中国药学杂志

No.	Molecular formula	t_R /min	Theoretical mass(m/z)	Experimental mass(m/z)	Adduct	Error (×10^-6)	m/z (MS²)	Component	Source	Category
1	C₂₁H₂₁NO₄	8.92	352.154 33	352.153 53	[M+H]⁺	0.086	336.122 25, 308.127 11	Palmatine	PC	Alkaloid
2	C₂₀H₂₀NO₇	3.80	387.131 25	387.189 94	[M+H]⁺	-0.428	145.028 03, 177.054 14	Trihydroxy jatrorrhizine	PC	Alkaloid
3	C₂₁H₁₃N₃O	8.30	324.113 13	324.121 95	[M+H]⁺	-0.754	308.090 70, 280.095 89	7,8-deoxyrutaecarpine	PC	Alkaloid
4	C₂₀H₁₇NO₅	7.52	352.117 94	352.116 85	[M+H]⁺	0.692	336.085 60, 308.090 73	Berlambine	PC	Alkaloid
5	C₂₀H₁₉NO₄	8.20	336.123 03	336.121 83	[M+H]⁺	0.41	320.090 70, 306.518 68	Berbine	PC	Alkaloid
6	C₂₁H₁₈N $O 4 +$	9.66	350.138 68	350.137 57	[M+H]⁺	0.651	334.106 29, 320.090 7	Chelerythrine	PC	Alkaloid
7	C₂₇H₃₀O₁₀	11.53	513.176 62	513.138 79	[M-H]^-	0.321	367.139 50	Baohuoside Ⅰ	RA	Flavonoids
8	C₁₉H₁₅NO₄	8.36	322.107 38	322.106 32	[M+H]⁺	-0.696	307.082 89, 279.087 95	Menisporphine	PC	Alkaloid
9	C₂₀H₂₁NO₄	8.03	339.146 5	339.140 87	[M+H]⁺	-0.79	323.113 01, 308.090 27	Flavonoids	PC	Alkaloid
10	C₂₁H₂₅NO₄	7.19	356.185 63	356.184 33	[M+H]⁺	0.106	311.126 56, 296.103 39	Menisperine	PC	Alkaloid
11	C₁₉H₁₇NO₄	7.36	324.123 03	324.122 16	[M+H]⁺	-0.754	308.090 70, 294.075 29	Berberrubine	PC	Alkaloid
12	C₁₇H₂₀O₉	5.44	367.103 45	367.103 27	[M-H]^-	-0.362	191.055 31, 173.044 59	3-O-Feruloylquinic acid	PC	Phenolic acids
13	C₁₆H₁₈O₉	3.22	353.087 8	353.086 06	[M-H]^-	-0.002	191.055 24, 179.034 03	Chlorogenic acid	PC	Phenolic acids
14	C₁₇H₁₇NO₅	4.14	316.117 94	316.283 69	[M+H]⁺	1.303	298.274 02, 280.262 12	Hippeastrine	PC	Alkaloid
15	C₂₆H₃₀O₁₂	6.88	533.166 44	533.151 55	[M-H]^-	0.218	371.100 46, 325.091 77	Aegineoside	PC	Flavonoids
16	C₂₃H₂₉NO₈	5.15	448.196 59	448.195 28	[M+H]⁺	0.066	107.049 18, 286.142 79	¹⁾	PC	Glycoside
17	C₁₅H₁₇NO₃	6.79	260.128 11	260.127 38	[M+H]⁺	2.317	242.116 73, 224.106 48	Pilocarpine	PC	Alkaloid
18	C1₉H₂₃NO₃	6.35	314.175 07	314.173 95	[M+H]⁺	0.517	299.112 82, 269.116 82	Isoliensinine	PC	Alkaloid
19	C₃₉H₆₄O₁₃	10.22	739.427 41	739.428 16	[M-H]^-	0.002	577.376 65, 529.609 62	Timosaponin AⅢ	RA	Saponins
20	C₂₅H₂₈O₁₆	3.86	583.130 45	583.129 39	[M-H]^-	0.050	421.077 61, 331.045 84	Neomangiferin	RA	Flavonoids
21	C₄₅H₇₄O₁₉	8.06	917.475 15	917.476 14	[M-H]^-	-0.085	71.012 41,89.023	Anemarsaponin D	RA	Saponins
22	C₁₃H₁₈O₈	1.66	301.092 89	301.093 66	[M-H]^-	1.756	123.007 48, 138.031 19	Tachioside	PC	Phenolic acids
23	C₁₉H₁₈O₁₁	4.88	421.077 63	421.077 97	[M-H]^-	0.225	301.035 10, 331.045 90	Isomangiferin	RA	Flavonoids
24	C₄₅H₇₄O₁₈	9.78	901.480 23	901.484 13	[M-H]^-	-0.026	101.023 06, 113.023 09	Anemarsaponin B	RA	Saponins
25	C₄₅H₇₆O₁₉	8.37	919.490 8	919.489 75	[M-H]^-	0.048	757.432 56, 595.383 79	Anemarsaponin BⅡ	RA	Saponins
26	C₂₀H₁₉NO₄	7.69	338.138 68	338.137 45	[M+H]⁺	0.320	323.113 83, 294.111 02	Jatrorrhizine	PC	Alkaloid
27	C₂₁H₂₀O₁₀	6.58	431.098 37	431.097 93	[M-H]^-	0.346	311.056 12, 283.061 00	Afzelin	RA	Flavonoids
28	C₂₀H₂₃NO₄	5.56	342.169 98	342.169 19	[M+H]⁺	0.133	297.111 05, 265.085 17	Magnoflorine	PC	Alkaloid
29	C₁₈H₃₄O₅	10.30	329.233 34	329.233 28	[M-H]^-	-0.936	311.222 41, 293.211 7	Sanleng acid	PC	Phenolic acids
30	C₁₁H₁₈NO	9.00	181.146 11	181.085 91	[M+H]⁺	-0.060	121.101 23, 93.070 31	N-Candicine	PC	Alkaloid
31	C₁₉H₁₈O₁₁	4.88	421.077 63	421.077 97	[M-H]^-	0.225	331.045 90, 301.035 10	Mangiferin	RA	Flavonoids
32	C₂₆H₃₄O₁₁	6.81	521.202 83	521.203 19	[M-H]^-	0.219	329.139 34, 175.075 52	²⁾	PC	Glycoside
33	C₁₅H₂₄N₆O₉	4.17	431.153 19	431.155 64	[M-H]^-	0.811	191.054 50, 119.048 93	2-(p-hydroxyphenyl)ethanol1-O-β-D-glucoside	PC	Glycoside
34	C₄₅H₇₆O₂₀	7.74	935.485 71	935.485 41	[M-H]^-	-0.032	773.437 68, 449.330 32	Timosaponin N	RA	Saponins
35	C₃₉H₆₄O₁₄	11.91	755.422 32	755.422 3	[M-H]^-	0.089	325.255 52, 593.371 22	Timosaponin G	RA	Saponins
36	C₁₇H₁₆O₂	11.55	251.107 75	251.107 41	[M-H]^-	1.001	236.082 70, 157.064 74	Nyasol	RA	Phenylpropionin
37	C₁₄H₈N₂O	9.90	221.070 93	221.073 81	[M+H]⁺	-1.435	177.044 76, 150.036 79	6H-Indolo(3,2,1-de)(1,5)naphthyridin-6-one	PC	Alkaloid
38	C₁₆H₁₅NO₃	4.02	268.097 91	268.127 44	[M-H]^-	-1.491	250.099 72, 235.075 2	4-[N-(p-Methoxy-benzylidene)amino]-phenyl acetate	PC	Phenolic acids
39	C₁₃H₁₁NO₃	10.12	230.081 16	230.080 46	[M+H]⁺	2.751	215.057 01, 200.033 66	Γ-fagarine	PC	Alkaloid
40	C₁₅H₁₀O₇	4.83	303.045 62	303.050 17	[M+H]⁺	0.543	285.039 85, 275.055 94	Quercetin	PC	Flavonoids
41	C₂₆H₃₀O₈	11.00	469.195 67	469.189 45	[M-H]^-	-0.080	425.205 94	Limonin	PC	Lactones
42	C₁₄H₁₃NO₄	5.51	260.091 73	260.185 06	[M+H]⁺	-2.209	242.175 57, 237.908 63	Skimmianine	PC	Alkaloid
43	C₁₆H₁₄O₄	8.36	269.081 93	269.210 75	[M-H]^-	-1.379	253.216 60, 231.007 11	2'-O-methyl-isoliquiritigenin	RA	Flavonoids
44	C₂₆H₃₀O₉	13.53	485.181 70	485.282 71	[M-H]^-	-0.104	397.166 38, 383.149 08	Rutaevin	PC	Lactones
45	C₁₇H₂₄O₉	12.74	373.149 30	373.272 58	[M+H]⁺	0.539	355.262 42, 337.252 5	Syringin	PC	Phenols
46	C₂₁H₂₀O₁₂	7.86	463.088 19	463.306 52	[M-H]^-	-0.503	377.271 30, 359.261 08	Hyperoside	PC	Flavonoids

No.

Molecular
formula

t_R
/min

Theoretical
mass(m/z)

Experimental
mass(m/z)

Adduct

Error
(×10^-6)

m/z
(MS²)

Component

Source

No.	Molecular formula	t_R /min	Theoretical mass(m/z)	Experimental mass(m/z)	Adduct	Error (×10^-6)	m/z (MS²)	Component	Source	Category
1	C₂₁H₂₁NO₄	8.92	352.154 33	352.153 53	[M+H]⁺	0.086	336.122 25, 308.127 11	Palmatine	PC	Alkaloid
2	C₂₀H₂₀NO₇	3.80	387.131 25	387.189 94	[M+H]⁺	-0.428	145.028 03, 177.054 14	Trihydroxy jatrorrhizine	PC	Alkaloid
3	C₂₁H₁₃N₃O	8.30	324.113 13	324.121 95	[M+H]⁺	-0.754	308.090 70, 280.095 89	7,8-deoxyrutaecarpine	PC	Alkaloid
4	C₂₀H₁₇NO₅	7.52	352.117 94	352.116 85	[M+H]⁺	0.692	336.085 60, 308.090 73	Berlambine	PC	Alkaloid
5	C₂₀H₁₉NO₄	8.20	336.123 03	336.121 83	[M+H]⁺	0.41	320.090 70, 306.518 68	Berbine	PC	Alkaloid
6	C₂₁H₁₈N $O 4 +$	9.66	350.138 68	350.137 57	[M+H]⁺	0.651	334.106 29, 320.090 7	Chelerythrine	PC	Alkaloid
7	C₂₇H₃₀O₁₀	11.53	513.176 62	513.138 79	[M-H]^-	0.321	367.139 50	Baohuoside Ⅰ	RA	Flavonoids
8	C₁₉H₁₅NO₄	8.36	322.107 38	322.106 32	[M+H]⁺	-0.696	307.082 89, 279.087 95	Menisporphine	PC	Alkaloid
9	C₂₀H₂₁NO₄	8.03	339.146 5	339.140 87	[M+H]⁺	-0.79	323.113 01, 308.090 27	Flavonoids	PC	Alkaloid
10	C₂₁H₂₅NO₄	7.19	356.185 63	356.184 33	[M+H]⁺	0.106	311.126 56, 296.103 39	Menisperine	PC	Alkaloid
11	C₁₉H₁₇NO₄	7.36	324.123 03	324.122 16	[M+H]⁺	-0.754	308.090 70, 294.075 29	Berberrubine	PC	Alkaloid
12	C₁₇H₂₀O₉	5.44	367.103 45	367.103 27	[M-H]^-	-0.362	191.055 31, 173.044 59	3-O-Feruloylquinic acid	PC	Phenolic acids
13	C₁₆H₁₈O₉	3.22	353.087 8	353.086 06	[M-H]^-	-0.002	191.055 24, 179.034 03	Chlorogenic acid	PC	Phenolic acids
14	C₁₇H₁₇NO₅	4.14	316.117 94	316.283 69	[M+H]⁺	1.303	298.274 02, 280.262 12	Hippeastrine	PC	Alkaloid
15	C₂₆H₃₀O₁₂	6.88	533.166 44	533.151 55	[M-H]^-	0.218	371.100 46, 325.091 77	Aegineoside	PC	Flavonoids
16	C₂₃H₂₉NO₈	5.15	448.196 59	448.195 28	[M+H]⁺	0.066	107.049 18, 286.142 79	¹⁾	PC	Glycoside
17	C₁₅H₁₇NO₃	6.79	260.128 11	260.127 38	[M+H]⁺	2.317	242.116 73, 224.106 48	Pilocarpine	PC	Alkaloid
18	C1₉H₂₃NO₃	6.35	314.175 07	314.173 95	[M+H]⁺	0.517	299.112 82, 269.116 82	Isoliensinine	PC	Alkaloid
19	C₃₉H₆₄O₁₃	10.22	739.427 41	739.428 16	[M-H]^-	0.002	577.376 65, 529.609 62	Timosaponin AⅢ	RA	Saponins
20	C₂₅H₂₈O₁₆	3.86	583.130 45	583.129 39	[M-H]^-	0.050	421.077 61, 331.045 84	Neomangiferin	RA	Flavonoids
21	C₄₅H₇₄O₁₉	8.06	917.475 15	917.476 14	[M-H]^-	-0.085	71.012 41,89.023	Anemarsaponin D	RA	Saponins
22	C₁₃H₁₈O₈	1.66	301.092 89	301.093 66	[M-H]^-	1.756	123.007 48, 138.031 19	Tachioside	PC	Phenolic acids
23	C₁₉H₁₈O₁₁	4.88	421.077 63	421.077 97	[M-H]^-	0.225	301.035 10, 331.045 90	Isomangiferin	RA	Flavonoids
24	C₄₅H₇₄O₁₈	9.78	901.480 23	901.484 13	[M-H]^-	-0.026	101.023 06, 113.023 09	Anemarsaponin B	RA	Saponins
25	C₄₅H₇₆O₁₉	8.37	919.490 8	919.489 75	[M-H]^-	0.048	757.432 56, 595.383 79	Anemarsaponin BⅡ	RA	Saponins
26	C₂₀H₁₉NO₄	7.69	338.138 68	338.137 45	[M+H]⁺	0.320	323.113 83, 294.111 02	Jatrorrhizine	PC	Alkaloid
27	C₂₁H₂₀O₁₀	6.58	431.098 37	431.097 93	[M-H]^-	0.346	311.056 12, 283.061 00	Afzelin	RA	Flavonoids
28	C₂₀H₂₃NO₄	5.56	342.169 98	342.169 19	[M+H]⁺	0.133	297.111 05, 265.085 17	Magnoflorine	PC	Alkaloid
29	C₁₈H₃₄O₅	10.30	329.233 34	329.233 28	[M-H]^-	-0.936	311.222 41, 293.211 7	Sanleng acid	PC	Phenolic acids
30	C₁₁H₁₈NO	9.00	181.146 11	181.085 91	[M+H]⁺	-0.060	121.101 23, 93.070 31	N-Candicine	PC	Alkaloid
31	C₁₉H₁₈O₁₁	4.88	421.077 63	421.077 97	[M-H]^-	0.225	331.045 90, 301.035 10	Mangiferin	RA	Flavonoids
32	C₂₆H₃₄O₁₁	6.81	521.202 83	521.203 19	[M-H]^-	0.219	329.139 34, 175.075 52	²⁾	PC	Glycoside
33	C₁₅H₂₄N₆O₉	4.17	431.153 19	431.155 64	[M-H]^-	0.811	191.054 50, 119.048 93	2-(p-hydroxyphenyl)ethanol1-O-β-D-glucoside	PC	Glycoside
34	C₄₅H₇₆O₂₀	7.74	935.485 71	935.485 41	[M-H]^-	-0.032	773.437 68, 449.330 32	Timosaponin N	RA	Saponins
35	C₃₉H₆₄O₁₄	11.91	755.422 32	755.422 3	[M-H]^-	0.089	325.255 52, 593.371 22	Timosaponin G	RA	Saponins
36	C₁₇H₁₆O₂	11.55	251.107 75	251.107 41	[M-H]^-	1.001	236.082 70, 157.064 74	Nyasol	RA	Phenylpropionin
37	C₁₄H₈N₂O	9.90	221.070 93	221.073 81	[M+H]⁺	-1.435	177.044 76, 150.036 79	6H-Indolo(3,2,1-de)(1,5)naphthyridin-6-one	PC	Alkaloid
38	C₁₆H₁₅NO₃	4.02	268.097 91	268.127 44	[M-H]^-	-1.491	250.099 72, 235.075 2	4-[N-(p-Methoxy-benzylidene)amino]-phenyl acetate	PC	Phenolic acids
39	C₁₃H₁₁NO₃	10.12	230.081 16	230.080 46	[M+H]⁺	2.751	215.057 01, 200.033 66	Γ-fagarine	PC	Alkaloid
40	C₁₅H₁₀O₇	4.83	303.045 62	303.050 17	[M+H]⁺	0.543	285.039 85, 275.055 94	Quercetin	PC	Flavonoids
41	C₂₆H₃₀O₈	11.00	469.195 67	469.189 45	[M-H]^-	-0.080	425.205 94	Limonin	PC	Lactones
42	C₁₄H₁₃NO₄	5.51	260.091 73	260.185 06	[M+H]⁺	-2.209	242.175 57, 237.908 63	Skimmianine	PC	Alkaloid
43	C₁₆H₁₄O₄	8.36	269.081 93	269.210 75	[M-H]^-	-1.379	253.216 60, 231.007 11	2'-O-methyl-isoliquiritigenin	RA	Flavonoids
44	C₂₆H₃₀O₉	13.53	485.181 70	485.282 71	[M-H]^-	-0.104	397.166 38, 383.149 08	Rutaevin	PC	Lactones
45	C₁₇H₂₄O₉	12.74	373.149 30	373.272 58	[M+H]⁺	0.539	355.262 42, 337.252 5	Syringin	PC	Phenols
46	C₂₁H₂₀O₁₂	7.86	463.088 19	463.306 52	[M-H]^-	-0.503	377.271 30, 359.261 08	Hyperoside	PC	Flavonoids

Category

C₂₁H₂₁NO₄

8.92

352.154 33

352.153 53

[M+H]⁺

0.086

336.122 25, 308.127 11

Palmatine

Alkaloid

C₂₀H₂₀NO₇

3.80

387.131 25

387.189 94

[M+H]⁺

-0.428

145.028 03, 177.054 14

Trihydroxy jatrorrhizine

Alkaloid

C₂₁H₁₃N₃O

8.30

324.113 13

324.121 95

[M+H]⁺

-0.754

308.090 70, 280.095 89

7,8-deoxyrutaecarpine

Alkaloid

C₂₀H₁₇NO₅

7.52

352.117 94

352.116 85

[M+H]⁺

0.692

336.085 60, 308.090 73

Berlambine

Alkaloid

C₂₀H₁₉NO₄

8.20

336.123 03

336.121 83

[M+H]⁺

0.41

320.090 70, 306.518 68

Berbine

Alkaloid

C₂₁H₁₈N

O 4 +

9.66

350.138 68

350.137 57

[M+H]⁺

0.651

334.106 29, 320.090 7

Chelerythrine

Alkaloid

C₂₇H₃₀O₁₀

11.53

513.176 62

513.138 79

[M-H]^-

0.321

367.139 50

Baohuoside Ⅰ

Flavonoids

C₁₉H₁₅NO₄

8.36

322.107 38

322.106 32

[M+H]⁺

-0.696

307.082 89, 279.087 95

Menisporphine

Alkaloid

C₂₀H₂₁NO₄

8.03

339.146 5

339.140 87

[M+H]⁺

-0.79

323.113 01, 308.090 27

Flavonoids

Alkaloid

C₂₁H₂₅NO₄

7.19

356.185 63

356.184 33

[M+H]⁺

0.106

311.126 56, 296.103 39

Menisperine

Alkaloid

C₁₉H₁₇NO₄

7.36

324.123 03

324.122 16

[M+H]⁺

-0.754

308.090 70, 294.075 29

Berberrubine

Alkaloid

C₁₇H₂₀O₉

5.44

367.103 45

367.103 27

[M-H]^-

-0.362

191.055 31, 173.044 59

3-O-Feruloylquinic acid

Phenolic acids

C₁₆H₁₈O₉

3.22

353.087 8

353.086 06

[M-H]^-

-0.002

191.055 24, 179.034 03

Chlorogenic acid

Phenolic acids

C₁₇H₁₇NO₅

4.14

316.117 94

316.283 69

[M+H]⁺

1.303

298.274 02, 280.262 12

Hippeastrine

Alkaloid

C₂₆H₃₀O₁₂

6.88

533.166 44

533.151 55

[M-H]^-

0.218

371.100 46, 325.091 77

Aegineoside

Flavonoids

C₂₃H₂₉NO₈

5.15

448.196 59

448.195 28

[M+H]⁺

0.066

107.049 18, 286.142 79

¹⁾

Glycoside

C₁₅H₁₇NO₃

6.79

260.128 11

260.127 38

[M+H]⁺

2.317

242.116 73, 224.106 48

Pilocarpine

Alkaloid

C1₉H₂₃NO₃

6.35

314.175 07

314.173 95

[M+H]⁺

0.517

299.112 82, 269.116 82

Isoliensinine

Alkaloid

C₃₉H₆₄O₁₃

10.22

739.427 41

739.428 16

[M-H]^-

0.002

577.376 65, 529.609 62

Timosaponin AⅢ

Saponins

C₂₅H₂₈O₁₆

3.86

583.130 45

583.129 39

[M-H]^-

0.050

421.077 61, 331.045 84

Neomangiferin

Flavonoids

C₄₅H₇₄O₁₉

8.06

917.475 15

917.476 14

[M-H]^-

-0.085

71.012 41,89.023

Anemarsaponin D

Saponins

C₁₃H₁₈O₈

1.66

301.092 89

301.093 66

[M-H]^-

1.756

123.007 48, 138.031 19

Tachioside

Phenolic acids

C₁₉H₁₈O₁₁

4.88

421.077 63

421.077 97

[M-H]^-

0.225

301.035 10, 331.045 90

Isomangiferin

Flavonoids

C₄₅H₇₄O₁₈

9.78

901.480 23

901.484 13

[M-H]^-

-0.026

101.023 06, 113.023 09

Anemarsaponin B

Saponins

C₄₅H₇₆O₁₉

8.37

919.490 8

919.489 75

[M-H]^-

0.048

757.432 56, 595.383 79

Anemarsaponin BⅡ

Saponins

C₂₀H₁₉NO₄

7.69

338.138 68

338.137 45

[M+H]⁺

0.320

323.113 83, 294.111 02

Jatrorrhizine

Alkaloid

C₂₁H₂₀O₁₀

6.58

431.098 37

431.097 93

[M-H]^-

0.346

311.056 12, 283.061 00

Afzelin

Flavonoids

C₂₀H₂₃NO₄

5.56

342.169 98

342.169 19

[M+H]⁺

0.133

297.111 05, 265.085 17

Magnoflorine

Alkaloid

C₁₈H₃₄O₅

10.30

329.233 34

329.233 28

[M-H]^-

-0.936

311.222 41, 293.211 7

Sanleng acid

Phenolic acids

C₁₁H₁₈NO

9.00

181.146 11

181.085 91

[M+H]⁺

-0.060

121.101 23, 93.070 31

N-Candicine

Alkaloid

C₁₉H₁₈O₁₁

4.88

421.077 63

421.077 97

[M-H]^-

0.225

331.045 90, 301.035 10

Mangiferin

Flavonoids

C₂₆H₃₄O₁₁

6.81

521.202 83

521.203 19

[M-H]^-

0.219

329.139 34, 175.075 52

²⁾

Glycoside

C₁₅H₂₄N₆O₉

4.17

431.153 19

431.155 64

[M-H]^-

0.811

191.054 50, 119.048 93

2-(p-hydroxyphenyl)ethanol1-O-β-D-glucoside

Glycoside

C₄₅H₇₆O₂₀

7.74

935.485 71

935.485 41

[M-H]^-

-0.032

773.437 68, 449.330 32

Timosaponin N

Saponins

C₃₉H₆₄O₁₄

11.91

755.422 32

755.422 3

[M-H]^-

0.089

325.255 52, 593.371 22

Timosaponin G

Saponins

C₁₇H₁₆O₂

11.55

251.107 75

251.107 41

[M-H]^-

1.001

236.082 70, 157.064 74

Nyasol

Phenylpropionin

C₁₄H₈N₂O

9.90

221.070 93

221.073 81

[M+H]⁺

-1.435

177.044 76, 150.036 79

6H-Indolo(3,2,1-de)(1,5)naphthyridin-6-one

Alkaloid

C₁₆H₁₅NO₃

4.02

268.097 91

268.127 44

[M-H]^-

-1.491

250.099 72, 235.075 2

4-[N-(p-Methoxy-benzylidene)amino]-phenyl acetate

Phenolic acids

C₁₃H₁₁NO₃

10.12

230.081 16

230.080 46

[M+H]⁺

2.751

215.057 01, 200.033 66

Γ-fagarine

Alkaloid

C₁₅H₁₀O₇

4.83

303.045 62

303.050 17

[M+H]⁺

0.543

285.039 85, 275.055 94

Quercetin

Flavonoids

C₂₆H₃₀O₈

11.00

469.195 67

469.189 45

[M-H]^-

-0.080

425.205 94

Limonin

Lactones

C₁₄H₁₃NO₄

5.51

260.091 73

260.185 06

[M+H]⁺

-2.209

242.175 57, 237.908 63

Skimmianine

Alkaloid

C₁₆H₁₄O₄

8.36

269.081 93

269.210 75

[M-H]^-

-1.379

253.216 60, 231.007 11

2'-O-methyl-isoliquiritigenin

Flavonoids

C₂₆H₃₀O₉

13.53

485.181 70

485.282 71

[M-H]^-

-0.104

397.166 38, 383.149 08

Rutaevin

Lactones

C₁₇H₂₄O₉

12.74

373.149 30

373.272 58

[M+H]⁺

0.539

355.262 42, 337.252 5

Syringin

Phenols

C₂₁H₂₀O₁₂

7.86

463.088 19

463.306 52

[M-H]^-

-0.503

377.271 30, 359.261 08

Hyperoside

Flavonoids

No.	Molecular formula	t_R /min	Theoretical mass(m/z)	Experimental mass(m/z)	Adduct	Error (×10^-6)	m/z (MS²)	Identification	Prototype ingredients
M1	C₁₉H₁₆NO₄	11.70	322.107 38	322.190 95	[M+H]⁺	-0.439	307.166 87	Thalifendin or berberrubinen	Berberine
M2	C₂₅H₂₄NO₁₀	6.74	498.139 47	498.138 76	[M+H]⁺	0.231	322.106 78	Thalifendin-O-glucuronide or Berberrubinen-O-glucuronide	Berberine
M3	C₃₀H₃₀NO₁₆	16.36	660.155 91	660.423 46	[M+H]⁺	-0.328	308.710 30	Demethylenecoptisine-di-O-glucuronide	Berberine
M4	C₂₅H₂₆NO₁₀	6.26	500.155 12	500.153 84	[M+H]⁺	0.121	324.122 44	Demethyleneepiberberine-O-glucuronide	Jatrorrhizine
M5	C₂₆H₂₈NO₁₀	6.52	514.170 77	514.169 49	[M+H]⁺	0.117	338.138 12	Jateorhizine-O-glucuronide	Jatrorrhizine
M6	C₂₀H₂₀O₁₁	5.86	435.092 18	435.092 96	[M-H]^-	-0.565	259.061 07	Methylation	Mangiferin
M7	C₁₉H₁₅O₁₂	1.4	435.092 18	436.953 00	[M-H]^-	-0.351	315.041 02	Monomethylated Mangiferin	Mangiferin
M8	C₁₃H₈O₆	5.59	260.031 53	261.030 09	[M-H]^-	0.436	221.841 96	Norathyriol	Mangiferin
M9	C₂₅H₂₄O₁₀N	6.74	498.139 47	498.138 76	[M+H]⁺	0.123	322.106 78	9-O-β-D-Glucuronic acid berberine	Berberrubine

No.

Molecular
formula

t_R
/min

Theoretical
mass(m/z)

Experimental
mass(m/z)

Adduct

Error
(×10^-6)

m/z
(MS²)

Identification

Prototype
ingredients

C₁₉H₁₆NO₄

11.70

322.107 38

322.190 95

[M+H]⁺

-0.439

307.166 87

Thalifendin or berberrubinen

Berberine

C₂₅H₂₄NO₁₀

6.74

498.139 47

498.138 76

[M+H]⁺

0.231

322.106 78

Thalifendin-O-glucuronide or Berberrubinen-O-glucuronide

Berberine

C₃₀H₃₀NO₁₆

16.36

660.155 91

660.423 46

[M+H]⁺

-0.328

308.710 30

Demethylenecoptisine-di-O-glucuronide

Berberine

C₂₅H₂₆NO₁₀

6.26

500.155 12

500.153 84

[M+H]⁺

0.121

324.122 44

Demethyleneepiberberine-O-glucuronide

Jatrorrhizine

C₂₆H₂₈NO₁₀

6.52

514.170 77

514.169 49

[M+H]⁺

0.117

338.138 12

Jateorhizine-O-glucuronide

Jatrorrhizine

C₂₀H₂₀O₁₁

5.86

435.092 18

435.092 96

[M-H]^-

-0.565

259.061 07

Methylation

Mangiferin

C₁₉H₁₅O₁₂

1.4

435.092 18

436.953 00

[M-H]^-

-0.351

315.041 02

Monomethylated Mangiferin

Mangiferin

C₁₃H₈O₆

5.59

260.031 53

261.030 09

[M-H]^-

0.436

221.841 96

Norathyriol

Mangiferin

C₂₅H₂₄O₁₀N

6.74

498.139 47

498.138 76

[M+H]⁺

0.123

322.106 78

9-O-β-D-Glucuronic acid berberine

Berberrubine

No.	Molecular formula	t_R /min	Theoretical mass(m/z)	Experimental mass(m/z)	Adduct	Error (×10^-6)	m/z (MS²)	Identification	Prototype ingredients
M1	C₁₉H₁₆NO₄	11.70	322.107 38	322.190 95	[M+H]⁺	-0.439	307.166 87	Thalifendin or berberrubinen	Berberine
M2	C₂₅H₂₄NO₁₀	6.74	498.139 47	498.138 76	[M+H]⁺	0.231	322.106 78	Thalifendin-O-glucuronide or Berberrubinen-O-glucuronide	Berberine
M3	C₃₀H₃₀NO₁₆	16.36	660.155 91	660.423 46	[M+H]⁺	-0.328	308.710 30	Demethylenecoptisine-di-O-glucuronide	Berberine
M4	C₂₅H₂₆NO₁₀	6.26	500.155 12	500.153 84	[M+H]⁺	0.121	324.122 44	Demethyleneepiberberine-O-glucuronide	Jatrorrhizine
M5	C₂₆H₂₈NO₁₀	6.52	514.170 77	514.169 49	[M+H]⁺	0.117	338.138 12	Jateorhizine-O-glucuronide	Jatrorrhizine
M6	C₂₀H₂₀O₁₁	5.86	435.092 18	435.092 96	[M-H]^-	-0.565	259.061 07	Methylation	Mangiferin
M7	C₁₉H₁₅O₁₂	1.4	435.092 18	436.953 00	[M-H]^-	-0.351	315.041 02	Monomethylated Mangiferin	Mangiferin
M8	C₁₃H₈O₆	5.59	260.031 53	261.030 09	[M-H]^-	0.436	221.841 96	Norathyriol	Mangiferin
M9	C₂₅H₂₄O₁₀N	6.74	498.139 47	498.138 76	[M+H]⁺	0.123	322.106 78	9-O-β-D-Glucuronic acid berberine	Berberrubine

No.

Molecular
formula

t_R
/min

Theoretical
mass(m/z)

Experimental
mass(m/z)

Adduct

Error
(×10^-6)

m/z
(MS²)

Identification

Prototype
ingredients

C₁₉H₁₆NO₄

11.70

322.107 38

322.190 95

[M+H]⁺

-0.439

307.166 87

Thalifendin or berberrubinen

Berberine

C₂₅H₂₄NO₁₀

6.74

498.139 47

498.138 76

[M+H]⁺

0.231

322.106 78

Thalifendin-O-glucuronide or Berberrubinen-O-glucuronide

Berberine

C₃₀H₃₀NO₁₆

16.36

660.155 91

660.423 46

[M+H]⁺

-0.328

308.710 30

Demethylenecoptisine-di-O-glucuronide

Berberine

C₂₅H₂₆NO₁₀

6.26

500.155 12

500.153 84

[M+H]⁺

0.121

324.122 44

Demethyleneepiberberine-O-glucuronide

Jatrorrhizine

C₂₆H₂₈NO₁₀

6.52

514.170 77

514.169 49

[M+H]⁺

0.117

338.138 12

Jateorhizine-O-glucuronide

Jatrorrhizine

C₂₀H₂₀O₁₁

5.86

435.092 18

435.092 96

[M-H]^-

-0.565

259.061 07

Methylation

Mangiferin

C₁₉H₁₅O₁₂

1.4

435.092 18

436.953 00

[M-H]^-

-0.351

315.041 02

Monomethylated Mangiferin

Mangiferin

C₁₃H₈O₆

5.59

260.031 53

261.030 09

[M-H]^-

0.436

221.841 96

Norathyriol

Mangiferin

C₂₅H₂₄O₁₀N

6.74

498.139 47

498.138 76

[M+H]⁺

0.123

322.106 78

9-O-β-D-Glucuronic acid berberine

Berberrubine

基于UPLC-Q-Exactive Orbitrap-MS的知母-黄柏药对入血成分分析

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夏天利 ¹ , 王岩 ² , 王迪 ¹ , 赵德萍 ³ , 苏婷 ¹ , 赵继会 ² , 雷霞 ⁴^,^* , 张宁 ¹^,³^,^*

中国药学杂志 | 论著 2024,59(4): 330-337

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中国药学杂志 | 论著 2024, 59(4): 330-337

基于UPLC-Q-Exactive Orbitrap-MS的知母-黄柏药对入血成分分析

全屏

夏天利¹, 王岩², 王迪¹, 赵德萍³, 苏婷¹, 赵继会², 雷霞⁴^,^*, 张宁¹^,³^,^*

作者信息

¹ 佳木斯大学, 黑龙江佳木斯 154007

² 湖南医药学院药学院, 湖南怀化 418000

³ 黑龙江中医药大学药学院, 哈尔滨 150040

⁴ 无锡市中医医院中医药研究所, 江苏无锡 214071

夏天利,女,硕士研究生研究方向:药物分析

通讯作者:

*雷霞,女,博士,副教授研究方向:药理学 Tel:(0510)81008810;

张宁,男,博士,教授研究方向:药物分析 Tel:(0451)82114400

Analysis of Absorbed Components of Rhizoma Anemarrhenae-Cortex Phellodendri Chinesis Herb-Pair Based on UPLC-Q-Exactive Orbitrap-MS

XIA Tianli¹, WANG Yan², WANG Di¹, ZHAO Deping³, SU Ting¹, ZHAO Jihui², LEI Xia⁴^,^*, ZHANG Ning¹^,³^,^*

Affiliations

¹ Jiamusi University, Jiamusi 154007, China

² School of Pharmaceutical Sciences, Hunan University of Medicine, Huaihua 418000, China

³ School of Pharmacy, Heilongjiang University of Traditional Chinese Medicine, Harbin 150040, China

⁴ Institute of Traditional Chinese Medicine, Wuxi Traditional Chinese Medicine Hospital, Wuxi 214071, China

出版时间: 2024-02-22 doi: 10.11669/cpj.2024.04.006

文章导航

摘要

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目的以正常家兔为实验对象,探讨知母-黄柏药对口服吸收后的入血成分。方法采用相当于生药量4.5 g·kg^-1的知母-黄柏水煎液灌胃家兔,每日1次,连续7 d,末次给药后取家兔含药血清,利用超高效液相色谱-四级杆-静电场轨道阱高分辨质谱联用技术(UPLC-Q-Exactive Orbitrap-MS),在正、负两种离子模式下,根据保留时间、精确相对分子质量、二级质谱裂解碎片,确定入血成分。结果家兔含药血清中共鉴别出54个入血成分,包括46个原型入血成分,包括生物碱类、黄酮类、皂苷类、糖苷类、酚酸类、苯丙素类、内酯类、酚类衍生物;9个代谢产物,分别来源于小檗碱、药根碱、小檗红碱、芒果苷。结论知母-黄柏药对中含有46个可以入血的化学成分,还可能含有4个化学成分在体内代谢为9个代谢产物。

关键词

知母 / 黄柏 / 药对 / 超高效液相色谱-四极杆-静电场轨道阱高分辨质谱联用技术 / 入血成分

Abstract

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OBJECTIVE To study the components of Rhizoma Anemarrhenae-Cortex Phellodendri Chinesis (RA-PC) herb-pair absorbed into blood in normal rabbits. METHODS Rabbits were fed with the decoction of RA-PC herb pair (4.5 g·kg^-1) once a day for 7 days, then blood samples were taken. High-performance liquid chromatography-quadrupole-electrostatic field orbital trap (UPLC-Q-Exactive Orbitrap-MS) technique was applied for the analysis, and fragments were split in both positive and negative ion modes. The compositions into blood were determined according to retention time, accurate relative molecular mass, and secondary mass spectra. RESULTS A total of 54 components were identified in the drug-containing serum of rabbits, including 46 prototype components, which were alkaloids, flavonoids, saponins, glycosides, phenolic acids, phenylpropanoids, lactones, and phenolic derivatives. The nine metabolites were derived from berberine, jatrorrhizine, berberrubine, and mangiferin, respectively. CONCLUSION RA-PC contains 46 chemical components that can enter blood of normal rabbits, and may also contains four chemical components which are metabolized into nine metabolites.

Key words

Rhizoma Anemarrhenae / Cortex Phellodendri Chinesis / herb pair / UPLC-Q-Exactive Orbitrap-MS / absorbed component

引用本文

夏天利, 王岩, 王迪, 赵德萍, 苏婷, 赵继会, 雷霞, 张宁. 基于UPLC-Q-Exactive Orbitrap-MS的知母-黄柏药对入血成分分析. 中国药学杂志, 2024 , 59 (4) : 330 -337 . DOI: 10.11669/cpj.2024.04.006

XIA Tianli, WANG Yan, WANG Di, ZHAO Deping, SU Ting, ZHAO Jihui, LEI Xia, ZHANG Ning. Analysis of Absorbed Components of Rhizoma Anemarrhenae-Cortex Phellodendri Chinesis Herb-Pair Based on UPLC-Q-Exactive Orbitrap-MS[J]. Chinese Pharmaceutical Journal, 2024 , 59 (4) : 330 -337 . DOI: 10.11669/cpj.2024.04.006

正文

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知母-黄柏药对(以下简称“知柏”)是中医临床治疗阴虚内热证的经典药对,最早出自金李杲的《兰室秘藏》^[1],在知柏地黄丸、滋肾丸、大补阴丸等方剂中都用到了知母和黄柏的配伍。知母甘寒上清肺金而泻火,下润肾燥而滋阴;黄柏苦寒坚阴,清热燥湿,泻火解毒,善退虚热。二药伍用,发挥滋肾阴之亏损,兼泻妄动之相火的功效,临床常用其治疗糖尿病^[2]。

目前国内外对于知母、黄柏的研究较多,但大多集中于对药理作用的研究。研究表明,知母中的主要成分为皂苷及黄酮类化合物,其中知母皂苷类物质具有改善老年痴呆症状、降血糖、抗肿瘤等药理作用^[3];黄柏中多以生物碱类物质为主,具有抗炎、抗氧化、抑菌等药理作用^[4]。对于知柏的体内吸收成分研究较少,对于知母,大多集中于皂苷类物质及以芒果苷为代表的黄酮类物质的研究,知母中苯丙素类物质的体内情况研究较少;对于黄柏,更多侧重于对其药理活性的研究,或是以生物碱类物质为代表的某一大类成分的研究,有关黄柏酚酸类、糖苷类、内酯类及酚类衍生物的体内情况研究较少。目前缺乏对知柏入血成分的整体分析药对配伍是通过协同作用发挥药效,因此有必要对入血成分进行全面研究,以明确药效成分,以期阐明药效物质基础^[5]。

1 材料

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1.1 动物

家兔12只,雄性,黑龙江中医药大学实验中心,合格证号:SCXK(黑)2018-003。SPF级环境饲养,饲养环境温度范围-22~20 ℃,相对湿度(50±10)%,自由采食、饮水,分笼饲养,每笼1只。实验流程均符合黑龙江中医药大学实验动物管理委员会对实验动物管理和保护的相关规定。

1.2 仪器

Dionex UltiMate 3000超高液相色谱与Thermo Q-Exactive Focus四极杆-静电场轨道阱高分辨质谱仪(赛默飞世尔科技公司);Milli-Q纯水器(密理博上海贸易有限公司);Mikro 200R离心机(德国Hettich公司)。

1.3 药材与试剂

知母饮片、黄柏饮片均购自北京同仁堂药店(哈尔滨店),并经黑龙江中医药大学佳木斯学院陈效忠副教授鉴定为百合科植物知母(Anemarrhena asphodeloides Bge.)的干燥根茎以及芸香科植物黄皮树(Phellodendron chinense Schneid.)的干燥树皮,符合2020年版《中国药典》标准。甲醇、乙腈、甲酸(色谱纯)。

2 方法

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2.1 知母-黄柏药液的制备

将知母饮片、黄柏饮片按1∶1混合,加入10倍量水,煎煮1 h,过滤,收集滤液,残渣加入8倍量水煎煮1 h,合并两次滤液,浓缩至相当于饮片1 g·mL^-1,即得。

2.2 动物给药与含药血清样品采集

将家兔随机分为空白组和给药组,每组6只,实验前12 h禁食不禁水,给药组按饮片量4.5 g·kg^-1的剂量灌胃给予知母-黄柏药液,空白组灌胃给予等量水,分别于给药后30、60、90、120 min心脏取血(左手放在右胸处,轻轻向左推,将心脏固定。拇指在兔胸左侧由下向上数第 3 至第 4肋骨间探测心脏搏动最剧烈处,右手持注射器,在此搏动最强烈处刺入心脏,缓慢抽取血液,达足量时,拔出注射器),室温静置30 min后,离心,取血清,用0.22 μm过滤器无菌过滤,-80 ℃保存备用。

2.3 血清样本处理

血清样品置于4 ℃解冻后,取500 μL血清样品,加入2 mL甲醇溶液涡旋混合1 min,超声2 min,以13 000 r·min^-1力、4 ℃下离心10 min,取上清液置真空浓缩仪中,40 ℃下吹干。复溶:加100 μL 体积分数70%的甲醇溶液复溶,0.22 μm微孔滤膜过滤,进样5 μL。

2.4 超高效液相色谱-四极杆-静电场轨道阱高分辨质谱联用(UPLC-Q-Exactive Orbitrap-MS)条件

2.4.1 色谱条件

色谱柱:ACQUITY UPLC^TM HSS T₃柱(2.1 mm×100 mm,1.8 μm);柱温:40 ℃;流动相:0.1%甲酸的水溶液(流动相A)、0.1%甲酸的乙腈溶液(流动相B);梯度洗脱:0~3.5 min,0%~15%B;3.5~6 min,15%~30%B;6~12.5 min,30%~70%B;12.5~18 min,70%~100%B;流速:0.4 mL·min^-1。

2.4.2 质谱条件

电喷雾离子源,正、负离子模式下扫描。质谱扫描范围:m/z 50~1500,离子传输管和辅助气温度分别为320 ℃和350 ℃,正离子模式下:喷雾电压为3.5 kV;负离子模式下:喷雾电压为3.2 kV;鞘气压力:275 800 Pa;碰撞能量:20,40,60 eV;分辨率:一级高分辨质谱70000 FWHM,二级高分辨质谱17500 FWHM;动态排除时间:5 s。

3 结果

收起

3.1 知母-黄柏入血成分鉴定

使用 UPLC-Q-Exactive Orbitrap-MS 技术,对知母-黄柏家兔入血成分进行定性分析,在正、负离子模式下的质谱高分辨提取离子流图见图1。入血成分总离子流图见图2。质谱分析软件 Thermo Scientific Xcalibu对主要出峰化合物进行辨认。根据保留时间和质谱特征、化合物 MS 一级质谱和 MS/MS 二级质谱信息,同时结合体外成分分析结果进行成分确认,结果见表1。

3.2 知母-黄柏入血成分质谱裂解规律

3.2.1 生物碱类

以化合物28为例,保留时间5.56 min,在正离子模式下,准分子离子峰为m/z 42.17 [M+H]⁺,分子式为C₂₀H₂₃NO₄,MS²产生碎片离子m/z 297.11 [M+H-C₂H₇N]⁺、m/z 265.09 [M+H-C₂H₇N-CH₃OH]⁺、m/z 237.09 [M+H-C₂H₇N-CH₃OH-CO]⁺,与参考文献[6]对比,初步鉴定为木兰花碱。裂解途径见图3。

3.2.2 酚酸类

以化合物13为例,保留时间为3.22 min,在负离子模式下,准分子离子峰为m/z 353.09 [M-H]^-,分子式为C₁₆H₁₈O₉,MS²产生碎片离子m/z 191.06 [M-H-caffeoul]^-,奎宁取代基位置可为1、3或5位,通过与文献[7]对比,初步鉴定为绿原酸。裂解途径见图4。

3.2.3 黄酮类

以化合物31为例,化合物23和31互为同分异构体,分子式均为C₁₉H₁₈O₁₁,准分子离子峰均为m/z 421.08 [M-H]^-,MS²均产生碎片离子m/z 331.05 [M-H-C₃H₆O₃]^-、m/z 301.04 [M-H-C₄H₈O₄]^-,这种方式为碳苷类化合物典型的裂解形式,结合文献[8-9]对比,初步鉴定化合物31为芒果苷,化合物23为异芒果苷。裂解途径见图5。

3.2.4 皂苷类

以化合物25为例,保留时间为8.37 min,在负离子模式下,准分子离子峰为m/z 919.49 [M-H]^-,分子式为C₄₅H₇₆O₁₉,MS²产生碎片离子m/z 757.43 [M-H-Glc]^-、m/z 595.38 [M-H-Glc-Gal]^-,通过与文献对比^[10],初步鉴定为知母皂苷BⅡ。推测裂解途径见图6。

3.2.5 内酯类

以化合物41为例,保留时间为11.00 min,在负离子模式下,准分子离子峰为m/z 469.19 [M-H]^-,分子式为C₂₆H₃₀O₈,MS²产生碎片离子m/z 425.20 [M-H-CO₂-H₂O]⁺、通过与文献[11]对比,初步鉴定为柠檬苦素。推测裂解途径见图7。

3.2.6 糖苷类

以化合物32为例,保留时间为6.81 min,在负离子模式下,准分子离子峰为m/z 431.16[M-H]^-,分子式为C₁₅H₂₄N₆O₉,MS²产生碎片离子m/z 269.05 [M-H-C₆H₁₀O₅]^-,通过与文献[12]对比,初步鉴定为2-(p-hydroxyphenyl)ethanol1-O-β-D-glucoside。

3.2.7 其他类成分

化合物45为酚类衍生物,通过与文献[13]对比,初步鉴定为丁香苷。化合物36为苯丙素类化合物,通过与文献[14]对比,初步鉴定为nyasol。

代谢产物结构鉴定:表2显示,家兔血清中共鉴定出小檗碱代谢产物3个,代谢途径分别为去甲基化、葡萄糖醛酸化;药根碱代谢产物2个,代谢途径分别为去甲基化、葡萄糖醛酸化;芒果苷代谢产物3个,代谢途径分别为甲基化、去糖基化;小檗红碱代谢产物1个,代谢途径为葡萄糖醛酸化。

4 讨论

收起

过往对知柏成分的研究主要集中于体外,而忽视了体内环境对中药成分的影响。目前,已有研究报道了知柏水煎液在大鼠体内的吸收情况,Ma等^[15]使用高效液相色谱-二极管阵列-串联质谱技术(HPLC-DAD-MS/MS)方法评估吸收入血的原型成分,包括4个生物碱类(木兰花碱、巴马丁、小檗碱、蝙蝠葛碱)、5个皂苷类(知母皂苷N或知母皂苷E1、知母皂苷D、知母皂苷C或知母皂苷B、知母皂苷BⅡ、知母皂苷AⅢ)。Sun等^[16]使用超高效液相色谱-离子阱-静电场轨道阱质谱技术(UHPLC-LTQ-Orbitrap MS)方法在大鼠体内发现16个原型入血成分,包括3个黄酮类(新芒果苷、芒果苷、异芒果苷)、5个皂苷类(知母皂苷N、知母皂苷C、知母皂苷BⅡ、知母皂苷AⅢ、知母皂苷E1)、6个生物碱类(小檗碱、巴马丁、黄柏碱、木兰花碱、四氢巴马丁、药根碱)、2个有机酸类(3-O-阿魏酰奎宁酸、5-O-阿魏酰奎宁酸),这些研究为揭示知柏的药效物质基础提供依据。现有研究表明^[17-18],知母和黄柏中至少有270个化合物,在众多化合物中筛选到真正的药效物质难度巨大。通过分析知柏口服后的入血成分,确定其体内直接作用物质,已成为快速、准确地确定知柏药效物质基础的有效途径。

家兔的代谢和免疫方面与人类接近,且家兔的血流量更为充足,避免了SD大鼠多时间点、重复采血的技术局限性,因此选择家兔作为实验动物。结合前期预实验结果,选择30、60、90、120 min 4个时间点取血,等体积混匀,检测离子数目最多,能最大程度反应知柏的入血成分。结果显示,初步检测出46个原型入血成分,包括皂苷类6个、生物碱类19个、酚酸类5个、糖苷类3个、苯丙素类1个、黄酮类9个、内酯类2个以及1个酚类衍生物。研究表明,丁香苷作为酚类衍生物,具有抗癌、抗氧化、保护肝脏等作用^[19];金丝桃苷具有改善心血管功能、抗肿瘤、抗抑郁、抗炎等药理活性^[20];吴茱萸苦素具有抑制血管生成和扩张血管的作用,提示其可能具有开发成为抗高血压药物的潜在药物^[21]。

在鉴别出的9个代谢产物中,3个是以知母中芒果苷为母体代谢产生的;6个是以黄柏中生物碱类物质为母体代谢而来。小檗碱、药根碱和小檗红碱代谢物类型主要为葡萄糖醛酸结合的产物,芒果苷的代谢产物主要以甲基化产物和葡萄糖醛酸化产物为主。其中小檗碱的口服生物利用度很低^[22],口服给药后生物利用度不足1%,提示小檗碱吸收入血量较少,进入体内的小檗碱绝大部分经肝脏代谢转化呈葡萄糖醛酸结合物,血中剩余少量的原型被检出。

只有被吸收入血的药物才可能有效,因此本研究采用准确度高、灵敏度高的UPLC-Q-Exactive Orbitrap-MS技术,对知柏体内成分进行全面评估,为更准确地筛选到知柏清热作用的潜在药效物质基础、新药研发及临床研究提供数据参考。

基金

收起

黑龙江省自然科学基金项目资助(LH2020H097)
黑龙江省中医药科研项目资助(ZHY202088)
江苏省中医退行性骨关节病临床医学创新中心资助项目资助(苏中科教[2021]4号)

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2024年第59卷第4期

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doi: 10.11669/cpj.2024.04.006

接收时间：2023-02-13
首发时间：2025-11-13
出版时间：2024-02-22

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收稿日期：2023-02-13

基金

黑龙江省自然科学基金项目资助(LH2020H097)

黑龙江省中医药科研项目资助(ZHY202088)

江苏省中医退行性骨关节病临床医学创新中心资助项目资助(苏中科教[2021]4号)

作者信息

¹ 佳木斯大学, 黑龙江佳木斯 154007

² 湖南医药学院药学院, 湖南怀化 418000

³ 黑龙江中医药大学药学院, 哈尔滨 150040

⁴ 无锡市中医医院中医药研究所, 江苏无锡 214071

通讯作者:

*雷霞,女,博士,副教授研究方向:药理学 Tel:(0510)81008810;

张宁,男,博士,教授研究方向:药物分析 Tel:(0451)82114400

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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.	Molecular formula	t_R /min	Theoretical mass(m/z)	Experimental mass(m/z)	Adduct	Error (×10^-6)	m/z (MS²)	Component	Source	Category
1	C₂₁H₂₁NO₄	8.92	352.154 33	352.153 53	[M+H]⁺	0.086	336.122 25, 308.127 11	Palmatine	PC	Alkaloid
2	C₂₀H₂₀NO₇	3.80	387.131 25	387.189 94	[M+H]⁺	-0.428	145.028 03, 177.054 14	Trihydroxy jatrorrhizine	PC	Alkaloid
3	C₂₁H₁₃N₃O	8.30	324.113 13	324.121 95	[M+H]⁺	-0.754	308.090 70, 280.095 89	7,8-deoxyrutaecarpine	PC	Alkaloid
4	C₂₀H₁₇NO₅	7.52	352.117 94	352.116 85	[M+H]⁺	0.692	336.085 60, 308.090 73	Berlambine	PC	Alkaloid
5	C₂₀H₁₉NO₄	8.20	336.123 03	336.121 83	[M+H]⁺	0.41	320.090 70, 306.518 68	Berbine	PC	Alkaloid
6	C₂₁H₁₈N $O 4 +$	9.66	350.138 68	350.137 57	[M+H]⁺	0.651	334.106 29, 320.090 7	Chelerythrine	PC	Alkaloid
7	C₂₇H₃₀O₁₀	11.53	513.176 62	513.138 79	[M-H]^-	0.321	367.139 50	Baohuoside Ⅰ	RA	Flavonoids
8	C₁₉H₁₅NO₄	8.36	322.107 38	322.106 32	[M+H]⁺	-0.696	307.082 89, 279.087 95	Menisporphine	PC	Alkaloid
9	C₂₀H₂₁NO₄	8.03	339.146 5	339.140 87	[M+H]⁺	-0.79	323.113 01, 308.090 27	Flavonoids	PC	Alkaloid
10	C₂₁H₂₅NO₄	7.19	356.185 63	356.184 33	[M+H]⁺	0.106	311.126 56, 296.103 39	Menisperine	PC	Alkaloid
11	C₁₉H₁₇NO₄	7.36	324.123 03	324.122 16	[M+H]⁺	-0.754	308.090 70, 294.075 29	Berberrubine	PC	Alkaloid
12	C₁₇H₂₀O₉	5.44	367.103 45	367.103 27	[M-H]^-	-0.362	191.055 31, 173.044 59	3-O-Feruloylquinic acid	PC	Phenolic acids
13	C₁₆H₁₈O₉	3.22	353.087 8	353.086 06	[M-H]^-	-0.002	191.055 24, 179.034 03	Chlorogenic acid	PC	Phenolic acids
14	C₁₇H₁₇NO₅	4.14	316.117 94	316.283 69	[M+H]⁺	1.303	298.274 02, 280.262 12	Hippeastrine	PC	Alkaloid
15	C₂₆H₃₀O₁₂	6.88	533.166 44	533.151 55	[M-H]^-	0.218	371.100 46, 325.091 77	Aegineoside	PC	Flavonoids
16	C₂₃H₂₉NO₈	5.15	448.196 59	448.195 28	[M+H]⁺	0.066	107.049 18, 286.142 79	¹⁾	PC	Glycoside
17	C₁₅H₁₇NO₃	6.79	260.128 11	260.127 38	[M+H]⁺	2.317	242.116 73, 224.106 48	Pilocarpine	PC	Alkaloid
18	C1₉H₂₃NO₃	6.35	314.175 07	314.173 95	[M+H]⁺	0.517	299.112 82, 269.116 82	Isoliensinine	PC	Alkaloid
19	C₃₉H₆₄O₁₃	10.22	739.427 41	739.428 16	[M-H]^-	0.002	577.376 65, 529.609 62	Timosaponin AⅢ	RA	Saponins
20	C₂₅H₂₈O₁₆	3.86	583.130 45	583.129 39	[M-H]^-	0.050	421.077 61, 331.045 84	Neomangiferin	RA	Flavonoids
21	C₄₅H₇₄O₁₉	8.06	917.475 15	917.476 14	[M-H]^-	-0.085	71.012 41,89.023	Anemarsaponin D	RA	Saponins
22	C₁₃H₁₈O₈	1.66	301.092 89	301.093 66	[M-H]^-	1.756	123.007 48, 138.031 19	Tachioside	PC	Phenolic acids
23	C₁₉H₁₈O₁₁	4.88	421.077 63	421.077 97	[M-H]^-	0.225	301.035 10, 331.045 90	Isomangiferin	RA	Flavonoids
24	C₄₅H₇₄O₁₈	9.78	901.480 23	901.484 13	[M-H]^-	-0.026	101.023 06, 113.023 09	Anemarsaponin B	RA	Saponins
25	C₄₅H₇₆O₁₉	8.37	919.490 8	919.489 75	[M-H]^-	0.048	757.432 56, 595.383 79	Anemarsaponin BⅡ	RA	Saponins
26	C₂₀H₁₉NO₄	7.69	338.138 68	338.137 45	[M+H]⁺	0.320	323.113 83, 294.111 02	Jatrorrhizine	PC	Alkaloid
27	C₂₁H₂₀O₁₀	6.58	431.098 37	431.097 93	[M-H]^-	0.346	311.056 12, 283.061 00	Afzelin	RA	Flavonoids
28	C₂₀H₂₃NO₄	5.56	342.169 98	342.169 19	[M+H]⁺	0.133	297.111 05, 265.085 17	Magnoflorine	PC	Alkaloid
29	C₁₈H₃₄O₅	10.30	329.233 34	329.233 28	[M-H]^-	-0.936	311.222 41, 293.211 7	Sanleng acid	PC	Phenolic acids
30	C₁₁H₁₈NO	9.00	181.146 11	181.085 91	[M+H]⁺	-0.060	121.101 23, 93.070 31	N-Candicine	PC	Alkaloid
31	C₁₉H₁₈O₁₁	4.88	421.077 63	421.077 97	[M-H]^-	0.225	331.045 90, 301.035 10	Mangiferin	RA	Flavonoids
32	C₂₆H₃₄O₁₁	6.81	521.202 83	521.203 19	[M-H]^-	0.219	329.139 34, 175.075 52	²⁾	PC	Glycoside
33	C₁₅H₂₄N₆O₉	4.17	431.153 19	431.155 64	[M-H]^-	0.811	191.054 50, 119.048 93	2-(p-hydroxyphenyl)ethanol1-O-β-D-glucoside	PC	Glycoside
34	C₄₅H₇₆O₂₀	7.74	935.485 71	935.485 41	[M-H]^-	-0.032	773.437 68, 449.330 32	Timosaponin N	RA	Saponins
35	C₃₉H₆₄O₁₄	11.91	755.422 32	755.422 3	[M-H]^-	0.089	325.255 52, 593.371 22	Timosaponin G	RA	Saponins
36	C₁₇H₁₆O₂	11.55	251.107 75	251.107 41	[M-H]^-	1.001	236.082 70, 157.064 74	Nyasol	RA	Phenylpropionin
37	C₁₄H₈N₂O	9.90	221.070 93	221.073 81	[M+H]⁺	-1.435	177.044 76, 150.036 79	6H-Indolo(3,2,1-de)(1,5)naphthyridin-6-one	PC	Alkaloid
38	C₁₆H₁₅NO₃	4.02	268.097 91	268.127 44	[M-H]^-	-1.491	250.099 72, 235.075 2	4-[N-(p-Methoxy-benzylidene)amino]-phenyl acetate	PC	Phenolic acids
39	C₁₃H₁₁NO₃	10.12	230.081 16	230.080 46	[M+H]⁺	2.751	215.057 01, 200.033 66	Γ-fagarine	PC	Alkaloid
40	C₁₅H₁₀O₇	4.83	303.045 62	303.050 17	[M+H]⁺	0.543	285.039 85, 275.055 94	Quercetin	PC	Flavonoids
41	C₂₆H₃₀O₈	11.00	469.195 67	469.189 45	[M-H]^-	-0.080	425.205 94	Limonin	PC	Lactones
42	C₁₄H₁₃NO₄	5.51	260.091 73	260.185 06	[M+H]⁺	-2.209	242.175 57, 237.908 63	Skimmianine	PC	Alkaloid
43	C₁₆H₁₄O₄	8.36	269.081 93	269.210 75	[M-H]^-	-1.379	253.216 60, 231.007 11	2'-O-methyl-isoliquiritigenin	RA	Flavonoids
44	C₂₆H₃₀O₉	13.53	485.181 70	485.282 71	[M-H]^-	-0.104	397.166 38, 383.149 08	Rutaevin	PC	Lactones
45	C₁₇H₂₄O₉	12.74	373.149 30	373.272 58	[M+H]⁺	0.539	355.262 42, 337.252 5	Syringin	PC	Phenols
46	C₂₁H₂₀O₁₂	7.86	463.088 19	463.306 52	[M-H]^-	-0.503	377.271 30, 359.261 08	Hyperoside	PC	Flavonoids

No.

Molecular
formula

t_R
/min

Theoretical
mass(m/z)

Experimental
mass(m/z)

Adduct

Error
(×10^-6)

m/z
(MS²)

Component

Source