药学学报

No.	Compound	t_R /min	Formula	Experimental (m/z)	Mode	Mass error	Fragment	Type	FC
1	Rhamnocitrin	23.14	C₁₆H₁₂O₆	301.070 7	[M+H]⁺	0.1	216.045 0, 203.068 3, 188.044 7, 175.075 4, 171.043 8, 167.037 9, 139.030 6	A	3.75
2	Pectolinarigenin	20.77	C₁₇H₁₄O₆	313.071 8	[M-H]^-	0.1	297.041 5, 283.026 1, 269.047 1, 225.029 6, 227.035 3, 183.044 6, 171.044 6	A	1.57
3	Rhamnocitrin-Hex	14.79	C₂₂H₂₂O₁₁	461.109 3	[M+H]^-	0.8	299.056 4, 284.032 8, 255.030 6, 135.008 9, 503.119 1	A	2.45
4	Trihydroxy-methoxypterocarpan-Glc-Mal	19.44	C₂₅H₂₆O₁₄	551.137 3	[M+H]⁺	-4.1	439.357 4, 302.072 3, 204.086 0, 186.073 2, 138.058 8	B	1.79
5	Astrapterocarpan-3-O-Glc-Mal-Api	20.54	C₃₁H₃₆O₁₇	698.229 0	[M+NH₄]⁺	-0.2	549.160 0, 371.097 0, 325.108 9, 301.107 5, 191.069 3, 167.070 2	B	1.62
6	Astrapterocarpan-O-Glc-Mal isomer 1	20.54	C₂₆H₂₈O₁₃	549.160 3	[M+H]⁺	0.0	301.107 2, 191.070 0, 167.070 2, 123.044 4	B	1.69
7	Tetrahydroxy-pterocarpan-Glc-Mal	16.54	C₂₄H₂₄O₁₄	537.123 9	[M+H]⁺	0.1	289.069 6, 281.049 3, 289.082 6, 243.058 7, 137.022 1, 127.038 1, 73.045 7	B	1.56
8	Astrapterocarpan-3-O-Glc-Ace	20.80	C₂₅H₂₈O₁₁	505.169 7	[M+H]⁺	-2.4	181.047 5, 167.071 3, 191.367 3, 255.090 2, 301.069 9, 445.154 7	B	1.28
9	Isomucronulatol-7-O-Glc-Mal-Glc-Mal	21.26	C₃₅H₄₂O₂₁	816.255 7	[M+NH₄]⁺	-1.5	257.044 1, 239.006 7, 165.057 1, 150.033 4, 149.032 2, 121.028 6, 109.030 1, 91.063 3	C	1.93
10	Isomucronulatol-Glc-Glc	17.98	C₂₉H₃₈O₁₅	644.254 8	[M+NH₄]⁺	-0.2	465.175 3, 429.170 9, 369.169 6, 369.134 0, 345.145 0, 303.122 8, 167.070 0, 123.044 3	C	2.63
11	Trihydroxy-dimethoxyisoflavan-Glc-Glc	13.80	C₂₉H₃₈O₁₆	660.249 6	[M+NH₄]⁺	-0.4	660.248 4, 557.295 5, 445.143 5, 427.165 0, 343.112 6, 319.120 2, 235.069 0, 183.070 0, 123.042 5	C	2.55
12	Isomucronulatol-7-O-Glc-Glc-Mal	18.73	C₃₂H₄₀O₁₈	730.255 3	[M+NH₄]⁺	-0.4	429.160 5, 393.127 0, 369.139 4, 303.121 5, 249.076 5, 239.067 1, 193.085 5, 189.055 5, 181.085 8	C	2.37
13	Isomucronulatol-7-O-Glc-Mal-Api	18.21	C₃₁H₃₈O₁₇	700.244 7	[M+NH₄]⁺	-0.6	339.121 6, 339.134 5, 297.108 0, 273.111 1, 177.052 4, 153.051 1, 133.066 3	C	1.33
14	Isomucronulatol	20.32	C₁₇H₁₈O₅	301.108 2	[M-H]^-	0.1	301.111 3, 286.087 3, 271.062 6, 256.038 7, 253.051 8, 149.025 0, 135.045 9	C	2.34
15	Isomucronulatol-7, 2′-di-O-Glc	17.56	C₂₉H₃₈O₁₅	625.213 0	[M-H]^-	-1.2	463.159 7, 301.107 9, 301.107 3, 135.044 9, 286.084 4	C	2.51
16	Trihydroxy-dimethoxyisoflavan-Glc	19.48	C₂₂H₂₆O₁₀	449.144 0	[M-H]^-	-2.9	287.093 8, 272.071 7, 257.044 1, 239.006 7, 165.057 1, 150.033 4, 149.032 2, 121.028 6, 109.030 1	C	3.71
17	Trihydroxy-methoxyisoflavan	20.69	C₁₆H₁₆O₅	287.091 9	[M-H]^-	-0.1	287.094 6, 272.069 3, 257.045 4, 254.045 4, 254.056 9, 165.056 4, 163.040 8, 149.023 8, 150.032 2	C	1.87
18	Isomucronulatol-O-Glc-Ace isomer	20.73	C₂₅H₃₀O₁₁	505.171 2	[M-H]^-	-0.8	301.108 5, 286.085 0, 271.061 5, 179.071 9, 164.048 5, 135.045 6, 121.029 7, 122.037 7	C	1.44
19	Isomucronulatol-7-O-Glc	20.61	C₂₃H₂₈O₁₀	465.175 1	[M+H]⁺	-1.0	303.123 6, 165.053 2, 167.070 1, 167.085 9, 123.043 3, 167.085 9, 83.012 7	C	3.08
20	Isomucronulatol isomer	23.56	C₁₇H₁₈O₅	303.126 6	[M+H]⁺	-0.4	193.084 9, 167.069 1, 161.058 7, 133.064 0, 123.043 0, 118.040 5, 106.040 2	C	2.57
21	Isomucronulatol-O-Glc isomer	20.61	C₂₃H₂₈O₁₀	465.175 0	[M+H]⁺	-0.2	429.150 1, 369.139 3, 339.124 0, 303.125 0, 303.138 8, 249.080 9, 189.053 6, 167.069 2, 123.044 5	C	3.08
22	Calycosin-7-O-Glc-6″-O-Mal isomer	18.13	C₂₅H₂₄O₁₃	533.128 5	[M+H]⁺	-0.2	285.075 3, 270.052 5, 253.049 9, 225.055 2, 137.023 4	D	1.45
23	Calycosin-7-O-Glc-6″-O-Mal	18.44	C₂₅H₂₄O₁₃	533.129 3	[M+H]⁺	-1.0	285.075 2, 285.092 8, 270.052 2, 253.050 2, 225.055 0, 137.023 4	D	1.38
24	Odoratin-7-O-Glc-6″-O-Mal	18.53	C₂₆H₂₆O₁₄	563.139 6	[M+H]⁺	0.1	315.086 5, 300.064 0, 283.060 1, 255.066 2	D	2.23
25	Pratensein-7-O-Glc	14.79	C₂₂H₂₂O₁₁	461.109 3	[M-H]^-	0.8	299.057 1, 284.033 6, 255.031 4, 240.045 5	D	2.46
26	Isoafrormosin	19.29	C₁₇H₁₄O₅	297.076 4	[M-H]^-	-1.6	297.076 4, 281.044 2, 267.029 9, 253.050 5, 239.034 5	D	1.42
27	Pratensein-Glc	16.29	C₂₄H₂₄O₁₂	503.119 6	[M-H]^-	0.3	443.091 6, 299.057 6, 256.038 1	D	1.86
28	3′-Methoxy-5′-hydroxyisoflavone-7-O-Glc	17.79	C₂₂H₂₂O₁₀	445.111 8	[M-H]^-	-4.9	297.042 6, 283.061 8, 268.038 3, 239.035 6	D	2.11
29	Pratensein-7-O-Glc isomer	18.58	C₂₂H₂₂O₁₁	463.123 5	[M+H]⁺	0.3	301.071 0, 286.040 5, 269.045 4, 241.045 2, 153.016 5	D	1.50
30	Calycosin-7-O-Glc	16.78	C₂₂H₂₂O₁₀	447.128 5	[M+H]⁺	-1.7	286.384 0, 285.075 1, 270.051 6, 253.050 1, 225.055 4, 214.061 1, 137.021 7	D	1.79
31	Ononin	19.71	C₂₂H₂₂O₉	431.133 1	[M+H]⁺	-1.3	269.080 4, 254.057 4, 213.092 0, 197.061 8, 136.014 5	D	1.62
32	Odoratin-7-O-Glc	17.05	C₂₃H₂₄O₁₁	477.139 1	[M+H]⁺	0.0	315.086 7, 300.064 8, 272.070 3, 255.094 1, 200.046 3, 175.035 4	D	2.73
33	Pratensein isomer 1	22.29	C₁₆H₁₂O₆	301.071 3	[M+H]⁺	2.2	151.078 8, 153.018 5, 163.038 4, 229.045 3, 241.046 5, 301.068 6, 301.097 9	D	1.99
34	Afrormosin-7-O-Glc-6″-O-Mal	20.65	C₂₆H₂₆O₁₃	547.144 7	[M+H]⁺	0.1	299.091 5, 284.068 5, 243.111 7, 183.084 9	D	2.03
35	Pratensein	23.15	C₁₆H₁₂O₆	301.070 7	[M+H]⁺	-0.5	123.044 5, 171.039 3, 167.060 1, 167.032 9, 188.044 5, 203.068 1, 217.054 8, 231.065 2, 301.070 8	D	3.40
36	3′, 5′-Dihydroxy-4′-methoxyisoflavone	23.88	C₁₇H₁₆O₄	285.112 2	[M+H]⁺	0.1	285.001 6, 243.101 2, 239.106 8, 191.070 6, 176.046 6, 137.059 6, 121.027 8	D	2.50
37	Tetrahydroxy-methoxy-isoflavone-Glc-Mal	17.98	C₂₅H₂₄O₁₅	565.118 9	[M+H]⁺	0.1	565.112 6, 302.045 8, 317.065 2, 285.041 3, 299.052 1, 191.035 5	D	1.54
38	Afrormosin-Glc-Mal isomer	19.71	C₂₆H₂₆O₁₃	547.144 7	[M+H]⁺	0.1	547.148 8, 299.092 1, 300.095 7, 284.068 0, 239.068 9, 211.074 1	D	1.44
39	Calycosin-Glc-Glc-Mal	17.99	C₃₁H₃₄O₁₈	695.181 8	[M+H]⁺	-0.3	533.135 5, 285.075 3, 270.067 9, 270.043 9, 225.053 7, 158.075 2	D	2.83
40	Pratensein-Glc-Mal-Glc-Mal	15.35	C₃₄H₃₆O₂₂	797.177 1	[M+H]⁺	0.3	547.406 1, 549.123 4, 301.070 5, 269.054 1	D	2.48
41	Guanosine	7.84	C₁₀H₁₃N₅O₅	284.099 1	[M+H]⁺	0.4	152.056 3, 135.029 7, 110.034 4	E	4.30
42	Adenosine	7.77	C₁₀H₁₃N₅O₄	268.104 6	[M+H]⁺	2.2	136.062 4, 57.035 8	E	2.53
43	Tryptophan	10.25	C₁₁H₁₂N₂O₂	203.082 8	[M-H]^-	0.8	159.095 0, 142.067 8, 116.051 7, 74.025 7	F	1.34
44	Salicylic acid	18.02	C₇H₆O₃	137.024 4	[M-H]^-	-0.7	137.027 1, 93.035 7, 93.051 0, 78.963 7, 65.039 4	G	3.31
45	Acetylastragaloside I isomer	27.12	C₄₇H₇₄O₁₇	911.498 3	[M+H]⁺	-1.7	731.428 8, 713.424 5, 695.411 0, 437.348 3, 419.335 9, 199.059 9, 157.050 9, 143.107 9, 125.098 0	H	2.83
46	Argentatin A-Glc-Ara	22.58	C₄₁H₆₆O₁₃	767.456 1	[M+H]⁺	-1.9	587.398 0, 599.388 5, 569.369 3, 473.364 2, 455.348 4, 437.342 9, 419.328 4, 253.193 1, 187.145 2	H	0.67
47	Astragaloside II-Mal	23.82	C₄₆H₇₂O₁₈	913.479 4	[M+H]⁺	0.2	599.403 4, 473.368 5, 455.353 6, 437.341 0, 419.335 8, 243.049 3, 261.061 4, 143.105 4	H	0.41
48	Astragaloside II-Mal-Ace	24, 91	C₄₈H₇₄O₁₉	955.490 7	[M+H]⁺	1.5	739.405 8, 721.399 8, 455.352 0, 437.340 8, 419.328 7, 303.070 8, 157.049 1, 143.105 5	H	0.63
49	Astragaloside I-Glc-Mal-Mal	23.37	C₅₇H₈₆O₂₇	1203.538 4	[M+H]⁺	-3.7	455.356 1, 437.344 1, 419.331 9, 401.310 7, 199.058 8, 157.049 4, 143.106 6, 97.028 3	H	0.64
50	HuangqiyeninA-Ara	22.48	C₄₁H₆₆O₁₄	783.451 2	[M+H]⁺	-1.7	453.334 5, 435.325 6, 417.313 4, 271.213 3, 143.107 9	H	0.53
51	Agroastragaloside IV	22.04	C₄₉H₈₀O₂₀	987.518 1	[M-H]^-	1.1	941.521 9	H	0.60
52	SoyasapogenolB-Glc-Rha	22.44	C₄₁H₆₆O₁₃	765.441 2	[M-H]^-	-2.4	603.394 5, 157.009 9, 113.020 5, 85.029 5	H	0.72
53	Vitamin B₂	14.66	C₁₇H₂₀N₄O₆	377.145 6	[M+H]⁺	0.0	243.087 0, 172.086 8, 57.032 9, 198.066 7, 145.076 2, 81.032 9, 57.033 1	I	1.99
54	FA 16:0	30.88	C₁₆H₃₂O₂	255.232 91	[M-H]^-	-0.5	255.232 4, 256.235 8, 257.239 1	J	1.69
55	FA 18:2	29.94	C₁₈H₃₂O₂	279.232 97	[M-H]^-	0.2	279.232 0, 280.235 5, 281.238 9	J	2.00
56	FA 18:2;O	26.43	C₁₈H₃₂O₃	295.224 7	[M-H]^-	-0.8	295.228 1, 296.231 0, 297.234 8	J	1.82
57	ST 28:3;O	24.90	C₂₈H₄₄O	419.328 92	[M+Na]⁺	0.5	419.329 5, 420.332 8, 421.336 2	K	0.74
58	DG 20:0	27.20	C₂₃H₄₄O₅	423.312 71	[M+Na]⁺	4.0	423.309 0, 331.266 2	L	2.65
59	LPG 16:0	31.98	C₂₂H₄₅O₉P	483.270 11	[M-H]^-	-0.2	483.272 5, 484.275 8, 485.279 2	M	1.97
60	LPC 16:0	26.61	C₂₄H₅₀NO₇P	496.339 48	[M+H]⁺	0.9	496.339 2, 497.342 5, 498.345 9	N	5.21
61	LPC 18:0	28.38	C₂₆H₅₄NO₇P	524.367 92	[M+H]⁺	0.4	524.371 2, 525.374 5, 526.377 9	N	2.61
62	PI 19:0	25.33	C₂₈H₅₃O₁₃P	627.317 32	[M-H]^-	2.3	627.317 3, 628.320 6, 629.324 0	O	3.89

No.	Compound	t_R /min	Formula	Experimental (m/z)	Mode	Mass error	Fragment	Type	FC
1	Rhamnocitrin	23.14	C₁₆H₁₂O₆	301.070 7	[M+H]⁺	0.1	216.045 0, 203.068 3, 188.044 7, 175.075 4, 171.043 8, 167.037 9, 139.030 6	A	3.75
2	Pectolinarigenin	20.77	C₁₇H₁₄O₆	313.071 8	[M-H]^-	0.1	297.041 5, 283.026 1, 269.047 1, 225.029 6, 227.035 3, 183.044 6, 171.044 6	A	1.57
3	Rhamnocitrin-Hex	14.79	C₂₂H₂₂O₁₁	461.109 3	[M+H]^-	0.8	299.056 4, 284.032 8, 255.030 6, 135.008 9, 503.119 1	A	2.45
4	Trihydroxy-methoxypterocarpan-Glc-Mal	19.44	C₂₅H₂₆O₁₄	551.137 3	[M+H]⁺	-4.1	439.357 4, 302.072 3, 204.086 0, 186.073 2, 138.058 8	B	1.79
5	Astrapterocarpan-3-O-Glc-Mal-Api	20.54	C₃₁H₃₆O₁₇	698.229 0	[M+NH₄]⁺	-0.2	549.160 0, 371.097 0, 325.108 9, 301.107 5, 191.069 3, 167.070 2	B	1.62
6	Astrapterocarpan-O-Glc-Mal isomer 1	20.54	C₂₆H₂₈O₁₃	549.160 3	[M+H]⁺	0.0	301.107 2, 191.070 0, 167.070 2, 123.044 4	B	1.69
7	Tetrahydroxy-pterocarpan-Glc-Mal	16.54	C₂₄H₂₄O₁₄	537.123 9	[M+H]⁺	0.1	289.069 6, 281.049 3, 289.082 6, 243.058 7, 137.022 1, 127.038 1, 73.045 7	B	1.56
8	Astrapterocarpan-3-O-Glc-Ace	20.80	C₂₅H₂₈O₁₁	505.169 7	[M+H]⁺	-2.4	181.047 5, 167.071 3, 191.367 3, 255.090 2, 301.069 9, 445.154 7	B	1.28
9	Isomucronulatol-7-O-Glc-Mal-Glc-Mal	21.26	C₃₅H₄₂O₂₁	816.255 7	[M+NH₄]⁺	-1.5	257.044 1, 239.006 7, 165.057 1, 150.033 4, 149.032 2, 121.028 6, 109.030 1, 91.063 3	C	1.93
10	Isomucronulatol-Glc-Glc	17.98	C₂₉H₃₈O₁₅	644.254 8	[M+NH₄]⁺	-0.2	465.175 3, 429.170 9, 369.169 6, 369.134 0, 345.145 0, 303.122 8, 167.070 0, 123.044 3	C	2.63
11	Trihydroxy-dimethoxyisoflavan-Glc-Glc	13.80	C₂₉H₃₈O₁₆	660.249 6	[M+NH₄]⁺	-0.4	660.248 4, 557.295 5, 445.143 5, 427.165 0, 343.112 6, 319.120 2, 235.069 0, 183.070 0, 123.042 5	C	2.55
12	Isomucronulatol-7-O-Glc-Glc-Mal	18.73	C₃₂H₄₀O₁₈	730.255 3	[M+NH₄]⁺	-0.4	429.160 5, 393.127 0, 369.139 4, 303.121 5, 249.076 5, 239.067 1, 193.085 5, 189.055 5, 181.085 8	C	2.37
13	Isomucronulatol-7-O-Glc-Mal-Api	18.21	C₃₁H₃₈O₁₇	700.244 7	[M+NH₄]⁺	-0.6	339.121 6, 339.134 5, 297.108 0, 273.111 1, 177.052 4, 153.051 1, 133.066 3	C	1.33
14	Isomucronulatol	20.32	C₁₇H₁₈O₅	301.108 2	[M-H]^-	0.1	301.111 3, 286.087 3, 271.062 6, 256.038 7, 253.051 8, 149.025 0, 135.045 9	C	2.34
15	Isomucronulatol-7, 2′-di-O-Glc	17.56	C₂₉H₃₈O₁₅	625.213 0	[M-H]^-	-1.2	463.159 7, 301.107 9, 301.107 3, 135.044 9, 286.084 4	C	2.51
16	Trihydroxy-dimethoxyisoflavan-Glc	19.48	C₂₂H₂₆O₁₀	449.144 0	[M-H]^-	-2.9	287.093 8, 272.071 7, 257.044 1, 239.006 7, 165.057 1, 150.033 4, 149.032 2, 121.028 6, 109.030 1	C	3.71
17	Trihydroxy-methoxyisoflavan	20.69	C₁₆H₁₆O₅	287.091 9	[M-H]^-	-0.1	287.094 6, 272.069 3, 257.045 4, 254.045 4, 254.056 9, 165.056 4, 163.040 8, 149.023 8, 150.032 2	C	1.87
18	Isomucronulatol-O-Glc-Ace isomer	20.73	C₂₅H₃₀O₁₁	505.171 2	[M-H]^-	-0.8	301.108 5, 286.085 0, 271.061 5, 179.071 9, 164.048 5, 135.045 6, 121.029 7, 122.037 7	C	1.44
19	Isomucronulatol-7-O-Glc	20.61	C₂₃H₂₈O₁₀	465.175 1	[M+H]⁺	-1.0	303.123 6, 165.053 2, 167.070 1, 167.085 9, 123.043 3, 167.085 9, 83.012 7	C	3.08
20	Isomucronulatol isomer	23.56	C₁₇H₁₈O₅	303.126 6	[M+H]⁺	-0.4	193.084 9, 167.069 1, 161.058 7, 133.064 0, 123.043 0, 118.040 5, 106.040 2	C	2.57
21	Isomucronulatol-O-Glc isomer	20.61	C₂₃H₂₈O₁₀	465.175 0	[M+H]⁺	-0.2	429.150 1, 369.139 3, 339.124 0, 303.125 0, 303.138 8, 249.080 9, 189.053 6, 167.069 2, 123.044 5	C	3.08
22	Calycosin-7-O-Glc-6″-O-Mal isomer	18.13	C₂₅H₂₄O₁₃	533.128 5	[M+H]⁺	-0.2	285.075 3, 270.052 5, 253.049 9, 225.055 2, 137.023 4	D	1.45
23	Calycosin-7-O-Glc-6″-O-Mal	18.44	C₂₅H₂₄O₁₃	533.129 3	[M+H]⁺	-1.0	285.075 2, 285.092 8, 270.052 2, 253.050 2, 225.055 0, 137.023 4	D	1.38
24	Odoratin-7-O-Glc-6″-O-Mal	18.53	C₂₆H₂₆O₁₄	563.139 6	[M+H]⁺	0.1	315.086 5, 300.064 0, 283.060 1, 255.066 2	D	2.23
25	Pratensein-7-O-Glc	14.79	C₂₂H₂₂O₁₁	461.109 3	[M-H]^-	0.8	299.057 1, 284.033 6, 255.031 4, 240.045 5	D	2.46
26	Isoafrormosin	19.29	C₁₇H₁₄O₅	297.076 4	[M-H]^-	-1.6	297.076 4, 281.044 2, 267.029 9, 253.050 5, 239.034 5	D	1.42
27	Pratensein-Glc	16.29	C₂₄H₂₄O₁₂	503.119 6	[M-H]^-	0.3	443.091 6, 299.057 6, 256.038 1	D	1.86
28	3′-Methoxy-5′-hydroxyisoflavone-7-O-Glc	17.79	C₂₂H₂₂O₁₀	445.111 8	[M-H]^-	-4.9	297.042 6, 283.061 8, 268.038 3, 239.035 6	D	2.11
29	Pratensein-7-O-Glc isomer	18.58	C₂₂H₂₂O₁₁	463.123 5	[M+H]⁺	0.3	301.071 0, 286.040 5, 269.045 4, 241.045 2, 153.016 5	D	1.50
30	Calycosin-7-O-Glc	16.78	C₂₂H₂₂O₁₀	447.128 5	[M+H]⁺	-1.7	286.384 0, 285.075 1, 270.051 6, 253.050 1, 225.055 4, 214.061 1, 137.021 7	D	1.79
31	Ononin	19.71	C₂₂H₂₂O₉	431.133 1	[M+H]⁺	-1.3	269.080 4, 254.057 4, 213.092 0, 197.061 8, 136.014 5	D	1.62
32	Odoratin-7-O-Glc	17.05	C₂₃H₂₄O₁₁	477.139 1	[M+H]⁺	0.0	315.086 7, 300.064 8, 272.070 3, 255.094 1, 200.046 3, 175.035 4	D	2.73
33	Pratensein isomer 1	22.29	C₁₆H₁₂O₆	301.071 3	[M+H]⁺	2.2	151.078 8, 153.018 5, 163.038 4, 229.045 3, 241.046 5, 301.068 6, 301.097 9	D	1.99
34	Afrormosin-7-O-Glc-6″-O-Mal	20.65	C₂₆H₂₆O₁₃	547.144 7	[M+H]⁺	0.1	299.091 5, 284.068 5, 243.111 7, 183.084 9	D	2.03
35	Pratensein	23.15	C₁₆H₁₂O₆	301.070 7	[M+H]⁺	-0.5	123.044 5, 171.039 3, 167.060 1, 167.032 9, 188.044 5, 203.068 1, 217.054 8, 231.065 2, 301.070 8	D	3.40
36	3′, 5′-Dihydroxy-4′-methoxyisoflavone	23.88	C₁₇H₁₆O₄	285.112 2	[M+H]⁺	0.1	285.001 6, 243.101 2, 239.106 8, 191.070 6, 176.046 6, 137.059 6, 121.027 8	D	2.50
37	Tetrahydroxy-methoxy-isoflavone-Glc-Mal	17.98	C₂₅H₂₄O₁₅	565.118 9	[M+H]⁺	0.1	565.112 6, 302.045 8, 317.065 2, 285.041 3, 299.052 1, 191.035 5	D	1.54
38	Afrormosin-Glc-Mal isomer	19.71	C₂₆H₂₆O₁₃	547.144 7	[M+H]⁺	0.1	547.148 8, 299.092 1, 300.095 7, 284.068 0, 239.068 9, 211.074 1	D	1.44
39	Calycosin-Glc-Glc-Mal	17.99	C₃₁H₃₄O₁₈	695.181 8	[M+H]⁺	-0.3	533.135 5, 285.075 3, 270.067 9, 270.043 9, 225.053 7, 158.075 2	D	2.83
40	Pratensein-Glc-Mal-Glc-Mal	15.35	C₃₄H₃₆O₂₂	797.177 1	[M+H]⁺	0.3	547.406 1, 549.123 4, 301.070 5, 269.054 1	D	2.48
41	Guanosine	7.84	C₁₀H₁₃N₅O₅	284.099 1	[M+H]⁺	0.4	152.056 3, 135.029 7, 110.034 4	E	4.30
42	Adenosine	7.77	C₁₀H₁₃N₅O₄	268.104 6	[M+H]⁺	2.2	136.062 4, 57.035 8	E	2.53
43	Tryptophan	10.25	C₁₁H₁₂N₂O₂	203.082 8	[M-H]^-	0.8	159.095 0, 142.067 8, 116.051 7, 74.025 7	F	1.34
44	Salicylic acid	18.02	C₇H₆O₃	137.024 4	[M-H]^-	-0.7	137.027 1, 93.035 7, 93.051 0, 78.963 7, 65.039 4	G	3.31
45	Acetylastragaloside I isomer	27.12	C₄₇H₇₄O₁₇	911.498 3	[M+H]⁺	-1.7	731.428 8, 713.424 5, 695.411 0, 437.348 3, 419.335 9, 199.059 9, 157.050 9, 143.107 9, 125.098 0	H	2.83
46	Argentatin A-Glc-Ara	22.58	C₄₁H₆₆O₁₃	767.456 1	[M+H]⁺	-1.9	587.398 0, 599.388 5, 569.369 3, 473.364 2, 455.348 4, 437.342 9, 419.328 4, 253.193 1, 187.145 2	H	0.67
47	Astragaloside II-Mal	23.82	C₄₆H₇₂O₁₈	913.479 4	[M+H]⁺	0.2	599.403 4, 473.368 5, 455.353 6, 437.341 0, 419.335 8, 243.049 3, 261.061 4, 143.105 4	H	0.41
48	Astragaloside II-Mal-Ace	24, 91	C₄₈H₇₄O₁₉	955.490 7	[M+H]⁺	1.5	739.405 8, 721.399 8, 455.352 0, 437.340 8, 419.328 7, 303.070 8, 157.049 1, 143.105 5	H	0.63
49	Astragaloside I-Glc-Mal-Mal	23.37	C₅₇H₈₆O₂₇	1203.538 4	[M+H]⁺	-3.7	455.356 1, 437.344 1, 419.331 9, 401.310 7, 199.058 8, 157.049 4, 143.106 6, 97.028 3	H	0.64
50	HuangqiyeninA-Ara	22.48	C₄₁H₆₆O₁₄	783.451 2	[M+H]⁺	-1.7	453.334 5, 435.325 6, 417.313 4, 271.213 3, 143.107 9	H	0.53
51	Agroastragaloside IV	22.04	C₄₉H₈₀O₂₀	987.518 1	[M-H]^-	1.1	941.521 9	H	0.60
52	SoyasapogenolB-Glc-Rha	22.44	C₄₁H₆₆O₁₃	765.441 2	[M-H]^-	-2.4	603.394 5, 157.009 9, 113.020 5, 85.029 5	H	0.72
53	Vitamin B₂	14.66	C₁₇H₂₀N₄O₆	377.145 6	[M+H]⁺	0.0	243.087 0, 172.086 8, 57.032 9, 198.066 7, 145.076 2, 81.032 9, 57.033 1	I	1.99
54	FA 16:0	30.88	C₁₆H₃₂O₂	255.232 91	[M-H]^-	-0.5	255.232 4, 256.235 8, 257.239 1	J	1.69
55	FA 18:2	29.94	C₁₈H₃₂O₂	279.232 97	[M-H]^-	0.2	279.232 0, 280.235 5, 281.238 9	J	2.00
56	FA 18:2;O	26.43	C₁₈H₃₂O₃	295.224 7	[M-H]^-	-0.8	295.228 1, 296.231 0, 297.234 8	J	1.82
57	ST 28:3;O	24.90	C₂₈H₄₄O	419.328 92	[M+Na]⁺	0.5	419.329 5, 420.332 8, 421.336 2	K	0.74
58	DG 20:0	27.20	C₂₃H₄₄O₅	423.312 71	[M+Na]⁺	4.0	423.309 0, 331.266 2	L	2.65
59	LPG 16:0	31.98	C₂₂H₄₅O₉P	483.270 11	[M-H]^-	-0.2	483.272 5, 484.275 8, 485.279 2	M	1.97
60	LPC 16:0	26.61	C₂₄H₅₀NO₇P	496.339 48	[M+H]⁺	0.9	496.339 2, 497.342 5, 498.345 9	N	5.21
61	LPC 18:0	28.38	C₂₆H₅₄NO₇P	524.367 92	[M+H]⁺	0.4	524.371 2, 525.374 5, 526.377 9	N	2.61
62	PI 19:0	25.33	C₂₈H₅₃O₁₃P	627.317 32	[M-H]^-	2.3	627.317 3, 628.320 6, 629.324 0	O	3.89

基于代谢组学分析尖孢镰刀菌侵染对黄芪代谢的影响

PDF下载

王富鑫 ¹ , 宋海清 ¹ , 赵利 ¹ , 秦雪梅 ¹ , 高芬 ²^,^* , 李震宇 ¹^,^*

药学学报 | 研究论文 2023,58(7): 1971-1980

收起

药学学报 | 研究论文 2023, 58(7): 1971-1980

基于代谢组学分析尖孢镰刀菌侵染对黄芪代谢的影响

全屏

王富鑫¹, 宋海清¹, 赵利¹, 秦雪梅¹, 高芬²^,^*, 李震宇¹^,^*

作者信息

1.山西大学中医药现代研究中心, 山西太原 030006

2.山西大学应用化学研究所, 山西太原 030006

通讯作者:

*高芬, Tel: 86-351-7018390, E-mail: gaofen@sxu.edu.cn;

李震宇, Tel: 86-351-7011202, E-mail: lizhenyu@sxu.edu.cn

Analysis of Fusarium oxysporum infection on changes of Astragalus metabolism by metabolomic approach

Fu-xin WANG¹, Hai-qing SONG¹, Li ZHAO¹, Xue-mei QIN¹, Fen GAO²^,^*, Zhen-yu LI¹^,^*

Affiliations

1. Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan 030006, China

2. Institute of Applied Chemistry, Shanxi University, Taiyuan 030006, China

出版时间: 2023-07-12 doi: 10.16438/j.0513-4870.2022-1377

文章导航

摘要

收起

黄芪是中医临床常用中药材, 随着近年来种植面积的增加, 黄芪根腐病危害逐年加重, 严重影响其质量和产量。尖孢镰刀菌(Fusarium oxysporum) 是引起黄芪根腐病的主要病原菌之一。本研究采用基于UPLC-Q-TOF-MS的代谢组学技术结合多元统计分析, 解析黄芪响应尖孢镰刀菌侵染的代谢产物变化。结果表明, 黄芪接种尖孢镰刀菌后, 有62种代谢物发生显著的变化。极性代谢物包括40个黄酮、8个皂苷、2个核苷、1个维生素、1个有机酸、1个氨基酸; 脂质代谢物包括3个脂肪酸、1个甘油二酯、2个溶血磷脂酰胆碱、1个溶血磷脂甘油、1个磷脂酰肌醇、1个固醇脂。尖孢镰刀菌接种组和对照组的差异代谢物中, 黄酮类以及维生素B₂、色氨酸、水杨酸的相对含量增加, 而皂苷类的相对含量减少。相关性分析表明, 黄酮类差异代谢物之间都互为正相关, 且与大部分脂类物质也呈正相关, 但与大部分的皂苷类物质呈负相关。另外, 研究显示尖孢镰刀菌侵染不是黄酮类化合物丙二酰取代发生的影响因子。本研究从植物代谢角度, 阐释根腐病菌尖孢镰刀菌侵染对黄芪代谢的影响, 为探索二者的互作机制, 辅助分子抗病育种提供依据。

关键词

黄芪 / 根腐病 / 代谢物 / 尖孢镰刀菌 / 黄酮类

Abstract

收起

Astragalus is a commonly used Chinese medicinal material in traditional Chinese medicine (TCM), and with the increase of planting area in recent years, the damage of Astragalus root rot has worsened year by year, which seriously affecting its quality and yield. Fusarium oxysporum is one of the main pathogens causing root rot in astragalus. In this study, UPLC-Q-TOF-MS based metabolomic approach combined with multivariate statistical analysis were used to analyze the metabolite changes of Astragalus in response to F. oxysporum infection. The results showed that 62 metabolites in the Astragalus had significant changes after inoculation of F. oxysporum. Polar metabolites included 40 flavonoids, 8 saponins, 2 nucleosides, 1 vitamin, 1 organic acid, 1 amino acid; while lipid metabolites included 3 fatty acids, 1 diradylglycerols, 2 lysophosphatidylcholine, 1 lysophosphatidylglycerol, 1 phosphatidylinositol, 1 sterol lipid. Among these differential metabolites, the relative content of flavonoids, vitamin B₂, tryptophan and salicylic acid were increased, while the relative content of saponins were decreased. Correlation analysis showed that the flavonoids were positively correlated with each other, and positively correlated with most lipids, but negatively correlated with most saponins. In addition, studies have shown that F. oxysporum infection is not an influencing factor for the generation of malonyl substitution of flavonoid. This study elucidates the effect of F. oxysporum infection on Astragalus from the perspective of plant metabolism, which provides a basis for exploring the interaction mechanism between the Astragalus and F. oxysporum and further promoting molecular breeding.

Key words

Astragalus membranaceus var. mongholicus / root rot / metabolite / Fusarium oxysporum / flavonoid

引用本文

王富鑫, 宋海清, 赵利, 秦雪梅, 高芬, 李震宇. 基于代谢组学分析尖孢镰刀菌侵染对黄芪代谢的影响. 药学学报, 2023 , 58 (7) : 1971 -1980 . DOI: 10.16438/j.0513-4870.2022-1377

Fu-xin WANG, Hai-qing SONG, Li ZHAO, Xue-mei QIN, Fen GAO, Zhen-yu LI. Analysis of Fusarium oxysporum infection on changes of Astragalus metabolism by metabolomic approach[J]. Acta Pharmaceutica Sinica, 2023 , 58 (7) : 1971 -1980 . DOI: 10.16438/j.0513-4870.2022-1377

正文

收起

2020版《中华人民共和国药典》收载的黄芪为豆科植物蒙古黄芪Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao或膜荚黄芪Astragalus membranaceus (Fisch.) Bge.的干燥根。黄芪性温、味甘, 具有补气升阳, 利尿托毒, 排腺和敛疮生肌的功效^[1]。蒙古黄芪是市场主流商品, 生产中以人工种植为主。近年来, 随着种植面积的不断增加, 根腐病在各主产区普遍发生, 黄芪的产量和质量受到了严重的影响, 限制了黄芪产业的可持续发展^[2]。

病原真菌在侵染植物的过程中, 会严重干扰植物的正常代谢以满足自身的营养吸收和利用, 同时, 植物也会调整自身代谢途径抵御病菌的侵染。鉴定与环境胁迫相关的代谢物, 对提高作物产量和作物改良遗传育种有着至关重要的作用^{[3, 4]}。代谢组学研究有助于明确植物感染病菌前后代谢分子的变化规律, 进而找到相关的代谢路径和关键蛋白/基因, 为抗病功能机制研究提供思路^[5]。目前, 代谢组学分析在镰刀菌属真菌侵染农作物后代谢物谱的变化, 抗病相关代谢物指认及互作机制解析方面已有大量相关报道。如Hamzehzarghani等^{[6, 7]}基于气质联用技术检测小麦被禾谷镰刀菌侵染后的代谢谱, 发现抗性品种与易感基因型品种相比, 多种代谢物的含量发生了显著变化, 且多种代谢物可能与植物抗病相关。Bollina和Kumaraswamy等^{[8, 9]}基于液质联用技术分析大麦与禾谷镰刀菌的互作机制, 发现苯丙素类、酚类、黄酮类、脂肪酸和萜类等代谢物与大麦抵抗病菌侵染相关。Du等^[10]分析黄瓜对尖孢镰刀菌侵染响应的代谢组变化, 发现尖孢镰刀菌处理导致74种代谢物的含量升高, 26种代谢物的含量降低, 涉及的代谢途径如磷酸戊糖、糖酵解、三羧酸循环也发生了显著变化。

尖孢镰刀菌是引起黄芪根腐病的主要病原菌之一, 在山西、甘肃、内蒙古、宁夏等黄芪种植区均有报道^{[2, 11]}。本课题组前期采用基于NMR的代谢组学技术研究发现, 尖孢镰刀菌侵染导致黄芪的代谢物谱发生了明显变化, 并指认出13种差异代谢物^[12]。研究表明, 植物主要通过次生代谢物来应对胁迫^[13], 而前期研究中指认出的差异代谢物绝大多数属于初级代谢产物, 无法全面反映黄芪被尖孢镰刀菌侵染后代谢物谱的变化。

UPLC-Q-TOF-MS质谱技术具有高分辨、高灵敏度的特点, 更加适合对植物次级代谢物的分析指认^{[14, 15]}。因此, 本研究拟采用基于UPLC-Q-TOF-MS的代谢组学技术, 结合多元统计分析, 解析黄芪响应尖孢镰刀菌侵染的代谢谱变化。以期更加全面、深入地从代谢物角度, 阐释根腐病菌尖孢镰刀菌侵染对黄芪的影响, 为探索二者的互作机制, 辅助分子抗病育种提供依据。

材料与方法

收起

仪器与试剂 Agilent 1290超高效液相色谱仪(美国Agilent Technologies公司); Q-TOF-MS/MS 5600+四极杆飞行时间质谱仪(美国AB SCIEX公司); KQ5200E超声波清洗器(昆山市超声仪器有限公司)。甲酸、乙腈(质谱级, 美国Thermo公司), 超纯水由Milli-Q纯净水系统(美国Millipore公司) 制备。

黄芪与病原菌 一年生粗细均匀的健康黄芪根, 由山西省应县黄芪种植基地提供。经山西大学中医药现代研究中心秦雪梅教授鉴定为蒙古黄芪Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao。尖孢镰刀菌(Fusarium oxysporum), 由本课题组分离自根腐病典型病根, 通过形态学和分子鉴定确定其分类地位, 菌株保存于本实验室^[2]。

病菌的培养、接种及样本培育 参照本课题组建立的方法进行^[16]。将活化后的尖孢镰刀菌接入PDA平板, 25 ℃下培养5~7天后, 打取5个菌碟(直径5 mm) 接入100 mL PD培养液, 振荡培养2天(25 ℃、180 r·min^-1); 滤除菌丝后用无菌水调整其终浓度至1.0~1.5×10⁶ cfu·mL^-1。选择粗细一致的黄芪根样本, 清水洗净后, 先用无菌水清洗1次, 再于75%酒精中浸泡10~15 s, 最后无菌水清洗2或3次。用灭菌接种针在各黄芪根段上平滑健康的相同部位作梅花状五点刺伤, 刺伤间隔2 cm, 每株刺伤5处; 接种组在尖孢镰刀菌孢子悬浮液中浸泡接种1 h后取出, 对照组以无菌水处理。对照组和接种组均为每组6次重复, 每个重复6株; 对照组和接种组均在人工气候室恒温培养[(25 ± 1) ℃, 光暗交替12 h/12 h], 28天时整根采样, 记录病斑面积和颜色, 并计算病情指数, 分级标准参照文献^[12]。样品经液氮速冻淬灭后, -80 ℃保存, 用于后续分析。

$ \mathrm{病}\mathrm{情}\mathrm{指}\mathrm{数}=\frac{\sum (\mathrm{各}\mathrm{级}\mathrm{病}\mathrm{株}\mathrm{数}\times \mathrm{代}\mathrm{表}\mathrm{数}\mathrm{值})}{(\mathrm{发}\mathrm{病}\mathrm{最}\mathrm{高}\mathrm{级}\mathrm{代}\mathrm{表}\mathrm{数}\mathrm{值}\times \mathrm{调}\mathrm{查}\mathrm{总}\mathrm{株}\mathrm{数})}\times 100 $

基于UPLC-Q-TOF-MS高分辨质谱的代谢组分析

样品前处理样品用液氮充分研磨后, 置于冻干机中冷冻干燥24 h。取黄芪粉末(过四号筛) 约0.10 g于玻璃离心管中, 加入2 mL 50%甲醇, 超声提取20 min, 3 500 r·min^-1离心10 min, 静置, 取上清液, 过0.22 μm的微孔滤膜, 置于4 ℃冰箱中备用。

色谱条件色谱柱Waters Acquity UPLC T3 (150 mm × 2.1 mm, 1.8 µm), 高效液相色谱检测条件: 流动相为0.1%甲酸水(A)-乙腈(B), 梯度洗脱: 0~5 min, 5%~20% B; 5~10 min, 20%~30% B; 10~15 min, 30%~43% B; 15~20 min, 43%~60% B; 20~30 min, 60%~100% B; 30~30.5 min, 100%~5% B; 30.5~35 min, 5% B; 柱温40 ℃; 流速0.3 mL·min^-1, 进样体积2 μL。

质谱条件电喷雾离子源(ESI), 采用正负离子模式分别扫描, 离子源参数如下: 喷雾电压(IS) -4 500 V (负离子), +5 500 V (正离子), 离子源温度550 ℃; 去簇裂解电压(DP) 为60 V; 二级碰撞能量(CE) 为40 eV; 碰撞能量滚动区间(CES) 为20 eV; 检测模式为IDA (信息关联采集模式) 触发二级的条件。

数据处理 将采集到的高分辨质谱数据导入XCMS软件(版本3.6.3), 获取包含RT、m/z、PI等信息的样本峰表数据矩阵。对于脂类成分, 进一步采用MSDIAL软件进行匹配得到包含RT、m/z、PI等信息的样本峰表数据矩阵。将样本峰表数据矩阵导入SIMCA-P软件(版本14.1), 通过无监督的主成分分析(principal component analysis, PCA) 观察样本原始离散程度, 通过FC值(FC > 1.2或FC < 0.8) 结合t检验(P < 0.05) 为标准确定组间差异代谢物。将所采集的高分辨质谱数据导入SCIEX OS软件, 通过分析质谱裂解规律进行差异代谢物的鉴定, 设置质量误差小于等于5 ppm, 并通过在线质谱数据库(Mass Bank database, https://massbank.eu/MassBank/Index; HMDB database, https://hmdb.ca/metabolites, mz Cloud database, https://www.mzcloud.org/) 及文献中质谱数据比对, 进一步确认化学结构。将鉴定的差异代谢物导入MetaboAnalyst 5.0 (https://www.metaboanalyst.ca/) 在线分析网站生成热图, 通过悟空(https://omicsolution.org/wkomics/main/) 在线分析网站生成Spearman相关性分析。

结果

收起

1 样本发病情况及病情指数

黄芪接种尖孢镰刀菌28天后, 接种部位出现大小约5~9 mm的病斑, 呈褐色或者深褐色(图 1), 个别黄芪根出现轻微腐烂症状。对照组无病斑出现, 少数接种部位为浅褐色。经计算, 尖孢镰刀菌接种组的病情指数为79.24。

2 极性代谢物差异分析

如图 2A、B所示, 正负离子模式下, 质控样本(quality control, QC) 分布在2倍SD以内, 说明方法重复性及稳定性良好, 可用于批量样本的分析。主成分分析结果如图C、D所示, 正离子模式下第一主成分(PC1) 的贡献率为36.4%, 第二主成分(PC2) 的贡献率为18.9%, 对照组和接种组两组样本虽然距离较近, 但仍能明显区分。说明尖孢镰刀菌接种后, 黄芪代谢物发生变化。在负离子模式下, 接种组和对照组也有明显分离趋势。

根据t检验(P < 0.05)、倍数变化值(FC > 1.2或FC < 0.8) 的标准, 筛选出尖孢镰刀菌接种组与对照组间的差异代谢物53个, 包括40个黄酮类、8个皂苷类、2个核苷类、1个维生素、1个有机酸、1个氨基酸(表 1)。相比对照组, 接种组有46个差异代谢物显著上调, 7个差异代谢物显著下调, 差异代谢物上调的数目大于下调, 差异代谢物中占比较多的是黄酮类成分。为了更加直观地比较黄芪在接种尖孢镰刀菌前后差异代谢物相对含量的变化情况, 进一步对53个差异代谢物进行层次聚类并绘制热图(图 3)。

3 脂质代谢物差异分析

由于XCMS得到的峰表中脂类物质较少, 因此进一步采用Msdial软件得到脂类物质的峰表后进行多元统计分析。如图 4A、B所示, 正负离子模式下, 质控样本分布在2SD以内, 说明对于脂类成分, 仪器采样过程同样稳定可靠, 可用于批量样本的分析。如图 4C、D所示, 从脂类物质来看, 两组样本虽未完全分离, 但均有分离趋势。

根据t检验(P < 0.05)、倍数变化值(FC > 1.2或FC < 0.8) 的标准, 筛选出尖孢镰刀菌接种组与对照组中9个差异脂质代谢物(表 1), 包括3个脂肪酸、1个甘油二酯、2个溶血磷脂酰胆碱、1个溶血磷脂甘油、1个磷脂酰肌醇、1个固醇脂。从差异代谢物的热图(图 5) 可见, 黄芪接种尖孢镰刀菌后有8个差异代谢物显著上调, 1个差异代谢物显著下调, 差异代谢物上调的数目大于下调, 差异代谢物中占比较多的是脂肪酸。

4 差异代谢物通路分析

葡萄糖在莽草酸途径相关酶的催化下, 依次经3-脱氧-D-阿拉伯糖-庚酮酸-7-磷酸(3-deoxy-D-arabinose-heptulosonate-7-phosphate, DAHP)、预苯酸(prephenic acid) 等重要中间体, 转化生成芳香族氨基酸如L-苯丙氨酸、L-酪氨酸等。L-苯丙氨酸可在苯丙氨酸解氨酶的催化下生成反式肉桂酸(transcinnamic acid), 随即经肉桂酸-4-羟化酶催化, 生成对香豆酸(p-coumaric acid), 香豆酸是合成黄酮类物质重要中间体^[17]。葡萄糖经糖酵解途径生成乙酰辅酶A (acetyl-CoA), 乙酰辅酶A可作为皂苷类与脂肪酸类生物合成的共同前体物质^[18]。本研究表明, 黄芪受到尖孢镰刀菌侵染后, 在植物体内葡萄糖总量不变的情况下, 可能是黄酮类和脂肪酸类物质合成途径激活而含量升高, 进一步导致皂苷合成减少而含量降低(图 6)。

5 差异代谢物相关性分析

对62个差异代谢物特征成分进行相关性分析(图 7)。在3 844组相关中, 其中2 538组呈现显著相关(P < 0.05), 其中正相关2 099个、负相关439个。正相关中包括1 698组强的正相关(r > 0.7), 383组中等强度的正相关(0.5 ≤ r ≤ 0.7)。负相关中包括249组强的负相关(-0.7 > r), 183组中等强度的负相关(-0.7 ≤ r ≤ -0.5)。

所有的黄酮类差异代谢物之间都互为正相关, 此外, 黄酮类还与大部分脂类物质呈现正相关, 与大部分的皂苷类物质呈现负相关。水杨酸为植物激素类成分, 与14个黄酮、2个脂类成分呈现显著正相关。维生素B₂与35个黄酮、7个脂类成分、1个皂苷类成分呈现显著正相关, 与3个皂苷类成分呈现显著负相关。鸟嘌呤与32个黄酮、7个脂质成分呈现显著正相关。腺苷与15个黄酮、6个脂质成分呈现显著正相关、与3个皂苷类成分呈现显著负相关。

黄芪中的黄酮类物质会发生丙二酰取代, 如calycosin-7-O-glucoside-6″-O-malonate (calycosin-7-O-Glc-6″-O-Mal)。为了分析丙二酰取代是否与尖孢镰刀菌侵染相关, 进一步在差异代谢物中选择4个黄酮类成分及其相应的丙二酰取代物进行比较(图 8)。Calycosin-7-O-Glc (FC = 1.75) 和calycosin-7-O-Glc-6″-O-Mal (FC = 1.83) 在尖孢镰刀菌侵染后含量均升高; 此外, odoratin-7-O-Glc和odoratin-7-O-Glc-6″-O-Mal, pratensein-Glc和pratensein-Glc-Mal-Glc-Mal, 以及isomucronulatol-7-O-Glc和isomucronulatol-7-O-Glc-Mal-Glc-Mal在尖孢镰刀菌侵染后含量也均升高, 说明尖孢镰刀菌侵染不是丙二酰取代发生的影响因子。

讨论

收起

本研究采用UPLC-Q-TOF-MS代谢组学技术发现, 黄芪接种尖孢镰刀菌后, 有62种代谢物发生显著的变化, 其中黄酮类、维生素B₂、水杨酸、色氨酸的相对含量明显增加, 而皂苷类的相对含量则明显减少。本课题组前期^[19]研究表明, 锐顶镰刀菌侵染黄芪后, 苯丙氨酸解氨酶、肉桂酸-4-羟化酶和辅酶A连接酶等苯丙烷途径关键酶活性显著升高, 这些酶与黄酮类化合物的合成相关。靶向代谢组学技术研究发现, 锐顶镰刀菌侵染黄芪后21天, 阿魏酸、对香豆酸、甘草素、芒柄花素显示出上调趋势, 而毛蕊异黄酮葡萄糖苷、芒柄花苷、柚皮素显著下调。本研究则发现黄芪被尖孢镰刀菌接种侵染28天后, 大部分黄酮类成分显著上调。上述结果提示, 根腐病致病镰刀菌侵染能导致黄芪中黄酮类成分发生上调或下调, 但不同致病菌引起的黄酮上调/下调种类、变化程度存在差异。同时, 侵染时间和发病程度也是黄酮类成分发生不同变化的重要影响因子。此外, 大麦受到禾谷镰刀菌侵染后, 也发现16种黄酮和异黄酮类代谢物显著上调^{[8, 9]}。Sun^[20]采用代谢组学技术研究表明, 腐皮镰刀菌侵染人参后, 其中的人参皂苷类物质显著下调。与本研究中尖孢镰刀菌侵染黄芪后, 皂苷类代谢物同样显著下调的结果一致。皂苷类是黄芪中重要的药效成分, 本研究结果表明尖孢镰刀菌侵染可能导致黄芪质量的降低。

相对于对照大麦组, 禾谷镰刀菌接种组中亚油酸含量上调, 体外实验也验证了亚油酸对禾谷镰刀菌菌丝体生长的抑制作用^[9]。水杨酸介导的信号通路在植物先天免疫反应中发挥重要作用, 拟南芥和小麦在受到禾谷镰刀菌侵染后会积累高水平的水杨酸, 表明水杨酸可能在防御禾谷镰刀菌的侵染方面具有重要作用^[21]。维生素B₂具有强的抗氧化潜力, 在保护植物免受胁迫方面发挥重要作用^{[22, 23]}, 如黄瓜根可通过上调维生素B₂防御尖孢镰刀菌的侵染。氨基酸是许多功能代谢物的前体, 通过促进相容渗透剂的积累, 有助于提高植物的抗逆耐受性。尖孢镰刀菌接种后黄瓜根部色氨酸、谷氨酸等4种氨基酸含量升高^[10]。在本研究中, 黄芪受到尖孢镰刀菌侵染后, 亚油酸、水杨酸、维生素B₂、色氨酸显著上调, 与上述文献的结果一致, 表明这些成分可能与黄芪对尖孢镰刀菌的抗性有关, 但具体的机制还需要深入研究。

本研究采用植物代谢组学技术初步揭示了黄芪受到尖孢镰刀菌侵染后初级和次级代谢产物的变化, 为深入研究黄芪根腐病的抗性标志物和抗性品种选育鉴定了基础。

作者贡献: 宋海清、赵利负责病菌接种及样品采集; 王富鑫负责样本测定及数据分析处理, 并撰写稿件; 秦雪梅参与黄芪样品品种的鉴定; 李震宇、高芬对整个研究工作进行设计、监督和指导, 以及对稿件的修改。

利益冲突: 本文的研究无任何利益冲突。

基金

收起

国家重点研发计划项目(2019YFC1710805)
山西省基础研究计划资助项目(202103021224029)
山西省现代农业产业技术体系建设专项资金资助(2021-11)

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

[1]

Guo YX, Li RR, Qin XM, et al. Characteristic components of Astragali Radix from Hengshan Mountains based on multiindex components determination[J]. Mod Chin Med (中国现代中药), 2022, 24: 1473-1480 https://www.cnki.com.cn/Article/CJFDTOTAL-YJXX202305011.htm

[2]

Gao F, Zhao XX, Qin XM, et al. Analysis of dominant pathogen community causing Astragalus membranaceus var. mongholicus root rot in Shanxi Province[J]. Acta Phytophylacica Sin (植物保护学报), 2018, 45: 878-885 https://www.cnki.com.cn/Article/CJFDTOTAL-ZWBF202303022.htm

[3]

Mitchell-Olds T, Schmitt J. Genetic mechanisms and evolutionary significance of natural variation in Arabidopsis[J]. Nature, 2006, 441: 947-952.

https://doi.org/10.1038/nature04878

[4]

Sumner LW, Lei Z, Nikolau BJ, et al. Modern plant metabolomics: advanced natural product gene discoveries, improved technologies, and future prospects[J]. Nat Prod Rep, 2015, 32: 212-229.

https://doi.org/10.1039/C4NP00072B

[5]

Fiehn O. Metabolomics-the link between genotypes and phenotypes[J]. Plant Mol Biol, 2002, 48: 155-171.

https://doi.org/10.1023/A:1013713905833

[6]

Hamzehzarghani H, Kushalappa A, Dion Y, et al. Metabolic profiling and factor analysis to discriminate quantitative resistance in wheat cultivars against Fusarium head blight[J]. Physiol Mol Plant Pathol, 2005, 66: 119-133.

https://doi.org/10.1016/j.pmpp.2005.05.005

[7]

Hamzehzarghani H, Paranidharan V, Abu-Nada Y, et al. Metabolite profiling coupled with statistical analyses for potential high-throughput screening of quantitative resistance to Fusarium head blight in wheat[J]. Can J Plant Pathol, 2008, 30: 24-36.

https://doi.org/10.1080/07060660809507493

[8]

Bollina V, Kumaraswamy GK, Kushalappa AC, et al. Mass spectrometry-based metabolomics application to identify quantitative resistance-related metabolites in barley against Fusarium head blight[J]. Mol Plant Pathol, 2010, 11: 769-782

[9]

Kumaraswamy GK, Bollina V, Kushalappa AC, et al. Metabolomics technology to phenotype resistance in barley against Gibberella zeae[J]. Eur J Plant Pathol, 2011, 130: 29-43.

https://doi.org/10.1007/s10658-010-9729-3

[10]

Du N, Yang Q, Guo H, et al. Dissection of Paenibacillus polymyxa NSY50-induced defense in cucumber roots against Fusarium oxysporum f. sp. Cucumerinum by target metabolite profiling[J]. Biology (Basel), 2022, 11: 1028

[11]

Niu SQ, Geng H, Han CH, et al. Isolation and identification of pathogens causing Astragalus membranaceus root rot in Gansu Longxi[J]. Northwest Norm Univ Nat Sci (西北师范大学学报), 2016, 52: 75-78, 83, 83. https://www.cnki.com.cn/Article/CJFDTOTAL-XBLX202303017.htm

[12]

Zhao L. Discovery and Functional Verification of Resistance-Related Metabolites of Astragalus membranaceus var. mongholicus Aganist Mixed Infestations of Root Rot Pathogens (黄芪抗根腐病菌复合侵染的相关代谢物挖掘及验证) [D]. Taiyuan: Shanxi University, 2022.

[13]

Nakabayashi R, Saito K. Integrated metabolomics for abiotic stress responses in plants[J]. Curr Opin Plant Biol, 2015, 24: 10-16.

https://doi.org/10.1016/j.pbi.2015.01.003

[14]

Wei R, Yang LJ, Qin XM et al. Rapid identification of chemical constituents in the dried stem bark of Asparagus officinalis L. based on UPLC-Q-TOF-MS/MS[J]. Acta Pharm Sin (药学学报), 2022, 57: 2839-2850 https://www.cnki.com.cn/Article/CJFDTOTAL-ZYXB202306022.htm

[15]

Lou GG, Xia J, Yang J, et al. Differences in the chemical composition of Dendrobium officinale Kimura et Migo and Dendrobium crepidatum Lindl based on UPLC-Q-TOF-MS/MS and metabolomics[J]. Acta Pharm Sin (药学学报), 2021, 56: 3331-3344 https://www.cnki.com.cn/Article/CJFDTOTAL-HNZK202307001.htm

[16]

Gao F, Chao JB, Guo J, et al. 1H NMR-based metabolomics to identify resistance-related metabolites in Astragalus membranaceus var. mongholicus against Fusarium root rot[J]. Int J Agric Biol, 2021, 26: 69-78.

https://doi.org/10.17957/IJAB/15.1810

[17]

Lin CC, Chen DW, Dai JG. Advances of synthetic biology of flavonoids[J]. Acta Pharm Sin (药学学报), 2022, 57: 1322-1335 https://www.cnki.com.cn/Article/CJFDTOTAL-YXBY202305025.htm

[18]

Ming QL, Han H, Huang F, et al. Advances in studies on ginsenoside biosynthesis and its related enzymes[J]. Chin Tradit Herb Drugs (中草药), 2010, 41: 1913-1917 https://www.cnki.com.cn/Article/CJFDTOTAL-YYPF202306013.htm

[19]

Zhao XJ. RT-qPCR Detection of Fusarium acuminatum and Metabolomic Analysis of Astragalas membranaceus Challenged with the Pathogen (锐顶镰刀菌的RT-qPCR检测及黄芪对其侵染响应的代谢组分析) [D]. Taiyuan: Shanxi University, 2020.

[20]

Sun R. Study on Enzyme Activity and Metabonomic Changes of Panax ginseng to Fusarium solani Infection (人参对腐皮镰刀菌侵染的酶活响应及代谢组变化研究) [D]. Changchun: Changchun University of Chinese Medicine, 2022.

[21]

Makandar R, Nalam VJ, Chowdhury Z, et al. The combined action of enhanced disease susceptibility 1, phytoalexin deficient 4, and senescence-associated 101 promotes salicylic acid-mediated defenses to limit Fusarium graminearum infection in Arabidopsis thaliana[J]. Mol Plant Microbe Interact, 2015, 28: 943-953.

https://doi.org/10.1094/MPMI-04-15-0079-R

[22]

Boubakri H, Gargouri M, Mliki A, et al. Vitamins for enhancing plant resistance[J]. Planta, 2016, 244: 529-543.

https://doi.org/10.1007/s00425-016-2552-0

[23]

Saikia R, Yadav M, Varghesh S, et al. Role of riboflavin in induced resistance against Fusarium wilt and charcoal rot diseases of chickpea[J]. Plant Pathol J, 2006, 22: 339-347.

https://doi.org/10.5423/PPJ.2006.22.4.339

2023年第58卷第7期

PDF下载

180

引用本文

BibTeX

文章信息

doi: 10.16438/j.0513-4870.2022-1377

接收时间：2022-12-17
首发时间：2025-11-21
出版时间：2023-07-12

补充材料

相关文章

文章信息

作者

出版历史

收稿日期：2022-12-17
修回日期：2023-02-10

基金

国家重点研发计划项目(2019YFC1710805)

山西省基础研究计划资助项目(202103021224029)

山西省现代农业产业技术体系建设专项资金资助(2021-11)

作者信息

1.山西大学中医药现代研究中心, 山西太原 030006

2.山西大学应用化学研究所, 山西太原 030006

通讯作者:

*高芬, Tel: 86-351-7018390, E-mail: gaofen@sxu.edu.cn;

李震宇, Tel: 86-351-7011202, E-mail: lizhenyu@sxu.edu.cn

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/yxxb/CN/10.16438/j.0513-4870.2022-1377

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

No.	Compound	t_R /min	Formula	Experimental (m/z)	Mode	Mass error	Fragment	Type	FC
1	Rhamnocitrin	23.14	C₁₆H₁₂O₆	301.070 7	[M+H]⁺	0.1	216.045 0, 203.068 3, 188.044 7, 175.075 4, 171.043 8, 167.037 9, 139.030 6	A	3.75
2	Pectolinarigenin	20.77	C₁₇H₁₄O₆	313.071 8	[M-H]^-	0.1	297.041 5, 283.026 1, 269.047 1, 225.029 6, 227.035 3, 183.044 6, 171.044 6	A	1.57
3	Rhamnocitrin-Hex	14.79	C₂₂H₂₂O₁₁	461.109 3	[M+H]^-	0.8	299.056 4, 284.032 8, 255.030 6, 135.008 9, 503.119 1	A	2.45
4	Trihydroxy-methoxypterocarpan-Glc-Mal	19.44	C₂₅H₂₆O₁₄	551.137 3	[M+H]⁺	-4.1	439.357 4, 302.072 3, 204.086 0, 186.073 2, 138.058 8	B	1.79
5	Astrapterocarpan-3-O-Glc-Mal-Api	20.54	C₃₁H₃₆O₁₇	698.229 0	[M+NH₄]⁺	-0.2	549.160 0, 371.097 0, 325.108 9, 301.107 5, 191.069 3, 167.070 2	B	1.62
6	Astrapterocarpan-O-Glc-Mal isomer 1	20.54	C₂₆H₂₈O₁₃	549.160 3	[M+H]⁺	0.0	301.107 2, 191.070 0, 167.070 2, 123.044 4	B	1.69
7	Tetrahydroxy-pterocarpan-Glc-Mal	16.54	C₂₄H₂₄O₁₄	537.123 9	[M+H]⁺	0.1	289.069 6, 281.049 3, 289.082 6, 243.058 7, 137.022 1, 127.038 1, 73.045 7	B	1.56
8	Astrapterocarpan-3-O-Glc-Ace	20.80	C₂₅H₂₈O₁₁	505.169 7	[M+H]⁺	-2.4	181.047 5, 167.071 3, 191.367 3, 255.090 2, 301.069 9, 445.154 7	B	1.28
9	Isomucronulatol-7-O-Glc-Mal-Glc-Mal	21.26	C₃₅H₄₂O₂₁	816.255 7	[M+NH₄]⁺	-1.5	257.044 1, 239.006 7, 165.057 1, 150.033 4, 149.032 2, 121.028 6, 109.030 1, 91.063 3	C	1.93
10	Isomucronulatol-Glc-Glc	17.98	C₂₉H₃₈O₁₅	644.254 8	[M+NH₄]⁺	-0.2	465.175 3, 429.170 9, 369.169 6, 369.134 0, 345.145 0, 303.122 8, 167.070 0, 123.044 3	C	2.63
11	Trihydroxy-dimethoxyisoflavan-Glc-Glc	13.80	C₂₉H₃₈O₁₆	660.249 6	[M+NH₄]⁺	-0.4	660.248 4, 557.295 5, 445.143 5, 427.165 0, 343.112 6, 319.120 2, 235.069 0, 183.070 0, 123.042 5	C	2.55
12	Isomucronulatol-7-O-Glc-Glc-Mal	18.73	C₃₂H₄₀O₁₈	730.255 3	[M+NH₄]⁺	-0.4	429.160 5, 393.127 0, 369.139 4, 303.121 5, 249.076 5, 239.067 1, 193.085 5, 189.055 5, 181.085 8	C	2.37
13	Isomucronulatol-7-O-Glc-Mal-Api	18.21	C₃₁H₃₈O₁₇	700.244 7	[M+NH₄]⁺	-0.6	339.121 6, 339.134 5, 297.108 0, 273.111 1, 177.052 4, 153.051 1, 133.066 3	C	1.33
14	Isomucronulatol	20.32	C₁₇H₁₈O₅	301.108 2	[M-H]^-	0.1	301.111 3, 286.087 3, 271.062 6, 256.038 7, 253.051 8, 149.025 0, 135.045 9	C	2.34
15	Isomucronulatol-7, 2′-di-O-Glc	17.56	C₂₉H₃₈O₁₅	625.213 0	[M-H]^-	-1.2	463.159 7, 301.107 9, 301.107 3, 135.044 9, 286.084 4	C	2.51
16	Trihydroxy-dimethoxyisoflavan-Glc	19.48	C₂₂H₂₆O₁₀	449.144 0	[M-H]^-	-2.9	287.093 8, 272.071 7, 257.044 1, 239.006 7, 165.057 1, 150.033 4, 149.032 2, 121.028 6, 109.030 1	C	3.71
17	Trihydroxy-methoxyisoflavan	20.69	C₁₆H₁₆O₅	287.091 9	[M-H]^-	-0.1	287.094 6, 272.069 3, 257.045 4, 254.045 4, 254.056 9, 165.056 4, 163.040 8, 149.023 8, 150.032 2	C	1.87
18	Isomucronulatol-O-Glc-Ace isomer	20.73	C₂₅H₃₀O₁₁	505.171 2	[M-H]^-	-0.8	301.108 5, 286.085 0, 271.061 5, 179.071 9, 164.048 5, 135.045 6, 121.029 7, 122.037 7	C	1.44
19	Isomucronulatol-7-O-Glc	20.61	C₂₃H₂₈O₁₀	465.175 1	[M+H]⁺	-1.0	303.123 6, 165.053 2, 167.070 1, 167.085 9, 123.043 3, 167.085 9, 83.012 7	C	3.08
20	Isomucronulatol isomer	23.56	C₁₇H₁₈O₅	303.126 6	[M+H]⁺	-0.4	193.084 9, 167.069 1, 161.058 7, 133.064 0, 123.043 0, 118.040 5, 106.040 2	C	2.57
21	Isomucronulatol-O-Glc isomer	20.61	C₂₃H₂₈O₁₀	465.175 0	[M+H]⁺	-0.2	429.150 1, 369.139 3, 339.124 0, 303.125 0, 303.138 8, 249.080 9, 189.053 6, 167.069 2, 123.044 5	C	3.08
22	Calycosin-7-O-Glc-6″-O-Mal isomer	18.13	C₂₅H₂₄O₁₃	533.128 5	[M+H]⁺	-0.2	285.075 3, 270.052 5, 253.049 9, 225.055 2, 137.023 4	D	1.45
23	Calycosin-7-O-Glc-6″-O-Mal	18.44	C₂₅H₂₄O₁₃	533.129 3	[M+H]⁺	-1.0	285.075 2, 285.092 8, 270.052 2, 253.050 2, 225.055 0, 137.023 4	D	1.38
24	Odoratin-7-O-Glc-6″-O-Mal	18.53	C₂₆H₂₆O₁₄	563.139 6	[M+H]⁺	0.1	315.086 5, 300.064 0, 283.060 1, 255.066 2	D	2.23
25	Pratensein-7-O-Glc	14.79	C₂₂H₂₂O₁₁	461.109 3	[M-H]^-	0.8	299.057 1, 284.033 6, 255.031 4, 240.045 5	D	2.46
26	Isoafrormosin	19.29	C₁₇H₁₄O₅	297.076 4	[M-H]^-	-1.6	297.076 4, 281.044 2, 267.029 9, 253.050 5, 239.034 5	D	1.42
27	Pratensein-Glc	16.29	C₂₄H₂₄O₁₂	503.119 6	[M-H]^-	0.3	443.091 6, 299.057 6, 256.038 1	D	1.86
28	3′-Methoxy-5′-hydroxyisoflavone-7-O-Glc	17.79	C₂₂H₂₂O₁₀	445.111 8	[M-H]^-	-4.9	297.042 6, 283.061 8, 268.038 3, 239.035 6	D	2.11
29	Pratensein-7-O-Glc isomer	18.58	C₂₂H₂₂O₁₁	463.123 5	[M+H]⁺	0.3	301.071 0, 286.040 5, 269.045 4, 241.045 2, 153.016 5	D	1.50
30	Calycosin-7-O-Glc	16.78	C₂₂H₂₂O₁₀	447.128 5	[M+H]⁺	-1.7	286.384 0, 285.075 1, 270.051 6, 253.050 1, 225.055 4, 214.061 1, 137.021 7	D	1.79
31	Ononin	19.71	C₂₂H₂₂O₉	431.133 1	[M+H]⁺	-1.3	269.080 4, 254.057 4, 213.092 0, 197.061 8, 136.014 5	D	1.62
32	Odoratin-7-O-Glc	17.05	C₂₃H₂₄O₁₁	477.139 1	[M+H]⁺	0.0	315.086 7, 300.064 8, 272.070 3, 255.094 1, 200.046 3, 175.035 4	D	2.73
33	Pratensein isomer 1	22.29	C₁₆H₁₂O₆	301.071 3	[M+H]⁺	2.2	151.078 8, 153.018 5, 163.038 4, 229.045 3, 241.046 5, 301.068 6, 301.097 9	D	1.99
34	Afrormosin-7-O-Glc-6″-O-Mal	20.65	C₂₆H₂₆O₁₃	547.144 7	[M+H]⁺	0.1	299.091 5, 284.068 5, 243.111 7, 183.084 9	D	2.03
35	Pratensein	23.15	C₁₆H₁₂O₆	301.070 7	[M+H]⁺	-0.5	123.044 5, 171.039 3, 167.060 1, 167.032 9, 188.044 5, 203.068 1, 217.054 8, 231.065 2, 301.070 8	D	3.40
36	3′, 5′-Dihydroxy-4′-methoxyisoflavone	23.88	C₁₇H₁₆O₄	285.112 2	[M+H]⁺	0.1	285.001 6, 243.101 2, 239.106 8, 191.070 6, 176.046 6, 137.059 6, 121.027 8	D	2.50
37	Tetrahydroxy-methoxy-isoflavone-Glc-Mal	17.98	C₂₅H₂₄O₁₅	565.118 9	[M+H]⁺	0.1	565.112 6, 302.045 8, 317.065 2, 285.041 3, 299.052 1, 191.035 5	D	1.54
38	Afrormosin-Glc-Mal isomer	19.71	C₂₆H₂₆O₁₃	547.144 7	[M+H]⁺	0.1	547.148 8, 299.092 1, 300.095 7, 284.068 0, 239.068 9, 211.074 1	D	1.44
39	Calycosin-Glc-Glc-Mal	17.99	C₃₁H₃₄O₁₈	695.181 8	[M+H]⁺	-0.3	533.135 5, 285.075 3, 270.067 9, 270.043 9, 225.053 7, 158.075 2	D	2.83
40	Pratensein-Glc-Mal-Glc-Mal	15.35	C₃₄H₃₆O₂₂	797.177 1	[M+H]⁺	0.3	547.406 1, 549.123 4, 301.070 5, 269.054 1	D	2.48
41	Guanosine	7.84	C₁₀H₁₃N₅O₅	284.099 1	[M+H]⁺	0.4	152.056 3, 135.029 7, 110.034 4	E	4.30
42	Adenosine	7.77	C₁₀H₁₃N₅O₄	268.104 6	[M+H]⁺	2.2	136.062 4, 57.035 8	E	2.53
43	Tryptophan	10.25	C₁₁H₁₂N₂O₂	203.082 8	[M-H]^-	0.8	159.095 0, 142.067 8, 116.051 7, 74.025 7	F	1.34
44	Salicylic acid	18.02	C₇H₆O₃	137.024 4	[M-H]^-	-0.7	137.027 1, 93.035 7, 93.051 0, 78.963 7, 65.039 4	G	3.31
45	Acetylastragaloside I isomer	27.12	C₄₇H₇₄O₁₇	911.498 3	[M+H]⁺	-1.7	731.428 8, 713.424 5, 695.411 0, 437.348 3, 419.335 9, 199.059 9, 157.050 9, 143.107 9, 125.098 0	H	2.83
46	Argentatin A-Glc-Ara	22.58	C₄₁H₆₆O₁₃	767.456 1	[M+H]⁺	-1.9	587.398 0, 599.388 5, 569.369 3, 473.364 2, 455.348 4, 437.342 9, 419.328 4, 253.193 1, 187.145 2	H	0.67
47	Astragaloside II-Mal	23.82	C₄₆H₇₂O₁₈	913.479 4	[M+H]⁺	0.2	599.403 4, 473.368 5, 455.353 6, 437.341 0, 419.335 8, 243.049 3, 261.061 4, 143.105 4	H	0.41
48	Astragaloside II-Mal-Ace	24, 91	C₄₈H₇₄O₁₉	955.490 7	[M+H]⁺	1.5	739.405 8, 721.399 8, 455.352 0, 437.340 8, 419.328 7, 303.070 8, 157.049 1, 143.105 5	H	0.63
49	Astragaloside I-Glc-Mal-Mal	23.37	C₅₇H₈₆O₂₇	1203.538 4	[M+H]⁺	-3.7	455.356 1, 437.344 1, 419.331 9, 401.310 7, 199.058 8, 157.049 4, 143.106 6, 97.028 3	H	0.64
50	HuangqiyeninA-Ara	22.48	C₄₁H₆₆O₁₄	783.451 2	[M+H]⁺	-1.7	453.334 5, 435.325 6, 417.313 4, 271.213 3, 143.107 9	H	0.53
51	Agroastragaloside IV	22.04	C₄₉H₈₀O₂₀	987.518 1	[M-H]^-	1.1	941.521 9	H	0.60
52	SoyasapogenolB-Glc-Rha	22.44	C₄₁H₆₆O₁₃	765.441 2	[M-H]^-	-2.4	603.394 5, 157.009 9, 113.020 5, 85.029 5	H	0.72
53	Vitamin B₂	14.66	C₁₇H₂₀N₄O₆	377.145 6	[M+H]⁺	0.0	243.087 0, 172.086 8, 57.032 9, 198.066 7, 145.076 2, 81.032 9, 57.033 1	I	1.99
54	FA 16:0	30.88	C₁₆H₃₂O₂	255.232 91	[M-H]^-	-0.5	255.232 4, 256.235 8, 257.239 1	J	1.69
55	FA 18:2	29.94	C₁₈H₃₂O₂	279.232 97	[M-H]^-	0.2	279.232 0, 280.235 5, 281.238 9	J	2.00
56	FA 18:2;O	26.43	C₁₈H₃₂O₃	295.224 7	[M-H]^-	-0.8	295.228 1, 296.231 0, 297.234 8	J	1.82
57	ST 28:3;O	24.90	C₂₈H₄₄O	419.328 92	[M+Na]⁺	0.5	419.329 5, 420.332 8, 421.336 2	K	0.74
58	DG 20:0	27.20	C₂₃H₄₄O₅	423.312 71	[M+Na]⁺	4.0	423.309 0, 331.266 2	L	2.65
59	LPG 16:0	31.98	C₂₂H₄₅O₉P	483.270 11	[M-H]^-	-0.2	483.272 5, 484.275 8, 485.279 2	M	1.97
60	LPC 16:0	26.61	C₂₄H₅₀NO₇P	496.339 48	[M+H]⁺	0.9	496.339 2, 497.342 5, 498.345 9	N	5.21
61	LPC 18:0	28.38	C₂₆H₅₄NO₇P	524.367 92	[M+H]⁺	0.4	524.371 2, 525.374 5, 526.377 9	N	2.61
62	PI 19:0	25.33	C₂₈H₅₃O₁₃P	627.317 32	[M-H]^-	2.3	627.317 3, 628.320 6, 629.324 0	O	3.89

No.

Compound

t_R /min

Formula

Experimental (m/z)

Mode

Mass error

Fragment

Type

Rhamnocitrin

23.14

C₁₆H₁₂O₆

301.070 7

[M+H]⁺

0.1

216.045 0, 203.068 3, 188.044 7, 175.075 4, 171.043 8, 167.037 9, 139.030 6

3.75

Pectolinarigenin

20.77

C₁₇H₁₄O₆

313.071 8

[M-H]^-

0.1

297.041 5, 283.026 1, 269.047 1, 225.029 6, 227.035 3, 183.044 6, 171.044 6

1.57

Rhamnocitrin-Hex

14.79

C₂₂H₂₂O₁₁

461.109 3

[M+H]^-

0.8

299.056 4, 284.032 8, 255.030 6, 135.008 9, 503.119 1

2.45

Trihydroxy-methoxypterocarpan-Glc-Mal

19.44

C₂₅H₂₆O₁₄

551.137 3

[M+H]⁺

-4.1

439.357 4, 302.072 3, 204.086 0, 186.073 2, 138.058 8

1.79

Astrapterocarpan-3-O-Glc-Mal-Api

20.54

C₃₁H₃₆O₁₇

698.229 0

[M+NH₄]⁺

-0.2

549.160 0, 371.097 0, 325.108 9, 301.107 5, 191.069 3, 167.070 2

1.62

Astrapterocarpan-O-Glc-Mal isomer 1

20.54

C₂₆H₂₈O₁₃

549.160 3

[M+H]⁺

0.0

301.107 2, 191.070 0, 167.070 2, 123.044 4

1.69

Tetrahydroxy-pterocarpan-Glc-Mal

16.54

C₂₄H₂₄O₁₄

537.123 9

[M+H]⁺

0.1

289.069 6, 281.049 3, 289.082 6, 243.058 7, 137.022 1, 127.038 1, 73.045 7

1.56

Astrapterocarpan-3-O-Glc-Ace

20.80

C₂₅H₂₈O₁₁

505.169 7

[M+H]⁺

-2.4

181.047 5, 167.071 3, 191.367 3, 255.090 2, 301.069 9, 445.154 7

1.28

Isomucronulatol-7-O-Glc-Mal-Glc-Mal

21.26

C₃₅H₄₂O₂₁

816.255 7

[M+NH₄]⁺

-1.5

257.044 1, 239.006 7, 165.057 1, 150.033 4, 149.032 2, 121.028 6, 109.030 1, 91.063 3

1.93

Isomucronulatol-Glc-Glc

17.98

C₂₉H₃₈O₁₅

644.254 8

[M+NH₄]⁺

-0.2

465.175 3, 429.170 9, 369.169 6, 369.134 0, 345.145 0, 303.122 8, 167.070 0, 123.044 3

2.63

Trihydroxy-dimethoxyisoflavan-Glc-Glc

13.80

C₂₉H₃₈O₁₆

660.249 6

[M+NH₄]⁺

-0.4

660.248 4, 557.295 5, 445.143 5, 427.165 0, 343.112 6, 319.120 2, 235.069 0, 183.070 0, 123.042 5

2.55

Isomucronulatol-7-O-Glc-Glc-Mal

18.73

C₃₂H₄₀O₁₈

730.255 3

[M+NH₄]⁺

-0.4

429.160 5, 393.127 0, 369.139 4, 303.121 5, 249.076 5, 239.067 1, 193.085 5, 189.055 5, 181.085 8

2.37

Isomucronulatol-7-O-Glc-Mal-Api

18.21

C₃₁H₃₈O₁₇

700.244 7

[M+NH₄]⁺

-0.6

339.121 6, 339.134 5, 297.108 0, 273.111 1, 177.052 4, 153.051 1, 133.066 3

1.33

Isomucronulatol

20.32

C₁₇H₁₈O₅

301.108 2

[M-H]^-

0.1

301.111 3, 286.087 3, 271.062 6, 256.038 7, 253.051 8, 149.025 0, 135.045 9

2.34

Isomucronulatol-7, 2′-di-O-Glc

17.56

C₂₉H₃₈O₁₅

625.213 0

[M-H]^-

-1.2

463.159 7, 301.107 9, 301.107 3, 135.044 9, 286.084 4

2.51

Trihydroxy-dimethoxyisoflavan-Glc

19.48

C₂₂H₂₆O₁₀

449.144 0

[M-H]^-

-2.9

287.093 8, 272.071 7, 257.044 1, 239.006 7, 165.057 1, 150.033 4, 149.032 2, 121.028 6, 109.030 1

3.71

Trihydroxy-methoxyisoflavan

20.69

C₁₆H₁₆O₅

287.091 9

[M-H]^-

-0.1

287.094 6, 272.069 3, 257.045 4, 254.045 4, 254.056 9, 165.056 4, 163.040 8, 149.023 8, 150.032 2

1.87

Isomucronulatol-O-Glc-Ace isomer

20.73

C₂₅H₃₀O₁₁

505.171 2

[M-H]^-

-0.8

301.108 5, 286.085 0, 271.061 5, 179.071 9, 164.048 5, 135.045 6, 121.029 7, 122.037 7

1.44

Isomucronulatol-7-O-Glc

20.61

C₂₃H₂₈O₁₀

465.175 1

[M+H]⁺

-1.0

303.123 6, 165.053 2, 167.070 1, 167.085 9, 123.043 3, 167.085 9, 83.012 7

3.08

Isomucronulatol isomer

23.56

C₁₇H₁₈O₅

303.126 6

[M+H]⁺

-0.4

193.084 9, 167.069 1, 161.058 7, 133.064 0, 123.043 0, 118.040 5, 106.040 2

2.57

Isomucronulatol-O-Glc isomer

20.61

C₂₃H₂₈O₁₀

465.175 0

[M+H]⁺

-0.2

429.150 1, 369.139 3, 339.124 0, 303.125 0, 303.138 8, 249.080 9, 189.053 6, 167.069 2, 123.044 5

3.08

Calycosin-7-O-Glc-6″-O-Mal isomer

18.13

C₂₅H₂₄O₁₃

533.128 5

[M+H]⁺

-0.2

285.075 3, 270.052 5, 253.049 9, 225.055 2, 137.023 4

1.45

Calycosin-7-O-Glc-6″-O-Mal

18.44

C₂₅H₂₄O₁₃

533.129 3

[M+H]⁺

-1.0

285.075 2, 285.092 8, 270.052 2, 253.050 2, 225.055 0, 137.023 4

1.38

Odoratin-7-O-Glc-6″-O-Mal

18.53

C₂₆H₂₆O₁₄

563.139 6

[M+H]⁺

0.1

315.086 5, 300.064 0, 283.060 1, 255.066 2

2.23

Pratensein-7-O-Glc

14.79

C₂₂H₂₂O₁₁

461.109 3

[M-H]^-

0.8

299.057 1, 284.033 6, 255.031 4, 240.045 5

2.46

Isoafrormosin

19.29

C₁₇H₁₄O₅

297.076 4

[M-H]^-

-1.6

297.076 4, 281.044 2, 267.029 9, 253.050 5, 239.034 5

1.42

Pratensein-Glc

16.29

C₂₄H₂₄O₁₂

503.119 6

[M-H]^-

0.3

443.091 6, 299.057 6, 256.038 1

1.86

3′-Methoxy-5′-hydroxyisoflavone-7-O-Glc

17.79

C₂₂H₂₂O₁₀

445.111 8

[M-H]^-

-4.9

297.042 6, 283.061 8, 268.038 3, 239.035 6

2.11

Pratensein-7-O-Glc isomer

18.58

C₂₂H₂₂O₁₁

463.123 5

[M+H]⁺

0.3

301.071 0, 286.040 5, 269.045 4, 241.045 2, 153.016 5

1.50

Calycosin-7-O-Glc

16.78

C₂₂H₂₂O₁₀

447.128 5

[M+H]⁺

-1.7

286.384 0, 285.075 1, 270.051 6, 253.050 1, 225.055 4, 214.061 1, 137.021 7

1.79

Ononin

19.71

C₂₂H₂₂O₉

431.133 1

[M+H]⁺

-1.3

269.080 4, 254.057 4, 213.092 0, 197.061 8, 136.014 5

1.62

Odoratin-7-O-Glc

17.05

C₂₃H₂₄O₁₁

477.139 1

[M+H]⁺

0.0

315.086 7, 300.064 8, 272.070 3, 255.094 1, 200.046 3, 175.035 4

2.73

Pratensein isomer 1

22.29

C₁₆H₁₂O₆

301.071 3

[M+H]⁺

2.2

151.078 8, 153.018 5, 163.038 4, 229.045 3, 241.046 5, 301.068 6, 301.097 9

1.99

Afrormosin-7-O-Glc-6″-O-Mal

20.65

C₂₆H₂₆O₁₃

547.144 7

[M+H]⁺

0.1

299.091 5, 284.068 5, 243.111 7, 183.084 9

2.03

Pratensein

23.15

C₁₆H₁₂O₆

301.070 7

[M+H]⁺

-0.5

123.044 5, 171.039 3, 167.060 1, 167.032 9, 188.044 5, 203.068 1, 217.054 8, 231.065 2, 301.070 8

3.40

3′, 5′-Dihydroxy-4′-methoxyisoflavone

23.88

C₁₇H₁₆O₄

285.112 2

[M+H]⁺

0.1

285.001 6, 243.101 2, 239.106 8, 191.070 6, 176.046 6, 137.059 6, 121.027 8

2.50

Tetrahydroxy-methoxy-isoflavone-Glc-Mal

17.98

C₂₅H₂₄O₁₅

565.118 9

[M+H]⁺

0.1

565.112 6, 302.045 8, 317.065 2, 285.041 3, 299.052 1, 191.035 5

1.54

Afrormosin-Glc-Mal isomer

19.71

C₂₆H₂₆O₁₃

547.144 7

[M+H]⁺

0.1

547.148 8, 299.092 1, 300.095 7, 284.068 0, 239.068 9, 211.074 1

1.44

Calycosin-Glc-Glc-Mal

17.99

C₃₁H₃₄O₁₈

695.181 8

[M+H]⁺

-0.3

533.135 5, 285.075 3, 270.067 9, 270.043 9, 225.053 7, 158.075 2

2.83

Pratensein-Glc-Mal-Glc-Mal

15.35

C₃₄H₃₆O₂₂

797.177 1

[M+H]⁺

0.3

547.406 1, 549.123 4, 301.070 5, 269.054 1

2.48

Guanosine

7.84

C₁₀H₁₃N₅O₅

284.099 1

[M+H]⁺

0.4

152.056 3, 135.029 7, 110.034 4

4.30

Adenosine

7.77

C₁₀H₁₃N₅O₄

268.104 6

[M+H]⁺

2.2

136.062 4, 57.035 8

2.53

Tryptophan

10.25

C₁₁H₁₂N₂O₂

203.082 8

[M-H]^-

0.8

159.095 0, 142.067 8, 116.051 7, 74.025 7

1.34

Salicylic acid

18.02

C₇H₆O₃

137.024 4

[M-H]^-

-0.7

137.027 1, 93.035 7, 93.051 0, 78.963 7, 65.039 4

3.31

Acetylastragaloside I isomer

27.12

C₄₇H₇₄O₁₇

911.498 3

[M+H]⁺

-1.7

731.428 8, 713.424 5, 695.411 0, 437.348 3, 419.335 9, 199.059 9, 157.050 9, 143.107 9, 125.098 0

2.83

Argentatin A-Glc-Ara

22.58

C₄₁H₆₆O₁₃

767.456 1

[M+H]⁺

-1.9

587.398 0, 599.388 5, 569.369 3, 473.364 2, 455.348 4, 437.342 9, 419.328 4, 253.193 1, 187.145 2

0.67

Astragaloside II-Mal

23.82

C₄₆H₇₂O₁₈

913.479 4

[M+H]⁺

0.2

599.403 4, 473.368 5, 455.353 6, 437.341 0, 419.335 8, 243.049 3, 261.061 4, 143.105 4

0.41

Astragaloside II-Mal-Ace

24, 91

C₄₈H₇₄O₁₉

955.490 7

[M+H]⁺

1.5

739.405 8, 721.399 8, 455.352 0, 437.340 8, 419.328 7, 303.070 8, 157.049 1, 143.105 5

0.63

Astragaloside I-Glc-Mal-Mal

23.37

C₅₇H₈₆O₂₇

1203.538 4

[M+H]⁺

-3.7

455.356 1, 437.344 1, 419.331 9, 401.310 7, 199.058 8, 157.049 4, 143.106 6, 97.028 3

0.64

HuangqiyeninA-Ara

22.48

C₄₁H₆₆O₁₄

783.451 2

[M+H]⁺

-1.7

453.334 5, 435.325 6, 417.313 4, 271.213 3, 143.107 9

0.53

Agroastragaloside IV

22.04

C₄₉H₈₀O₂₀

987.518 1

[M-H]^-

1.1

941.521 9

0.60

SoyasapogenolB-Glc-Rha

22.44

C₄₁H₆₆O₁₃

765.441 2

[M-H]^-

-2.4

603.394 5, 157.009 9, 113.020 5, 85.029 5

0.72

Vitamin B₂

14.66

C₁₇H₂₀N₄O₆

377.145 6

[M+H]⁺

0.0

243.087 0, 172.086 8, 57.032 9, 198.066 7, 145.076 2, 81.032 9, 57.033 1

1.99

FA 16:0

30.88

C₁₆H₃₂O₂

255.232 91

[M-H]^-

-0.5

255.232 4, 256.235 8, 257.239 1

1.69

FA 18:2

29.94

C₁₈H₃₂O₂

279.232 97

[M-H]^-

0.2

279.232 0, 280.235 5, 281.238 9

2.00

FA 18:2;O

26.43

C₁₈H₃₂O₃

295.224 7

[M-H]^-

-0.8

295.228 1, 296.231 0, 297.234 8

1.82

ST 28:3;O

24.90

C₂₈H₄₄O

419.328 92

[M+Na]⁺

0.5

419.329 5, 420.332 8, 421.336 2

0.74

DG 20:0

27.20

C₂₃H₄₄O₅

423.312 71

[M+Na]⁺

4.0

423.309 0, 331.266 2

2.65

LPG 16:0

31.98

C₂₂H₄₅O₉P

483.270 11

[M-H]^-

-0.2

483.272 5, 484.275 8, 485.279 2

1.97

LPC 16:0

26.61

C₂₄H₅₀NO₇P

496.339 48

[M+H]⁺

0.9

496.339 2, 497.342 5, 498.345 9

5.21

LPC 18:0

28.38

C₂₆H₅₄NO₇P

524.367 92

[M+H]⁺

0.4

524.371 2, 525.374 5, 526.377 9

2.61

PI 19:0

25.33

C₂₈H₅₃O₁₃P

627.317 32

[M-H]^-

2.3

627.317 3, 628.320 6, 629.324 0

3.89