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Peak ID	t_R/min	Formula	Measured (m/z)	Error (ppm)	Ion mode	MS and MS^E data (m/z)	Xenobiotics or metabolites	Source
1^a	0.95	C₁₃H₁₆O₁₀	331.066 5	-0.6	[M-H]^-	331.066 5, 169.014 1	Gallic acid-O-glc	C
2^a	1.01	C₇H₆O₅	169.013 6	-0.6	[M-H]^-	169.013 6, 125.024 4	Gallic acid	C, MG
3 (M1)	1.37	C₂₆H₃₁NO₁₀	518.202 8	+0.4	[M+H]⁺	518.202 8, 342.169 2, 192.101 3	Magnoflorine-O- glucuronidation	C, MG
4 (M2)	1.58	C₁₉H₂₂NO₄	328.158 1	+9.8	[M]⁺	283.095 4, 265.089 8, 237.091 5	Desmethyl magnoflorine	MG
5^a	1.59	C₉H₁₀O₅	197.045 3	+1.5	[M-H]^-	197.045 3, 124.045 4	Progallin A	C, MG
6 (M3)	1.68	C₂₀H₂₃NO₅	358.165 0	-1.1	[M]⁺	358.165 0, 281.077 8, 253.087 7	Hydroxylated magnoflorine	MG
7^a	1.70	C₂₁H₂₄O₁₁	451.124 3	+0.7	[M-H]^-	451.124 3, 289.082 1	(+)-Catechin-5-O-β-D-glc	C
8^a	2.41	C₁₆H₁₈O₉	353.087 3	0.0	[M-H]^-	353.087 3, 191.089 9	Chlorogenic acid	C, MG
9^a	2.57	C₁₅H₁₄O₆	289.071 0	-0.7	[M-H]^-	289.071 0, 245.082 1	(+)-Catechin	C, MG
10^a	2.78	C₂₀H₂₃NO₄	342.171 5	+2.9	[M+H]⁺	342.171 5, 296.113 0	Isocorydine	C, MG
11 (M4)	2.98	C₂₆H₂₈NO₁₀	514.172 3	+1.9	[M]⁺	514.172 3, 338.137 7, 323.114 4	Jateorhizine-O-glucuronide	C, MG
12 (M5)	3.09	C₂₆H₂₆NO₁₀	512.155 7	0.0	[M]⁺	512.155 7, 336.123 5, 321.098 2	Berberine-O-glucuronide	MG
13^a	3.20	C₂₀H₂₄NO₄	342.170 0	-1.5	[M]⁺	342.170 0, 297.112 2, 282.087 5, 265.086 0	Magnoflorine	C, MG
14 (M6)	3.87	C₂₁H₁₈O₁₁	445.077 0	-0.2	[M-H]^-	445.077 0, 269.045 2, 225.055 6	Emodin-O-glucuronide	C, MG
15 (M7)	4.03	C₂₁H₁₆O₁₂	459.056 3	-0.2	[M-H]^-	459.056 3, 283.024 3, 239.035 9, 211.040 8	Rhein-O-glucuronide	C, MG
16^a	4.56	C₂₁H₂₀O₁₀	431.098 5	+1.6	[M-H]^-	431.098 5, 269.046 0, 241.049 2	Emodin 8-O-β-D-glc	C, MG
17 (M8)	4.68	C₂₅H₂₆NO₁₀	500.157 7	+4.0	[M]⁺	500.157 7, 324.154 9, 309.079 0	Demethyleneberberine-O- glucuronide	C, MG
18 (M9)	4.70	C₂₀H₁₈NO₇S	416.148 9	-3.6	[M]⁺	416.148 9, 336.123 8	Berberine-O-sulfate	C, MG
19^a	4.78	C₂₁H₁₈O₁₁	445.077 7	+1.3	[M-H]^-	445.077 7, 283.025 3, 239.035 3, 211.040 0	Rhein 1-O-β-D-glc	C, MG
20 (M10)	4.90	C₂₀H₁₈NO₅	352.116 5	-5.7	[M]⁺	352.116 5, 336.127 3, 320.097 3	Hydroxylated berberine	MG
21 (M11)	4.96	C₁₅H₈O₉S	362.984 3	+8.8	[M-H]^-	362.984 3, 283.025 5, 239.035 7, 211.040 0	Rhein-O-sulfate	C, MG
22 (M12)	4.98	C₂₆H₂₆NO₁₀	512.155 7	0.0	[M]⁺	512.155 7, 336.123 0, 321.098 0	Berberine-O-glucuronide	C, MG
23^a	5.15	C₂₁H₂₀O₁₂	463.087 5	-0.4	[M-H]^-	463.087 5, 301.003 8	Isoquercitrin	C
24 (M13)	5.52	C₁₅H₈O₉S	362.984 3	+8.8	[M-H]^-	362.984 3, 283.025 3, 239.035 5	Rhein-O-sulfate	MG
25 (M14)	5.75	C₂₅H₂₄NO₁₀	498.140 4	+0.8	[M]⁺	498.140 4, 322.109 8, 307.085 4	Berberrubinen-O-glucuronide	C, MG
26 (M15)	5.85	C₂₁H₁₈O₁₁	445.077 0	-0.2	[M-H]^-	445.077 0, 269.045 4, 225.054 9	Emodin-O-glucuronide	C, MG
27 (M16)	6.01	C₂₇H₂₉NO₁₀	528.188 0	+1.9	[M]⁺	528.188 0, 352.123 0, 337.121 3	Palmatine-O-glucuronide	C, MG
28 (M17)	6.19	C₁₉H₁₆NSO₇	402.066 1	+3.5	[M]⁺	402.066 1, 322.125 1, 307.085 0	Berberrubinen-O-sulfate	C, MG
29 (M18)	6.55	C₂₀H₂₀NO₅	354.135 0	+2.5	[M+H]⁺	354.135 0, 338.139 9	Hydroxylated jateorhizine	C, MG
30^a	6.86	C₂₁H₂₀O₁₁	447.092 7	-2.9	[M-H]^-	447.092 0, 357.091 7	Homoorientin	C, MG
31^a	7.15	C₂₀H₂₀NO₄	338.140 8	+4.7	[M]⁺	338.140 8, 322.099 7, 308.130 9	Jatrorrhizine	C, MG
32 (M19)	7.35	C₁₉H₁₈NO₄	324.120 6	-9.3	[M]⁺	324.120 6, 294.114 6	Demethyleneberberine	C, MG
33 (M20)	7.47	C₁₉H₁₆NO₄	322.109 1	+3.7	[M]⁺	322.109 1, 307.063 8	Berberrubinen	C, MG
34^{a, b}	7.69	C₄₇H₈₀O₁₈	931.529 6	+3.2	[M-H]^-	977.540 0, 931.529 6, 799.485 4, 637.431 2, 475.378 4	Notoginsenoside R₁	C, MG
35 (M21)	7.81	C₁₅H₁₀O₇S	333.0067	-0.6	[M-H]^-	333.006 7, 253.050 5, 225.056 1	Chrysophanol-O-sulfate	C, MG
36 (M22)	8.17	C₁₅H₁₀O₈S	349.001 8	0.0	[M-H]^-	349.001 8, 269.045 9, 241.044 2	Emodin-O-sulfate	C, MG
37 (M23)	8.25	C₁₅H₁₀O₇S	333.007 3	+1.2	[M-H]^-	333.007 3, 253.050 4, 225.056 0	Chrysophanol-O-sulfate	C, MG
38^{a, b}	8.36	C₂₀H₁₈NO₄	336.123 0	-1.8	[M]⁺	336.123 0, 321.098 4, 320.091 6, 306.076 3, 292.096 7, 278.081 4	Berberine	C, MG
39^a	8.39	C₄₂H₇₂O₁₄	799.486 2	+2.3	[M-H]^-	845.492 0, 799.486 2, 637.432 2, 475.378 6	Ginsenoside Rg₁	C, MG
40^a	8.48	C₂₁H₂₂NO₄	352.154 0	-2.6	[M]⁺	352.154 0, 337.143 1, 336.098 4	Palmatine	C, MG
41 (M24)	8.55	C₂₁H₁₈O₁₀	429.083 8	+3.7	[M-H]^-	429.083 8, 253.064 3, 225.065 5	Chrysophanol-O-glucuronide	C, MG
42 (M25)	8.78	C₁₅H₁₀O₈S	349.001 6	+0.6	[M-H]^-	349.001 6, 269.045 9, 225.045 9	Emodin-O-sulfate	C, MG
43 (M26)	9.15	C₂₆H₂₈NO₁₁	530.168 1	+3.6	[M]⁺	530.168 1, 338.104 3	Hydroxylated Jateorhizine- O-glucuronide	C, MG
44^a	9.23	C₁₅H₁₀O₆	285.039 0	-3.2	[M-H]^-	285.039 0, 229.045 0, 169.014 5	Kaempferol	C, MG
45^{a, b}	9.46	C₁₅H₁₀O₇	301.036 0	+4.0	[M-H]^-	301.036 0, 151.045 2, 107.015 2	Quercetin	C, MG
46^a	9.61	C₂₁H₂₂O₉	417.119 5	+2.2	[M-H]^-	417.119 5, 255.066 6, 227.071 8	Aloin	C, MG
47 (M27)	10.01	C₂₁H₁₈O₁₁	445.077 0	-0.2	[M-H]^-	445.077 0, 269.045 3, 241.055 0	Emodin-O-glucuronide	C, MG
48 (M28)	10.37	C₂₁H₁₈O₁₀	429.081 5	-1.6	[M-H]^-	429.081 5, 253.064 3, 225.055 5	Chrysophanol-O-glucuronide	C, MG
49^a	10.56	C₁₅H₁₂O₅	271.061 5	+3.3	[M-H]^-	271.061 5, 151.066 6	Naringenin	C, MG
50 (M29)	10.65	C₁₅H₁₀O₇S	333.007 2	-0.9	[M-H]^-	333.007 2, 253.050 1, 225.056 6	Chrysophanol-O-sulfate	C, MG
51 (M30)	10.80	C₁₆H₉O₉S	376.997 3	+1.1	[M-H]^-	376.997 3, 297.040 3	6-Methyl-rhein-O-sulfate	C, MG
52^a	11.76	C₂₂H₂₂O₁₀	445.113 8	+0.7	[M-H]^-	445.113 8, 283.061 7, 239.043 3	Physcion 8-O-β-D-glc	C, MG
53 (M31)	11.94	C₂₁H₁₆O₁₂	459.056 3	-0.2	[M-H]^-	459.056 3, 283.024 2, 211.041 0	Rhein-O-glucuronide	C, MG
54^a	12.53	C₅₄H₉₂O₂₃	1 107.605 0	+8.7	[M-H]^-	1 153.601 2, 1 107.605 0, 945.546 9, 783.491 7	Ginsenoside Rb1	C, MG
55^a	13.15	C₁₆H₁₀O₆	297.040 4	+1.7	[M-H]^-	297.040 4, 267.051 1, 239.055 7	6-Methyl-rhein	C, MG
56^a	13.36	C₃₆H₆₂O₉	637.434 1	+3.9	[M-H]^-	683.439 1, 637.434 1	Ginsenoside Rh₁	C, MG
57^a	13.56	C₂₄H₂₄O₁₁	487.125 2	+2.5	[M-H]^-	487.125 2, 283.061 5, 253.051 0	Physcion 8-O-β-D-(6" -O- acetyl)-glc	C, MG
58 (M32)	13.98	C₁₅H₁₀O₈S	349.002 9	+3.2	[M-H]^-	349.002 9, 269.045 3, 241.045 5	Emodin-O-sulfate	C, MG
59^a	14.13	C₃₀H₄₈O₆	505.352 7	-0.4	[M+H]⁺	505.352 7, 487.349 0, 451.349 7	16-Oxo-alisol-A	C, MG
60^{a, b}	14.52	C₁₅H₁₀O₅	271.060 7	+0.7	[M+H]⁺	271.060 7, 241.025 8	Aloe-emodin	C, MG
61^{a, b}	15.30	C₁₅H₈O₆	283.024 9	+2.1	[M-H]^-	283.024 9, 239.034 9, 211.040 8, 183.035 6	Rhein	C, MG
62^a	16.70	C₂₀H₁₈NO₅	351.119 2	+2.0	[M]⁺	352.119 2, 337.094 8	8-Oxobererine	C, MG
63^a	17.05	C₃₀H₄₆O₅	487.341 0	-2.7	[M+H]⁺	487.341 0, 469.326 5, 451.321 5	Alisol C	C, MG
64^{a, b}	17.25	C₁₆H₁₂O₆	299.055 5	-0.3	[M-H]^-	299.055 5, 284.049 2	Kaempferide	C, MG
65^a	17.46	C₁₉H₁₆O₄	307.097 3	+1.0	[M-H]^-	307.097 3, 187.040 6	Bisdemethoxy curcumin	C, MG
66^a	17.93	C₂₀H₁₈O₅	337.107 7	+0.3	[M-H]^-	337.107 7, 217.050 8	Demethoxy curcumin	C, MG
67^{a, b}	18.38	C₂₁H₂₀O₆	367.119 1	+2.5	[M-H]^-	367.119 1, 217.052 4	Curcumin	C, MG
68^{a, b}	19.25	C₁₅H₁₀O₅	269.045 4	+1.5	[M-H]^-	269.045 4, 241.050 5, 225.055 7	Emodin	C, MG
69^a	19.71	C₃₀H₄₈O₅	511.339 3	-1.2	[M+Na]⁺	511.339 3, 471.269 6, 383.268 7	Alisol F	C, MG
70^a	20.04	C₃₂H₄₆O₅	511.342 8	+1.0	[M+H]⁺	511.342 8, 467.315 7	Alisol L 23-acetate	C, MG
71^{a, b}	21.71	C₁₅H₁₀O₄	253.051 5	+5.5	[M-H]^-	253.051 5, 225.053 5	Chrysophanol	C, MG
72^a	22.90	C₃₂H₅₂O₆	555.365 9	-0.5	[M+Na]⁺	555.365 9, 515.352 6, 383.279 9	Alisol A 23-acetate	C, MG
73^a	23.54	C₃₀H₅₀O₅	491.373 1	-1.0	[M+H]⁺	491.373 1, 473.353 7, 455.352 3	Alisol A	C, MG
74^a	23.79	C₃₀H₄₈O₄	473.363 5	+0.8	[M+H]⁺	473.363 5, 455.353 5, 437.352 8	Alisol B	C, MG
75^a	23.93	C₃₀H₄₄O₄	469.330 0	-3.8	[M+H]⁺	469.330 0, 451.318 8, 433.312 3	Alisol L	C, MG
76^a	25.28	C₃₂H₅₂O₆	555.364 8	-2.5	[M+Na]⁺	555.364 8, 515.341 5, 383.278 7	Alisol A 24-acetate	C, MG
77^{a, b}	25.32	C₃₂H₅₀O₅	515.373 0	-1.2	[M+H]⁺	515.373 0, 497.362 1, 419.329 3	Alisol B 23-acetate	C, MG

Peak ID	t_R/min	Formula	Measured (m/z)	Error (ppm)	Ion mode	MS and MS^E data (m/z)	Xenobiotics or metabolites	Source
1^a	0.95	C₁₃H₁₆O₁₀	331.066 5	-0.6	[M-H]^-	331.066 5, 169.014 1	Gallic acid-O-glc	C
2^a	1.01	C₇H₆O₅	169.013 6	-0.6	[M-H]^-	169.013 6, 125.024 4	Gallic acid	C, MG
3 (M1)	1.37	C₂₆H₃₁NO₁₀	518.202 8	+0.4	[M+H]⁺	518.202 8, 342.169 2, 192.101 3	Magnoflorine-O- glucuronidation	C, MG
4 (M2)	1.58	C₁₉H₂₂NO₄	328.158 1	+9.8	[M]⁺	283.095 4, 265.089 8, 237.091 5	Desmethyl magnoflorine	MG
5^a	1.59	C₉H₁₀O₅	197.045 3	+1.5	[M-H]^-	197.045 3, 124.045 4	Progallin A	C, MG
6 (M3)	1.68	C₂₀H₂₃NO₅	358.165 0	-1.1	[M]⁺	358.165 0, 281.077 8, 253.087 7	Hydroxylated magnoflorine	MG
7^a	1.70	C₂₁H₂₄O₁₁	451.124 3	+0.7	[M-H]^-	451.124 3, 289.082 1	(+)-Catechin-5-O-β-D-glc	C
8^a	2.41	C₁₆H₁₈O₉	353.087 3	0.0	[M-H]^-	353.087 3, 191.089 9	Chlorogenic acid	C, MG
9^a	2.57	C₁₅H₁₄O₆	289.071 0	-0.7	[M-H]^-	289.071 0, 245.082 1	(+)-Catechin	C, MG
10^a	2.78	C₂₀H₂₃NO₄	342.171 5	+2.9	[M+H]⁺	342.171 5, 296.113 0	Isocorydine	C, MG
11 (M4)	2.98	C₂₆H₂₈NO₁₀	514.172 3	+1.9	[M]⁺	514.172 3, 338.137 7, 323.114 4	Jateorhizine-O-glucuronide	C, MG
12 (M5)	3.09	C₂₆H₂₆NO₁₀	512.155 7	0.0	[M]⁺	512.155 7, 336.123 5, 321.098 2	Berberine-O-glucuronide	MG
13^a	3.20	C₂₀H₂₄NO₄	342.170 0	-1.5	[M]⁺	342.170 0, 297.112 2, 282.087 5, 265.086 0	Magnoflorine	C, MG
14 (M6)	3.87	C₂₁H₁₈O₁₁	445.077 0	-0.2	[M-H]^-	445.077 0, 269.045 2, 225.055 6	Emodin-O-glucuronide	C, MG
15 (M7)	4.03	C₂₁H₁₆O₁₂	459.056 3	-0.2	[M-H]^-	459.056 3, 283.024 3, 239.035 9, 211.040 8	Rhein-O-glucuronide	C, MG
16^a	4.56	C₂₁H₂₀O₁₀	431.098 5	+1.6	[M-H]^-	431.098 5, 269.046 0, 241.049 2	Emodin 8-O-β-D-glc	C, MG
17 (M8)	4.68	C₂₅H₂₆NO₁₀	500.157 7	+4.0	[M]⁺	500.157 7, 324.154 9, 309.079 0	Demethyleneberberine-O- glucuronide	C, MG
18 (M9)	4.70	C₂₀H₁₈NO₇S	416.148 9	-3.6	[M]⁺	416.148 9, 336.123 8	Berberine-O-sulfate	C, MG
19^a	4.78	C₂₁H₁₈O₁₁	445.077 7	+1.3	[M-H]^-	445.077 7, 283.025 3, 239.035 3, 211.040 0	Rhein 1-O-β-D-glc	C, MG
20 (M10)	4.90	C₂₀H₁₈NO₅	352.116 5	-5.7	[M]⁺	352.116 5, 336.127 3, 320.097 3	Hydroxylated berberine	MG
21 (M11)	4.96	C₁₅H₈O₉S	362.984 3	+8.8	[M-H]^-	362.984 3, 283.025 5, 239.035 7, 211.040 0	Rhein-O-sulfate	C, MG
22 (M12)	4.98	C₂₆H₂₆NO₁₀	512.155 7	0.0	[M]⁺	512.155 7, 336.123 0, 321.098 0	Berberine-O-glucuronide	C, MG
23^a	5.15	C₂₁H₂₀O₁₂	463.087 5	-0.4	[M-H]^-	463.087 5, 301.003 8	Isoquercitrin	C
24 (M13)	5.52	C₁₅H₈O₉S	362.984 3	+8.8	[M-H]^-	362.984 3, 283.025 3, 239.035 5	Rhein-O-sulfate	MG
25 (M14)	5.75	C₂₅H₂₄NO₁₀	498.140 4	+0.8	[M]⁺	498.140 4, 322.109 8, 307.085 4	Berberrubinen-O-glucuronide	C, MG
26 (M15)	5.85	C₂₁H₁₈O₁₁	445.077 0	-0.2	[M-H]^-	445.077 0, 269.045 4, 225.054 9	Emodin-O-glucuronide	C, MG
27 (M16)	6.01	C₂₇H₂₉NO₁₀	528.188 0	+1.9	[M]⁺	528.188 0, 352.123 0, 337.121 3	Palmatine-O-glucuronide	C, MG
28 (M17)	6.19	C₁₉H₁₆NSO₇	402.066 1	+3.5	[M]⁺	402.066 1, 322.125 1, 307.085 0	Berberrubinen-O-sulfate	C, MG
29 (M18)	6.55	C₂₀H₂₀NO₅	354.135 0	+2.5	[M+H]⁺	354.135 0, 338.139 9	Hydroxylated jateorhizine	C, MG
30^a	6.86	C₂₁H₂₀O₁₁	447.092 7	-2.9	[M-H]^-	447.092 0, 357.091 7	Homoorientin	C, MG
31^a	7.15	C₂₀H₂₀NO₄	338.140 8	+4.7	[M]⁺	338.140 8, 322.099 7, 308.130 9	Jatrorrhizine	C, MG
32 (M19)	7.35	C₁₉H₁₈NO₄	324.120 6	-9.3	[M]⁺	324.120 6, 294.114 6	Demethyleneberberine	C, MG
33 (M20)	7.47	C₁₉H₁₆NO₄	322.109 1	+3.7	[M]⁺	322.109 1, 307.063 8	Berberrubinen	C, MG
34^{a, b}	7.69	C₄₇H₈₀O₁₈	931.529 6	+3.2	[M-H]^-	977.540 0, 931.529 6, 799.485 4, 637.431 2, 475.378 4	Notoginsenoside R₁	C, MG
35 (M21)	7.81	C₁₅H₁₀O₇S	333.0067	-0.6	[M-H]^-	333.006 7, 253.050 5, 225.056 1	Chrysophanol-O-sulfate	C, MG
36 (M22)	8.17	C₁₅H₁₀O₈S	349.001 8	0.0	[M-H]^-	349.001 8, 269.045 9, 241.044 2	Emodin-O-sulfate	C, MG
37 (M23)	8.25	C₁₅H₁₀O₇S	333.007 3	+1.2	[M-H]^-	333.007 3, 253.050 4, 225.056 0	Chrysophanol-O-sulfate	C, MG
38^{a, b}	8.36	C₂₀H₁₈NO₄	336.123 0	-1.8	[M]⁺	336.123 0, 321.098 4, 320.091 6, 306.076 3, 292.096 7, 278.081 4	Berberine	C, MG
39^a	8.39	C₄₂H₇₂O₁₄	799.486 2	+2.3	[M-H]^-	845.492 0, 799.486 2, 637.432 2, 475.378 6	Ginsenoside Rg₁	C, MG
40^a	8.48	C₂₁H₂₂NO₄	352.154 0	-2.6	[M]⁺	352.154 0, 337.143 1, 336.098 4	Palmatine	C, MG
41 (M24)	8.55	C₂₁H₁₈O₁₀	429.083 8	+3.7	[M-H]^-	429.083 8, 253.064 3, 225.065 5	Chrysophanol-O-glucuronide	C, MG
42 (M25)	8.78	C₁₅H₁₀O₈S	349.001 6	+0.6	[M-H]^-	349.001 6, 269.045 9, 225.045 9	Emodin-O-sulfate	C, MG
43 (M26)	9.15	C₂₆H₂₈NO₁₁	530.168 1	+3.6	[M]⁺	530.168 1, 338.104 3	Hydroxylated Jateorhizine- O-glucuronide	C, MG
44^a	9.23	C₁₅H₁₀O₆	285.039 0	-3.2	[M-H]^-	285.039 0, 229.045 0, 169.014 5	Kaempferol	C, MG
45^{a, b}	9.46	C₁₅H₁₀O₇	301.036 0	+4.0	[M-H]^-	301.036 0, 151.045 2, 107.015 2	Quercetin	C, MG
46^a	9.61	C₂₁H₂₂O₉	417.119 5	+2.2	[M-H]^-	417.119 5, 255.066 6, 227.071 8	Aloin	C, MG
47 (M27)	10.01	C₂₁H₁₈O₁₁	445.077 0	-0.2	[M-H]^-	445.077 0, 269.045 3, 241.055 0	Emodin-O-glucuronide	C, MG
48 (M28)	10.37	C₂₁H₁₈O₁₀	429.081 5	-1.6	[M-H]^-	429.081 5, 253.064 3, 225.055 5	Chrysophanol-O-glucuronide	C, MG
49^a	10.56	C₁₅H₁₂O₅	271.061 5	+3.3	[M-H]^-	271.061 5, 151.066 6	Naringenin	C, MG
50 (M29)	10.65	C₁₅H₁₀O₇S	333.007 2	-0.9	[M-H]^-	333.007 2, 253.050 1, 225.056 6	Chrysophanol-O-sulfate	C, MG
51 (M30)	10.80	C₁₆H₉O₉S	376.997 3	+1.1	[M-H]^-	376.997 3, 297.040 3	6-Methyl-rhein-O-sulfate	C, MG
52^a	11.76	C₂₂H₂₂O₁₀	445.113 8	+0.7	[M-H]^-	445.113 8, 283.061 7, 239.043 3	Physcion 8-O-β-D-glc	C, MG
53 (M31)	11.94	C₂₁H₁₆O₁₂	459.056 3	-0.2	[M-H]^-	459.056 3, 283.024 2, 211.041 0	Rhein-O-glucuronide	C, MG
54^a	12.53	C₅₄H₉₂O₂₃	1 107.605 0	+8.7	[M-H]^-	1 153.601 2, 1 107.605 0, 945.546 9, 783.491 7	Ginsenoside Rb1	C, MG
55^a	13.15	C₁₆H₁₀O₆	297.040 4	+1.7	[M-H]^-	297.040 4, 267.051 1, 239.055 7	6-Methyl-rhein	C, MG
56^a	13.36	C₃₆H₆₂O₉	637.434 1	+3.9	[M-H]^-	683.439 1, 637.434 1	Ginsenoside Rh₁	C, MG
57^a	13.56	C₂₄H₂₄O₁₁	487.125 2	+2.5	[M-H]^-	487.125 2, 283.061 5, 253.051 0	Physcion 8-O-β-D-(6" -O- acetyl)-glc	C, MG
58 (M32)	13.98	C₁₅H₁₀O₈S	349.002 9	+3.2	[M-H]^-	349.002 9, 269.045 3, 241.045 5	Emodin-O-sulfate	C, MG
59^a	14.13	C₃₀H₄₈O₆	505.352 7	-0.4	[M+H]⁺	505.352 7, 487.349 0, 451.349 7	16-Oxo-alisol-A	C, MG
60^{a, b}	14.52	C₁₅H₁₀O₅	271.060 7	+0.7	[M+H]⁺	271.060 7, 241.025 8	Aloe-emodin	C, MG
61^{a, b}	15.30	C₁₅H₈O₆	283.024 9	+2.1	[M-H]^-	283.024 9, 239.034 9, 211.040 8, 183.035 6	Rhein	C, MG
62^a	16.70	C₂₀H₁₈NO₅	351.119 2	+2.0	[M]⁺	352.119 2, 337.094 8	8-Oxobererine	C, MG
63^a	17.05	C₃₀H₄₆O₅	487.341 0	-2.7	[M+H]⁺	487.341 0, 469.326 5, 451.321 5	Alisol C	C, MG
64^{a, b}	17.25	C₁₆H₁₂O₆	299.055 5	-0.3	[M-H]^-	299.055 5, 284.049 2	Kaempferide	C, MG
65^a	17.46	C₁₉H₁₆O₄	307.097 3	+1.0	[M-H]^-	307.097 3, 187.040 6	Bisdemethoxy curcumin	C, MG
66^a	17.93	C₂₀H₁₈O₅	337.107 7	+0.3	[M-H]^-	337.107 7, 217.050 8	Demethoxy curcumin	C, MG
67^{a, b}	18.38	C₂₁H₂₀O₆	367.119 1	+2.5	[M-H]^-	367.119 1, 217.052 4	Curcumin	C, MG
68^{a, b}	19.25	C₁₅H₁₀O₅	269.045 4	+1.5	[M-H]^-	269.045 4, 241.050 5, 225.055 7	Emodin	C, MG
69^a	19.71	C₃₀H₄₈O₅	511.339 3	-1.2	[M+Na]⁺	511.339 3, 471.269 6, 383.268 7	Alisol F	C, MG
70^a	20.04	C₃₂H₄₆O₅	511.342 8	+1.0	[M+H]⁺	511.342 8, 467.315 7	Alisol L 23-acetate	C, MG
71^{a, b}	21.71	C₁₅H₁₀O₄	253.051 5	+5.5	[M-H]^-	253.051 5, 225.053 5	Chrysophanol	C, MG
72^a	22.90	C₃₂H₅₂O₆	555.365 9	-0.5	[M+Na]⁺	555.365 9, 515.352 6, 383.279 9	Alisol A 23-acetate	C, MG
73^a	23.54	C₃₀H₅₀O₅	491.373 1	-1.0	[M+H]⁺	491.373 1, 473.353 7, 455.352 3	Alisol A	C, MG
74^a	23.79	C₃₀H₄₈O₄	473.363 5	+0.8	[M+H]⁺	473.363 5, 455.353 5, 437.352 8	Alisol B	C, MG
75^a	23.93	C₃₀H₄₄O₄	469.330 0	-3.8	[M+H]⁺	469.330 0, 451.318 8, 433.312 3	Alisol L	C, MG
76^a	25.28	C₃₂H₅₂O₆	555.364 8	-2.5	[M+Na]⁺	555.364 8, 515.341 5, 383.278 7	Alisol A 24-acetate	C, MG
77^{a, b}	25.32	C₃₂H₅₀O₅	515.373 0	-1.2	[M+H]⁺	515.373 0, 497.362 1, 419.329 3	Alisol B 23-acetate	C, MG

UPLC-Q-TOF-MS^E结合外源性代谢组学方法分析鉴定肝豆汤在大鼠尿液中的代谢产物

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张雪燕 ¹ , 刘艺 ³ , 吴欢 ¹ , 曹仕健 ² , 吴鹏 ¹ , 周安 ¹^,^*

药学学报 | 研究论文 2020,55(5): 971-978

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药学学报 | 研究论文 2020, 55(5): 971-978

UPLC-Q-TOF-MS^E结合外源性代谢组学方法分析鉴定肝豆汤在大鼠尿液中的代谢产物

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张雪燕¹, 刘艺³, 吴欢¹, 曹仕健², 吴鹏¹, 周安¹^,^*

作者信息

1.安徽中医药大学科研实验中心, 中药复方安徽省重点实验室, 安徽合肥 230038

2.安徽中医药大学第一附属医院, 安徽合肥 230031

3.沃特世科技 (上海) 有限公司, 上海 201206

通讯作者:

*周安, Tel:86-551-65169752, E-mail:anzhou@ahtcm.edu.cn

Identification of major bioactive components and metabolites of Gandou decoction in rat urine by an integrative approach based on UPLC-Q-TOF-MS^E coupled with xenometabolomics analytical platform

Xue-yan ZHANG¹, Yi LIU³, Huan WU¹, Shi-jian CAO², Peng WU¹, An ZHOU¹^,^*

Affiliations

1. The Experimental Research Center, Anhui Province Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine, Hefei 230038, China

2. The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230031, China

3. Waters Corporation(China), Shanghai 201206, China

出版时间: 2020-05-15 doi: 10.16438/j.0513-4870.2019-0796

文章导航

摘要

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采用UPLC-Q-TOF-MS^E技术结合外源性代谢组学方法，对肝豆汤给药后正常和铜负荷模型大鼠尿液中的原形成分及代谢产物进行定性分析，在正、负离子扫描模式下获取大鼠尿液样品的质谱信息，包括保留时间、精确分子质量及离子强度。应用主成分分析（PCA）、正交偏最小二乘法-判别分析（OPLS-DA）外源性代谢组学方法筛选鉴定肝豆汤原形成分和相关代谢产物，最终从大鼠尿液中共检测到77个外源性成分，包括45个原形成分和32个代谢产物，10个原形成分经对照品确证；肝豆汤在大鼠尿液中原形成分主要有蒽醌、生物碱、原萜烷型萜和黄酮；这些成分在大鼠尿液中代谢途径有羟基化、去甲基化、硫酸化、葡萄糖醛酸化等。结果表明UPLC-Q-TOF-MS^E结合外源性代谢组学方法准确便捷，可作为筛选鉴定体内微量代谢成分的有效工具。动物实验经安徽中医药大学实验动物伦理委员会批准（批准号：2019025）。

关键词

肝豆汤 / 外源性代谢组学 / UPLC-Q-TOF-MS^E / 威尔逊病 / 代谢产物

Abstract

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To identify major bioactive components and metabolites of Gandou decoction (GDD) in urine of normal and copper-laden rats, an integrative approach that ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS^E) coupled with xenometabolomics analytical platform was established. Mass spectral data information about retention time, accurate m/z and ionic strength of rat urine samples was performed under positive and negative ion modes. Unsupervised principal components analysis (PCA) and supervised orthogonal partial least-squared discriminant analysis (OPLS-DA) were used to reveal the differential ions. As a result, a total of 77 compounds including 45 prototypes and 32 metabolites in urine were detected. Results indicated that anthraquinones, alkaloids and tetracyclic triterpenoids and flavonoids were the main chemical components of GDD in rat urine; the main metabolic pathways of these compounds in rat urine mainly include hydroxyl, methylation, sulfating, glucuronidation, and so on. UPLC-QTOF-MS^E coupled with xenometabolomics analytical platform is fast and efficient so that facilitates authentication of the material basis of Chinese herb compound in vivo, can also be used as an effective tool for ascertaining trace bioactive components in vivo. The animal experiments were approved by the Experimental Animal Ethics Committee of Anhui University of Chinese Medicine (No. 2019025).

Key words

Gandou decoction / xenometabolomics / UPLC-Q-TOF-MS^E / Wilson disease / metabolite

引用本文

张雪燕, 刘艺, 吴欢, 曹仕健, 吴鹏, 周安. UPLC-Q-TOF-MS^E结合外源性代谢组学方法分析鉴定肝豆汤在大鼠尿液中的代谢产物. 药学学报, 2020 , 55 (5) : 971 -978 . DOI: 10.16438/j.0513-4870.2019-0796

Xue-yan ZHANG, Yi LIU, Huan WU, Shi-jian CAO, Peng WU, An ZHOU. Identification of major bioactive components and metabolites of Gandou decoction in rat urine by an integrative approach based on UPLC-Q-TOF-MS^E coupled with xenometabolomics analytical platform[J]. Acta Pharmaceutica Sinica, 2020 , 55 (5) : 971 -978 . DOI: 10.16438/j.0513-4870.2019-0796

正文

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肝豆状核变性(hepatolenticular degeneration, HLD)又称Wilson病(Wilson disease, WD), 是常染色体隐性遗传的铜代谢障碍性疾病。其病因是患者体内缺少铜蓝蛋白后不能清除体内多余的铜, 导致铜在体内大量蓄积, 进而严重损伤组织器官^[1]。青霉胺等铜离子络合剂驱铜是目前HLD基本治疗方法, 该类药物可在体内与Cu²⁺配位形成配合物, 再经尿液排出体外, 从而达到治疗目的, 但由于该类药物不良反应严重患者不能长期服用^{[2, 3]}。肝豆汤(Gandou decoction, GDD)由“大黄、黄连、姜黄、金钱草、泽泻、三七”六味中药组成, 具有清热解毒、利尿通淋的功效。由于临床疗效确切, 该方被《肝豆状核变性诊断与治疗指南》推荐为治疗HLD的基本方^[4]。临床研究表明肝豆汤可显著提高HLD患者尿液中铜含量, 通过促进尿液排铜改善患者临床症状和生活质量^{[5, 6]}。课题组前期研究发现肝豆汤可有效促进铜负荷HLD模型大鼠尿液排铜^[7], 但是该方促进尿液排铜的化学成分及其代谢产物尚不明确, 分析鉴定肝豆汤在大鼠尿液中的原形成分和及其代谢产物有助于阐明该方排铜作用物质基础。

超高效液相色谱-四级杆飞行时间质谱(UPLC-Q-TOF-MS)是中药代谢产物分析鉴定工作有力工具, 具有分析速度快、高分辨率、高灵敏度等优势^{[8, 9]}。UPLC-Q-TOF-MS鉴定代谢产物时通常先设定目标原形成分结构, 再根据原形成分裂解规律设定可能的中性丢失碎片, 选择可能发生Ⅰ相和Ⅱ相的代谢途径来筛选代谢产物, 但由于生物基质背景复杂及代谢物响应低, 该方法可能造成代谢物遗漏^[10]。UPLC-Q-TOF-MS结合外源性代谢组学方法可以从全局角度分析生物样本中所有的代谢物轮廓, 结合主成分分析(principal component analysis, PCA), 正交偏最小二乘法判别分析(orthogonal partial least-squares discriminant analysis, OPLS-DA)等化学计量学方法可找出差异性外源性物质, 整体辨识生物体中外源性成分, 目前已成功应用于单体化合物外源性代谢物的鉴定^{[11, 12]}, 但该方法用于鉴定中药复方外源性代谢产物研究鲜有报道。前期研究采用UPLC-Q-TOF-MS^E技术鉴别了肝豆汤中96种化学成分, 并对该方主要成分肠吸收特征进行了考察^{[13, 14]}。为探究该方促尿液排铜作用物质基础, 本实验采用UPLC-Q-TOF-MS^E技术结合外源性代谢组学方法对正常大鼠和铜负荷HLD模型大鼠口服豆汤后尿液中的原形成分与代谢产物进行了系统研究, 结果表明该方法简便快捷, 可全面了解肝豆汤在大鼠尿液中的主要存在形式, 为肝豆汤促尿液排铜药效物质的阐明提供了参考。

材料与方法

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仪器 Waters Acquity UPLC超高效液相色谱系统, Waters Xevo G2 Q-TOF高分辨率飞行时间质谱仪, 配有电喷雾离子源(ESI) (美国Waters公司); Milli-Q Gradient A10超纯水器(美国Millipore公司); HP-5016SY氮吹仪(上海济成分析仪器有限公司)。

试剂对照品盐酸小檗碱(批号: 110713-201212)、大黄素甲醚(批号: 110758-201415)、大黄素(批号: 110756-201512)、23-乙酰泽泻醇B (批号: 111846-201504)、三七皂苷R₁ (批号: 110745-201318)、槲皮素(批号: 100081-200406)、姜黄素(批号: 110823-201404)均购于中国食品药品检定研究院; 大黄酸(批号: 150904, 纯度: 99.3%)、大黄酚(批号: 151028, 纯度大于98%)、芦荟大黄素(批号: 150730, 纯度大于97%)、山柰素(批号: 150122, 纯度大于98%)均购于北京北纳创联生物技术研究院, 色谱纯乙腈(TIEDA公司); 色谱纯甲酸(天津市光复精细化工研究所), 其他试剂均为分析纯。

实验动物 清洁级Wistar雄性大鼠, 体重(220 ± 20) g, 由扬州大学比较医学中心提供, 许可证号: SCXK (苏) 2017-0007。饲养室温度25 ± 2 ℃, 湿度50% ± 10%, 12 h人工昼夜交替。动物实验经安徽中医药大学动物实验伦理委员会批准(批准号: 2019025)。

肝豆汤冻干粉的制备 大黄(Rheum palmatum L.)、黄连(Coptis chinensis Franch.)、姜黄(Curcuma longa L.)、金钱草(Lysimachia christinae Hance.)、泽泻(Alisma orientale (Sam.) Juzep.)和三七(Parmx notoginseng (Burk.) F.H.Chen)均购于合肥市同仁堂大药房, 经安徽中医药大学刘先华鉴定为合格药材。参照课题组前期方法^[7]制备肝豆汤冻干粉, 经HPLC多成分定量分析冻干粉中主要成分含量, 符合课题组对肝豆汤冻干粉的质量控制要求。

对照品溶液的配制 精密称取盐酸小檗碱6.90 mg、大黄素甲醚5.74 mg、大黄素8.94 mg、23-乙酰泽泻醇B 4.12 mg、三七皂苷R₁ 9.78 mg、槲皮素6.27 mg、大黄酸5.79 mg、大黄酚6.15 mg、芦荟大黄素4.68 mg和山柰素5.21 mg的对照品置于5 mL量瓶中, 甲醇定容, 精密移取1.0 mL置于25 mL量瓶中, 甲醇定容, 经0.22 µm滤膜滤过作为对照品试液。

实验动物分组 Wistar雄性大鼠24只适应性饲养1周后, 随机分为4组:正常对照组、正常给药组、模型对照组、模型给药组, 每组6只。按照课题组前期方法^[7]复制铜负荷HLD大鼠模型, 临用前将肝豆汤冻干粉溶于0.5% CMC-Na溶液中配置成1.2 g·mL^-1冻干粉溶液, 正常给药组和模型给药组分别按11.5 g·kg^-1 (相当于生药量80 g·kg^-1)给药, 每天1次, 连续灌胃给药3天, 其余2组灌胃给予等体积的CMC-Na溶液。

样品采集与处理 末次给药前禁食12 h, 自由饮水, 待给药后将大鼠置于代谢笼中, 收集合并0~24 h尿液, 16 000 ×g离心5 min取上清液, 置于-80 ℃储存备用。检测前将尿液于4 ℃复融, 精密吸取样品1.0 mL加入4倍量甲醇, 涡旋混匀2 min, 离心15 min, 取上清液于EP管中, 30 ℃条件下氮气吹干, 0.5 mL甲醇复溶, 0.22 μm微孔滤膜过滤后进样分析。

色谱检测条件 色谱柱Agilent Eclipse Plus C₁₈ (2.1 mm×100 mm, 1.8 μm); 柱温30 ℃; 流动相: 0.1%甲酸水(A)-乙腈(B); 梯度洗脱: 0~1 min, 10% B; 1~4 min, 10%~20% B; 4~10 min, 20%~30% B; 10~15 min, 30%~40% B; 15~18 min, 40%~50% B; 18~23 min, 50%~75% B; 23~25 min, 75%~85% B; 25~27 min, 85%~100% B。流速0.3 mL·min^-1; 进样体积1 μL。

质谱检测条件 电喷雾离子源(ESI)正、负离子模式分别进行MS^E扫描; 低碰撞电压: 6 V; 高碰撞电压: 20~80 V; 离子源温度: 120 ℃ (ESI⁺)和110 ℃ (ESI^-); 锥孔电压: 40 V; 锥孔气流量: 50 L·h^-1; 电喷雾电压: 3.0 kV (ESI⁺)与2.5 kV (ESI^-); 雾化气体积流量: 600 L·h^-1; 脱溶剂气温度: 350 ℃; 扫描范围: m/z 50~1 200;亮氨酸脑啡肽实时校准质量数。

外源性代谢组学技术分析原形及代谢物策略 ① Progenesis QI V2.0软件将各组质谱数据进行峰对齐、峰识别, 提取特征峰离子强度, 获得序号、保留时间、质荷比及对应离子强度的三维数据集; ②将三维数据集导入EZinfo 3.0软件中进行主成分分析(PCA)无监督数据分析, 聚类各组数据提出离群样本; ③使用正交偏最小二乘法判别分析(OPLS-DA)进行有监督的数据分析, 得到S-plot载荷图筛选得到差异离子; ④结合化学成分库和裂解规律, 对差异离子一级质谱信息、二级碎片信息、相关的特征丢失碎片信息进行分析鉴定原形成分和外源性代谢物。

结果

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1 肝豆汤原形及代谢产物的识别与鉴定

采用有监督的OPLS-DA模型分别对正、负离子数据集进行分析, 得到的OPLS-DA得分图由交叉验证参数R²X和Q²描述, 正常对照组和正常给药组在正离子模式下, R²X和Q²分别为0.98和0.95, 负离子模式下, 分别为0.99和0.97。模型对照组和模型给药组在正离子模式下, R²X和Q²分别为0.99和0.98, 负离子模式下, 分别为0.99和0.98, 均接近1, 远大于0.5, 提示模型具有很好的稳定性和预测能力。图 1和图 2分别为正负离子模式下空白组与给药组(正常给药组、模型给药组) PCA得分图, 结合OPLS-DA中S-plot载荷图(图 3、4, VIP > 1)筛选给药组中对分类起主要作用的离子, S-plot载荷图中离原点越远, 对应VIP值越大, 对分类影响也就越大, 这些差异较大离子为肝豆汤原形成分或者代谢物, 也有可能是浓度差异大的内源性成分, 剔除内源性成分, 再将原形成分和代谢物提取得到提取离子流图(EICs), 见图 5。

2 肝豆汤大鼠尿液原形及代谢产物的分析鉴定

将上述提取得到的差异性离子导入Masslynx V4.1软件分别对各个组别尿液样品的UPLC-Q-TOF-MS数据进行分析。参照上述“外源性代谢组学技术分析原形及代谢物策略”最终从大鼠尿液样品中筛选出45个潜在的原形成分和32个潜在的代谢产物, 结果见表 1。

2.1 肝豆汤大鼠尿液原形成分鉴定

肝豆汤尿液共筛选出45个原形成分(蒽醌类10个、生物碱类6个、原萜烷型三萜类10个、黄酮类6个、姜黄素类3个、皂苷类4个、鞣质类5个、有机酸类1个), 其中10个化学成分经标准品进一步确证。原形成分提取离子峰、质谱信息和碎片信息见图 5和表 1。

图 5和表 1中看出正常和模型大鼠尿液样本中共计检测出10个蒽醌类原形成分, 分别为峰16、19、46、52、55、57、60、61、68和71, 其中峰60、61、68和71经对照品归属分别为芦荟大黄素、大黄酸、大黄素和大黄酚。大黄素的分子离子峰为m/z 269.045 4 [M-H]^-, 在高能量MS/MS中逐级失去CO碎片得到m/z 241.050 5 [M-H-CO]^-, 并且出现m/z 225.055 7 [M-H-CO₂]^-的CO₂中性丢失。在裂解过程中, 大黄酸会出现连续的CO丢失, 在负离子模式下, 形成m/z 283.024 9、m/z 239.034 9、m/z 211.040 8、m/z 183.035 6的主要碎片离子。尿液样本中检测到6个生物碱类原形成分, 分别为峰10、13、31、38、40和62, 其中峰38经对照品归属为小檗碱。正离子模式下, 小檗碱分子离子峰为m/z 336.123 0 [M]⁺, MS/MS显示主要碎片为m/z 321.098 4、m/z 320.091 6、m/z 306.076 3、m/z 292.096 7、m/z 278.081 4等。峰59、63、69、70、72~77为三萜类成分, 以峰74为例, t_R=23.79 min, 正离子模式下, 其母核结构稳定但其支链容易发生脱水断链, m/z 473.363 1 [M+H]⁺脱去一分子H₂O形成m/z 455.353 5 [M+H-H₂O]⁺的碎片离子, 接着失去一分子H₂O形成m/z 437.352 8 [M+H-2H₂O]⁺的碎片离子, 结合对照品保留时间和质谱信息, 鉴定该化合物为泽泻醇B。峰34、39、54和56为皂苷类成分, 其中峰34经对照品归属为三七皂苷R₁。负离子模式下, 三七皂苷R₁的m/z 977.540 0 [M-H+HCOO]^-和m/z 931.529 6 [M-H]^-的基峰, 高能量时碎片离子m/z 799.485 4、m/z 637.431 2、m/z 475.378 4分别代表[M-H-Xyl]^-、[M-H-Xyl-Glu]^-以及[M-H-Xyl-2Glu]^-。峰65~67为姜黄素类成分, 该类成分负离子响应较好, 峰65和峰66的保留时间分别为17.46和17.93 min, 低能量时分别得到m/z 307.097 3和m/z 337.107 6的分子离子峰, 高能量下得到相应的碎片离子, 可推测峰65和峰66分别为双去甲氧基姜黄素和去甲氧基姜黄素。峰23、30、44、45、49和64为黄酮类成分, 其中峰45和64分别为槲皮素和山柰素。

2.2 肝豆汤大鼠尿液中代谢产物鉴定

结合原形产物及其相应的裂解规律, 从大鼠尿液中共检测到32个代谢产物(表 1中M1~32), 经质谱鉴定主要为生物碱类代谢物和蒽醌类代谢物。以M5为例, M5分子离子峰为m/z 512.155 7 [M]⁺, 分子式为C₂₆H₂₅NO₅, 在高能量质谱图中, 特征丢失葡萄糖醛酸分子(176 Da), 同时也产生小檗碱的特征离子, 推测M5为小檗碱Ⅱ相葡萄糖醛酸化的产物。M4的分子离子峰[M]⁺为m/z 514.172 3, 低能量下有m/z 388.137 7的产物离子, 表明其具有GluA单元连接到苷元部分, 推测M4为药根碱的葡萄糖醛酸化产物, 然而葡萄糖醛酸化的具体位置不能被确认。根据类似的方法对其他生物碱类代谢物进行鉴别, 其中M1、M2、M3分别是木兰花碱的葡萄糖醛酸化产物、去甲基后产物和羟基化产物; M8和M14分别为葡萄糖醛酸化的去亚甲基小檗碱和去甲基小檗碱, M10为小檗碱的羟基化产物; M17为硫酸化的去甲基小檗碱; M9、M19和M20分别为硫酸化小檗碱、去亚甲基小檗碱和去甲基小檗碱; M4、M18、M26分别为药根碱的葡萄糖醛酸化产物、药根碱羟基化和羟基化后葡萄糖醛酸结合物; M16为巴马汀的葡萄糖醛酸结合物。16个生物碱代谢物中, M2、M3、M10、M18~M20为Ⅰ相代谢产物, 其他为硫酸化和葡萄糖醛酸化的Ⅱ相代谢产物。

鉴定出的16个蒽醌类代谢产物均为硫酸化和葡萄糖醛酸化的Ⅱ相代谢产物。如M15的分子离子峰为m/z 445.077 0 [M-H]^-, 在高能量下, 特征丢失葡萄糖醛酸分子(176 Da), 产生m/z 269.045 4子离子, 同时也产生m/z 241.050 6和m/z 225.054 9等一系列大黄素的特征离子。M6、M27与M15的分子离子峰相同, 采用类似的方法进行鉴别, 推测M6、M15、M27为大黄素的葡萄糖醛酸化产物。M22的分子离子峰为m/z 349.001 8 [M-H]^-, 在高能量质谱图中, 特征丢失硫酸化分子(80 Da), 产生m/z 269.045 9的子离子, 同时也产生m/z 241.044 2大黄素的特征离子。按照类似方法对其他蒽醌类代谢物进行了推测, 推测M22、M25、M32为大黄素硫酸化结合物; M7、M31为大黄酸葡萄糖醛酸化结合物, M11、M13为大黄酸硫酸化结合物; M24、M28为大黄酚葡萄糖醛酸化结合物, M21、M23、M29为大黄酚硫酸化结合物; M30为6-甲基大黄酸硫酸化结合物。模型给药组大鼠与正常给药组在药物的吸收代谢稍有差异, 课题组前期对铜负荷大鼠病理切片可清晰看出模型大鼠存在一定的肝损伤^[7], 可能影响到代谢酶水平。

3 肝豆汤大鼠尿液代谢轮廓分析

从上述结果中可看出, 蒽醌类、生物碱类化合物的原形成分及其相关的代谢物为肝豆汤主要原形及代谢物, 这些成分能经尿液代谢排至体外, 在尿液样品中共计检测到22个生物碱类成分, 葡萄糖醛酸化和硫酸化、去甲基化、羟基化等代谢途径是生物碱类成分的重要的代谢方式。在尿液中共计检测到26个蒽醌类成分, 葡萄糖醛酸化和硫酸化为蒽醌类化合物的主要代谢途径。除此之外, 共检测到10种原萜烷型萜类原形成分、4种皂苷原形成分和3种姜黄素类原形成分。

讨论

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全面有效地检测和鉴定中药复方代谢产物目前仍然存在较大技术挑战^{[15, 16]}, 本研究使用UPLC-Q-TOF-MS^E技术结合外源性代谢组学技术可快速全面有效筛选出大鼠尿液中原形成分和代谢产物。通过比较直接质谱检测分析发现一些质谱响应较低的峰, 尤其是代谢产物很难进行鉴定, 在直接采用质谱检测时同样因为峰度太低造成遗漏。外源性代谢组学结合UPLC-Q-TOF-MS方法能去除内源性干扰离子, 数据分析过程更加准确便捷, 简化了化合物解析过程, 可作为筛选鉴定体内微量代谢成分的有效工具。

文献报道中药(复方)可以通过配位金属离子促进重金属排出体外, 发挥驱铅、驱铜、排结石等作用。本研究从尿液中检测得到的45个原形成分主要为蒽醌类、黄酮类、生物碱类和原萜烷型萜类化合物, 这几类化合物含有酚羟基、羰基、氨基等配位基团, 能提供一对或多对孤对电子与含有空轨道的金属元素离子发生配位, 如黄酮类化合物槲皮素中的3位羟基和4位羰基与Cu²⁺络合形成络合物^[17], 蒽醌类化合物大黄素^[18]、大黄酚^[19]、大黄素甲醚^[20]均可以与Cu²⁺发生络合, 生成不同类型的络合物。研究结果表明, 肝豆汤在正常和模型大鼠尿液中的黄酮和蒽醌类化合物可能是其发挥尿液排铜的药效物质基础。

基金

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国家自然科学基金项目(81872976)
安徽省重点研究与开发计划(201904a07020051)
安徽省自然科学基金项目(1908085MH266)
安徽省高校优秀青年骨干人才国外访学研修项目(gxgwfx2018042)

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2020年第55卷第5期

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文章信息

doi: 10.16438/j.0513-4870.2019-0796

接收时间：2019-10-08
首发时间：2026-01-21
出版时间：2020-05-15

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收稿日期：2019-10-08
修回日期：2019-12-20

基金

国家自然科学基金项目(81872976)

安徽省重点研究与开发计划(201904a07020051)

安徽省自然科学基金项目(1908085MH266)

安徽省高校优秀青年骨干人才国外访学研修项目(gxgwfx2018042)

作者信息

1.安徽中医药大学科研实验中心, 中药复方安徽省重点实验室, 安徽合肥 230038

2.安徽中医药大学第一附属医院, 安徽合肥 230031

3.沃特世科技 (上海) 有限公司, 上海 201206

通讯作者:

*周安, Tel:86-551-65169752, E-mail:anzhou@ahtcm.edu.cn

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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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TxT

Peak ID	t_R/min	Formula	Measured (m/z)	Error (ppm)	Ion mode	MS and MS^E data (m/z)	Xenobiotics or metabolites	Source
1^a	0.95	C₁₃H₁₆O₁₀	331.066 5	-0.6	[M-H]^-	331.066 5, 169.014 1	Gallic acid-O-glc	C
2^a	1.01	C₇H₆O₅	169.013 6	-0.6	[M-H]^-	169.013 6, 125.024 4	Gallic acid	C, MG
3 (M1)	1.37	C₂₆H₃₁NO₁₀	518.202 8	+0.4	[M+H]⁺	518.202 8, 342.169 2, 192.101 3	Magnoflorine-O- glucuronidation	C, MG
4 (M2)	1.58	C₁₉H₂₂NO₄	328.158 1	+9.8	[M]⁺	283.095 4, 265.089 8, 237.091 5	Desmethyl magnoflorine	MG
5^a	1.59	C₉H₁₀O₅	197.045 3	+1.5	[M-H]^-	197.045 3, 124.045 4	Progallin A	C, MG
6 (M3)	1.68	C₂₀H₂₃NO₅	358.165 0	-1.1	[M]⁺	358.165 0, 281.077 8, 253.087 7	Hydroxylated magnoflorine	MG
7^a	1.70	C₂₁H₂₄O₁₁	451.124 3	+0.7	[M-H]^-	451.124 3, 289.082 1	(+)-Catechin-5-O-β-D-glc	C
8^a	2.41	C₁₆H₁₈O₉	353.087 3	0.0	[M-H]^-	353.087 3, 191.089 9	Chlorogenic acid	C, MG
9^a	2.57	C₁₅H₁₄O₆	289.071 0	-0.7	[M-H]^-	289.071 0, 245.082 1	(+)-Catechin	C, MG
10^a	2.78	C₂₀H₂₃NO₄	342.171 5	+2.9	[M+H]⁺	342.171 5, 296.113 0	Isocorydine	C, MG
11 (M4)	2.98	C₂₆H₂₈NO₁₀	514.172 3	+1.9	[M]⁺	514.172 3, 338.137 7, 323.114 4	Jateorhizine-O-glucuronide	C, MG
12 (M5)	3.09	C₂₆H₂₆NO₁₀	512.155 7	0.0	[M]⁺	512.155 7, 336.123 5, 321.098 2	Berberine-O-glucuronide	MG
13^a	3.20	C₂₀H₂₄NO₄	342.170 0	-1.5	[M]⁺	342.170 0, 297.112 2, 282.087 5, 265.086 0	Magnoflorine	C, MG
14 (M6)	3.87	C₂₁H₁₈O₁₁	445.077 0	-0.2	[M-H]^-	445.077 0, 269.045 2, 225.055 6	Emodin-O-glucuronide	C, MG
15 (M7)	4.03	C₂₁H₁₆O₁₂	459.056 3	-0.2	[M-H]^-	459.056 3, 283.024 3, 239.035 9, 211.040 8	Rhein-O-glucuronide	C, MG
16^a	4.56	C₂₁H₂₀O₁₀	431.098 5	+1.6	[M-H]^-	431.098 5, 269.046 0, 241.049 2	Emodin 8-O-β-D-glc	C, MG
17 (M8)	4.68	C₂₅H₂₆NO₁₀	500.157 7	+4.0	[M]⁺	500.157 7, 324.154 9, 309.079 0	Demethyleneberberine-O- glucuronide	C, MG
18 (M9)	4.70	C₂₀H₁₈NO₇S	416.148 9	-3.6	[M]⁺	416.148 9, 336.123 8	Berberine-O-sulfate	C, MG
19^a	4.78	C₂₁H₁₈O₁₁	445.077 7	+1.3	[M-H]^-	445.077 7, 283.025 3, 239.035 3, 211.040 0	Rhein 1-O-β-D-glc	C, MG
20 (M10)	4.90	C₂₀H₁₈NO₅	352.116 5	-5.7	[M]⁺	352.116 5, 336.127 3, 320.097 3	Hydroxylated berberine	MG
21 (M11)	4.96	C₁₅H₈O₉S	362.984 3	+8.8	[M-H]^-	362.984 3, 283.025 5, 239.035 7, 211.040 0	Rhein-O-sulfate	C, MG
22 (M12)	4.98	C₂₆H₂₆NO₁₀	512.155 7	0.0	[M]⁺	512.155 7, 336.123 0, 321.098 0	Berberine-O-glucuronide	C, MG
23^a	5.15	C₂₁H₂₀O₁₂	463.087 5	-0.4	[M-H]^-	463.087 5, 301.003 8	Isoquercitrin	C
24 (M13)	5.52	C₁₅H₈O₉S	362.984 3	+8.8	[M-H]^-	362.984 3, 283.025 3, 239.035 5	Rhein-O-sulfate	MG
25 (M14)	5.75	C₂₅H₂₄NO₁₀	498.140 4	+0.8	[M]⁺	498.140 4, 322.109 8, 307.085 4	Berberrubinen-O-glucuronide	C, MG
26 (M15)	5.85	C₂₁H₁₈O₁₁	445.077 0	-0.2	[M-H]^-	445.077 0, 269.045 4, 225.054 9	Emodin-O-glucuronide	C, MG
27 (M16)	6.01	C₂₇H₂₉NO₁₀	528.188 0	+1.9	[M]⁺	528.188 0, 352.123 0, 337.121 3	Palmatine-O-glucuronide	C, MG
28 (M17)	6.19	C₁₉H₁₆NSO₇	402.066 1	+3.5	[M]⁺	402.066 1, 322.125 1, 307.085 0	Berberrubinen-O-sulfate	C, MG
29 (M18)	6.55	C₂₀H₂₀NO₅	354.135 0	+2.5	[M+H]⁺	354.135 0, 338.139 9	Hydroxylated jateorhizine	C, MG
30^a	6.86	C₂₁H₂₀O₁₁	447.092 7	-2.9	[M-H]^-	447.092 0, 357.091 7	Homoorientin	C, MG
31^a	7.15	C₂₀H₂₀NO₄	338.140 8	+4.7	[M]⁺	338.140 8, 322.099 7, 308.130 9	Jatrorrhizine	C, MG
32 (M19)	7.35	C₁₉H₁₈NO₄	324.120 6	-9.3	[M]⁺	324.120 6, 294.114 6	Demethyleneberberine	C, MG
33 (M20)	7.47	C₁₉H₁₆NO₄	322.109 1	+3.7	[M]⁺	322.109 1, 307.063 8	Berberrubinen	C, MG
34^{a, b}	7.69	C₄₇H₈₀O₁₈	931.529 6	+3.2	[M-H]^-	977.540 0, 931.529 6, 799.485 4, 637.431 2, 475.378 4	Notoginsenoside R₁	C, MG
35 (M21)	7.81	C₁₅H₁₀O₇S	333.0067	-0.6	[M-H]^-	333.006 7, 253.050 5, 225.056 1	Chrysophanol-O-sulfate	C, MG
36 (M22)	8.17	C₁₅H₁₀O₈S	349.001 8	0.0	[M-H]^-	349.001 8, 269.045 9, 241.044 2	Emodin-O-sulfate	C, MG
37 (M23)	8.25	C₁₅H₁₀O₇S	333.007 3	+1.2	[M-H]^-	333.007 3, 253.050 4, 225.056 0	Chrysophanol-O-sulfate	C, MG
38^{a, b}	8.36	C₂₀H₁₈NO₄	336.123 0	-1.8	[M]⁺	336.123 0, 321.098 4, 320.091 6, 306.076 3, 292.096 7, 278.081 4	Berberine	C, MG
39^a	8.39	C₄₂H₇₂O₁₄	799.486 2	+2.3	[M-H]^-	845.492 0, 799.486 2, 637.432 2, 475.378 6	Ginsenoside Rg₁	C, MG
40^a	8.48	C₂₁H₂₂NO₄	352.154 0	-2.6	[M]⁺	352.154 0, 337.143 1, 336.098 4	Palmatine	C, MG
41 (M24)	8.55	C₂₁H₁₈O₁₀	429.083 8	+3.7	[M-H]^-	429.083 8, 253.064 3, 225.065 5	Chrysophanol-O-glucuronide	C, MG
42 (M25)	8.78	C₁₅H₁₀O₈S	349.001 6	+0.6	[M-H]^-	349.001 6, 269.045 9, 225.045 9	Emodin-O-sulfate	C, MG
43 (M26)	9.15	C₂₆H₂₈NO₁₁	530.168 1	+3.6	[M]⁺	530.168 1, 338.104 3	Hydroxylated Jateorhizine- O-glucuronide	C, MG
44^a	9.23	C₁₅H₁₀O₆	285.039 0	-3.2	[M-H]^-	285.039 0, 229.045 0, 169.014 5	Kaempferol	C, MG
45^{a, b}	9.46	C₁₅H₁₀O₇	301.036 0	+4.0	[M-H]^-	301.036 0, 151.045 2, 107.015 2	Quercetin	C, MG
46^a	9.61	C₂₁H₂₂O₉	417.119 5	+2.2	[M-H]^-	417.119 5, 255.066 6, 227.071 8	Aloin	C, MG
47 (M27)	10.01	C₂₁H₁₈O₁₁	445.077 0	-0.2	[M-H]^-	445.077 0, 269.045 3, 241.055 0	Emodin-O-glucuronide	C, MG
48 (M28)	10.37	C₂₁H₁₈O₁₀	429.081 5	-1.6	[M-H]^-	429.081 5, 253.064 3, 225.055 5	Chrysophanol-O-glucuronide	C, MG
49^a	10.56	C₁₅H₁₂O₅	271.061 5	+3.3	[M-H]^-	271.061 5, 151.066 6	Naringenin	C, MG
50 (M29)	10.65	C₁₅H₁₀O₇S	333.007 2	-0.9	[M-H]^-	333.007 2, 253.050 1, 225.056 6	Chrysophanol-O-sulfate	C, MG
51 (M30)	10.80	C₁₆H₉O₉S	376.997 3	+1.1	[M-H]^-	376.997 3, 297.040 3	6-Methyl-rhein-O-sulfate	C, MG
52^a	11.76	C₂₂H₂₂O₁₀	445.113 8	+0.7	[M-H]^-	445.113 8, 283.061 7, 239.043 3	Physcion 8-O-β-D-glc	C, MG
53 (M31)	11.94	C₂₁H₁₆O₁₂	459.056 3	-0.2	[M-H]^-	459.056 3, 283.024 2, 211.041 0	Rhein-O-glucuronide	C, MG
54^a	12.53	C₅₄H₉₂O₂₃	1 107.605 0	+8.7	[M-H]^-	1 153.601 2, 1 107.605 0, 945.546 9, 783.491 7	Ginsenoside Rb1	C, MG
55^a	13.15	C₁₆H₁₀O₆	297.040 4	+1.7	[M-H]^-	297.040 4, 267.051 1, 239.055 7	6-Methyl-rhein	C, MG
56^a	13.36	C₃₆H₆₂O₉	637.434 1	+3.9	[M-H]^-	683.439 1, 637.434 1	Ginsenoside Rh₁	C, MG
57^a	13.56	C₂₄H₂₄O₁₁	487.125 2	+2.5	[M-H]^-	487.125 2, 283.061 5, 253.051 0	Physcion 8-O-β-D-(6" -O- acetyl)-glc	C, MG
58 (M32)	13.98	C₁₅H₁₀O₈S	349.002 9	+3.2	[M-H]^-	349.002 9, 269.045 3, 241.045 5	Emodin-O-sulfate	C, MG
59^a	14.13	C₃₀H₄₈O₆	505.352 7	-0.4	[M+H]⁺	505.352 7, 487.349 0, 451.349 7	16-Oxo-alisol-A	C, MG
60^{a, b}	14.52	C₁₅H₁₀O₅	271.060 7	+0.7	[M+H]⁺	271.060 7, 241.025 8	Aloe-emodin	C, MG
61^{a, b}	15.30	C₁₅H₈O₆	283.024 9	+2.1	[M-H]^-	283.024 9, 239.034 9, 211.040 8, 183.035 6	Rhein	C, MG
62^a	16.70	C₂₀H₁₈NO₅	351.119 2	+2.0	[M]⁺	352.119 2, 337.094 8	8-Oxobererine	C, MG
63^a	17.05	C₃₀H₄₆O₅	487.341 0	-2.7	[M+H]⁺	487.341 0, 469.326 5, 451.321 5	Alisol C	C, MG
64^{a, b}	17.25	C₁₆H₁₂O₆	299.055 5	-0.3	[M-H]^-	299.055 5, 284.049 2	Kaempferide	C, MG
65^a	17.46	C₁₉H₁₆O₄	307.097 3	+1.0	[M-H]^-	307.097 3, 187.040 6	Bisdemethoxy curcumin	C, MG
66^a	17.93	C₂₀H₁₈O₅	337.107 7	+0.3	[M-H]^-	337.107 7, 217.050 8	Demethoxy curcumin	C, MG
67^{a, b}	18.38	C₂₁H₂₀O₆	367.119 1	+2.5	[M-H]^-	367.119 1, 217.052 4	Curcumin	C, MG
68^{a, b}	19.25	C₁₅H₁₀O₅	269.045 4	+1.5	[M-H]^-	269.045 4, 241.050 5, 225.055 7	Emodin	C, MG
69^a	19.71	C₃₀H₄₈O₅	511.339 3	-1.2	[M+Na]⁺	511.339 3, 471.269 6, 383.268 7	Alisol F	C, MG
70^a	20.04	C₃₂H₄₆O₅	511.342 8	+1.0	[M+H]⁺	511.342 8, 467.315 7	Alisol L 23-acetate	C, MG
71^{a, b}	21.71	C₁₅H₁₀O₄	253.051 5	+5.5	[M-H]^-	253.051 5, 225.053 5	Chrysophanol	C, MG
72^a	22.90	C₃₂H₅₂O₆	555.365 9	-0.5	[M+Na]⁺	555.365 9, 515.352 6, 383.279 9	Alisol A 23-acetate	C, MG
73^a	23.54	C₃₀H₅₀O₅	491.373 1	-1.0	[M+H]⁺	491.373 1, 473.353 7, 455.352 3	Alisol A	C, MG
74^a	23.79	C₃₀H₄₈O₄	473.363 5	+0.8	[M+H]⁺	473.363 5, 455.353 5, 437.352 8	Alisol B	C, MG
75^a	23.93	C₃₀H₄₄O₄	469.330 0	-3.8	[M+H]⁺	469.330 0, 451.318 8, 433.312 3	Alisol L	C, MG
76^a	25.28	C₃₂H₅₂O₆	555.364 8	-2.5	[M+Na]⁺	555.364 8, 515.341 5, 383.278 7	Alisol A 24-acetate	C, MG
77^{a, b}	25.32	C₃₂H₅₀O₅	515.373 0	-1.2	[M+H]⁺	515.373 0, 497.362 1, 419.329 3	Alisol B 23-acetate	C, MG

Peak ID

t_R/min

Formula

Measured (m/z)

Error (ppm)

Ion mode

MS and MS^E data (m/z)

Xenobiotics or metabolites

Source

1^a

0.95

C₁₃H₁₆O₁₀

331.066 5

-0.6

[M-H]^-

331.066 5, 169.014 1

Gallic acid-O-glc

2^a

1.01

C₇H₆O₅

169.013 6

-0.6

[M-H]^-

169.013 6, 125.024 4

Gallic acid

C, MG

3 (M1)

1.37

C₂₆H₃₁NO₁₀

518.202 8

+0.4

[M+H]⁺

518.202 8, 342.169 2, 192.101 3

Magnoflorine-O-
glucuronidation

C, MG

4 (M2)

1.58

C₁₉H₂₂NO₄

328.158 1

+9.8

[M]⁺

283.095 4, 265.089 8, 237.091 5

Desmethyl magnoflorine

5^a

1.59

C₉H₁₀O₅

197.045 3

+1.5

[M-H]^-

197.045 3, 124.045 4

Progallin A

C, MG

6 (M3)

1.68

C₂₀H₂₃NO₅

358.165 0

-1.1

[M]⁺

358.165 0, 281.077 8, 253.087 7

Hydroxylated magnoflorine

7^a

1.70

C₂₁H₂₄O₁₁

451.124 3

+0.7

[M-H]^-

451.124 3, 289.082 1

(+)-Catechin-5-O-β-D-glc

8^a

2.41

C₁₆H₁₈O₉

353.087 3

0.0

[M-H]^-

353.087 3, 191.089 9

Chlorogenic acid

C, MG

9^a

2.57

C₁₅H₁₄O₆

289.071 0

-0.7

[M-H]^-

289.071 0, 245.082 1

(+)-Catechin

C, MG

10^a

2.78

C₂₀H₂₃NO₄

342.171 5

+2.9

[M+H]⁺

342.171 5, 296.113 0

Isocorydine

C, MG

11 (M4)

2.98

C₂₆H₂₈NO₁₀

514.172 3

+1.9

[M]⁺

514.172 3, 338.137 7, 323.114 4

Jateorhizine-O-glucuronide

C, MG

12 (M5)

3.09

C₂₆H₂₆NO₁₀

512.155 7

0.0

[M]⁺

512.155 7, 336.123 5, 321.098 2

Berberine-O-glucuronide

13^a

3.20

C₂₀H₂₄NO₄

342.170 0

-1.5

[M]⁺

342.170 0, 297.112 2, 282.087 5, 265.086 0

Magnoflorine

C, MG

14 (M6)

3.87

C₂₁H₁₈O₁₁

445.077 0

-0.2

[M-H]^-

445.077 0, 269.045 2, 225.055 6

Emodin-O-glucuronide

C, MG

15 (M7)

4.03

C₂₁H₁₆O₁₂

459.056 3

-0.2

[M-H]^-

459.056 3, 283.024 3, 239.035 9, 211.040 8

Rhein-O-glucuronide

C, MG

16^a

4.56

C₂₁H₂₀O₁₀

431.098 5

+1.6

[M-H]^-

431.098 5, 269.046 0, 241.049 2

Emodin 8-O-β-D-glc

C, MG

17 (M8)

4.68

C₂₅H₂₆NO₁₀

500.157 7

+4.0

[M]⁺

500.157 7, 324.154 9, 309.079 0

Demethyleneberberine-O-
glucuronide

C, MG

18 (M9)

4.70

C₂₀H₁₈NO₇S

416.148 9

-3.6

[M]⁺

416.148 9, 336.123 8

Berberine-O-sulfate

C, MG

19^a

4.78

C₂₁H₁₈O₁₁

445.077 7

+1.3

[M-H]^-

445.077 7, 283.025 3, 239.035 3, 211.040 0

Rhein 1-O-β-D-glc

C, MG

20 (M10)

4.90

C₂₀H₁₈NO₅

352.116 5

-5.7

[M]⁺

352.116 5, 336.127 3, 320.097 3

Hydroxylated berberine

21 (M11)

4.96

C₁₅H₈O₉S

362.984 3

+8.8

[M-H]^-

362.984 3, 283.025 5, 239.035 7, 211.040 0

Rhein-O-sulfate

C, MG

22 (M12)

4.98

C₂₆H₂₆NO₁₀

512.155 7

0.0

[M]⁺

512.155 7, 336.123 0, 321.098 0

Berberine-O-glucuronide

C, MG

23^a

5.15

C₂₁H₂₀O₁₂

463.087 5

-0.4

[M-H]^-

463.087 5, 301.003 8

Isoquercitrin

24 (M13)

5.52

C₁₅H₈O₉S

362.984 3

+8.8

[M-H]^-

362.984 3, 283.025 3, 239.035 5

Rhein-O-sulfate

25 (M14)

5.75

C₂₅H₂₄NO₁₀

498.140 4

+0.8

[M]⁺

498.140 4, 322.109 8, 307.085 4

Berberrubinen-O-glucuronide

C, MG

26 (M15)

5.85

C₂₁H₁₈O₁₁

445.077 0

-0.2

[M-H]^-

445.077 0, 269.045 4, 225.054 9

Emodin-O-glucuronide

C, MG

27 (M16)

6.01

C₂₇H₂₉NO₁₀

528.188 0

+1.9

[M]⁺

528.188 0, 352.123 0, 337.121 3

Palmatine-O-glucuronide

C, MG

28 (M17)

6.19

C₁₉H₁₆NSO₇

402.066 1

+3.5

[M]⁺

402.066 1, 322.125 1, 307.085 0

Berberrubinen-O-sulfate

C, MG

29 (M18)

6.55

C₂₀H₂₀NO₅

354.135 0

+2.5

[M+H]⁺

354.135 0, 338.139 9

Hydroxylated jateorhizine

C, MG

30^a

6.86

C₂₁H₂₀O₁₁

447.092 7

-2.9

[M-H]^-

447.092 0, 357.091 7

Homoorientin

C, MG

31^a

7.15

C₂₀H₂₀NO₄

338.140 8

+4.7

[M]⁺

338.140 8, 322.099 7, 308.130 9

Jatrorrhizine

C, MG

32 (M19)

7.35

C₁₉H₁₈NO₄

324.120 6

-9.3

[M]⁺

324.120 6, 294.114 6

Demethyleneberberine

C, MG

33 (M20)

7.47

C₁₉H₁₆NO₄

322.109 1

+3.7

[M]⁺

322.109 1, 307.063 8

Berberrubinen

C, MG

34^{a, b}

7.69

C₄₇H₈₀O₁₈

931.529 6

+3.2

[M-H]^-

977.540 0, 931.529 6, 799.485 4, 637.431 2, 475.378 4

Notoginsenoside R₁

C, MG

35 (M21)

7.81

C₁₅H₁₀O₇S

333.0067

-0.6

[M-H]^-

333.006 7, 253.050 5, 225.056 1

Chrysophanol-O-sulfate

C, MG

36 (M22)

8.17

C₁₅H₁₀O₈S

349.001 8

0.0

[M-H]^-

349.001 8, 269.045 9, 241.044 2

Emodin-O-sulfate

C, MG

37 (M23)

8.25

C₁₅H₁₀O₇S

333.007 3

+1.2

[M-H]^-

333.007 3, 253.050 4, 225.056 0

Chrysophanol-O-sulfate

C, MG

38^{a, b}

8.36

C₂₀H₁₈NO₄

336.123 0

-1.8

[M]⁺

336.123 0, 321.098 4, 320.091 6, 306.076 3, 292.096 7, 278.081 4

Berberine

C, MG

39^a

8.39

C₄₂H₇₂O₁₄

799.486 2

+2.3

[M-H]^-

845.492 0, 799.486 2, 637.432 2, 475.378 6

Ginsenoside Rg₁

C, MG

40^a

8.48

C₂₁H₂₂NO₄

352.154 0

-2.6

[M]⁺

352.154 0, 337.143 1, 336.098 4

Palmatine

C, MG

41 (M24)

8.55

C₂₁H₁₈O₁₀

429.083 8

+3.7

[M-H]^-

429.083 8, 253.064 3, 225.065 5

Chrysophanol-O-glucuronide

C, MG

42 (M25)

8.78

C₁₅H₁₀O₈S

349.001 6

+0.6

[M-H]^-

349.001 6, 269.045 9, 225.045 9

Emodin-O-sulfate

C, MG

43 (M26)

9.15

C₂₆H₂₈NO₁₁

530.168 1

+3.6

[M]⁺

530.168 1, 338.104 3

Hydroxylated Jateorhizine-
O-glucuronide

C, MG

44^a

9.23

C₁₅H₁₀O₆

285.039 0

-3.2

[M-H]^-

285.039 0, 229.045 0, 169.014 5

Kaempferol

C, MG

45^{a, b}

9.46

C₁₅H₁₀O₇

301.036 0

+4.0

[M-H]^-

301.036 0, 151.045 2, 107.015 2

Quercetin

C, MG

46^a

9.61

C₂₁H₂₂O₉

417.119 5

+2.2

[M-H]^-

417.119 5, 255.066 6, 227.071 8

Aloin

C, MG

47 (M27)

10.01

C₂₁H₁₈O₁₁

445.077 0

-0.2

[M-H]^-

445.077 0, 269.045 3, 241.055 0

Emodin-O-glucuronide

C, MG

48 (M28)

10.37

C₂₁H₁₈O₁₀

429.081 5

-1.6

[M-H]^-

429.081 5, 253.064 3, 225.055 5

Chrysophanol-O-glucuronide

C, MG

49^a

10.56

C₁₅H₁₂O₅

271.061 5

+3.3

[M-H]^-

271.061 5, 151.066 6

Naringenin

C, MG

50 (M29)

10.65

C₁₅H₁₀O₇S

333.007 2

-0.9

[M-H]^-

333.007 2, 253.050 1, 225.056 6

Chrysophanol-O-sulfate

C, MG

51 (M30)

10.80

C₁₆H₉O₉S

376.997 3

+1.1

[M-H]^-

376.997 3, 297.040 3

6-Methyl-rhein-O-sulfate

C, MG

52^a

11.76

C₂₂H₂₂O₁₀

445.113 8

+0.7

[M-H]^-

445.113 8, 283.061 7, 239.043 3

Physcion 8-O-β-D-glc

C, MG

53 (M31)

11.94

C₂₁H₁₆O₁₂

459.056 3

-0.2

[M-H]^-

459.056 3, 283.024 2, 211.041 0

Rhein-O-glucuronide

C, MG

54^a

12.53

C₅₄H₉₂O₂₃

1 107.605 0

+8.7

[M-H]^-

1 153.601 2, 1 107.605 0, 945.546 9, 783.491 7

Ginsenoside Rb1

C, MG

55^a

13.15

C₁₆H₁₀O₆

297.040 4

+1.7

[M-H]^-

297.040 4, 267.051 1, 239.055 7

6-Methyl-rhein

C, MG

56^a

13.36

C₃₆H₆₂O₉

637.434 1

+3.9

[M-H]^-

683.439 1, 637.434 1

Ginsenoside Rh₁

C, MG

57^a

13.56

C₂₄H₂₄O₁₁

487.125 2

+2.5

[M-H]^-

487.125 2, 283.061 5, 253.051 0

Physcion 8-O-β-D-(6" -O-
acetyl)-glc

C, MG

58 (M32)

13.98

C₁₅H₁₀O₈S

349.002 9

+3.2

[M-H]^-

349.002 9, 269.045 3, 241.045 5

Emodin-O-sulfate

C, MG

59^a

14.13

C₃₀H₄₈O₆

505.352 7

-0.4

[M+H]⁺

505.352 7, 487.349 0, 451.349 7

16-Oxo-alisol-A

C, MG

60^{a, b}

14.52

C₁₅H₁₀O₅

271.060 7

+0.7

[M+H]⁺

271.060 7, 241.025 8

Aloe-emodin

C, MG

61^{a, b}

15.30

C₁₅H₈O₆

283.024 9

+2.1

[M-H]^-

283.024 9, 239.034 9, 211.040 8, 183.035 6

Rhein

C, MG

62^a

16.70

C₂₀H₁₈NO₅

351.119 2

+2.0

[M]⁺

352.119 2, 337.094 8

8-Oxobererine

C, MG

63^a

17.05

C₃₀H₄₆O₅

487.341 0

-2.7

[M+H]⁺

487.341 0, 469.326 5, 451.321 5

Alisol C

C, MG

64^{a, b}

17.25

C₁₆H₁₂O₆

299.055 5

-0.3

[M-H]^-

299.055 5, 284.049 2

Kaempferide

C, MG

65^a

17.46

C₁₉H₁₆O₄

307.097 3

+1.0

[M-H]^-

307.097 3, 187.040 6

Bisdemethoxy curcumin

C, MG

66^a

17.93

C₂₀H₁₈O₅

337.107 7

+0.3

[M-H]^-

337.107 7, 217.050 8

Demethoxy curcumin

C, MG

67^{a, b}

18.38

C₂₁H₂₀O₆

367.119 1

+2.5

[M-H]^-

367.119 1, 217.052 4

Curcumin

C, MG

68^{a, b}

19.25

C₁₅H₁₀O₅

269.045 4

+1.5

[M-H]^-

269.045 4, 241.050 5, 225.055 7

Emodin

C, MG

69^a

19.71

C₃₀H₄₈O₅

511.339 3

-1.2

[M+Na]⁺

511.339 3, 471.269 6, 383.268 7

Alisol F

C, MG

70^a

20.04

C₃₂H₄₆O₅

511.342 8

+1.0

[M+H]⁺

511.342 8, 467.315 7

Alisol L 23-acetate

C, MG

71^{a, b}

21.71

C₁₅H₁₀O₄

253.051 5

+5.5

[M-H]^-

253.051 5, 225.053 5

Chrysophanol

C, MG

72^a

22.90

C₃₂H₅₂O₆

555.365 9

-0.5

[M+Na]⁺

555.365 9, 515.352 6, 383.279 9

Alisol A 23-acetate

C, MG

73^a

23.54

C₃₀H₅₀O₅

491.373 1

-1.0

[M+H]⁺

491.373 1, 473.353 7, 455.352 3

Alisol A

C, MG

74^a

23.79

C₃₀H₄₈O₄

473.363 5

+0.8

[M+H]⁺

473.363 5, 455.353 5, 437.352 8

Alisol B

C, MG

75^a

23.93

C₃₀H₄₄O₄

469.330 0

-3.8

[M+H]⁺

469.330 0, 451.318 8, 433.312 3

Alisol L

C, MG

76^a

25.28

C₃₂H₅₂O₆

555.364 8

-2.5

[M+Na]⁺

555.364 8, 515.341 5, 383.278 7

Alisol A 24-acetate

C, MG

77^{a, b}

25.32

C₃₂H₅₀O₅

515.373 0

-1.2

[M+H]⁺

515.373 0, 497.362 1, 419.329 3

Alisol B 23-acetate

C, MG