285.076 3	1.89	135.007 4, 241.122 1, 270.052 0, 285.076 2	D	JX
28	Dehydrodiisoeugenol^[20]	C₂₀H₂₂O₄	26.16	[M+H]⁺	327.159 0	327.153 8	-6.12	137.057 6, 171.077 7, 203.103 3, 329.131 4	B	RDK
29	Machilin A^{[20, 21]}	C₂₀H₂₂O₄	26.16	[M+H]⁺	327.159 0	327.153 8	-6.12	137.057 6, 151.072 9, 171.077 7, 203.103 3	B	RDK
30	Ristiphenol (±)^[22]	C₁₆H₁₆O₄	26.58	[M-H]^-	271.097 5	271.096 7	0.82	109.027 9, 135.043 4, 256.109 1, 271.096 7	D	JX
31	Methyl eugenol^[22]	C₁₁H₁₄O₂	26.91	[M+H]⁺	179.106 6	179.103 6	-6.95	119.083 9, 151.072 8, 164.076 5, 179.103 6	C	RDK
32	Luteolin^[22]	C₁₅H₁₀O₆	27.13	[M-H]^-	285.040 4	285.039 4	0.01	241.122 7, 285.039 3	D	JX
33	2′-Methoxyisoliquiriti-genin^[22]	C₁₆H₁₄O₄	27.96	[M-H]^-	269.081 9	269.080 6	-0.76	226.094 4, 239.107 1, 254.057 2, 269.080 6	D	JX
34	Pterolinus B^[22]	C₁₆H₁₄O₄	27.96	[M-H]^-	269.081 9	269.080 6	-0.76	225.090 7, 239.107 1, 254.057 2, 269.080 6	F	JX
35^*	Cryptotanshinone^{[17, 18]}	C₁₉H₂₀O₃	28.31	[M+H]⁺	297.148 5	297.148 5	-5.98	279.133 4, 297.143 7	E	DS
36	Galbacin^{[18, 19]}	C₂₀H₂₀O₅	28.44	[M+H]⁺	341.138 3	341.130 6	-2.66	151.073 0, 163.052 8, 203.203 4, 341.130 6	B	RDK
37	Methylenedihydrotanshinquinone^[17]	C₁₈H₁₆O₃	28.45	[M-H]^-	279.102 6	279.101 4	-0.46	261.092 1, 279.101 4	E	DS
38	Dan shen spiroketal lactone^[17]	C₁₈H₂₂O₃	29	[M-H]^-	285.149 6	285.148 6	0.24	239.143 9, 285.148 5	E	DS
39	Tanshinone Ⅱ B^{[17, 18]}	C₁₉H₁₈O₄	29.12	[M+H]⁺	311.127 7	311.123 0	-5.31	247.107 0, 250.094 3, 275.103 1, 293.112 3, 311.123 0	E	DS
40	Danshenxinkun A^[17]	C₁₈H₁₆O₄	31.45	[M-H]^-	295.097 5	295.095 8	-2.22	277.180 7, 295.095 8	E	DS
41^*	Tanshinone I^{[17, 18]}	C₁₈H₁₂O₃	32.21	[M+H]⁺	277.085 9	277.081 5	-5.91	193.118 8, 203.103 5, 221.150 7, 249.087 0, 277.081 5	E	DS
42	Tanshinone IIA^{[17, 18]}	C₁₉H₁₈O₃	33.21	[M+H]⁺	295.132 8	295.128 3	-5.48	249.123 5, 262.095 0, 266.089 8, 277.118 1, 280.105 2, 295.128 3	E	DS

No.

Compound

Formula

t_R/min

Ion mode

Theoretical value

Measured value

Error (ppm)

Ion fragment

Classification

Source

5-Hydroxymethylfurfural^[21]

C₆H₆O₃

0.93

[M+H]⁺

127.038 9

127.037 2

-4.25

81.032 7, 109.026 9, 127.037 1

2^*

Quinic acid^[21]

C₇H₁₂O₆

1.09

[M-H]^-

191.056 1

191.054 6

-2.27

93.032 9, 128.038 3

Malic acid^[21]

C₄H₆O₅

1.28

[M-H]^-

133.014 2

133.012 5

4.81

71.012 1, 115.001 8

Trimethyl citrate^[21]

C₇H₁₀O₇

2.09

[M-H]^-

205.035 3

205.033 9

-1.99

87.068 8, 111.006 9, 143.033 6

Triethyl citrate^[21]

C₈H₁₂O₇

2.27

[M-H]^-

219.051 0

219.049 4

-2.59

87.001 0, 111.007 2, 157.075 3

2-Isopropylmalic acid^[21]

C₇H₁₂O₅

11.77

[M-H]^-

175.061 1

175.059 5

-3.19

85.064 1, 113.059 0, 115.038 3

7^*

Protocatechuic acid^[21]

C₇H₆O₄

13.83

[M-H]^-

153.019 3

153.017 6

-4.28

91.016 8, 108.051 4, 109.027 7

8^*

Ferulic acid^[21]

C₁₀H₁₀O₄

15.53

[M-H]^-

193.050 6

193.047 4

-7.11

134.035 6, 178.025 5, 193.047 3

9^*

Caffeic acid^[17]

C₉H₈O₄

18.32

[M-H]^-

179.034 9

179.033 4

-2.54

135.043 4, 179.033 4

Danshensu^[17]

C₉H₁₀O₅

19.01

[M-H]^-

197.049 9

197.044 1

-1.72

178.996 9, 197.044 1

Isorhamnetin^[23]

C₇H₆O₄

19.43

[M-H]^-

153.019 3

315.052 3

7.36

151.038 3, 315.052 2

SJ/SN

Scopolamine^[21]

C₁₀H₈O₄

19.63

[M+H]⁺

193.049 5

193.046 5

-5.77

122.058 2, 133.099 2, 149.958 7, 178.047 0, 193.046 4

Xenognoside B^[22]

C₁₆H₁₂O₅

20.14

[M-H]^-

283.061 1

283.060 1

-0.03

195.137 2, 224.046 2, 268.036 5, 283.060 0

3-Hydroxyvestitone^[22]

C₁₆H₁₄O₆

20.17

[M-H]^-

301.071 7

301.070 5

-0.64

135.007 1, 301.070 4

Homoeriodictyol^[22]

C₁₆H₁₄O₆

20.17

[M-H]^-

301.071 7

301.070 5

-0.64

121.027 2, 135.071 3, 177.017 6, 178.020 7, 268.036 5, 301.070 4

2-2-(2, 4-Dimethoxy-phenyl)-2-oxoe-thoxy-4-hydrobenzoic acid^[22]

C₁₇H₁₆O₇

20.21

[M-H]^-

331.082 3

331.081 5

0.84

176.010 5, 255.030 3, 301.069 7, 331.081 5

Myristicin^{[19, 20]}

C₁₁H₁₂O₃

20.7

[M+H]⁺

193.085 9

193.082 3

-8.54

105.068 7, 133.062 6, 165.087 8, 193.082 3

RDK

Ellagic acid^[23]

C₁₄H₆O₈

20.78

[M-H]^-

300.998 9

300.997 8

-0.41

185.059 4, 229.049 1, 257.116 3, 283.059 8

SN/SJ

19^*

Kaempferol^[23]

C₁₅H₁₀O₆

21.3

[M+H]⁺

287.055 0

287.054 1

-3.25

153.052 1, 165.052 1, 286.074 3, 287.054 0, 289.136 5

α-Curcumene^[19]

C₁₅H₂₂

22.47

[M+H]⁺

203.179 4

203.176 0

-6.72

81.069 2, 95.084 6, 109.099 6, 147.114 6, 203.176 0

(3R)-7, 3′-Dihydroxy-6, 2′, 4′-trime-thoxyisoflavanone^[22]

C₁₈H₁₈O₇

23.48

[M-H]^-

345.097 9

345.098 9

5.71

149.095 3, 271.135 6, 301.143 3, 315.049 9, 330.072 6, 345.098 8

5, 7-Dihydroxy-2′, 3′, 4′-trimethoxyisofla-vanone^[22]

C₁₈H₁₈O₇

23.67

[M-H]^-

345.097 9

345.096 8

-0.23

135.007 6, 151.038 4, 177.017 9, 295.060 7, 315.049 3, 330.072 7, 345.096 8

Fisetin^[22]

C₁₅H₁₀O₆

23.68

[M-H]^-

285.040 4

285.042 7

6.56

121.064 3, 149.022 4, 162.098 6, 239.164 1, 257.080 3, 285.042 6

Fustin^[22]

C₁₅H₁₂O₆

24.18

[M-H]^-

287.056 1

287.058 2

4.09

109.027 5, 149.058 7, 225.111 3

Genistein^[22]

C₁₅H₁₀O₅

24.34

[M-H]^-

269.045 5

269.044 5

0.14

133.064 3, 135.079 3, 159.100 6, 201.021 0, 269.044 4

3′-O-Methylviolanone^[22]

C₁₈H₁₈O₆

24.38

[M-H]^-

329.103 0

329.101 6

-1.04

135.006 9, 148.016 4, 161.022 5, 284.099 4, 299.054 7, 314.005 4, 329.101 6

Vestitone^[22]

C₁₆H₁₄O₅

25.77

[M-H]^-

285.076 8

285.076 3

1.89

135.007 4, 241.122 1, 270.052 0, 285.076 2

Dehydrodiisoeugenol^[20]

C₂₀H₂₂O₄

26.16

[M+H]⁺

327.159 0

327.153 8

-6.12

137.057 6, 171.077 7, 203.103 3, 329.131 4

RDK

Machilin A^{[20, 21]}

C₂₀H₂₂O₄

26.16

[M+H]⁺

327.159 0

327.153 8

-6.12

137.057 6, 151.072 9, 171.077 7, 203.103 3

RDK

Ristiphenol (±)^[22]

C₁₆H₁₆O₄

26.58

[M-H]^-

271.097 5

271.096 7

0.82

109.027 9, 135.043 4, 256.109 1, 271.096 7

Methyl eugenol^[22]

C₁₁H₁₄O₂

26.91

[M+H]⁺

179.106 6

179.103 6

-6.95

119.083 9, 151.072 8, 164.076 5, 179.103 6

RDK

Luteolin^[22]

C₁₅H₁₀O₆

27.13

[M-H]^-

285.040 4

285.039 4

0.01

241.122 7, 285.039 3

2′-Methoxyisoliquiriti-genin^[22]

C₁₆H₁₄O₄

27.96

[M-H]^-

269.081 9

269.080 6

-0.76

226.094 4, 239.107 1, 254.057 2, 269.080 6

Pterolinus B^[22]

C₁₆H₁₄O₄

27.96

[M-H]^-

269.081 9

269.080 6

-0.76

225.090 7, 239.107 1, 254.057 2, 269.080 6

35^*

Cryptotanshinone^{[17, 18]}

C₁₉H₂₀O₃

28.31

[M+H]⁺

297.148 5

-5.98

279.133 4, 297.143 7

Galbacin^{[18, 19]}

C₂₀H₂₀O₅

28.44

[M+H]⁺

341.138 3

341.130 6

-2.66

151.073 0, 163.052 8, 203.203 4, 341.130 6

RDK

Methylenedihydrotanshinquinone^[17]

C₁₈H₁₆O₃

28.45

[M-H]^-

279.102 6

279.101 4

-0.46

261.092 1, 279.101 4

Dan shen spiroketal lactone^[17]

C₁₈H₂₂O₃

[M-H]^-

285.149 6

285.148 6

0.24

239.143 9, 285.148 5

Tanshinone Ⅱ B^{[17, 18]}

C₁₉H₁₈O₄

29.12

[M+H]⁺

311.127 7

311.123 0

-5.31

247.107 0, 250.094 3, 275.103 1, 293.112 3, 311.123 0

Danshenxinkun A^[17]

C₁₈H₁₆O₄

31.45

[M-H]^-

295.097 5

295.095 8

-2.22

277.180 7, 295.095 8

41^*

Tanshinone I^{[17, 18]}

C₁₈H₁₂O₃

32.21

[M+H]⁺

277.085 9

277.081 5

-5.91

193.118 8, 203.103 5, 221.150 7, 249.087 0, 277.081 5

Tanshinone IIA^{[17, 18]}

C₁₉H₁₈O₃

33.21

[M+H]⁺

295.132 8

295.128 3

-5.48

249.123 5, 262.095 0, 266.089 8, 277.118 1, 280.105 2, 295.128 3

基于HPLC-Q-Exactive-MS/MS及网络药理学的冠心七味片入血成分抗动脉粥样硬化机制研究

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廖园红 ¹ , 陆景坤 ² , 牛燕 ² , 李君 ¹ , 布仁 ¹ , 张彭鹏 ³ , 康跃 ³ , 王跃武 ¹^,^*

药学学报 | 研究论文 2025,60(2): 449-458

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药学学报 | 研究论文 2025, 60(2): 449-458

基于HPLC-Q-Exactive-MS/MS及网络药理学的冠心七味片入血成分抗动脉粥样硬化机制研究

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廖园红¹, 陆景坤², 牛燕², 李君¹, 布仁¹, 张彭鹏³, 康跃³, 王跃武¹^,^*

作者信息

1.内蒙古医科大学药学院, 内蒙古呼和浩特 010110

2.内蒙古医科大学基础医学院, 内蒙古呼和浩特 010110

3.包头中药有限公司, 内蒙古包头 014040

通讯作者:

^*王跃武, Tel: 15847177513, E-mail: wywimmu@163.com

Study on anti-atherosclerosis mechanism of blood components of Guanxin Qiwei tablets based on HPLC-Q-Exactive-MS/MS and network pharmacology

Yuan-hong LIAO¹, Jing-kun LU², Yan NIU², Jun LI¹, Ren BU¹, Peng-peng ZHANG³, Yue KANG³, Yue-wu WANG¹^,^*

Affiliations

1. College of Pharmacy, Inner Mongolia Medical University, Hohhot 010110, China

2. School of Basic Medicine, Inner Mongolia Medical University, Hohhot 010110, China

3. Baotou Traditional Chinese Medicine Co., Ltd., Baotou 014040, China

出版时间: 2025-02-12 doi: 10.16438/j.0513-4870.2024-0741

文章导航

摘要

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以冠心七味片入血成分为基础, 结合网络药理学筛选冠心七味片抗动脉粥样硬化(atherosclerosis, AS) 的关键通路, 明确冠心七味片抗AS的作用机制, 并通过细胞实验加以验证。采用HPLC-Q-Exactive-MS/MS技术对冠心七味片入血成分进行分析, 确定化合物的精确质荷比, 并利用二级质谱裂解碎片、文献比对所含成分进行全面分析, 最终共鉴定出冠心七味片入血成分42个。利用Swiss Target Prediction数据库对其成分靶点预测。在疾病数据库Genecard、OMIM、Disgent中搜索AS靶点, 用Venny 2.1.0软件绘制Venny图得到疾病靶点与成分靶点的交集靶点181个, 利用String数据库分析蛋白质相互作用, 并用Cytscape软件筛选得到核心靶点32个。DAVID数据库进行核心靶点的GO (Gene Ontology) 富集分析和KEGG (Kyoto Encyclopedia of Genes and Genomes) 通路分析, 发现冠心七味片抗AS的通路主要包括脂质代谢与动脉粥样硬化(lipid metabolism and atherosclerosis)、高级糖基化终末产物-受体信号通路(AGE-RAGE signaling pathway in diabetic complications) 等。将核心靶点和核心化合物进行分子对接发现, 冠心七味片中隐丹参酮、丹参酮ⅡA与TNF、PPARγ、AKT1、PTG2等靶点结合较好。使用人hl-7702肝细胞构建脂滴模型, 验证冠心七味片调控脂质代谢与动脉粥样硬化的关键通路。这项研究初步鉴定了冠心七味片治疗AS的潜在药效成分并预测了作用通路, 对冠心七味片调节脂质代谢与动脉粥样硬化的关系进行体外实验验证, 为该方药效物质基础及作用机制的深入研究提供参考。本实验经过内蒙古医科大学医学伦理委员会批准(编号: YKD202401262)。

关键词

冠心七味片 / 网络药理学 / 动脉粥样硬化 / 入血成分 / 脂代谢

Abstract

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The analysis presented here is based on the blood components of Guanxin Qiwei tablets, the key anti-atherosclerosis pathway of Guanxin Qiwei tablets was screened by network pharmacology, and the anti-atherosclerosis mechanism of Guanxin Qiwei tablets was clarified and verified by cell experiments. HPLC-Q-Exactive-MS/MS technique was used to analyze the components of Guanxin Qiwei tablets into blood, to determine the precise mass charge ratio of the compounds, and to conduct a comprehensive analysis of the components by using secondary mass spectrometry fragments and literature comparison. Finally, a total of 42 components of Guanxin Qiwei tablets into blood were identified. To better understand the interactions, we employed the Swiss Target Prediction database to predict the associated targets. Atherosclerosis (AS) disease targets were searched in disease databases Genecard, OMIM and Disgent, and 181 intersection targets of disease targets and component targets were obtained by Venny 2.1.0 software. Protein interactions were analyzed by String database. The 32 core targets were selected by Cytscape software. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed in DAVID database. It was found that the anti-atherosclerosis pathways of Guanxin Qiwei tablets mainly include lipid metabolism and atherosclerosis and AGE-RAGE signaling pathway in diabetic complications and other signal pathways. The core targets and the core compounds were interlinked, and it was found that cryptotanshinone and tanshinone ⅡA in Guanxin Qiwei tablets were well bound to TNF, PPARγ, AKT1, PTG2 and other targets. The lipid metabolism and atherosclerotic pathway was verified using human hl-7702 hepatocytes. This study preliminarily identified the potential pharmacodynamic components of Guanxin Qiwei tablets in the treatment of AS diseases and predicted their pathways of action, and verified the relationship between regulating lipid metabolism and atherosclerosis of Guanxin Qiwei tablets in vitro, providing a reference for the further study of the pharmacodynamic material basis and mechanism of action of this prescription. This experiment was approved by the Medical Ethics Committee of Inner Mongolia Medical University (No. YKD202401262).

Key words

Guanxin Qiwei tablet / network pharmacology / atherosclerosis / blood component / lipid metabolism

引用本文

廖园红, 陆景坤, 牛燕, 李君, 布仁, 张彭鹏, 康跃, 王跃武. 基于HPLC-Q-Exactive-MS/MS及网络药理学的冠心七味片入血成分抗动脉粥样硬化机制研究. 药学学报, 2025 , 60 (2) : 449 -458 . DOI: 10.16438/j.0513-4870.2024-0741

Yuan-hong LIAO, Jing-kun LU, Yan NIU, Jun LI, Ren BU, Peng-peng ZHANG, Yue KANG, Yue-wu WANG. Study on anti-atherosclerosis mechanism of blood components of Guanxin Qiwei tablets based on HPLC-Q-Exactive-MS/MS and network pharmacology[J]. Acta Pharmaceutica Sinica, 2025 , 60 (2) : 449 -458 . DOI: 10.16438/j.0513-4870.2024-0741

正文

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动脉粥样硬化(atherosclerosis, AS) 是迄今为止最常见的冠状动脉疾病、颈动脉疾病和外周动脉疾病的潜在原因。AS是一种慢性疾病^[1], 其诱因主要有高血压、高血脂、糖尿病及生活习惯等。AS一般不是独立发病的, 通常和其他心血管疾病伴随发生^[2]。

冠心七味片由丹参、肉豆蔻、檀香、广枣、山柰、降香、沙棘七味药材组成, 蒙药名为“乌兰温都顺-7”, 临床用于治疗冠心病、心烦心悸、心绞痛^[3-7]。其复方中丹参作为君药含有多种活性成分, 具有促进血液循环, 减少血小板聚集, 降低血液黏稠度^{[8, 9]}, 从而改善微循环, 防治血栓形成, 抗氧化, 舒张血管, 抗炎等作用^{[10, 11]}。在临床上用于治疗心血管疾病的丹参复方药物有很多, 如丹参滴丸、丹参益心胶囊、丹参保心茶等^[12]。药理学研究表明, 冠心七味片可通过降血脂、抗炎、改善血管弹性等途径对实验性AS起到延缓或治疗作用^{[13, 14]}, 但目前对于冠心七味片抗AS的物质基础和作用机制尚不明确。药物成分如何经消化道吸收入血到达靶器官或靶点在体内发挥相应的药效尚不清楚^[15]。本项目在冠心七味片入血成分的基础上, 结合网络药理学筛选出冠心七味片治疗AS的相关靶点和通路, 对网络药理学预测结果通过体外实验进行验证, 进一步明确冠心七味片治疗AS的物质基础和作用机制, 探索冠心七味片治疗AS疾病的科学内涵, 以期为临床用药提供理论支撑。

材料与方法

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仪器 HPLC-Q-Exactive型高效液相色谱-质谱联用系统和全自动酶标仪(美国Thermo Fisher公司); CVE-3110真空离心浓缩仪(日本东京理化公司); 3-18K高速温控离心机(德国Sigma公司)。

药品与试剂 冠心七味片(C24017, 片重0.31 g, 包头中药有限公司); 甲醇(210386, 色谱纯)、乙腈(203096, 色谱纯), 美国Thermo Fisher公司; 胆固醇试剂盒(cholesterol, TC, ATWH25101)、甘油三酯试剂盒(triglyceride, TG, ATWH22101), Abbkine生物技术有限公司; 饱和油红O染液(G2013, 武汉塞维尔生物科技有限公司); 纯净水(屈臣氏)。

实验动物 雄性SD大鼠, 体质量(230 ± 50 g), 购自于斯贝福(北京) 生物技术有限公司, 许可证号SCXK (蒙) 2020-0003。经过内蒙古医科大学医学伦理委员会批准(编号: YKD202401262)。动物实验严格按照动物伦理学进行操作, 动物饲养于内蒙古医科大学实验动物研究中心, 适应性喂养5 d, 给予标准饲料和饮用水, 实验前12 h禁食, 正常饮水。

细胞人hl-7702细胞系为本实验室保存。

冠心七味片入血成分分析

色谱条件色谱柱为Agilent ZORB-AX SB-Aq (4.6 mm × 150 mm, 5 μm), 流动相: 乙腈(A)-0.1%甲酸水(B), 梯度洗脱(0~5 min, 5% A; 5~12 min, 5%→34% A; 12~16 min, 34%→65% A; 16~20 min, 65%→70% A; 20~25 min, 70%→85% A; 25~35 min, 85%→95% A; 35~40 min, 95% A); 流速为1.0 mL·min^-1, 4 ℃自动进样; 柱温25 ℃, 进样10 μL。

质谱条件采用电喷雾离子源(ESI), 检测方式为Full MS/dd-MS2, Full MS分辨率为70 000, dd-MS2分辨率为17 500, 正、负离子模式下获取质谱信息。条件如下: ①鞘气流速: 35 L·min^-1 (+)、40 L·min^-1 (-); ②辅助气流速: 5 L·min^-1; ③锥孔电压: 3.5 kV (-), 4.0 kV (+); ④毛细管离子传输管温度: 320 ℃; ⑤ S-lens电压: 50 kV; ⑥加热温度: 350 ℃, 质谱扫描范围为m/z 100~1 200^[16]。

药物处理冠心七味片去包衣后研钵研细, 过60目筛得粗粉, 加入0.1%羧甲基纤维素钠溶液, 配制成0.36 g·mL^-1的药液备用。

血清样本的制备与处理 SD大鼠随机分为空白组(n = 6) 和给药组(n = 6)。给药组灌胃给予冠心七味片药液10 mL·kg^-1 (相当于冠心七味片药量3.6 g·kg^-1, 临床给药剂量的15倍), 空白组给予相同体积的0.1%羧甲基纤维素钠溶液。在给药15、30、45 min和1、1.5、2 h后分别麻醉腹主动脉取血, 放室温静置1 h, 3 500 r·min^-1离心15 min, 取上清, 加4倍甲醇涡旋5 min, 4 ℃、12 000 r·min^-1离心15 min除蛋白, 取上清真空离心浓缩仪浓缩干燥, 用100 μL甲醇溶解, 涡旋15 min, 4 ℃、12 000 r·min^-1离心15 min, 取上清液于进样瓶中, 待进样^[16]。

数据处理通过中国知网、PubMed数据平台查找冠心七味片中丹参、肉豆蔻、檀香、广枣、山奈、降香、沙棘的化学成分、质谱碎片信息等文献资料, 并创建包括化合物名称、分子式、多级质谱碎片等信息的数据库。通过Xcalibur 3.0提取冠心七味片离子碎片信息, 通过与相关文献比对进行入血成分鉴定。

网络药理学分析

成分靶点、疾病靶点的获取在入血成分的基础上, 运用ChemicalBook (https://www.chemicalbook.com/) 数据库搜索成分CAS号, 在Pubchem (https://pubchem.ncbi.nlm.nih.gov) 数据库根据CAS号得到Canonial SMILES号, 通过Swiss Target Prediction (http://www.swisstargetprediction.ch/) 数据库获取成分靶点, 去除重复得冠心七味片入血成分相关靶点。利用疾病数据库Gencard (https://www.genecards.org/)、OMIM (https://www.omim.org/)、Disgent (https://www.disgenet.org/search) 以AS为关键词搜索疾病靶点, 去除重复靶点, 得到疾病靶点。将成分靶点与疾病靶点用Venny 2.1.0 (http://www.liuxiaoyuyuan.cn/) 取交集, 得到交集靶点。

PPI网络构建与核心靶点筛选将潜在靶点导入String (http://www.string-db.org/) 数据库, 获取蛋白相互作用信息, 导入Cytoscape3.9.1软件, 以平均节点度值(degree) 大于30.177 777 78、平均介数(betweenness) 大于170.933 333 3、最短路径(clossness) 大于0.002 906 131为标准筛选, 得到核心靶点。

GO和KEGG富集分析将核心靶点导入DAVID (https://david.ncifcrf.gov/) 数据库中, 得到GO和KEGG富集分析结果, 用微生信可视化平台进行可视化。

分子对接在TCMSP (https://old.tcmsp-e.com/tcmsp.php) 数据库中搜索入血成分中度值排名前10的化合物, 下载其mol 2形式结构作为小分子配体, 通过RCSB PDB (https://www.rcsb.org/) 数据库查找度值排名前10的核心靶点的3D结构, 用Autodock1.5.6软件进行分子对接, 用Pymol 2.6软件进行可视化。

体外验证

脂滴模型的建立复苏人hl-7702肝细胞, 调整细胞悬液浓度并分布于T25培养瓶中, 待细胞长至80%左右, 转移至12孔板中, 每孔1 mL, 约有2.0×10⁵个细胞。将细胞孔板放置于37 ℃、5% CO₂培养箱中培养过夜, 12 h后换无胎牛血清培养基培养24 h, 后向细胞培养液中分别加入终浓度为0、0.2、0.4、0.6、0.8、1.0 mmol·L^-1油酸(用10% NaOH+10% BSA配制), 继续培养24 h。油红O脂质染色, 拍照, 后用异丙醇溶解油红, 检测吸光度。

最佳给药浓度确定将人hl-7702肝细胞接种于96孔板, 37 ℃、5% CO₂培养箱培养至对数生长期, 分别用含0%、2.5%、5%、7.5%、10%、12.5%、15%含药血清的培养基培养细胞24、48 h。最后用CCK-8检测含药血清对细胞增殖能力的影响, 细胞存活率(%) = (实验组的吸光度-空白组吸光度)/(对照组的吸光度-空白组吸光度) × 100%。细胞存活率越大, 说明药物对细胞的损伤越小。最后筛选出含药血清的最佳作用浓度。

冠心七味片含药血清对肝细胞内TC、TG及脂质含量的影响将人hl-7702肝细胞分为3组: 空白对照组(培养液中加入等量普通培养基); 模型组(培养基中加入0.6 mmol·L^-1油酸处理); 治疗组(加入0.6 mmol·L^-1油酸24 h, 再给予12.5%含药血清预处理24和48 h)。根据试剂盒说明书制备标准曲线并利用无水乙醇制备细胞悬液, 通过8 000 r·min^-1离心5 min后取上清, 在96孔板中加入上清和TC、TG工作液进行37 ℃孵育15 min, 500 nm测定吸光值, 将吸光度值代入标准曲线中计算得到TC、TG浓度值。油红O脂质染色, 拍照。后用异丙醇溶解油红, 检测吸光度。

冠心七味片含药血清对人hl-7702肝细胞ABCA1、ABCG1、PPARγ蛋白表达水平的影响将各组细胞用RIPA缓冲液(1% PMSF) 裂解30 min提取蛋白质, 经蛋白定量后行蛋白变性。配制十二烷基硫酸钠-聚丙烯酰胺凝胶, 凝胶固定后上样进行电泳、转膜, 5%脱脂奶粉封闭90 min, 加一抗ABCA1 (1∶1 000)、ABCG1 (1∶1 000)、PPARγ (1∶1 000)、β-actin (1∶2 000) 4 ℃孵育过夜, 洗膜, 加二抗(1∶1 000) 室温孵育1 h, 显影并拍照, Image J 6.0软件分析其灰度值。

统计学分析所有数据均使用GraphPad Prism 6.0软件进行数据处理, 多组之间比较采用单因素方差分析, 两组之间比较采用t检验, P < 0.05认为差异具有统计学意义。

结果

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1 入血成分鉴定

制备含药血清, 分析鉴定后得到42个入血成分, 其中有机酸12个、木酚素3个、苯丙素类3个、黄酮类14个、萜类8个、其他类2个。正、负离子模式下血清样本总离子流图见图 1所示, 化合物具体信息见表 1^[17-23]。

2 网络药理学分析

2.1 核心靶点网络构建

从成分数据库中检索得到冠心七味片入血成分的作用靶点1 900个, 通过去重后得到565个。通过疾病数据库检索并去除重复后获得疾病靶点2 015个。将成分靶点和疾病靶点做韦恩图, 得到交集靶点181个, 以交集靶点作为冠心七味片抗AS的潜在靶点, 在String数据库中导入潜在靶点, 获取蛋白互作关系, 在Cytoscape3.9.1中筛选平均度值(30.177 777 78), 平均最短路径(170.933 333 3) 和平均介数(0.002 906 131) 得到核心靶点32个, 结果见图 2A、D。

2.2 核心靶点GO功能富集和KEGG信号通路分析

对32个核心靶点导入DAVID数据库进行GO富集分析和KEGG信号通路分析, 删除P > 0.05的数据, 结果得到生物学过程(biology process, BP) 共涉及有229个, 主要有RNA聚合酶Ⅱ启动子正调控、细胞凋亡过程的负调控、基因表达的正调控等。分子功能(molecular function, MF) 共涉及46个, 主要是关于蛋白结合、酶结合、DNA结合等。细胞组分(cellcular function, CC) 共涉及30个, 主要有细胞核、质膜、细胞质等见图 2B。KEGG信号通路分析共得到110条信号通路, 主要包括参与AS斑块形成的脂质和AS通路、介导糖尿病的AGE-RAGE信号通路、参与细胞凋亡的PI3K-AKT信号通路、参与阿尔茨海默症(Alzheimer disease, AD) 通路、IL-17信号通路等见图 2C。

2.3 分子对接结果

为了进一步验证冠心七味片中药效成分与核心靶点的相互作用关系, 以靶点最多的隐丹参酮和丹参酮ⅡA为配体, 以TNF、AKT1、ALB、PPARγ、STAT3、PTGS2、EGFR、MMP9、CASP3、ESR1为受体进行结合能力预测(图 3)。对接结果只保留每对分子对接的predicted binding affinity绝对值最高的(一般认为结合能小于-4.25 kJ·mol^-1提示配体与受体有一定的结合活性, 小于-5.0 kJ·mol^-1有较好的结合活性, 小于-7.0 kJ·mol^-1有强烈的结合活性)。实验结果显示, 隐丹参酮与PPARγ、丹参酮ⅡA与PPARγ结合活性最高, 与其结合较好的还有TNF、PTGS2、ESR1等。

3 体外实验

3.1 最佳油酸浓度

为了探究冠心七味片与AS脂代谢的作用关系、确定最佳油酸浓度, 首先采用油红O染色人hl-7702肝细胞, 结果显示见图 4A、B, 细胞内出现呈红色的脂肪滴, 随着油酸干预浓度升高, 出现红色脂滴的细胞增多, 在0.6 mmol·L^-1时脂滴最多, 而后脂滴减少, 且有部分细胞开始死亡。因此, 选择0.6 mmol·L^-1油酸干预hl-7702肝细胞来构建脂滴模型。

3.2 最佳含药血清浓度

为了确定最佳含药血清稀释倍数, 经过CCK8实验可知, 用10%的含药血清处理hl-7702肝细胞24或48 h后, 测得的细胞存活率均低于半数存活率(50%), 增加含药血清浓度, 细胞存活率均低于50%, 差异具有统计学意义(P < 0.01), 见图 4C。因此, 选用10%含药血清进行后续实验。

3.3 冠心七味片抑制肝细胞脂代谢水平

通过冠心七味片与AS的KEGG富集分析发现, 其抗AS与脂代谢通路密切相关。因此, 通过对人hl-7702肝细胞进行脂滴积累造模, 并给予10%含药血清, 收集细胞裂解后的上清液进行TC、TG的测定, 发现给予10%含药血清后大大降低了肝细胞内TC、TG水平。其次, 对细胞进行油红O染色并测量异丙醇溶解液的OD值, 结果显示给予含药血清可以降低肝细胞脂质积累, 见图 5。

4 冠心七味片含药血清对ABCA1、ABCG1、PPARγ的影响

通过KEGG通路富集和分子对接结果, 发现冠心七味片与AS的作用关系主要是经过脂代谢通路且主要靶点为PPARγ, 因此对相关靶点用蛋白免疫印迹法进行验证, 发现与空白组比较, 模型组ABCA1、ABCG1和PPARγ的蛋白表达水平显著下降(P < 0.01); 与模型组比较, 冠心七味组的蛋白表达水平显著上升(P < 0.05), 见图 6。

讨论

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AS是心血管疾病的一个重要致病因素, 是一种缓慢的终身进行性疾病^[24], 其发病率随着年龄的增长是呈正比增长的。目前的治疗手段包括手术治疗和药物治疗, 主要以药物治疗为主, 但都不能彻底解决问题。因此, 在斑块还未完全形成之前对其进行治疗是很有必要的^[25]。

冠心七味片是一种常见的蒙药, 主要由丹参、广枣、肉豆蔻等七味药物组成, 具有强心镇痛、活血化瘀的功效。其中, 丹参蒙古名为热贡巴, 为唇形科植物丹参的根和根茎, 有清血热、祛恶血之功效。肉豆蔻蒙古名为查干苏格莫勒, 有祛肾寒、抑赫依症、调胃火、消食等功效^[26]。

研究发现, 丹参中成分隐丹参酮、丹参酮ⅡA与PPARγ受体结合能较低。PPARγ为过氧化物酶体增生激活受体γ编码基因, 包括PPARγ1和PPERγ2两种亚型, 主要负责脂类的分解代谢等^[27]。PPARγ/ABCA1/ABCG1脂代谢通路是经典的抗AS通路之一^[28], 可以通过降低TC、TG含量, 从而降低血液血脂水平, 降低AS的发生^[29]。另外, 对排名前10的核心靶点进行分子对接发现, 隐丹参酮、丹参酮ⅡA和TNF、PTGS2、MMP9和ESR1结合能也较低, 而这些核心靶点都和血管内皮细胞炎症反应紧密相关^{[30, 31]}。而AS的发生与发展和慢性炎症反应密不可分, 在本实验中冠心七味片既可以通过调节脂质代谢来治疗AS, 同时也可以通过抗炎抗氧化而抗AS。

综上所述, 本研究以冠心七味片入血成分为基础, 通过网络药理学通路富集得到关键通路, 并在细胞水平得到了相应的验证, 为冠心七味片临床用药提供理论支撑。

作者贡献: 廖园红、陆景坤、牛燕、李君负责实验; 廖园红、张彭鹏、康跃负责数据整理并撰写文章; 布仁、王跃武负责文章修改。

利益冲突: 无任何利益冲突。

基金

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内蒙古自治区科技计划项目(2023YFHH0082)
国家自然科学基金项目(81960757)
内蒙古自治区科技成果转化专项(2019CG042)
内蒙古自然科学基金项目(2020MS08045)

参考文献

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文献

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2025年第60卷第2期

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doi: 10.16438/j.0513-4870.2024-0741

接收时间：2024-07-31
首发时间：2025-11-07
出版时间：2025-02-12

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收稿日期：2024-07-31
修回日期：2024-11-28

基金

内蒙古自治区科技计划项目(2023YFHH0082)

国家自然科学基金项目(81960757)

内蒙古自治区科技成果转化专项(2019CG042)

内蒙古自然科学基金项目(2020MS08045)

作者信息

1.内蒙古医科大学药学院, 内蒙古呼和浩特 010110

2.内蒙古医科大学基础医学院, 内蒙古呼和浩特 010110

3.包头中药有限公司, 内蒙古包头 014040

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^*王跃武, Tel: 15847177513, E-mail: wywimmu@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.	Compound	Formula	t_R/min	Ion mode	Theoretical value	Measured value	Error (ppm)	Ion fragment	Classification	Source
1	5-Hydroxymethylfurfural^[21]	C₆H₆O₃	0.93	[M+H]⁺	127.038 9	127.037 2	-4.25	81.032 7, 109.026 9, 127.037 1	F	GZ
2^*	Quinic acid^[21]	C₇H₁₂O₆	1.09	[M-H]^-	191.056 1	191.054 6	-2.27	93.032 9, 128.038 3	A	GZ
3	Malic acid^[21]	C₄H₆O₅	1.28	[M-H]^-	133.014 2	133.012 5	4.81	71.012 1, 115.001 8	A	GZ
4	Trimethyl citrate^[21]	C₇H₁₀O₇	2.09	[M-H]^-	205.035 3	205.033 9	-1.99	87.068 8, 111.006 9, 143.033 6	A	GZ
5	Triethyl citrate^[21]	C₈H₁₂O₇	2.27	[M-H]^-	219.051 0	219.049 4	-2.59	87.001 0, 111.007 2, 157.075 3	A	GZ
6	2-Isopropylmalic acid^[21]	C₇H₁₂O₅	11.77	[M-H]^-	175.061 1	175.059 5	-3.19	85.064 1, 113.059 0, 115.038 3	A	GZ
7^*	Protocatechuic acid^[21]	C₇H₆O₄	13.83	[M-H]^-	153.019 3	153.017 6	-4.28	91.016 8, 108.051 4, 109.027 7	A	GZ
8^*	Ferulic acid^[21]	C₁₀H₁₀O₄	15.53	[M-H]^-	193.050 6	193.047 4	-7.11	134.035 6, 178.025 5, 193.047 3	A	GZ
9^*	Caffeic acid^[17]	C₉H₈O₄	18.32	[M-H]^-	179.034 9	179.033 4	-2.54	135.043 4, 179.033 4	A	DS
10	Danshensu^[17]	C₉H₁₀O₅	19.01	[M-H]^-	197.049 9	197.044 1	-1.72	178.996 9, 197.044 1	A	DS
11	Isorhamnetin^[23]	C₇H₆O₄	19.43	[M-H]^-	153.019 3	315.052 3	7.36	151.038 3, 315.052 2	D	SJ/SN
12	Scopolamine^[21]	C₁₀H₈O₄	19.63	[M+H]⁺	193.049 5	193.046 5	-5.77	122.058 2, 133.099 2, 149.958 7, 178.047 0, 193.046 4	C	GZ
13	Xenognoside B^[22]	C₁₆H₁₂O₅	20.14	[M-H]^-	283.061 1	283.060 1	-0.03	195.137 2, 224.046 2, 268.036 5, 283.060 0	D	JX
14	3-Hydroxyvestitone^[22]	C₁₆H₁₄O₆	20.17	[M-H]^-	301.071 7	301.070 5	-0.64	135.007 1, 301.070 4	D	JX
15	Homoeriodictyol^[22]	C₁₆H₁₄O₆	20.17	[M-H]^-	301.071 7	301.070 5	-0.64	121.027 2, 135.071 3, 177.017 6, 178.020 7, 268.036 5, 301.070 4	D	JX
16	2-2-(2, 4-Dimethoxy-phenyl)-2-oxoe-thoxy-4-hydrobenzoic acid^[22]	C₁₇H₁₆O₇	20.21	[M-H]^-	331.082 3	331.081 5	0.84	176.010 5, 255.030 3, 301.069 7, 331.081 5	A	JX
17	Myristicin^{[19, 20]}	C₁₁H₁₂O₃	20.7	[M+H]⁺	193.085 9	193.082 3	-8.54	105.068 7, 133.062 6, 165.087 8, 193.082 3	C	RDK
18	Ellagic acid^[23]	C₁₄H₆O₈	20.78	[M-H]^-	300.998 9	300.997 8	-0.41	185.059 4, 229.049 1, 257.116 3, 283.059 8	A	SN/SJ
19^*	Kaempferol^[23]	C₁₅H₁₀O₆	21.3	[M+H]⁺	287.055 0	287.054 1	-3.25	153.052 1, 165.052 1, 286.074 3, 287.054 0, 289.136 5	D	SN
20	α-Curcumene^[19]	C₁₅H₂₂	22.47	[M+H]⁺	203.179 4	203.176 0	-6.72	81.069 2, 95.084 6, 109.099 6, 147.114 6, 203.176 0	E	TX
21	(3R)-7, 3′-Dihydroxy-6, 2′, 4′-trime-thoxyisoflavanone^[22]	C₁₈H₁₈O₇	23.48	[M-H]^-	345.097 9	345.098 9	5.71	149.095 3, 271.135 6, 301.143 3, 315.049 9, 330.072 6, 345.098 8	D	JX
22	5, 7-Dihydroxy-2′, 3′, 4′-trimethoxyisofla-vanone^[22]	C₁₈H₁₈O₇	23.67	[M-H]^-	345.097 9	345.096 8	-0.23	135.007 6, 151.038 4, 177.017 9, 295.060 7, 315.049 3, 330.072 7, 345.096 8	D	JX
23	Fisetin^[22]	C₁₅H₁₀O₆	23.68	[M-H]^-	285.040 4	285.042 7	6.56	121.064 3, 149.022 4, 162.098 6, 239.164 1, 257.080 3, 285.042 6	D	JX
24	Fustin^[22]	C₁₅H₁₂O₆	24.18	[M-H]^-	287.056 1	287.058 2	4.09	109.027 5, 149.058 7, 225.111 3	D	JX
25	Genistein^[22]	C₁₅H₁₀O₅	24.34	[M-H]^-	269.045 5	269.044 5	0.14	133.064 3, 135.079 3, 159.100 6, 201.021 0, 269.044 4	D	JX
26	3′-O-Methylviolanone^[22]	C₁₈H₁₈O₆	24.38	[M-H]^-	329.103 0	329.101 6	-1.04	135.006 9, 148.016 4, 161.022 5, 284.099 4, 299.054 7, 314.005 4, 329.101 6	A	JX
27	Vestitone^[22]	C₁₆H₁₄O₅	25.77	[M-H]^-	285.076 8	285.076 3	1.89	135.007 4, 241.122 1, 270.052 0, 285.076 2	D	JX
28	Dehydrodiisoeugenol^[20]	C₂₀H₂₂O₄	26.16	[M+H]⁺	327.159 0	327.153 8	-6.12	137.057 6, 171.077 7, 203.103 3, 329.131 4	B	RDK
29	Machilin A^{[20, 21]}	C₂₀H₂₂O₄	26.16	[M+H]⁺	327.159 0	327.153 8	-6.12	137.057 6, 151.072 9, 171.077 7, 203.103 3	B	RDK
30	Ristiphenol (±)^[22]	C₁₆H₁₆O₄	26.58	[M-H]^-	271.097 5	271.096 7	0.82	109.027 9, 135.043 4, 256.109 1, 271.096 7	D	JX
31	Methyl eugenol^[22]	C₁₁H₁₄O₂	26.91	[M+H]⁺	179.106 6	179.103 6	-6.95	119.083 9, 151.072 8, 164.076 5, 179.103 6	C	RDK
32	Luteolin^[22]	C₁₅H₁₀O₆	27.13	[M-H]^-	285.040 4	285.039 4	0.01	241.122 7, 285.039 3	D	JX
33	2′-Methoxyisoliquiriti-genin^[22]	C₁₆H₁₄O₄	27.96	[M-H]^-	269.081 9	269.080 6	-0.76	226.094 4, 239.107 1, 254.057 2, 269.080 6	D	JX
34	Pterolinus B^[22]	C₁₆H₁₄O₄	27.96	[M-H]^-	269.081 9	269.080 6	-0.76	225.090 7, 239.107 1, 254.057 2, 269.080 6	F	JX
35^*	Cryptotanshinone^{[17, 18]}	C₁₉H₂₀O₃	28.31	[M+H]⁺	297.148 5	297.148 5	-5.98	279.133 4, 297.143 7	E	DS
36	Galbacin^{[18, 19]}	C₂₀H₂₀O₅	28.44	[M+H]⁺	341.138 3	341.130 6	-2.66	151.073 0, 163.052 8, 203.203 4, 341.130 6	B	RDK
37	Methylenedihydrotanshinquinone^[17]	C₁₈H₁₆O₃	28.45	[M-H]^-	279.102 6	279.101 4	-0.46	261.092 1, 279.101 4	E	DS
38	Dan shen spiroketal lactone^[17]	C₁₈H₂₂O₃	29	[M-H]^-	285.149 6	285.148 6	0.24	239.143 9, 285.148 5	E	DS
39	Tanshinone Ⅱ B^{[17, 18]}	C₁₉H₁₈O₄	29.12	[M+H]⁺	311.127 7	311.123 0	-5.31	247.107 0, 250.094 3, 275.103 1, 293.112 3, 311.123 0	E	DS
40	Danshenxinkun A^[17]	C₁₈H₁₆O₄	31.45	[M-H]^-	295.097 5	295.095 8	-2.22	277.180 7, 295.095 8	E	DS
41^*	Tanshinone I^{[17, 18]}	C₁₈H₁₂O₃	32.21	[M+H]⁺	277.085 9	277.081 5	-5.91	193.118 8, 203.103 5, 221.150 7, 249.087 0, 277.081 5	E	DS
42	Tanshinone IIA^{[17, 18]}	C₁₉H₁₈O₃	33.21	[M+H]⁺	295.132 8	295.128 3	-5.48	249.123 5, 262.095 0, 266.089 8, 277.118 1, 280.105 2, 295.128 3	E	DS

No.

Compound

Formula

t_R/min

Ion mode

Theoretical value

Measured value

Error (ppm)

Ion fragment

Classification

Source

5-Hydroxymethylfurfural^[21]

C₆H₆O₃

0.93

[M+H]⁺

127.038 9

127.037 2

-4.25

81.032 7, 109.026 9, 127.037 1

2^*

Quinic acid^[21]

C₇H₁₂O₆

1.09

[M-H]^-

191.056 1

191.054 6

-2.27

93.032 9, 128.038 3

Malic acid^[21]

C₄H₆O₅

1.28

[M-H]^-

133.014 2

133.012 5

4.81

71.012 1, 115.001 8

Trimethyl citrate^[21]

C₇H₁₀O₇

2.09

[M-H]^-

205.035 3

205.033 9

-1.99

87.068 8, 111.006 9, 143.033 6

Triethyl citrate^[21]

C₈H₁₂O₇

2.27

[M-H]^-

219.051 0

219.049 4

-2.59

87.001 0, 111.007 2, 157.075 3

2-Isopropylmalic acid^[21]

C₇H₁₂O₅

11.77

[M-H]^-

175.061 1

175.059 5

-3.19

85.064 1, 113.059 0, 115.038 3

7^*

Protocatechuic acid^[21]

C₇H₆O₄

13.83

[M-H]^-

153.019 3

153.017 6

-4.28

91.016 8, 108.051 4, 109.027 7

8^*

Ferulic acid^[21]

C₁₀H₁₀O₄

15.53

[M-H]^-

193.050 6

193.047 4

-7.11

134.035 6, 178.025 5, 193.047 3

9^*

Caffeic acid^[17]

C₉H₈O₄

18.32

[M-H]^-

179.034 9

179.033 4

-2.54

135.043 4, 179.033 4

Danshensu^[17]

C₉H₁₀O₅

19.01

[M-H]^-

197.049 9

197.044 1

-1.72

178.996 9, 197.044 1

Isorhamnetin^[23]

C₇H₆O₄

19.43

[M-H]^-

153.019 3

315.052 3

7.36

151.038 3, 315.052 2

SJ/SN

Scopolamine^[21]

C₁₀H₈O₄

19.63

[M+H]⁺

193.049 5

193.046 5

-5.77

122.058 2, 133.099 2, 149.958 7, 178.047 0, 193.046 4

Xenognoside B^[22]

C₁₆H₁₂O₅

20.14

[M-H]^-

283.061 1

283.060 1

-0.03

195.137 2, 224.046 2, 268.036 5, 283.060 0

3-Hydroxyvestitone^[22]

C₁₆H₁₄O₆

20.17

[M-H]^-

301.071 7

301.070 5

-0.64

135.007 1, 301.070 4

Homoeriodictyol^[22]

C₁₆H₁₄O₆

20.17

[M-H]^-

301.071 7

301.070 5

-0.64

121.027 2, 135.071 3, 177.017 6, 178.020 7, 268.036 5, 301.070 4

2-2-(2, 4-Dimethoxy-phenyl)-2-oxoe-thoxy-4-hydrobenzoic acid^[22]

C₁₇H₁₆O₇

20.21

[M-H]^-

331.082 3

331.081 5

0.84

176.010 5, 255.030 3, 301.069 7, 331.081 5

Myristicin^{[19, 20]}

C₁₁H₁₂O₃

20.7

[M+H]⁺

193.085 9

193.082 3

-8.54

105.068 7, 133.062 6, 165.087 8, 193.082 3

RDK

Ellagic acid^[23]

C₁₄H₆O₈

20.78

[M-H]^-

300.998 9

300.997 8

-0.41

185.059 4, 229.049 1, 257.116 3, 283.059 8

SN/SJ

19^*

Kaempferol^[23]

C₁₅H₁₀O₆

21.3

[M+H]⁺

287.055 0

287.054 1

-3.25

153.052 1, 165.052 1, 286.074 3, 287.054 0, 289.136 5

α-Curcumene^[19]

C₁₅H₂₂

22.47

[M+H]⁺

203.179 4

203.176 0

-6.72

81.069 2, 95.084 6, 109.099 6, 147.114 6, 203.176 0

(3R)-7, 3′-Dihydroxy-6, 2′, 4′-trime-thoxyisoflavanone^[22]

C₁₈H₁₈O₇

23.48

[M-H]^-

345.097 9

345.098 9

5.71

149.095 3, 271.135 6, 301.143 3, 315.049 9, 330.072 6, 345.098 8

5, 7-Dihydroxy-2′, 3′, 4′-trimethoxyisofla-vanone^[22]

C₁₈H₁₈O₇

23.67

[M-H]^-

345.097 9

345.096 8

-0.23

135.007 6, 151.038 4, 177.017 9, 295.060 7, 315.049 3, 330.072 7, 345.096 8

Fisetin^[22]

C₁₅H₁₀O₆

23.68

[M-H]^-

285.040 4

285.042 7

6.56

121.064 3, 149.022 4, 162.098 6, 239.164 1, 257.080 3, 285.042 6

Fustin^[22]

C₁₅H₁₂O₆

24.18

[M-H]^-

287.056 1

287.058 2

4.09

109.027 5, 149.058 7, 225.111 3

Genistein^[22]

C₁₅H₁₀O₅

24.34

[M-H]^-

269.045 5

269.044 5

0.14

133.064 3, 135.079 3, 159.100 6, 201.021 0, 269.044 4

3′-O-Methylviolanone^[22]

C₁₈H₁₈O₆

24.38

[M-H]^-

329.103 0

329.101 6

-1.04

135.006 9, 148.016 4, 161.022 5, 284.099 4, 299.054 7, 314.005 4, 329.101 6

Vestitone^[22]

C₁₆H₁₄O₅

25.77

[M-H]^-

285.076 8

285.076 3

1.89

135.007 4, 241.122 1, 270.052 0, 285.076 2

Dehydrodiisoeugenol^[20]

C₂₀H₂₂O₄

26.16

[M+H]⁺

327.159 0

327.153 8

-6.12

137.057 6, 171.077 7, 203.103 3, 329.131 4

RDK

Machilin A^{[20, 21]}

C₂₀H₂₂O₄

26.16

[M+H]⁺

327.159 0

327.153 8

-6.12

137.057 6, 151.072 9, 171.077 7, 203.103 3

RDK

Ristiphenol (±)^[22]

C₁₆H₁₆O₄

26.58

[M-H]^-

271.097 5

271.096 7

0.82

109.027 9, 135.043 4, 256.109 1, 271.096 7

Methyl eugenol^[22]

C₁₁H₁₄O₂

26.91

[M+H]⁺

179.106 6

179.103 6

-6.95

119.083 9, 151.072 8, 164.076 5, 179.103 6

RDK

Luteolin^[22]

C₁₅H₁₀O₆

27.13

[M-H]^-

285.040 4

285.039 4

0.01

241.122 7, 285.039 3

2′-Methoxyisoliquiriti-genin^[22]

C₁₆H₁₄O₄

27.96

[M-H]^-

269.081 9

269.080 6

-0.76

226.094 4, 239.107 1, 254.057 2, 269.080 6

Pterolinus B^[22]

C₁₆H₁₄O₄

27.96

[M-H]^-

269.081 9

269.080 6

-0.76

225.090 7, 239.107 1, 254.057 2, 269.080 6

35^*

Cryptotanshinone^{[17, 18]}

C₁₉H₂₀O₃

28.31

[M+H]⁺

297.148 5

-5.98

279.133 4, 297.143 7

Galbacin^{[18, 19]}

C₂₀H₂₀O₅

28.44

[M+H]⁺

341.138 3

341.130 6

-2.66

151.073 0, 163.052 8, 203.203 4, 341.130 6

RDK

Methylenedihydrotanshinquinone^[17]

C₁₈H₁₆O₃

28.45

[M-H]^-

279.102 6

279.101 4

-0.46

261.092 1, 279.101 4

Dan shen spiroketal lactone^[17]

C₁₈H₂₂O₃

[M-H]^-

285.149 6

285.148 6

0.24

239.143 9, 285.148 5

Tanshinone Ⅱ B^{[17, 18]}

C₁₉H₁₈O₄

29.12

[M+H]⁺

311.127 7

311.123 0

-5.31

247.107 0, 250.094 3, 275.103 1, 293.112 3, 311.123 0

Danshenxinkun A^[17]

C₁₈H₁₆O₄

31.45

[M-H]^-

295.097 5

295.095 8

-2.22

277.180 7, 295.095 8

41^*

Tanshinone I^{[17, 18]}

C₁₈H₁₂O₃

32.21

[M+H]⁺

277.085 9

277.081 5

-5.91

193.118 8, 203.103 5, 221.150 7, 249.087 0, 277.081 5

Tanshinone IIA^{[17, 18]}

C₁₉H₁₈O₃

33.21

[M+H]⁺

295.132 8

295.128 3

-5.48

249.123 5, 262.095 0, 266.089 8, 277.118 1, 280.105 2, 295.128 3