药学学报

Compound No.	RT/min	Formula	Adduct (m/z)	Reference ion	m/z	Fragment ion (m/z)	Title
1	1.848	C₁₃H₁₁NO₅	261.061 55	[M+H]⁺	262.068 82	79.141 10, 131.248 10, 261.068 60	Oxolinic acid
2	2.013	C₆H₆O₃	126.030 27	[M-H]^-	125.022 99	69.032 91, 81.032 92, 97.027 93, 126.022 90	Pyrogallol
3	2.207	C₉ H₁₃N₃O₅	243.085 58	[M+H]⁺	244.092 86	112.050 71, 163.042 04, 228.747 22, 243.092 85	Cytarabine
4	2.343	C₇H₁₂O₆	192.062 15	[M-H]^-	191.054 87	133.028 08, 151.002 24	D-(-)-Quinic acid
5	2.555	C₇H₁₀O₅	174.051 70	[M-H]^-	173.044 37	71.012 23, 73.027 88, 83.048 61, 85.027 89, 99.007 19, 111.043 60, 137.022 98, 155.033 62	3, 4, 5-Trihydroxycyclohex-1-ene-1-carboxylic acid
6	4.04	C₅H₇NO₃	129.042 85	[M+H]⁺	130.050 13	84.044 98, 129.049 88	D-(+)-Pyroglutamic acid
7	4.046	C₅H₇NO₃	129.041 18	[M-H]^-	128.033 91	82.0282 2, 113.695 77	4-Oxoproline
8	6.048	C₁₀H₁₃N₅O₄	267.096 90	[M+H]⁺	268.104 19	57.034 41, 69.034 26, 97.028 97, 101.060 07, 136.061 66	Adenosine
9	6.277	C₁₀H₁₃N₅O₃	251.102 01	[M+H]⁺	252.109 28	73.029 08, 99.044 54, 177.054 73, 136.061 58 251.105 83	2′-Deoxyadenosine
10	6.727	C₁₁H₂₀N₂O₃	228.147 61	[M+H]⁺	229.154 91	70.065 90, 159.012 82, 173.028 43, 193.096 57, 211.107 77, 228.154 36	Prolylleucine
11	6.995	C₇H₆O₅	170.020 40	[M-H]^-	169.013 12	69.032 98, 81.032 94, 97.027 94, 126.026 20	Gallic acid
12	7.007	C₉ H₁₁NO₂	148.052 59	[M+NH₄]⁺	166.086 41	72.007 46, 75.007 18, 87.007 14, 93.032 95, 120.044 13, 121.028 05, 147.043 75	L-Phenylalanine
13	7.479	C₁₁H₁₅N₅O₄	281.112 66	[M+H]⁺	282.119 93	57.034 41, 69.034 26, 97.028 97, 119.035 14, 136.061 66, 281.119 45	2′-O-Methyladenosine
14	7.958	C₇H₆O₃	138.030 29	[M-H]^-	137.023 01	74.989 35, 93.032 97, 94.028 25, 109.027 81, 137.023 01	Salicylic acid
15	8.13	C₁₁H₁₅N₅O₄	281.112 68	[M+H]⁺	282.119 96	57.034 37, 69.034 22, 85.028 94, 136.061 65, 150.077 26, 281.119 20	N6-Me-Adenosine
16	8.692	C₈H₈O₂	136.050 95	[M-H]^-	135.043 67	75.007 17, 117.330 80, 134.035 90	2-Methylbenzoic acid
17	9.218	C₁₁H₉NO₂	187.063 48	[M+H]⁺	188.070 74	91.054 76, 115.054 44, 117.057 41, 118.065 31, 142.065 05, 143.072 86, 144.080 69, 146.059 88, 170.059 89, 187.070 51	Trans-3-indoleacrylic acid
20	9.956	C₉H₈O₄	180.040 51	[M-H]^-	179.033 20	59.012 25, 71.012 25, 85.027 85, 87.007 08, 89.022 83, 121.028 19, 134.035 96, 135.043 72, 136.047 10	Caffeic acid
18	10.183	C₉H₁₀O₅	198.053 03	[M+H]⁺	199.060 32	63.022 42, 78.009 43, 89.001 62, 91.017 28, 106.004 47, 121.027 98, 123.007 27, 138.030 81, 153.054 31, 166.997 36, 182.020 92, 198.047 99	Syringic acid
19	10.33	C₁₃H₁₃N₃	211.111 22	[M+H]⁺	212.118 50	92.049 96, 95.065 58, 119.060 49, 166.049 64, 194.044 86, 195.091 57, 211.117 86	N, N'-Diphenylguanidine
21	10.516	C₁₅H₁₆O₉	340.079 16	[M-H]^-	339.071 62	123.044 15, 133.028 37, 151.038 79, 179.033 65, 131.091 92	Esculin
23	11.118	C₉H₆O₄	178.026 51	[M+H]⁺	179.033 78	111.008 02, 137.022 99, 155.033 72, 178.033 66	Esculetin
24	11.179	C₁₀H₇NO₄	205.037 60	[M+H]⁺	206.044 88	132.044 45, 150.091 48, 178.049 67, 205.044 54	Xanthurenic acid
22	11.572	C₇H₆O₄	154.025 34	[M-H]^-	153.018 07	81.032 97, 85.027 90, 95.012 27, 108.020 13, 109.0279 7, 110.031 14, 123.007 23	Gentisic acid
25	11.745	C₁₆H₁₈O₉	354.094 37	[M-H]^-	353.086 98	85.027 85, 93.032 96, 111.043 62, 135.043 67, 137.023 03, 155.033 62, 173.044 30, 179.033 78, 191.055 02, 353.087 16	Chlorogenic acid
26	11.77	C₁₁H₁₂O₅	224.068 41	[M+H]⁺	225.075 68	91.054 70, 95.049 55, 119.049 26, 147.043 87, 149.023 24, 157.039 51, 175.038 80, 192.041 52, 207.064 79, 224.039 41	Sinapinic acid
27	12.258	C₁₀H₁₀O₄	194.058 06	[M+H]⁺	195.065 34	89.039 12, 107.857 80, 117.033 66, 135.043 98, 163.038 71, 177.090 87, 194.101 43	Methyl caffeate
28	13.221	C₂₆H₂₈O₁₄	564.147 82	[M+H]⁺	565.155 27	121.028 47, 203.033 58, 295.059 75, 307.059 91, 325.069 92, 379.080 44, 391.080 41, 409.091 13, 427.101 78, 511.123 14, 529.133 42, 564.149 90	Schaftoside
29	13.551	C₁₀H₈O₄	192.042 46	[M+H]⁺	193.049 74	133.028 38, 137.059 60, 161.059 54, 165.054 52, 178.025 92, 192.049 38	7-Hydroxy-6-methoxy-2H-chromen-2-one
30	13.633	C₈H₈O	120.055 99	[M-H]^-	119.048 70	59.009 26, 71.012 24, 119.048 70	Phenylacetaldehyde
31	13.636	C₉H₈O₃	164.04599	[M-H]^-	163.038 71	119.048 71, 120.052 01, 163.038 88	2, 3-Dihydro-1-benzofuran-2-carboxylic acid
32	13.638	C₉H₈O₃	164.047 49	[M+H]⁺	165.054 76	91.054 70, 119.049 22, 147.043 79, 164.054 64	4-Coumaric acid
33	13.837	C₂₇H₃₀O₁₆	610.153 26	[M+H]⁺	611.160 71	153.018 08, 287.054 41, 449.107 09, 610.159 97	Luteolin 30, 40-di-O-b-d-glucopyranoside
34	14.328	C₂₆H₂₈O₁₅	580.142 63	[M+H]⁺	581.149 90	85.028 98, 153.018 02, 171.028 58, 273.075 47, 287.054 47, 580.186 28	Kaempferol 3-O-vicianoside
35	14.364	C₁₅H₁₂O₅	272.068 15	[M+H]⁺	273.075 44	67.018 66, 119.049 33, 147.043 90, 153.018 05, 171.028 64, 272.075 44	Naringenin
36	14.364	C₂₇H₃₂O₁₄	580.178 17	[M-H]^-	579.170 84	65.001 66, 107.012 29, 119.048 70, 151.102 29, 271.060 85, 285.040 10, 459.114 50, 580.174 80	Naringin
38	14.418	C₁₆H₁₄O₆	302.079 00	[M+H]⁺	303.086 27	67.018 62, 117.033 66, 145.028 24, 153.018 01, 177.054 41, 179.033 62, 302.086 06	5, 7-Dihydroxy-2-(4-hydroxy-3-methoxyphenyl)-3, 4-dihydro-2H-1-benzopyran-4-one
39	14.501	C₂₁H₂₀O₁₁	448.100 19	[M+H]⁺	449.107 54	107.012 30, 133.028 08, 151.002 24, 284.032 26, 285.040 04, 447.092 53	Cynaroside
37	14.538	C₇H₆O₂	122.035 23	[M-H]^-	121.027 95	93.032 79, 121.027 97	Benzoic acid
40	15.156	C₁₆H₂₂O₇	343.163 04	[M+H]⁺	344.170 32	61.029 24, 69.034 24, 85.028 96, 97.028 82, 105.070 18, 137.059 59, 165.090 84, 180.086 27, 343.170 32	(2R, 3S, 4S, 5R, 6S)-2-(Hydroxymethyl)-6-[2-methoxy-4-(prop-2-en-1-yl)phenoxy]oxane-3, 4, 5-triol
41	15.252	C₂₁H₂₀O₁₀	432.105 44	[M+H]⁺	433.112 76	67.018 67, 119.049 28, 153.018 02, 271.059 63, 432.114 04	Apigetrin
42	15.514	C₉H₁₆O₄	188.103 73	[M-H]^-	187.096 45	57.032 96, 97.064 29, 123.080 09, 125.095 69, 169.085 62	Azelaic acid
43	15.626	C₁₅H₁₂O₆	288.062 96	[M+H]⁺	289.070 34	68.997 83, 89.039 07, 117.033 60, 135.043 98, 145.028 31, 153.017 99, 163.038 70, 179.037 75, 288.070 22	2-(3, 4-Dihydroxyphenyl)-5, 7-dihydroxy-3, 4-dihydro-2H-1-benzopyran-4-one
44	16.055	C₁₁H₆O₃	186.031 97	[M+H]⁺	187.039 25	69.034 27, 81.034 13, 109.101 53, 115.054 44, 131.049 10, 143.049 01, 159.116 41	Psoralen
45	16.25	C₃₆H₅₈O₁₁	712.403 03	[M-H]^-	711.395 75	419.310 67, 441.337 13, 453.300 17, 485.32 822, 503.337 49, 711.254 64	Sericoside
46	16.509	C₁₆H₁₂O₅	284.068 93	[M+H]⁺	285.076 20	81.070 51, 153.017 84, 242.056 88, 270.051 94, 284.075 38	Glycitein
48	17.054	C₁₅H₁₀O₆	286.047 40	[M+H]⁺	287.054 75	153.018 02, 241.049 03	Kaempferol
49	17.787	C₃₀H₄₈O₅	488.350 37	[M+H-H₂O]⁺	471.347 08	147.116 67, 201.163 56, 219.173 90, 271.059 63, 407.330 08, 425.340 76, 453.336 03	(3β, 5ξ, 9ξ)-3, 6, 19-Trihydroxyurs-12-en-28-oic acid
50	17.831	C₁₆H₁₂O₆	300.063 34	[M+H]⁺	301.070 65	258.051 88, 286.046 81	Diosmetin
47	17.993	C₃₆H₅₈O₁₀	696.407 79	[M-H]^-	695.400 51	59.011 43, 207.033 36, 469.331 85, 487.342 38	Rosamultin
51	21.237	C₃₀H₄₆O₃	436.334 49	[M+H]⁺	437.341 77	81.070 58, 95.086 05, 107.085 89, 119.085 74, 133.101 20, 145.101 12, 187.148 09, 205.158 69, 389.320 13, 407.331 09, 435.336 91	11, 12-Dehydroursolic acid lactone
52	21.829	C₃₀H₄₈O₅	488.349 86	[M-H]^-	487.342 19	443.353 09, 469.331 45	Asiatic acid
53	21.927	C₃₀H₄₆O₄	470.339 63	[M+H]⁺	471.347 02	81.070 54, 95.086 02, 107.085 88, 121.101 32, 189.163 71, 201.163 74, 407.330 20, 425.340 85	18-β-Glycyrrhetinic acid
54	24.008	C₁₆H₁₈O₉	354.094 53	[M-H]^-	353.087 25	85.027 89, 93.032 91, 111.043 87, 134.036 01, 135.043 72, 161.023 13, 179.033 83, 191.055 05	Neochlorogenic acid
55	24.014	C₃₀H₄₈O₃	456.359 88	[M+H-H₂O]⁺	439.356 84	81.070 54, 95.086 01, 107.085 85, 133.101 24, 147.116 59, 189.163 73, 411.319 27	Oleanolic acid
56	24.244	C₂₀H₃₂O₂	304.240 01	[M+H]⁺	305.247 28	67.055 05, 79.054 90, 93.070 38, 107.085 86, 121.101 34, 135.116 81, 149.132 43, 163.148 06, 259.205 35	Arachidonic acid
57	25.026	C₂₀H₃₄O₂	306.255 56	[M+H]⁺	307.262 85	67.055 02, 81.070 50, 95.085 98, 109.101 46, 123.116 92, 243.209 93, 261.221 13	Linolenic acid ethyl ester
58	25.548	C₃₅H₄₂N₂O₉	333.170 29	[M+H]⁺	334.170 56	269.371 87, 253.158 64, 161.042 73, 145.097 55, 133.283 19	Estriol
59	26.557	C₁₈H₃₇NO	283.287 55	[M+H]⁺	284.294 83	57.070 78, 60.045 28, 88.076 26, 102.091 72, 116.107 17	Stearamide

Compound No.	RT/min	Formula	Adduct (m/z)	Reference ion	m/z	Fragment ion (m/z)	Title
1	1.848	C₁₃H₁₁NO₅	261.061 55	[M+H]⁺	262.068 82	79.141 10, 131.248 10, 261.068 60	Oxolinic acid
2	2.013	C₆H₆O₃	126.030 27	[M-H]^-	125.022 99	69.032 91, 81.032 92, 97.027 93, 126.022 90	Pyrogallol
3	2.207	C₉ H₁₃N₃O₅	243.085 58	[M+H]⁺	244.092 86	112.050 71, 163.042 04, 228.747 22, 243.092 85	Cytarabine
4	2.343	C₇H₁₂O₆	192.062 15	[M-H]^-	191.054 87	133.028 08, 151.002 24	D-(-)-Quinic acid
5	2.555	C₇H₁₀O₅	174.051 70	[M-H]^-	173.044 37	71.012 23, 73.027 88, 83.048 61, 85.027 89, 99.007 19, 111.043 60, 137.022 98, 155.033 62	3, 4, 5-Trihydroxycyclohex-1-ene-1-carboxylic acid
6	4.04	C₅H₇NO₃	129.042 85	[M+H]⁺	130.050 13	84.044 98, 129.049 88	D-(+)-Pyroglutamic acid
7	4.046	C₅H₇NO₃	129.041 18	[M-H]^-	128.033 91	82.0282 2, 113.695 77	4-Oxoproline
8	6.048	C₁₀H₁₃N₅O₄	267.096 90	[M+H]⁺	268.104 19	57.034 41, 69.034 26, 97.028 97, 101.060 07, 136.061 66	Adenosine
9	6.277	C₁₀H₁₃N₅O₃	251.102 01	[M+H]⁺	252.109 28	73.029 08, 99.044 54, 177.054 73, 136.061 58 251.105 83	2′-Deoxyadenosine
10	6.727	C₁₁H₂₀N₂O₃	228.147 61	[M+H]⁺	229.154 91	70.065 90, 159.012 82, 173.028 43, 193.096 57, 211.107 77, 228.154 36	Prolylleucine
11	6.995	C₇H₆O₅	170.020 40	[M-H]^-	169.013 12	69.032 98, 81.032 94, 97.027 94, 126.026 20	Gallic acid
12	7.007	C₉ H₁₁NO₂	148.052 59	[M+NH₄]⁺	166.086 41	72.007 46, 75.007 18, 87.007 14, 93.032 95, 120.044 13, 121.028 05, 147.043 75	L-Phenylalanine
13	7.479	C₁₁H₁₅N₅O₄	281.112 66	[M+H]⁺	282.119 93	57.034 41, 69.034 26, 97.028 97, 119.035 14, 136.061 66, 281.119 45	2′-O-Methyladenosine
14	7.958	C₇H₆O₃	138.030 29	[M-H]^-	137.023 01	74.989 35, 93.032 97, 94.028 25, 109.027 81, 137.023 01	Salicylic acid
15	8.13	C₁₁H₁₅N₅O₄	281.112 68	[M+H]⁺	282.119 96	57.034 37, 69.034 22, 85.028 94, 136.061 65, 150.077 26, 281.119 20	N6-Me-Adenosine
16	8.692	C₈H₈O₂	136.050 95	[M-H]^-	135.043 67	75.007 17, 117.330 80, 134.035 90	2-Methylbenzoic acid
17	9.218	C₁₁H₉NO₂	187.063 48	[M+H]⁺	188.070 74	91.054 76, 115.054 44, 117.057 41, 118.065 31, 142.065 05, 143.072 86, 144.080 69, 146.059 88, 170.059 89, 187.070 51	Trans-3-indoleacrylic acid
20	9.956	C₉H₈O₄	180.040 51	[M-H]^-	179.033 20	59.012 25, 71.012 25, 85.027 85, 87.007 08, 89.022 83, 121.028 19, 134.035 96, 135.043 72, 136.047 10	Caffeic acid
18	10.183	C₉H₁₀O₅	198.053 03	[M+H]⁺	199.060 32	63.022 42, 78.009 43, 89.001 62, 91.017 28, 106.004 47, 121.027 98, 123.007 27, 138.030 81, 153.054 31, 166.997 36, 182.020 92, 198.047 99	Syringic acid
19	10.33	C₁₃H₁₃N₃	211.111 22	[M+H]⁺	212.118 50	92.049 96, 95.065 58, 119.060 49, 166.049 64, 194.044 86, 195.091 57, 211.117 86	N, N'-Diphenylguanidine
21	10.516	C₁₅H₁₆O₉	340.079 16	[M-H]^-	339.071 62	123.044 15, 133.028 37, 151.038 79, 179.033 65, 131.091 92	Esculin
23	11.118	C₉H₆O₄	178.026 51	[M+H]⁺	179.033 78	111.008 02, 137.022 99, 155.033 72, 178.033 66	Esculetin
24	11.179	C₁₀H₇NO₄	205.037 60	[M+H]⁺	206.044 88	132.044 45, 150.091 48, 178.049 67, 205.044 54	Xanthurenic acid
22	11.572	C₇H₆O₄	154.025 34	[M-H]^-	153.018 07	81.032 97, 85.027 90, 95.012 27, 108.020 13, 109.0279 7, 110.031 14, 123.007 23	Gentisic acid
25	11.745	C₁₆H₁₈O₉	354.094 37	[M-H]^-	353.086 98	85.027 85, 93.032 96, 111.043 62, 135.043 67, 137.023 03, 155.033 62, 173.044 30, 179.033 78, 191.055 02, 353.087 16	Chlorogenic acid
26	11.77	C₁₁H₁₂O₅	224.068 41	[M+H]⁺	225.075 68	91.054 70, 95.049 55, 119.049 26, 147.043 87, 149.023 24, 157.039 51, 175.038 80, 192.041 52, 207.064 79, 224.039 41	Sinapinic acid
27	12.258	C₁₀H₁₀O₄	194.058 06	[M+H]⁺	195.065 34	89.039 12, 107.857 80, 117.033 66, 135.043 98, 163.038 71, 177.090 87, 194.101 43	Methyl caffeate
28	13.221	C₂₆H₂₈O₁₄	564.147 82	[M+H]⁺	565.155 27	121.028 47, 203.033 58, 295.059 75, 307.059 91, 325.069 92, 379.080 44, 391.080 41, 409.091 13, 427.101 78, 511.123 14, 529.133 42, 564.149 90	Schaftoside
29	13.551	C₁₀H₈O₄	192.042 46	[M+H]⁺	193.049 74	133.028 38, 137.059 60, 161.059 54, 165.054 52, 178.025 92, 192.049 38	7-Hydroxy-6-methoxy-2H-chromen-2-one
30	13.633	C₈H₈O	120.055 99	[M-H]^-	119.048 70	59.009 26, 71.012 24, 119.048 70	Phenylacetaldehyde
31	13.636	C₉H₈O₃	164.04599	[M-H]^-	163.038 71	119.048 71, 120.052 01, 163.038 88	2, 3-Dihydro-1-benzofuran-2-carboxylic acid
32	13.638	C₉H₈O₃	164.047 49	[M+H]⁺	165.054 76	91.054 70, 119.049 22, 147.043 79, 164.054 64	4-Coumaric acid
33	13.837	C₂₇H₃₀O₁₆	610.153 26	[M+H]⁺	611.160 71	153.018 08, 287.054 41, 449.107 09, 610.159 97	Luteolin 30, 40-di-O-b-d-glucopyranoside
34	14.328	C₂₆H₂₈O₁₅	580.142 63	[M+H]⁺	581.149 90	85.028 98, 153.018 02, 171.028 58, 273.075 47, 287.054 47, 580.186 28	Kaempferol 3-O-vicianoside
35	14.364	C₁₅H₁₂O₅	272.068 15	[M+H]⁺	273.075 44	67.018 66, 119.049 33, 147.043 90, 153.018 05, 171.028 64, 272.075 44	Naringenin
36	14.364	C₂₇H₃₂O₁₄	580.178 17	[M-H]^-	579.170 84	65.001 66, 107.012 29, 119.048 70, 151.102 29, 271.060 85, 285.040 10, 459.114 50, 580.174 80	Naringin
38	14.418	C₁₆H₁₄O₆	302.079 00	[M+H]⁺	303.086 27	67.018 62, 117.033 66, 145.028 24, 153.018 01, 177.054 41, 179.033 62, 302.086 06	5, 7-Dihydroxy-2-(4-hydroxy-3-methoxyphenyl)-3, 4-dihydro-2H-1-benzopyran-4-one
39	14.501	C₂₁H₂₀O₁₁	448.100 19	[M+H]⁺	449.107 54	107.012 30, 133.028 08, 151.002 24, 284.032 26, 285.040 04, 447.092 53	Cynaroside
37	14.538	C₇H₆O₂	122.035 23	[M-H]^-	121.027 95	93.032 79, 121.027 97	Benzoic acid
40	15.156	C₁₆H₂₂O₇	343.163 04	[M+H]⁺	344.170 32	61.029 24, 69.034 24, 85.028 96, 97.028 82, 105.070 18, 137.059 59, 165.090 84, 180.086 27, 343.170 32	(2R, 3S, 4S, 5R, 6S)-2-(Hydroxymethyl)-6-[2-methoxy-4-(prop-2-en-1-yl)phenoxy]oxane-3, 4, 5-triol
41	15.252	C₂₁H₂₀O₁₀	432.105 44	[M+H]⁺	433.112 76	67.018 67, 119.049 28, 153.018 02, 271.059 63, 432.114 04	Apigetrin
42	15.514	C₉H₁₆O₄	188.103 73	[M-H]^-	187.096 45	57.032 96, 97.064 29, 123.080 09, 125.095 69, 169.085 62	Azelaic acid
43	15.626	C₁₅H₁₂O₆	288.062 96	[M+H]⁺	289.070 34	68.997 83, 89.039 07, 117.033 60, 135.043 98, 145.028 31, 153.017 99, 163.038 70, 179.037 75, 288.070 22	2-(3, 4-Dihydroxyphenyl)-5, 7-dihydroxy-3, 4-dihydro-2H-1-benzopyran-4-one
44	16.055	C₁₁H₆O₃	186.031 97	[M+H]⁺	187.039 25	69.034 27, 81.034 13, 109.101 53, 115.054 44, 131.049 10, 143.049 01, 159.116 41	Psoralen
45	16.25	C₃₆H₅₈O₁₁	712.403 03	[M-H]^-	711.395 75	419.310 67, 441.337 13, 453.300 17, 485.32 822, 503.337 49, 711.254 64	Sericoside
46	16.509	C₁₆H₁₂O₅	284.068 93	[M+H]⁺	285.076 20	81.070 51, 153.017 84, 242.056 88, 270.051 94, 284.075 38	Glycitein
48	17.054	C₁₅H₁₀O₆	286.047 40	[M+H]⁺	287.054 75	153.018 02, 241.049 03	Kaempferol
49	17.787	C₃₀H₄₈O₅	488.350 37	[M+H-H₂O]⁺	471.347 08	147.116 67, 201.163 56, 219.173 90, 271.059 63, 407.330 08, 425.340 76, 453.336 03	(3β, 5ξ, 9ξ)-3, 6, 19-Trihydroxyurs-12-en-28-oic acid
50	17.831	C₁₆H₁₂O₆	300.063 34	[M+H]⁺	301.070 65	258.051 88, 286.046 81	Diosmetin
47	17.993	C₃₆H₅₈O₁₀	696.407 79	[M-H]^-	695.400 51	59.011 43, 207.033 36, 469.331 85, 487.342 38	Rosamultin
51	21.237	C₃₀H₄₆O₃	436.334 49	[M+H]⁺	437.341 77	81.070 58, 95.086 05, 107.085 89, 119.085 74, 133.101 20, 145.101 12, 187.148 09, 205.158 69, 389.320 13, 407.331 09, 435.336 91	11, 12-Dehydroursolic acid lactone
52	21.829	C₃₀H₄₈O₅	488.349 86	[M-H]^-	487.342 19	443.353 09, 469.331 45	Asiatic acid
53	21.927	C₃₀H₄₆O₄	470.339 63	[M+H]⁺	471.347 02	81.070 54, 95.086 02, 107.085 88, 121.101 32, 189.163 71, 201.163 74, 407.330 20, 425.340 85	18-β-Glycyrrhetinic acid
54	24.008	C₁₆H₁₈O₉	354.094 53	[M-H]^-	353.087 25	85.027 89, 93.032 91, 111.043 87, 134.036 01, 135.043 72, 161.023 13, 179.033 83, 191.055 05	Neochlorogenic acid
55	24.014	C₃₀H₄₈O₃	456.359 88	[M+H-H₂O]⁺	439.356 84	81.070 54, 95.086 01, 107.085 85, 133.101 24, 147.116 59, 189.163 73, 411.319 27	Oleanolic acid
56	24.244	C₂₀H₃₂O₂	304.240 01	[M+H]⁺	305.247 28	67.055 05, 79.054 90, 93.070 38, 107.085 86, 121.101 34, 135.116 81, 149.132 43, 163.148 06, 259.205 35	Arachidonic acid
57	25.026	C₂₀H₃₄O₂	306.255 56	[M+H]⁺	307.262 85	67.055 02, 81.070 50, 95.085 98, 109.101 46, 123.116 92, 243.209 93, 261.221 13	Linolenic acid ethyl ester
58	25.548	C₃₅H₄₂N₂O₉	333.170 29	[M+H]⁺	334.170 56	269.371 87, 253.158 64, 161.042 73, 145.097 55, 133.283 19	Estriol
59	26.557	C₁₈H₃₇NO	283.287 55	[M+H]⁺	284.294 83	57.070 78, 60.045 28, 88.076 26, 102.091 72, 116.107 17	Stearamide

基于化学成分分析及网络药理学结合实验探讨岩青兰改善神经炎症缓解老年糖尿病小鼠脑神经病变的作用及机制研究

PDF下载

徐梦洁 ¹^,^* , 肖姣 ² , 李俊 ¹ , 高心茹 ¹ , 闫柯 ¹ , 和伟 ¹ , 颜廷旭 ³^,^*

药学学报 | 研究论文 2025,60(10): 3181-3193

收起

药学学报 |研究论文 2025 , 60 (10) : 3181 -3193

基于化学成分分析及网络药理学结合实验探讨岩青兰改善神经炎症缓解老年糖尿病小鼠脑神经病变的作用及机制研究

全屏

徐梦洁¹^,^*, 肖姣², 李俊¹, 高心茹¹, 闫柯¹, 和伟¹, 颜廷旭³^,^*

作者信息

¹忻州师范学院生物系, 山西忻州 034000

²沈阳药科大学无涯创新学院, 辽宁沈阳 110016

³沈阳药科大学功能食品与葡萄酒学院, 辽宁沈阳 110016

通讯作者:

徐梦洁, E-mail: xumj1019@126.com

颜廷旭, E-mail: ytxsyphu@163.com

作者简介:

作者贡献:

徐梦洁进行课题指导并撰写论文; 肖姣进行数据分析; 李俊、高心茹、闫柯进行整体实验和数据分析; 和伟参与整体实验; 颜廷旭参与指导实验方案设计并修改论文。

Exploring anti-neuroinflammation mechanism of Dracocephalum rupestre Hance in treating aging type 2 diabetes related encephalopathy based on chemical composition analysis and integration of network pharmacology combined with experimental validation

Meng-jie XU¹^,^*, Jiao XIAO², Jun LI¹, Xin-ru GAO¹, Ke YAN¹, Wei HE¹, Ting-xu YAN³^,^*

Affiliations

¹Department of Biological Sciences, Xinzhou Normal University, Xinzhou 034000, China

²Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang 110016, China

³School of Functional Food and Wine, Shenyang Pharmaceutical University, Shenyang 110016, China

出版时间: 2025-10-12 doi: 10.16438/j.0513-4870.2025-0177

文章导航

摘要

收起

以岩青兰化学成分为基础, 生信分析筛选岩青兰改善老年糖尿病小鼠脑神经病变的关键靶点及通路, 并通过小鼠实验进行验证, 明确其作用机制。通过Gene Cards搜集神经炎症靶点, GEO数据库筛选老年糖尿病患者脑相关差异基因(DEGs); 用Venny软件得到岩青兰的核心成分靶点与神经炎症交集靶点为56个、与DEGs交集靶点31个; 通过STRING数据库构建蛋白相互作用(PPI) 网络, 利用Cytoscape软件进行PPI及药物-成分-靶点网络的可视化处理, 并通过Maximum Clique Centrality (MCC) 筛选Top10核心靶点; 用DAVID数据库进行GO (gene ontology) 功能富集和KEGG (Kyoto encyclopedia of genes and genomes) 通路分析, 发现岩青兰主要通过神经炎症相关的丝裂原活化蛋白激酶(MAPK)、核转录因子(NF-κB) 等通路蛋白改善老年糖尿病神经病变; 使用分子对接软件对核心活性成分与核心靶点进行对接发现岩青兰中的绿原酸、香叶木素、山柰酚与蛋白激酶B (AKT1)、白细胞介素-1β (IL-1β)、B淋巴细胞瘤2 (BCL2)、血红素加氧酶1 (HMOX1) 等靶点均有较好的结合活性。动物实验显示, 岩青兰可以降低小鼠血糖、血脂水平, 改善肝脏组织细胞排列紊乱, 脂肪空泡的现象, 减少血清促炎细胞因子的水平; 下调脑肿瘤坏死因子α (TNF-α)、IL-1β、Toll样受体4 (TLR4)、核NF-κB蛋白的表达, 抑制小鼠脑离子钙结合衔接分子1 (Iba-1) 的过度激活, 降低NOD样受体热蛋白结构域蛋白3 (NLRP3) 炎症小体的激活。该研究初步揭示了岩青兰对老年糖尿病脑神经病变的治疗作用可能与调控TLR4/NF-κB、抑制炎症小体等炎症信号通路有关。本实验经过沈阳药科大学伦理委员会批准(No. SYPU-IACOC-2024-1016-103)。

关键词

岩青兰 / 2型糖尿病 / 老年脑病变 / 生信分析 / 神经炎症

Abstract

收起

Through network pharmacology and molecular docking technology, combined with in vivo experiment verification through aged db/db mice, we explored the anti-neuroinflammation mechanism of Dracocephalum rupestre Hance in aging type 2 diabetes. Neuroinflammatory targets were collected through Gene Cards, and nerve related differential genes (DEGs) in elderly patients with diabetes were screened by GEO NCBI database; 56 component targets and 31 component targets were obtained by Venny 2.1.0 software from core component targets of Dracocephalum rupestre Hance between neuroinflammatory targets and DEGs; protein-protein interaction (PPI) networks were constructed using STRING database, visualize PPI and medicine-composition-target network through Cytoscape software, and the Top 10 core targets were elected using Maximum Clique Centrality (MCC); GO (gene ontology) functional enrichment and KEGG (kyoto encyclopedia of genes and genes) pathway analysis were conducted by DAVID database. It was found that the neuroprotection effect of Dracocephalum rupestre Hance mainly include alleviated aging diabetes neuroinflammation through regulated MAPK, NF-κB and other pathway proteins. Molecular docking results showed that the core active ingredients of Dracocephalum rupestre Hance, chlorogenic acid, diosmetin, and kaempferol, have good binding activity with targets AKT1, IL-1β, BCL2, and HMOX1. In vivo experiments showed that Dracocephalum rupestre Hance can reduce blood glucose and lipid levels in aging db/db mice, ameliorate the disorder of liver tissue cell arrangement and lipid vacuoles, and reduce the level of serum pro-inflammatory cytokines; down-regulate the expression of brain TNF-α, IL-1β, TLR4, and nuclear NF-κB; inhibit the overactivation of Iba-1 in mouse brain. What's more, Dracocephalum rupestre Hance could decreased the activation of NLRP3 inflammasome. This study preliminarily revealed that the therapeutic effect of Dracocephalum rupestre Hance on aging diabetes encephalopathy may be related to improve the neuroinflammatory signal pathway, such as the regulation of TLR4/NF-κB, inhibition of NLRP3 inflammasome, etc. This experiment was approved by the Medical Ethics Committee of Shenyang Pharmaceutical University (No. SYPU-IACOC-2024-1016-103).

Key words

Dracocephalum rupestre Hance / type 2 diabetes / aging brain / bioinformatics analysis / neuroinflammation

引用本文

徐梦洁, 肖姣, 李俊, 高心茹, 闫柯, 和伟, 颜廷旭. 基于化学成分分析及网络药理学结合实验探讨岩青兰改善神经炎症缓解老年糖尿病小鼠脑神经病变的作用及机制研究. 药学学报, 2025 , 60 (10) : 3181 -3193 . DOI: 10.16438/j.0513-4870.2025-0177

Meng-jie XU, Jiao XIAO, Jun LI, Xin-ru GAO, Ke YAN, Wei HE, Ting-xu YAN. Exploring anti-neuroinflammation mechanism of Dracocephalum rupestre Hance in treating aging type 2 diabetes related encephalopathy based on chemical composition analysis and integration of network pharmacology combined with experimental validation[J]. Acta Pharmaceutica Sinica, 2025 , 60 (10) : 3181 -3193 . DOI: 10.16438/j.0513-4870.2025-0177

正文

收起

糖尿病是一种由于体内胰岛素(insulin, INS) 分泌不足导致的, 以高血糖为主要特征的全身性疾病^{[1, 2]}。2型糖尿病(type 2 diabetes mellitus, T2DM) 是最常见的糖尿病类型, 据国际糖尿病联盟方统计数字显示, 到2030年, 糖尿病人数将达到5.52亿, 其中T2DM患者所占比例达到了93.7%^{[3, 4]}。且T2DM和年龄之间的关系十分密切。35岁以上的糖尿病患者占总患者人数的90%以上, 而老年人群T2DM的发病率更比其他人群的患病率高出约20%^[5]。

T2DM会影响中枢神经系统, 包括海马异常、神经递质损伤、认知功能障碍等, 由于共同的生理机制, 衰老促进T2DM诱导的这些病理进程的发展, 加速脑病变的发生^{[6, 7]}。衰老型T2DM患者的脑病变是由多种因素引起的, 大量的临床及实验数据表明, 神经炎症的异常激活是其中一个重要的触发因素^[8]。近年来, 多项数据说明老年T2DM患者脑内炎症标志物水平升高, 并且存在与T2DM相关的先天免疫相关的风险基因, 这表明神经炎症在T2DM的发病机制中起着重要作用^{[9, 10]}。小胶质细胞是免疫细胞, 参与大脑的炎症反应, T2DM脑中促炎小胶质细胞的激活增加, 导致细胞因子大量分泌。此外, 高血糖和氧化应激会激活TLR4/NF-κB通路, 这是控制炎症级联反应的重要通路, 进而诱导炎性因子的分泌, 导致一系列神经病理的发生, 如神经元损伤、炎症小体的过度激活、细胞死亡等^{[11, 12]}。因此, 通过改善神经炎症以确定一些干预的治疗靶点对T2DM的治疗具有重要意义。

岩青兰(Dracocephalum rupestre Hance), 为唇形科(Labiatae) 青兰属(Dracocephalum) 多年生草本植物, 主要分布于我国河北、山西、辽宁、内蒙古、青海等地^{[13, 14]}。本草著作记载, 岩青兰具有疏风清热、凉肝止血的功效。民间常将岩青兰制成养生茶而饮用, 并发现其具有降低胆固醇、防止血压上升等作用, 在中老年人群中受到青睐^{[15, 16]}。前期有研究报道表明, 岩青兰的活性可能与抗炎、抗氧化、代谢调节等作用相关, 但其具体机制研究鲜有报道, 药效物质基础也尚不清楚。本项目在鉴定岩青兰化学成分的基础上, 利用生信技术筛选岩青兰活性成分, 并筛选出其相应改善老年T2DM神经炎症的相关靶点和通路; 通过db/db小鼠对网络药理学预测结果进行验证, 进一步明确岩青兰的物质基础和作用机制, 探索其治疗T2DM疾病的科学依据, 以期为临床用药和岩青兰的进一步开发提供理论支撑。

材料与方法

收起

实验动物 选取24只9月龄的SPF级雄性db/db小鼠, 体重35~45 g, 购自北京斯贝福生物科技股份有限公司, 质量许可证号SCXK (京) 2023-0008, 经过沈阳药科大学伦理委员会批准(No: SYPU-IACOC-2024-1016-103)。将小鼠适应性饲养1周于相对湿度45%~55%、温度(22 ± 1) ℃、12 h光暗循环的条件下。随后按血糖值随机分为模型组、盐酸二甲双胍组, 岩青兰低、高剂量组, 每组6只小鼠; 取db/m小鼠6只设为正常组。按体表面积法参考人体给药剂量计算小鼠给药剂量, 其中盐酸二甲双胍组剂量为200 mg·kg^-1, 岩青兰低剂量为300 mg·kg^-1, 高剂量为600 mg·kg^-1。各组灌胃给药, 每日1次, 连续3周, 模型组和空白组灌胃等体积0.5%羧甲基纤维素钠(CMC-Na)。

主要药品与试剂 岩青兰采集于在山西省宁武县(N38°15′, E113°20′), 标本保存于忻州师范学院生物系; 经沈阳药科大学颜廷旭教授鉴定为唇形科青兰属植物岩青兰Dracocephalum rupestre Hance。干燥后用10倍量的70%乙醇(天津方大同正, 20240302) 提取, 加热回流3次, 每次2 h, 将滤液浓缩后冷冻、干燥并储存在-4 ℃; 二甲双胍(德国默克制药, 500 mg规格); 重组Iba-1抗体(ET1705-78)、重组半胱天冬酶1 (caspase-1) 抗体(R1510-23)、重组NF-κB抗体(ET1603-12) 购自于杭州华安生物技术有限公司; 酶联免疫吸附试验(ELISA) 试剂盒白介素1β (IL-1β, EK201B) 和ELISA试剂盒肿瘤坏死因子-α (TNF-α, EK282) 购自于杭州联科生物股份有限公司; 重组TLR4抗体(GB11519-100)、重组NLRP3抗体(GB114320-50)、重组TMS1/ASC抗体(GB155270-50)、谷胱甘肽过氧化物酶(GSH) 检测试剂盒(G4305)、超氧化物歧化酶(SOD) 检测试剂盒(G4306)、血脂四项检测试剂盒(GM1113)、小鼠INS检测试剂盒(CA448Mu)、HRP标记的羊抗兔二抗(GB23303) 购自于武汉赛维尔生物科技有限公司。

岩青兰成分鉴定 通过Ultimate 3000 UPLC系统进行色谱分析。使用AQ-C18色谱柱(2.1 mm × 150 mm, 1.8 μm), 流动相: 甲醇溶液(A)-0.1%甲酸水(B), 梯度洗脱(0~5 min, 2%→20% A; 6~10 min, 20%→50% A; 11~15 min, 50%→80% A; 16~20 min, 80%→95% A; 21~27 min, 95% A; 28~30 min, 2% B); 流速为0.3 mL·min^-1, 柱温45 ℃, 进样10 μL。由Q Exactive^TM混合Q-Orbitrap质谱仪(赛默飞科技公司) 进行分析。采用电喷雾离子源(ESI), 正、负离子模式下获取质谱信息。条件如下: ①鞘气流速: 40 L·min^-1; ②辅助气流速: 15 L·min^-1; ③锥孔电压: ± 3.2 kV; ④毛细管离子传输管温度: 300 ℃; ⑤加热温度: 350 ℃。

成分靶点、疾病靶点的搜集 将鉴别出的成分信息导入SWISS ADME (http://www.swissadme.ch/) 进行分析, 结合文献报道, 确定具备生物活性的化合物作为岩青兰的活性成分, 利用SWISS Target (http://swisstargetprediction.ch/) 和DrugBank (https://go.drugbank.com/) 对活性成分靶点进行搜集。通过GEONCBI数据库获取GSE1613355基因表达数据集, 分析获得老年T2DM患者和健康志愿者之间的DEGs (http://sangerbox.com/)。通过GENECARDs (https://www.genecards.org/) 数据库筛选“neuroinflammation”靶点, 构建神经炎症疾病靶点数据库。利用Venny (http://www.bioinformatics.com.cn/) 分别绘制疾病靶点与岩青兰活性成分靶点的交集靶点。

PPI网络及疾病-药物-靶点网络图的构建、核心靶点的筛选 将交集靶点导入STRING11.0数据库(https://cn.stringdb.org) 进行分析, 模式设置为多种蛋白, 物种限定为“Homo sapiens”。通过Cytoscape 3.7.2可视化PPI网络, 构建“疾病-药物-靶点”网络图, 利用MCC算法, 分别筛选Top10靶点。

GO功能和KEGG通路富集分析 利用DAVID (https://david.ncifcrf.gov/) 数据库进行GO功能富集分析, 从细胞的分子功能(MF)、细胞成分(CC) 和生物过程(BP) 确定基因富集的功能, P值与富集程度呈正相关, 选取筛选校正后P < 0.05的富集结果。将结果绘制成相应气泡图; 利用KEGG途径富集分析获得枢纽靶点作用的途径, 将结果绘制成相应弦图。

分子对接 以“疾病-药物-靶点”图中的平均节点度值为标准, 选取Top3核心成分和Top5靶点, 用Autodock1.5.6软件进行分子对接计算结合值能量。

血糖、血脂、氧化因子及INS水平检测 给药干预3周后, 将小鼠禁食4 h, 尾尖采血, 测定小鼠血糖值; 隔天后将小鼠禁食且自由饮水12 h, 眼球取血, 根据生化试剂盒说明书对小鼠血脂、氧化因子及INS水平检测进行检测, 剩余的血样于-80 ℃下冷冻保存。

ELISA测定炎症因子水平 将小鼠处死后, 取小鼠脑组织, 于-80 ℃下冷冻保存。取小鼠血清及脑组织样本, 用生理盐水进行稀释或者超声破碎。按照ELISA试剂说明书进行操作, 然后于450 nm处测定吸光度值, 绘制标准曲线并计算血清及脑组织样本中的TNF-α及IL-1β的水平。

肝组织形态学观察 收集小鼠肝组织, 用4%多聚甲醛进行固定, 然后进行石蜡包埋, 制备成5 μm厚的组织切片, 用苏木精-伊红(hematoxylin eosin, HE) 染色, 然后在显微镜下观察分析肝细胞形态变化。

免疫组化检测 将石蜡包埋的脑组织切割成连续的5 μm横截面, 然后放入恒温60 ℃的烘箱中2 h。加热结束后, 脱蜡, 吸干水分。一抗TLR4、NF-κB、NLRP3、ASC、caspase-1或Iba-1孵育过夜, 37 ℃下用二抗孵育(荧光需避光处理)。使用AXIOCam 50相机获得显微照片, 并使用Image Pro Plus 6.0版软件进行分析。

统计学分析 采用GraphPad Prism 7统计软件分析, 结果用mean ± SEM表示, 组间均数用One-Way ANOVA方法进行方差齐性分析。认定P < 0.05为具有显著性差异。

结果

收起

1　岩青兰化学成分鉴定

分析鉴定岩青兰化学成分共59个, 其中包含有机酚酸17个、黄酮16个、生物碱11个、萜类8个、其他成分7个。正、负离子模式下岩青兰样品总离子流图见图1所示, 化合物具体信息见表1。

2　生信分析

2.1　核心靶点网络构建

通过Swiss ADME测试, 结合文献检索, 得到6个核心活性成分, 包括绿原酸^[17]、丁香酸^[18]、柚皮素^[19]、积雪草酸^[20]、山柰酚^[21]及香叶木素^[22], 并得到共计140个药物成分靶点。通过分析GSE1613355基因表达谱数据库, 共获取老年T2DM和健康志愿者脑DEGs 442个, 其中上调基因共有114个, 下调基因一共有328个; 通过Venny分析, 得到药物靶点与老年T2DM脑DEGs交集靶点共31个(图2A~C), 与神经炎症交集靶点共56个(图2D~F), 分别将交集靶点导入STRING数据库中作为潜在靶点, 获取PPI图。利用MCC算法识别互作网络中的Top10中心节点, 分别得到岩青兰治疗老年T2DM核心靶点为信号转导和转录激活因子3 (STAT3)、IL-1β、细胞周期蛋白依赖性激酶抑制因子1A (CDKN1A)、吞噬细胞糖蛋白1 (CD44)、血红素加氧酶1 (HMOX1)、生长停滞和DNA损伤诱导蛋白45α (GADD45α)、SQSTN1、早期生长反应因子1 (EGR1)、NCL1、分泌型磷蛋白1 (SPP1); 改善神经炎症的核心靶点为胱天蛋白酶3 (CASP3)、BCL2、TNF、IL-1β、MAPK3、STAT3、AKT1、半胱天冬酶9 (CASP9)、CASP8、MAPK8。运用Cytoscape 3.7.2软件构建出岩青兰联合分析老年T2DM、神经炎症的“药物-活性成分-疾病靶标”网络图, 见图2G。其中黄色节点代表岩青兰的活性成分, 紫色节点代表药物、老年T2DM以及神经炎症的共同靶点, 粉色的代表药物与老年T2DM的共同靶点, 蓝色的代表药物与神经炎症的共同靶点。节点之间的线代表活性成分与作用靶点之间的相互作用关系。共包含83个节点和154个连接, 见图2。

2.2　GO功能富集和KEGG信号通路分析

对交集靶点进行GO富集分析和KEGG信号通路分析, 筛选P < 0.05的数据, 结果见图3所示。分析药物靶点与老年T2DM脑DEGs交集靶点, 结果得到56个生物学过程, 主要包含铁下垂、对蛋白质磷酸化的正向调控、p38MAPK级联的正向调节、规范NF-κB信号转导的调节、凋亡过程等; 8个细胞组分, 主要有细胞核、细胞质、细胞外外泌体等; 10个分子功能, 涉及蛋白质的结合、蛋白激酶的结合、蛋白质螯合活性等。KEGG信号通路分析共得到28条信号通路, 主要包括P53信号通路、磷脂酰肌醇-3-激酶(PI3K)-AKT信号通路、MAPK信号通路等(图3A、C)。分析药物靶点与神经炎症交集靶点, 结果得到生物学过程共涉及有387个, 主要有凋亡过程、神经元凋亡过程的正向调控、IL-1β生成的正向调控、炎症反应的正向调节、细胞对肿瘤坏死因子的反应等; 45个细胞组分, 主要有线粒体、细胞质、细胞外泌体等; 分子功能53个, 涉及酶结合、蛋白丝氨酸激酶的活性、细胞因子活性等。KEGG信号通路分析共得到138条信号通路, 主要包括细胞凋亡、T2DM并发症中的高级糖基化终末产物-受体(AGE-RAGE) 信号通路、Toll样受体、NF-κB信号通路等(图3B、D)。

2.3　分子对接结果

为了进一步验证岩青兰中的核心活性药效成分与核心靶点之间的相互作用关系, 以“药物-活性成分-疾病靶标”网络图中成分中心性最大的成分绿原酸、香叶木素、山柰酚为配体, 以靶点中心性较高的AKT1、BCL2、HMOX1、IL-1β、HQO1, 与受体进行结合能力预测, 结果见图3E。当结合能小于-7.0 kJ·mol^-1, 表示配体和蛋白受体之间有强烈的结合活性。结果显示除AKT1靶点之外(AKT1与绿原酸、香叶木素、山柰酚的结合能分别为-5.8、-6.5、-6.1 kJ·mol^-1), BCL2、HMOX1、IL-1β、HQO1与3个化合物均表现出了极强的结合活性, 其中山柰酚与HMOX1、HQO1结合活性最高: BCL2与绿原酸、香叶木素、山柰酚结合能为-7.8、-7.8、-8.2 kJ·mol^-1; IL-1β与绿原酸、香叶木素、山柰酚的结合能为-7.1、-6.7、-7.7 kJ·mol^-1; HMOX1与绿原酸、香叶木素、山柰酚的结合能为-8.3、-8.8、-9.3 kJ·mol^-1; HQO1与绿原酸、香叶木素、山柰酚的结合能分别为-8.3、-8.5、-9.4 kJ·mol^-1。

3　体内实验验证

3.1　岩青兰可改善db/db小鼠的空腹血糖和体重的变化

如图4所示, 和空白组小鼠对比, 模型组小鼠空腹血糖值升高, 而高剂量的岩青兰(600 mg·kg^-1) 能显著降低小鼠的空腹血糖, 但低剂量的岩青兰(300 mg·kg^-1) 对小鼠的空腹血糖无显著的影响(图4A)。另外, 与模型组相比, 连续给予岩青兰21 d后, 高剂量组小鼠的体重有一定的下降趋势, 阳性化合物二甲双胍可显著降低小鼠的体重(图4B)。此外, 研究发现, 岩青兰对正常小鼠的体重无显著影响(图4C)。

3.2　岩青兰可降低db/db小鼠血脂水平及血INS水平

通过生化分析检测各组小鼠血清三酰甘油(TG)、总胆固醇(TC) 及丙氨酸氨基转移酶(ALT) 的水平, 如图4D~F所示, 结果表明, 模型组小鼠TG、TC、ALT水平显著升高; 而岩青兰高剂量(600 mg·kg^-1) 和二甲双胍给药后与空白组水平无显著性差异; 且与模型组相比, 给药后ALT水平显著下降, 结果见图4。这说明岩青兰可以有效降低db/db小鼠的高血脂水平, 这与前期的研究报道一致。此外, 对小鼠血清的INS水平进行检测, 结果如图4G所示, 表明岩青兰高剂量(600 mg·kg^-1) 和二甲双胍干预后, 小鼠INS水平相对模型组有所下降, 结合其对血糖水平的调节作用表明, 岩青兰可以有效改善小鼠的高胰岛素血症和高血糖、高血脂水平。

3.3　岩青兰降低小鼠血氧化因子水平并改善其肝组织的病理形态

通过生化试剂盒对小鼠血清SOD及GSH的水平进行分析, 结果见图4H、I。结果可知, 相较于空白小鼠, db/db小鼠血清氧化因子水平升高, 而岩青兰高剂量(600 mg·kg^-1) 和二甲双胍长时间给药后可以明显回调这两种氧化因子的水平。HE染色结果如图4J, 对照组小鼠肝细胞排列整齐, 几乎无脂滴空泡, 而db/db小鼠肝组织呈现严重的空泡变性, 少量肝细胞水样变性; 岩青兰高剂量(600 mg·kg^-1)、二甲双胍给药后可见肝组织脂肪变性不同程度减少, 胞质内可见圆形空泡现象有所改善, 细胞肿胀现象有一定减少。

3.4　岩青兰可以改善小鼠的血及脑中的炎症因子水平

检测小鼠促炎细胞因子的分泌, 血清TNF-α水平以及脑中IL-1β和TNF-α水平进行评估以初步确认DRH的抗炎活性。结果见图4K~M, 相较于空白组, 模型组小鼠血清及脑中的TNF-α水平及脑IL-1β水平均升高; 而与模型组比较, 岩青兰高剂量(600 mg·kg^-1) 可以有效降低TNF-α及IL-1β含量, 这提示岩青兰对db/db小鼠的促炎细胞因子的分泌有一定的抑制作用。

3.5　岩青兰可以抑制db/db小鼠脑小胶质细胞的激活

小胶质细胞是一种广泛分布于中枢神经系统的单核巨噬细胞, 是介导神经炎症发生的主要细胞。受刺激后, 小胶质细胞会分化为经典激活的M1型和代替激活的M2型, 其中, M1型小胶质细胞具有促炎功能, 会诱导神经损伤。Iba-1是M1型小胶质细胞标志蛋白。免疫荧光结果(图5A、B) 显示, 与空白组相比, 模型组小鼠Iba-1表达显著增加, 而高剂量(600 mg·kg^-1) 给药可以有效降低Iba-1的荧光强度。这表明长时间的高剂量(600 mg·kg^-1) 给药抑制了M1型小胶质细胞的过度激活。

3.6　岩青兰可以抑制db/db小鼠脑TLR4/NF-κB通路的激活

检测分析小鼠脑中神经炎症相关信号通路的变化, 结合网络药理学结果, 研究了岩青兰对TLR4/NF-κB通路的影响, TLR4/NF-κB是经典的炎症反应调控通路。结果如图5所示, 免疫组织化学结果显示, 模型组小鼠脑内, TLR4水平显著升高(图5C、D), NF-κB核移位现象明显(图5E~H), 这意味着TLR4/NF-κB通路的激活。NF-κB是一种重要的转录因子, 在免疫、炎症、细胞凋亡过程中发挥着关键作用, 如对TNF-α、IL-1β等细胞因子的释放有诱导性调节作用, 这是神经炎症发生的重要基础。而高剂量(600 mg·kg^-1) 给药治疗后, 可以在一定程度上降低TLR4的表达, 减少NF-κB的核移位现象。

3.7　岩青兰可以抑制db/db小鼠脑区炎症小体的激活

NLRP3作为细胞内关键的模式识别受体, 受到刺激时可以招募接头蛋白ASC和蛋白酶caspase-1形成NLRP3炎症小体, 在机体的免疫反应中发挥重要的作用。且研究表明, NLRP3炎症小体的异常激活与T2DM的炎症基础密切相关。免疫组化结果(图6) 显示, 与空白组相比, db/db小鼠NLRP3、ASC及caspase-1蛋白水平全部显著上调, 这表明老年T2DM小鼠脑内NLRP3炎症小体的异常激活, 且这一过程会诱导IL-1β的成熟和分泌, 这与前面的研究结果一致。而与db/db小鼠相比, 高剂量(600 mg·kg^-1) 给药后, 脑区NLRP3、ASC及caspase-1蛋白水平有回调趋势, 提示岩青兰高剂量(600 mg·kg^-1) 可以在一定程度上改善小鼠脑NLRP3的激活情况(图6)。

讨论

收起

本研究利用网络学分析整合实验验证以初步探索岩青兰的主要活性成分及其对老年T2DM脑神经炎症的改善作用。随着《“健康中国2030”规划纲要》中“坚持预防为主”要求的提出, 我国的医药的探索模式在向“治未病”转变。药食同源类植物是大健康产业发展的重要物质基础, 成为了近年来中医药领域的热点研究对象^[23]。岩青兰在我国资源丰富, 可全草入药, 且由于其特殊的香气, 在山西地区, 一直被制作为道地药茶进行冲泡饮用, 并根据不同的制茶工艺开发出了多种茶类产品。此外, 岩青兰在保护心肌细胞凋亡、抗氧化、清除自由基、抑菌、抗炎、抗肿瘤方面皆有报道。前期有研究者发现其乙酸乙酯萃取物能显著抑制小鼠血清中ALT及AST的水平, 且对肝细胞有一定的保护作用, 但其具体作用机制尚未阐明^[24]。本研究通过网络药理学预测了岩青兰具有减缓改善老年T2DM脑病的作用, 且发现其保护作用与其抗神经炎活性相关。

近年来, 糖尿病的炎症学说日益受到关注, T2DM被认为是一种自然免疫和低度炎症性疾病。有学者^[25]对老年db/db小鼠生命特征和疾病状态进行了研究, 发现4周龄的db/db小鼠逐渐出现高血糖、高血脂和胰岛素抵抗等特征, 后期出现慢性并发症和胰岛β细胞功能衰竭, 10个月左右死亡, 这种小鼠的发病过程与人类T2DM表现极为相似。35周龄的db/db小鼠存在显著过度的炎症水平^[25]。糖尿病脑病是由高糖导致的中枢神经元凋亡及中枢免疫系统紊乱引起的, 是糖尿病常见的并发症之一。鉴于全球范围内糖尿病发病率的不断上升和加剧的人口老龄化趋势, 寻找有效的治疗靶点对于糖尿病的预防及治疗十分重要。虽然糖尿病脑病的发病机制尚不明了, 但多项证据表明其与炎症因子的激活密切相关^[26]。临床数据显示, 许多促炎细胞因子, 如TNF-α、IL-1β、IL-6等在患者血液中的表达均升高, 而这些炎症标志物的异常表达可以损坏血脑屏障, 直接影响大脑的活动, 如通过TLR4/NF-κB通路激活小胶质细胞M1型的表达。本研究通过网络药理学分析了老年T2DM患者脑DEGs, 发现功能富集在p38MAPK级联的正向调节、规范NF-κB信号转导的调节等炎症相关生物学过程。KEGG通路进一步分析揭示了这些基因在PI3K-AKT信号通路、MAPK信号通路中高度聚集。研究表明, MAPK是NF-κB上游激活剂, 此外MAPK通路也对炎症因子的释放起着重要作用。而PI3K/AKT通路更是可以通过活化的AKT增加磷酸化IκB激酶-α的水平进而激活NF-κB途径。因此网络药理学结果与临床数据相一致, 不仅证明了炎症对于糖尿病脑病的关键作用, 而且揭示了岩青兰活性成分通过作用于改善炎症进而改善T2DM脑病的潜在机制。

小胶质细胞在中枢神经系统中发挥着重要的免疫调节作用, 如免疫应答、病原体或损伤细胞器的吞噬、炎症的调控等, 它的作用是双重的: 一方面其适当的激活可以加快细胞碎片和病原体的吞噬, 同时M2型小胶质细胞的激活会释放抗炎细胞因子, 以维持大脑的免疫平衡; 另一方面, 小胶质细胞过度激活, 尤其是M1型促炎小胶质细胞的异常激活会损害神经元并加剧神经病理的发生^[26]。TLR是小胶质细胞膜上的一种识别受体, 在脑免疫中作为“门户蛋白”启动机体炎症链式反应。TLR4被激活后, 可以经NF-κB信号转导激活相关蛋白, 促进细胞因子的大量分泌, 导致异常的致炎影响。有研究表明, 敲除TLR4基因后, 小鼠的认知水平得到了改善, 这表明了TLR4在神经系统中的重要作用^[27]。NLRP3炎症小体是小胶质细胞表达的一种多蛋白复合物, 小胶质细胞的激活也与NLRP3炎症小体有关。正常状态下, 炎性小体的激活可加快组织的修复, 而病理状态下, 炎性小体的激活则会引起过度的炎症反应, 导致组织损伤。首先, 免疫刺激物会通过膜识别受体激活NF-κB, 诱导NLRP3、caspase-1等组分的生成, 进而组装生成NLRP3炎症小体。同时激活的caspase-1可致使pro-IL-1β生成IL-1β, 启动下游的炎症级联反应^[28-30]。本研究表明, 岩青兰可以抑制老年T2DM小鼠脑中小胶质细胞的激活, 抑制神经炎症, 而这种作用是通过调节TLR4/NF-κB和NLRP3炎症小体实现的。其确切的药效物质基础以及直接作用的靶点尚需进一步研究。

综上所述, 本研究通过网络药理学的方法预测了岩青兰改善老年T2DM脑神经炎症的作用机制及药效物质基础, 并进一步通过体内实验验证其潜在的作用机制。结果表明, 岩青兰可以通过抑制TLR4/NF-κB通路及NLRP3炎症小体在一定程度上调节老年T2DM小鼠脑炎症的发生以及小胶质细胞的异常激活。本研究基于民间应用习惯对岩青兰的药效物质及作用机制进行了探讨, 为促进药食同源中药资源的进一步开发提供了科技数据和理论支撑。

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

Fei Z, Xu Y, Zhang G, et al. Natural products with potential hypoglycemic activity in T2DM: 2019-2023 [J]. Phytochemistry, 2024, 223: 114130.

Arnold SE, Arvanitakis Z, Macauley-Rambach SL, et al. Brain insulin resistance in type 2 diabetes and Alzheimer disease: concepts and conundrums [J]. Nat Rev Neurol, 2018, 14: 168-181.

Walker AF, Graham S, Maple-Brown L, et al. Interventions to address global inequity in diabetes: international progress [J]. Lancet, 2023, 402: 250-264.

DeFronzo RA, Ferrannini E, Groop L, et al. Type 2 diabetes mellitus [J]. Nat Rev Dis Primers, 2015, 1: 15019.

Cho SB. Comorbidity genes of Alzheimer's disease and type 2 diabetes associated with memory and cognitive function [J]. Int J Mol Sci, 2024, 25: 2211.

Lin C, Cai X, Yang W, et al. Age, sex, disease severity, and disease duration difference in placebo response: implications from a meta-analysis of diabetes mellitus [J]. BMC Med, 2020, 18: 233.

Doucet J, Gourdy P, Meyer L, et al. Management of glucose-lowering therapy in older adults with type 2 diabetes: challenges and opportunities [J]. Clin Interv Aging, 2023, 18: 1687-1703.

Chong ZZ, Menkes DL, Souayah N. Targeting neuroinflammation in distal symmetrical polyneuropathy in diabetes [J]. Drug Discov Today, 2024, 29: 104087.

Van BL, Lips MA, Visser A, et al. Increased systemic and adipose tissue inflammation differentiates obese women with T2DM from obese women with normal glucose tolerance [J]. Metabolism, 2014, 63: 492-501.

Liu X, Zhang Y, Liang H, et al. RETRACTED: overexpression of microRNA-216a-3p accelerates the inflammatory response in cardiomyocytes in type 2 diabetes mellitus by targeting IFN-α2 [J]. Front Endocrinol (Lausanne), 2020, 11: 522340.

Zhang Y, Li X, Xu S, et al. The acetylation of Ganoderma applanatum polysaccharides on ameliorating T2DM-induced hepatic and colonic injuries by modulating the Nrf2/keap1-TLR4/NFκB-Bax/Bcl-2 pathways [J]. Int J Biol Macromol, 2025, 294: 140055.

Han M, Wang G, Fu Y, et al. Ginsenoside Rg1 alleviates diabetic liver injury and fibrosis by inhibiting the PLC-NFAT2-NLRP3 signaling pathway in T2DM mice [J]. J Funct Foods, 2024, 121: 106455.

Wu L, Gao L, Jin X, et al. Ethanol extract of Mao Jian green tea attenuates gastrointestinal symptoms in a rat model of irritable bowel syndrome with constipation via the 5-hydroxytryptamine signaling pathway [J]. Foods, 2023, 12: 1101.

Wang F, Dong PZ, Pei XY, et al. Research progress on dracocephalum rupestre [J]. Chin J Ethnomed Ethnopharm (中国民族民间医药), 2022, 31: 50-55.

He N, Ma WW, Huang HJ, et al. Chemical constituents and antioxidant activity of Dracocephalum rupestre [J]. J Hebei Univ (河北大学学报), 2018, 38: 617-622.

Esmaeili MA, Sonboli A, Mirjalili MH. Oxidative stress protective effect of Dracocephalum multicaule essential oil against human cancer cell line [J]. Nat Prod Res, 2014, 28: 848-852.

Lin CH, Jiang WP, Itokazu N, et al. Chlorogenic acid attenuates 5-fluorouracil-induced intestinal mucositis in mice through SIRT1 signaling-mediated oxidative stress and inflammatory pathways [J]. Biomed Pharmacother, 2025, 186: 117982.

Wang WT, Zhang YY, Li ZR, et al. Syringic acid attenuates acute lung injury by modulating macrophage polarization in LPS-induced mice [J]. Phytomedicine, 2024, 129: 155591.

Abdelkawy YS, Elharoun M, Sheta E, et al. Liraglutide and naringenin relieve depressive symptoms in mice by enhancing neurogenesis and reducing inflammation [J]. Eur J Pharmacol, 2024, 971: 176525.

Bian D, Zhang J, Wu X, et al. Asiatic acid isolated from Centella asiatica inhibits TGF-β1-induced collagen expression in human keloid fibroblasts via PPAR-γ activation [J]. Int J Biol Sci, 2013, 9: 1032.

Imran M, Salehi B, Sharifi RJ, et al. Kaempferol: a key emphasis to its anticancer potential [J]. Molecules, 2019, 24: 2277.

Ge A, Ma Y, Liu YN, et al. Diosmetin prevents TGF-β1-induced epithelial-mesenchymal transition via ROS/MAPK signaling pathways [J]. Life Sci, 2016, 153: 1-8.

Guo S, Wang YY, Zhang F. Research and practical approaches of current status and development of medicinal-edible homologous Chinese medicinal resources [J]. J Nanjing Univ Chin Med (南京中医药大学学报), 2023, 39: 801-808.

Wang TF, Duan RB, Yang JL. Study on the mechanism of Maojian Tea in intervening hyperlipidemia based on network pharmacology and molecular docking [J]. Food Sci (食品科学), 2023, 44: 7-14.

Fan R, Hao YT, Liu XR, et al. Evaluation of disease status and inflammatory levels in aged db/db mice [J]. Chin J Gerontol (中国老年学杂志), 2021, 41: 4.

Dhamija R, Tewari S, Gill PS, et al. Association of apical periodontitis with glycated hemoglobin levels and systemic inflammatory markers in patients with type 2 diabetes: a cross-sectional study [J]. J Endod, 2025, 51: 124-131.

Liu L, Xu Y, Dai H, et al. Dynorphin activation of kappa opioid receptor promotes microglial polarization toward M2 phenotype via TLR4/NF-κB pathway [J]. Cell Biosci, 2020, 10: 42.

Li R, Zhou Y, Zhang S, et al. The natural (poly) phenols as modulators of microglia polarization via TLR4/NF-κB pathway exert anti-inflammatory activity in ischemic stroke [J]. Eur J Pharmacol, 2022, 914: 174660.

Cui W, Sun C, Ma Y, et al. Inhibition of TLR4 induces M2 microglial polarization and provides neuroprotection via the NLRP3 inflammasome in Alzheimer's disease [J]. Front Neurosci, 2020, 14: 444.

Yan Y, Jiang W, Liu L, et al. Dopamine controls systemic inflammation through inhibition of NLRP3 inflammasome [J]. Cell, 2015, 160: 62-73.

2025年第60卷第10期

PDF下载

235

124

引用本文

BibTeX

文章信息

doi: 10.16438/j.0513-4870.2025-0177

接收时间：2025-02-23
首发时间：2026-09-20
出版时间：2025-10-12

补充材料

相关文章

文章信息

作者

出版历史

收稿日期：2025-02-23
修回日期：2025-07-17

基金

作者信息

¹忻州师范学院生物系, 山西忻州 034000

²沈阳药科大学无涯创新学院, 辽宁沈阳 110016

³沈阳药科大学功能食品与葡萄酒学院, 辽宁沈阳 110016

通讯作者:

徐梦洁, E-mail: xumj1019@126.com

颜廷旭, E-mail: ytxsyphu@163.com

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/yxxb/CN/10.16438/j.0513-4870.2025-0177

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

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

关闭全屏

BibTeX
EndNote
RefWorks
TxT

Compound No.	RT/min	Formula	Adduct (m/z)	Reference ion	m/z	Fragment ion (m/z)	Title
1	1.848	C₁₃H₁₁NO₅	261.061 55	[M+H]⁺	262.068 82	79.141 10, 131.248 10, 261.068 60	Oxolinic acid
2	2.013	C₆H₆O₃	126.030 27	[M-H]^-	125.022 99	69.032 91, 81.032 92, 97.027 93, 126.022 90	Pyrogallol
3	2.207	C₉ H₁₃N₃O₅	243.085 58	[M+H]⁺	244.092 86	112.050 71, 163.042 04, 228.747 22, 243.092 85	Cytarabine
4	2.343	C₇H₁₂O₆	192.062 15	[M-H]^-	191.054 87	133.028 08, 151.002 24	D-(-)-Quinic acid
5	2.555	C₇H₁₀O₅	174.051 70	[M-H]^-	173.044 37	71.012 23, 73.027 88, 83.048 61, 85.027 89, 99.007 19, 111.043 60, 137.022 98, 155.033 62	3, 4, 5-Trihydroxycyclohex-1-ene-1-carboxylic acid
6	4.04	C₅H₇NO₃	129.042 85	[M+H]⁺	130.050 13	84.044 98, 129.049 88	D-(+)-Pyroglutamic acid
7	4.046	C₅H₇NO₃	129.041 18	[M-H]^-	128.033 91	82.0282 2, 113.695 77	4-Oxoproline
8	6.048	C₁₀H₁₃N₅O₄	267.096 90	[M+H]⁺	268.104 19	57.034 41, 69.034 26, 97.028 97, 101.060 07, 136.061 66	Adenosine
9	6.277	C₁₀H₁₃N₅O₃	251.102 01	[M+H]⁺	252.109 28	73.029 08, 99.044 54, 177.054 73, 136.061 58 251.105 83	2′-Deoxyadenosine
10	6.727	C₁₁H₂₀N₂O₃	228.147 61	[M+H]⁺	229.154 91	70.065 90, 159.012 82, 173.028 43, 193.096 57, 211.107 77, 228.154 36	Prolylleucine
11	6.995	C₇H₆O₅	170.020 40	[M-H]^-	169.013 12	69.032 98, 81.032 94, 97.027 94, 126.026 20	Gallic acid
12	7.007	C₉ H₁₁NO₂	148.052 59	[M+NH₄]⁺	166.086 41	72.007 46, 75.007 18, 87.007 14, 93.032 95, 120.044 13, 121.028 05, 147.043 75	L-Phenylalanine
13	7.479	C₁₁H₁₅N₅O₄	281.112 66	[M+H]⁺	282.119 93	57.034 41, 69.034 26, 97.028 97, 119.035 14, 136.061 66, 281.119 45	2′-O-Methyladenosine
14	7.958	C₇H₆O₃	138.030 29	[M-H]^-	137.023 01	74.989 35, 93.032 97, 94.028 25, 109.027 81, 137.023 01	Salicylic acid
15	8.13	C₁₁H₁₅N₅O₄	281.112 68	[M+H]⁺	282.119 96	57.034 37, 69.034 22, 85.028 94, 136.061 65, 150.077 26, 281.119 20	N6-Me-Adenosine
16	8.692	C₈H₈O₂	136.050 95	[M-H]^-	135.043 67	75.007 17, 117.330 80, 134.035 90	2-Methylbenzoic acid
17	9.218	C₁₁H₉NO₂	187.063 48	[M+H]⁺	188.070 74	91.054 76, 115.054 44, 117.057 41, 118.065 31, 142.065 05, 143.072 86, 144.080 69, 146.059 88, 170.059 89, 187.070 51	Trans-3-indoleacrylic acid
20	9.956	C₉H₈O₄	180.040 51	[M-H]^-	179.033 20	59.012 25, 71.012 25, 85.027 85, 87.007 08, 89.022 83, 121.028 19, 134.035 96, 135.043 72, 136.047 10	Caffeic acid
18	10.183	C₉H₁₀O₅	198.053 03	[M+H]⁺	199.060 32	63.022 42, 78.009 43, 89.001 62, 91.017 28, 106.004 47, 121.027 98, 123.007 27, 138.030 81, 153.054 31, 166.997 36, 182.020 92, 198.047 99	Syringic acid
19	10.33	C₁₃H₁₃N₃	211.111 22	[M+H]⁺	212.118 50	92.049 96, 95.065 58, 119.060 49, 166.049 64, 194.044 86, 195.091 57, 211.117 86	N, N'-Diphenylguanidine
21	10.516	C₁₅H₁₆O₉	340.079 16	[M-H]^-	339.071 62	123.044 15, 133.028 37, 151.038 79, 179.033 65, 131.091 92	Esculin
23	11.118	C₉H₆O₄	178.026 51	[M+H]⁺	179.033 78	111.008 02, 137.022 99, 155.033 72, 178.033 66	Esculetin
24	11.179	C₁₀H₇NO₄	205.037 60	[M+H]⁺	206.044 88	132.044 45, 150.091 48, 178.049 67, 205.044 54	Xanthurenic acid
22	11.572	C₇H₆O₄	154.025 34	[M-H]^-	153.018 07	81.032 97, 85.027 90, 95.012 27, 108.020 13, 109.0279 7, 110.031 14, 123.007 23	Gentisic acid
25	11.745	C₁₆H₁₈O₉	354.094 37	[M-H]^-	353.086 98	85.027 85, 93.032 96, 111.043 62, 135.043 67, 137.023 03, 155.033 62, 173.044 30, 179.033 78, 191.055 02, 353.087 16	Chlorogenic acid
26	11.77	C₁₁H₁₂O₅	224.068 41	[M+H]⁺	225.075 68	91.054 70, 95.049 55, 119.049 26, 147.043 87, 149.023 24, 157.039 51, 175.038 80, 192.041 52, 207.064 79, 224.039 41	Sinapinic acid
27	12.258	C₁₀H₁₀O₄	194.058 06	[M+H]⁺	195.065 34	89.039 12, 107.857 80, 117.033 66, 135.043 98, 163.038 71, 177.090 87, 194.101 43	Methyl caffeate
28	13.221	C₂₆H₂₈O₁₄	564.147 82	[M+H]⁺	565.155 27	121.028 47, 203.033 58, 295.059 75, 307.059 91, 325.069 92, 379.080 44, 391.080 41, 409.091 13, 427.101 78, 511.123 14, 529.133 42, 564.149 90	Schaftoside
29	13.551	C₁₀H₈O₄	192.042 46	[M+H]⁺	193.049 74	133.028 38, 137.059 60, 161.059 54, 165.054 52, 178.025 92, 192.049 38	7-Hydroxy-6-methoxy-2H-chromen-2-one
30	13.633	C₈H₈O	120.055 99	[M-H]^-	119.048 70	59.009 26, 71.012 24, 119.048 70	Phenylacetaldehyde
31	13.636	C₉H₈O₃	164.04599	[M-H]^-	163.038 71	119.048 71, 120.052 01, 163.038 88	2, 3-Dihydro-1-benzofuran-2-carboxylic acid
32	13.638	C₉H₈O₃	164.047 49	[M+H]⁺	165.054 76	91.054 70, 119.049 22, 147.043 79, 164.054 64	4-Coumaric acid
33	13.837	C₂₇H₃₀O₁₆	610.153 26	[M+H]⁺	611.160 71	153.018 08, 287.054 41, 449.107 09, 610.159 97	Luteolin 30, 40-di-O-b-d-glucopyranoside
34	14.328	C₂₆H₂₈O₁₅	580.142 63	[M+H]⁺	581.149 90	85.028 98, 153.018 02, 171.028 58, 273.075 47, 287.054 47, 580.186 28	Kaempferol 3-O-vicianoside
35	14.364	C₁₅H₁₂O₅	272.068 15	[M+H]⁺	273.075 44	67.018 66, 119.049 33, 147.043 90, 153.018 05, 171.028 64, 272.075 44	Naringenin
36	14.364	C₂₇H₃₂O₁₄	580.178 17	[M-H]^-	579.170 84	65.001 66, 107.012 29, 119.048 70, 151.102 29, 271.060 85, 285.040 10, 459.114 50, 580.174 80	Naringin
38	14.418	C₁₆H₁₄O₆	302.079 00	[M+H]⁺	303.086 27	67.018 62, 117.033 66, 145.028 24, 153.018 01, 177.054 41, 179.033 62, 302.086 06	5, 7-Dihydroxy-2-(4-hydroxy-3-methoxyphenyl)-3, 4-dihydro-2H-1-benzopyran-4-one
39	14.501	C₂₁H₂₀O₁₁	448.100 19	[M+H]⁺	449.107 54	107.012 30, 133.028 08, 151.002 24, 284.032 26, 285.040 04, 447.092 53	Cynaroside
37	14.538	C₇H₆O₂	122.035 23	[M-H]^-	121.027 95	93.032 79, 121.027 97	Benzoic acid
40	15.156	C₁₆H₂₂O₇	343.163 04	[M+H]⁺	344.170 32	61.029 24, 69.034 24, 85.028 96, 97.028 82, 105.070 18, 137.059 59, 165.090 84, 180.086 27, 343.170 32	(2R, 3S, 4S, 5R, 6S)-2-(Hydroxymethyl)-6-[2-methoxy-4-(prop-2-en-1-yl)phenoxy]oxane-3, 4, 5-triol
41	15.252	C₂₁H₂₀O₁₀	432.105 44	[M+H]⁺	433.112 76	67.018 67, 119.049 28, 153.018 02, 271.059 63, 432.114 04	Apigetrin
42	15.514	C₉H₁₆O₄	188.103 73	[M-H]^-	187.096 45	57.032 96, 97.064 29, 123.080 09, 125.095 69, 169.085 62	Azelaic acid
43	15.626	C₁₅H₁₂O₆	288.062 96	[M+H]⁺	289.070 34	68.997 83, 89.039 07, 117.033 60, 135.043 98, 145.028 31, 153.017 99, 163.038 70, 179.037 75, 288.070 22	2-(3, 4-Dihydroxyphenyl)-5, 7-dihydroxy-3, 4-dihydro-2H-1-benzopyran-4-one
44	16.055	C₁₁H₆O₃	186.031 97	[M+H]⁺	187.039 25	69.034 27, 81.034 13, 109.101 53, 115.054 44, 131.049 10, 143.049 01, 159.116 41	Psoralen
45	16.25	C₃₆H₅₈O₁₁	712.403 03	[M-H]^-	711.395 75	419.310 67, 441.337 13, 453.300 17, 485.32 822, 503.337 49, 711.254 64	Sericoside
46	16.509	C₁₆H₁₂O₅	284.068 93	[M+H]⁺	285.076 20	81.070 51, 153.017 84, 242.056 88, 270.051 94, 284.075 38	Glycitein
48	17.054	C₁₅H₁₀O₆	286.047 40	[M+H]⁺	287.054 75	153.018 02, 241.049 03	Kaempferol
49	17.787	C₃₀H₄₈O₅	488.350 37	[M+H-H₂O]⁺	471.347 08	147.116 67, 201.163 56, 219.173 90, 271.059 63, 407.330 08, 425.340 76, 453.336 03	(3β, 5ξ, 9ξ)-3, 6, 19-Trihydroxyurs-12-en-28-oic acid
50	17.831	C₁₆H₁₂O₆	300.063 34	[M+H]⁺	301.070 65	258.051 88, 286.046 81	Diosmetin
47	17.993	C₃₆H₅₈O₁₀	696.407 79	[M-H]^-	695.400 51	59.011 43, 207.033 36, 469.331 85, 487.342 38	Rosamultin
51	21.237	C₃₀H₄₆O₃	436.334 49	[M+H]⁺	437.341 77	81.070 58, 95.086 05, 107.085 89, 119.085 74, 133.101 20, 145.101 12, 187.148 09, 205.158 69, 389.320 13, 407.331 09, 435.336 91	11, 12-Dehydroursolic acid lactone
52	21.829	C₃₀H₄₈O₅	488.349 86	[M-H]^-	487.342 19	443.353 09, 469.331 45	Asiatic acid
53	21.927	C₃₀H₄₆O₄	470.339 63	[M+H]⁺	471.347 02	81.070 54, 95.086 02, 107.085 88, 121.101 32, 189.163 71, 201.163 74, 407.330 20, 425.340 85	18-β-Glycyrrhetinic acid
54	24.008	C₁₆H₁₈O₉	354.094 53	[M-H]^-	353.087 25	85.027 89, 93.032 91, 111.043 87, 134.036 01, 135.043 72, 161.023 13, 179.033 83, 191.055 05	Neochlorogenic acid
55	24.014	C₃₀H₄₈O₃	456.359 88	[M+H-H₂O]⁺	439.356 84	81.070 54, 95.086 01, 107.085 85, 133.101 24, 147.116 59, 189.163 73, 411.319 27	Oleanolic acid
56	24.244	C₂₀H₃₂O₂	304.240 01	[M+H]⁺	305.247 28	67.055 05, 79.054 90, 93.070 38, 107.085 86, 121.101 34, 135.116 81, 149.132 43, 163.148 06, 259.205 35	Arachidonic acid
57	25.026	C₂₀H₃₄O₂	306.255 56	[M+H]⁺	307.262 85	67.055 02, 81.070 50, 95.085 98, 109.101 46, 123.116 92, 243.209 93, 261.221 13	Linolenic acid ethyl ester
58	25.548	C₃₅H₄₂N₂O₉	333.170 29	[M+H]⁺	334.170 56	269.371 87, 253.158 64, 161.042 73, 145.097 55, 133.283 19	Estriol
59	26.557	C₁₈H₃₇NO	283.287 55	[M+H]⁺	284.294 83	57.070 78, 60.045 28, 88.076 26, 102.091 72, 116.107 17	Stearamide

Compound No.

RT/min

Formula

Adduct (m/z)

Reference ion

m/z

Fragment ion (m/z)

Title

1.848

C₁₃H₁₁NO₅

261.061 55

[M+H]⁺

262.068 82

79.141 10, 131.248 10, 261.068 60

Oxolinic acid

2.013

C₆H₆O₃

126.030 27

[M-H]^-

125.022 99

69.032 91, 81.032 92, 97.027 93, 126.022 90

Pyrogallol

2.207

C₉ H₁₃N₃O₅

243.085 58

[M+H]⁺

244.092 86

112.050 71, 163.042 04, 228.747 22, 243.092 85

Cytarabine

2.343

C₇H₁₂O₆

192.062 15

[M-H]^-

191.054 87

133.028 08, 151.002 24

D-(-)-Quinic acid

2.555

C₇H₁₀O₅

174.051 70

[M-H]^-

173.044 37

71.012 23, 73.027 88, 83.048 61, 85.027 89, 99.007 19, 111.043 60, 137.022 98, 155.033 62

3, 4, 5-Trihydroxycyclohex-1-ene-1-carboxylic acid

4.04

C₅H₇NO₃

129.042 85

[M+H]⁺

130.050 13

84.044 98, 129.049 88

D-(+)-Pyroglutamic acid

4.046

C₅H₇NO₃

129.041 18

[M-H]^-

128.033 91

82.0282 2, 113.695 77

4-Oxoproline

6.048

C₁₀H₁₃N₅O₄

267.096 90

[M+H]⁺

268.104 19

57.034 41, 69.034 26, 97.028 97, 101.060 07, 136.061 66

Adenosine

6.277

C₁₀H₁₃N₅O₃

251.102 01

[M+H]⁺

252.109 28

73.029 08, 99.044 54, 177.054 73, 136.061 58 251.105 83

2′-Deoxyadenosine

6.727

C₁₁H₂₀N₂O₃

228.147 61

[M+H]⁺

229.154 91

70.065 90, 159.012 82, 173.028 43, 193.096 57, 211.107 77, 228.154 36

Prolylleucine

6.995

C₇H₆O₅

170.020 40

[M-H]^-

169.013 12

69.032 98, 81.032 94, 97.027 94, 126.026 20

Gallic acid

7.007

C₉ H₁₁NO₂

148.052 59

[M+NH₄]⁺

166.086 41

72.007 46, 75.007 18, 87.007 14, 93.032 95, 120.044 13, 121.028 05, 147.043 75

L-Phenylalanine

7.479

C₁₁H₁₅N₅O₄

281.112 66

[M+H]⁺

282.119 93

57.034 41, 69.034 26, 97.028 97, 119.035 14, 136.061 66, 281.119 45

2′-O-Methyladenosine

7.958

C₇H₆O₃

138.030 29

[M-H]^-

137.023 01

74.989 35, 93.032 97, 94.028 25, 109.027 81, 137.023 01

Salicylic acid

8.13

C₁₁H₁₅N₅O₄

281.112 68

[M+H]⁺

282.119 96

57.034 37, 69.034 22, 85.028 94, 136.061 65, 150.077 26, 281.119 20

N6-Me-Adenosine

8.692

C₈H₈O₂

136.050 95

[M-H]^-

135.043 67

75.007 17, 117.330 80, 134.035 90

2-Methylbenzoic acid

9.218

C₁₁H₉NO₂

187.063 48

[M+H]⁺

188.070 74

91.054 76, 115.054 44, 117.057 41, 118.065 31, 142.065 05, 143.072 86, 144.080 69, 146.059 88, 170.059 89, 187.070 51

Trans-3-indoleacrylic acid

9.956

C₉H₈O₄

180.040 51

[M-H]^-

179.033 20

59.012 25, 71.012 25, 85.027 85, 87.007 08, 89.022 83, 121.028 19, 134.035 96, 135.043 72, 136.047 10

Caffeic acid

10.183

C₉H₁₀O₅

198.053 03

[M+H]⁺

199.060 32

63.022 42, 78.009 43, 89.001 62, 91.017 28, 106.004 47, 121.027 98, 123.007 27, 138.030 81, 153.054 31, 166.997 36, 182.020 92, 198.047 99

Syringic acid

10.33

C₁₃H₁₃N₃

211.111 22

[M+H]⁺

212.118 50

92.049 96, 95.065 58, 119.060 49, 166.049 64, 194.044 86, 195.091 57, 211.117 86

N, N'-Diphenylguanidine

10.516

C₁₅H₁₆O₉

340.079 16

[M-H]^-

339.071 62

123.044 15, 133.028 37, 151.038 79, 179.033 65, 131.091 92

Esculin

11.118

C₉H₆O₄

178.026 51

[M+H]⁺

179.033 78

111.008 02, 137.022 99, 155.033 72, 178.033 66

Esculetin

11.179

C₁₀H₇NO₄

205.037 60

[M+H]⁺

206.044 88

132.044 45, 150.091 48, 178.049 67, 205.044 54

Xanthurenic acid

11.572

C₇H₆O₄

154.025 34

[M-H]^-

153.018 07

81.032 97, 85.027 90, 95.012 27, 108.020 13, 109.0279 7, 110.031 14, 123.007 23

Gentisic acid

11.745

C₁₆H₁₈O₉

354.094 37

[M-H]^-

353.086 98

85.027 85, 93.032 96, 111.043 62, 135.043 67, 137.023 03, 155.033 62, 173.044 30, 179.033 78, 191.055 02, 353.087 16

Chlorogenic acid

11.77

C₁₁H₁₂O₅

224.068 41

[M+H]⁺

225.075 68

91.054 70, 95.049 55, 119.049 26, 147.043 87, 149.023 24, 157.039 51, 175.038 80, 192.041 52, 207.064 79, 224.039 41

Sinapinic acid

12.258

C₁₀H₁₀O₄

194.058 06

[M+H]⁺

195.065 34

89.039 12, 107.857 80, 117.033 66, 135.043 98, 163.038 71, 177.090 87, 194.101 43

Methyl caffeate

13.221

C₂₆H₂₈O₁₄

564.147 82

[M+H]⁺

565.155 27

121.028 47, 203.033 58, 295.059 75, 307.059 91, 325.069 92, 379.080 44, 391.080 41, 409.091 13, 427.101 78, 511.123 14, 529.133 42, 564.149 90

Schaftoside

13.551

C₁₀H₈O₄

192.042 46

[M+H]⁺

193.049 74

133.028 38, 137.059 60, 161.059 54, 165.054 52, 178.025 92, 192.049 38

7-Hydroxy-6-methoxy-2H-chromen-2-one

13.633

C₈H₈O

120.055 99

[M-H]^-

119.048 70

59.009 26, 71.012 24, 119.048 70

Phenylacetaldehyde

13.636

C₉H₈O₃

164.04599

[M-H]^-

163.038 71

119.048 71, 120.052 01, 163.038 88

2, 3-Dihydro-1-benzofuran-2-carboxylic acid

13.638

C₉H₈O₃

164.047 49

[M+H]⁺

165.054 76

91.054 70, 119.049 22, 147.043 79, 164.054 64

4-Coumaric acid

13.837

C₂₇H₃₀O₁₆

610.153 26

[M+H]⁺

611.160 71

153.018 08, 287.054 41, 449.107 09, 610.159 97

Luteolin 30, 40-di-O-b-d-glucopyranoside

14.328

C₂₆H₂₈O₁₅

580.142 63

[M+H]⁺

581.149 90

85.028 98, 153.018 02, 171.028 58, 273.075 47, 287.054 47, 580.186 28

Kaempferol 3-O-vicianoside

14.364

C₁₅H₁₂O₅

272.068 15

[M+H]⁺

273.075 44

67.018 66, 119.049 33, 147.043 90, 153.018 05, 171.028 64, 272.075 44

Naringenin

14.364

C₂₇H₃₂O₁₄

580.178 17

[M-H]^-

579.170 84

65.001 66, 107.012 29, 119.048 70, 151.102 29, 271.060 85, 285.040 10, 459.114 50, 580.174 80

Naringin

14.418

C₁₆H₁₄O₆

302.079 00

[M+H]⁺

303.086 27

67.018 62, 117.033 66, 145.028 24, 153.018 01, 177.054 41, 179.033 62, 302.086 06

5, 7-Dihydroxy-2-(4-hydroxy-3-methoxyphenyl)-3, 4-dihydro-2H-1-benzopyran-4-one

14.501

C₂₁H₂₀O₁₁

448.100 19

[M+H]⁺

449.107 54

107.012 30, 133.028 08, 151.002 24, 284.032 26, 285.040 04, 447.092 53

Cynaroside

14.538

C₇H₆O₂

122.035 23

[M-H]^-

121.027 95

93.032 79, 121.027 97

Benzoic acid

15.156

C₁₆H₂₂O₇

343.163 04

[M+H]⁺

344.170 32

61.029 24, 69.034 24, 85.028 96, 97.028 82, 105.070 18, 137.059 59, 165.090 84, 180.086 27, 343.170 32

(2R, 3S, 4S, 5R, 6S)-2-(Hydroxymethyl)-6-[2-methoxy-4-(prop-2-en-1-yl)phenoxy]oxane-3, 4, 5-triol

15.252

C₂₁H₂₀O₁₀

432.105 44

[M+H]⁺

433.112 76

67.018 67, 119.049 28, 153.018 02, 271.059 63, 432.114 04

Apigetrin

15.514

C₉H₁₆O₄

188.103 73

[M-H]^-

187.096 45

57.032 96, 97.064 29, 123.080 09, 125.095 69, 169.085 62

Azelaic acid

15.626

C₁₅H₁₂O₆

288.062 96

[M+H]⁺

289.070 34

68.997 83, 89.039 07, 117.033 60, 135.043 98, 145.028 31, 153.017 99, 163.038 70, 179.037 75, 288.070 22

2-(3, 4-Dihydroxyphenyl)-5, 7-dihydroxy-3, 4-dihydro-2H-1-benzopyran-4-one

16.055

C₁₁H₆O₃

186.031 97

[M+H]⁺

187.039 25

69.034 27, 81.034 13, 109.101 53, 115.054 44, 131.049 10, 143.049 01, 159.116 41

Psoralen

16.25

C₃₆H₅₈O₁₁

712.403 03

[M-H]^-

711.395 75

419.310 67, 441.337 13, 453.300 17, 485.32 822, 503.337 49, 711.254 64

Sericoside

16.509

C₁₆H₁₂O₅

284.068 93

[M+H]⁺

285.076 20

81.070 51, 153.017 84, 242.056 88, 270.051 94, 284.075 38

Glycitein

17.054

C₁₅H₁₀O₆

286.047 40

[M+H]⁺

287.054 75

153.018 02, 241.049 03

Kaempferol

17.787

C₃₀H₄₈O₅

488.350 37

[M+H-H₂O]⁺

471.347 08

147.116 67, 201.163 56, 219.173 90, 271.059 63, 407.330 08, 425.340 76, 453.336 03

(3β, 5ξ, 9ξ)-3, 6, 19-Trihydroxyurs-12-en-28-oic acid

17.831

C₁₆H₁₂O₆

300.063 34

[M+H]⁺

301.070 65

258.051 88, 286.046 81

Diosmetin

17.993

C₃₆H₅₈O₁₀

696.407 79

[M-H]^-

695.400 51

59.011 43, 207.033 36, 469.331 85, 487.342 38

Rosamultin

21.237

C₃₀H₄₆O₃

436.334 49

[M+H]⁺

437.341 77

81.070 58, 95.086 05, 107.085 89, 119.085 74, 133.101 20, 145.101 12, 187.148 09, 205.158 69, 389.320 13, 407.331 09, 435.336 91

11, 12-Dehydroursolic acid lactone

21.829

C₃₀H₄₈O₅

488.349 86

[M-H]^-

487.342 19

443.353 09, 469.331 45

Asiatic acid

21.927

C₃₀H₄₆O₄

470.339 63

[M+H]⁺

471.347 02

81.070 54, 95.086 02, 107.085 88, 121.101 32, 189.163 71, 201.163 74, 407.330 20, 425.340 85

18-β-Glycyrrhetinic acid

24.008

C₁₆H₁₈O₉

354.094 53

[M-H]^-

353.087 25

85.027 89, 93.032 91, 111.043 87, 134.036 01, 135.043 72, 161.023 13, 179.033 83, 191.055 05

Neochlorogenic acid

24.014

C₃₀H₄₈O₃

456.359 88

[M+H-H₂O]⁺

439.356 84

81.070 54, 95.086 01, 107.085 85, 133.101 24, 147.116 59, 189.163 73, 411.319 27

Oleanolic acid

24.244

C₂₀H₃₂O₂

304.240 01

[M+H]⁺

305.247 28

67.055 05, 79.054 90, 93.070 38, 107.085 86, 121.101 34, 135.116 81, 149.132 43, 163.148 06, 259.205 35

Arachidonic acid

25.026

C₂₀H₃₄O₂

306.255 56

[M+H]⁺

307.262 85

67.055 02, 81.070 50, 95.085 98, 109.101 46, 123.116 92, 243.209 93, 261.221 13

Linolenic acid ethyl ester

25.548

C₃₅H₄₂N₂O₉

333.170 29

[M+H]⁺

334.170 56

269.371 87, 253.158 64, 161.042 73, 145.097 55, 133.283 19

Estriol

26.557

C₁₈H₃₇NO

283.287 55

[M+H]⁺

284.294 83

57.070 78, 60.045 28, 88.076 26, 102.091 72, 116.107 17

Stearamide