中国药学杂志

No.	Medicinal part	Regions(in Chinese)	No.	Medicinal part	Regions(in Chinese)
S1	Rhizome	Wangqizhuang,Anguo,Hebei(河北安国王奇庄)	S12	Rhizome	Fucun,Anguo,Hebei(河北安国伏村)
S2	Rhizome	Wangqizhuang,Anguo,Hebei(河北安国王奇庄)	S13	Mother root	Mingguandian,Anguo,Hebei(河北安国明官店)
S3	Rhizome	Wangqizhuang,Anguo,Hebei(河北安国王奇庄)	S14	Mother root	Huozhuang,Anguo,Hebei(河北安国霍庄)
S4	Rhizome	Duancun,Anguo,Hebei(河北安国段村)	S15	Root	Dawunü,Anguo,Hebei(河北安国大五女)
S5	Rhizome	Duancun,Anguo,Hebei(河北安国段村)	S16	Root	Cuizhuang,Anguo,Hebei(河北安国崔庄)
S6	Rhizome	Huozhuang,Anguo,Hebei(河北安国霍庄)	S17	Root	Nanliu,Anguo,Hebei(河北安国南流)
S7	Rhizome	Huozhuang,Anguo,Hebei(河北安国霍庄)	S18	Root	Xihe,Anguo,Hebei(河北安国西河)
S8	Rhizome	Xishizhuang,Anguo,Hebei(河北安国西化庄)	S19	Root	Jiaozhuang,Anguo,Hebei(河北安国焦庄)
S9	Rhizome	Cuizhuang,Anguo,Hebei(河北安国崔庄)	S20	Root	Jiaozhuang,Anguo,Hebei(河北安国焦庄)
S10	Rhizome	Nanliu,Anguo,Hebei(河北安国南流)	S21	Root	Mingguandian,Anguo,Hebei(河北安国明官店)
S11	Rhizome	Mingguandian,Anguo,Hebei(河北安国明官店)	S22	Root	Mingguandian,Anguo,Hebei(河北安国明官店)

No.	Medicinal part	Regions(in Chinese)	No.	Medicinal part	Regions(in Chinese)
S1	Rhizome	Wangqizhuang,Anguo,Hebei(河北安国王奇庄)	S12	Rhizome	Fucun,Anguo,Hebei(河北安国伏村)
S2	Rhizome	Wangqizhuang,Anguo,Hebei(河北安国王奇庄)	S13	Mother root	Mingguandian,Anguo,Hebei(河北安国明官店)
S3	Rhizome	Wangqizhuang,Anguo,Hebei(河北安国王奇庄)	S14	Mother root	Huozhuang,Anguo,Hebei(河北安国霍庄)
S4	Rhizome	Duancun,Anguo,Hebei(河北安国段村)	S15	Root	Dawunü,Anguo,Hebei(河北安国大五女)
S5	Rhizome	Duancun,Anguo,Hebei(河北安国段村)	S16	Root	Cuizhuang,Anguo,Hebei(河北安国崔庄)
S6	Rhizome	Huozhuang,Anguo,Hebei(河北安国霍庄)	S17	Root	Nanliu,Anguo,Hebei(河北安国南流)
S7	Rhizome	Huozhuang,Anguo,Hebei(河北安国霍庄)	S18	Root	Xihe,Anguo,Hebei(河北安国西河)
S8	Rhizome	Xishizhuang,Anguo,Hebei(河北安国西化庄)	S19	Root	Jiaozhuang,Anguo,Hebei(河北安国焦庄)
S9	Rhizome	Cuizhuang,Anguo,Hebei(河北安国崔庄)	S20	Root	Jiaozhuang,Anguo,Hebei(河北安国焦庄)
S10	Rhizome	Nanliu,Anguo,Hebei(河北安国南流)	S21	Root	Mingguandian,Anguo,Hebei(河北安国明官店)
S11	Rhizome	Mingguandian,Anguo,Hebei(河北安国明官店)	S22	Root	Mingguandian,Anguo,Hebei(河北安国明官店)

Sample No.	Similarity	Sample No.	Similarity	Sample No.	Similarity
S1	0.926	S9	0.995	S17	0.975
S2	0.936	S10	0.976	S18	0.957
S3	0.954	S11	0.969	S19	0.920
S4	0.947	S12	0.951	S20	0.919
S5	0.929	S13	0.892	S21	0.955
S6	0.981	S14	0.898	S22	0.931
S7	0.985	S15	0.924
S8	0.978	S16	0.982

Sample No.	Similarity	Sample No.	Similarity	Sample No.	Similarity
S1	0.926	S9	0.995	S17	0.975
S2	0.936	S10	0.976	S18	0.957
S3	0.954	S11	0.969	S19	0.920
S4	0.947	S12	0.951	S20	0.919
S5	0.929	S13	0.892	S21	0.955
S6	0.981	S14	0.898	S22	0.931
S7	0.985	S15	0.924
S8	0.978	S16	0.982

Peak No.	Predicted compounds	Formula	m/z (Observe)	Molecular ion peak	Error /×10^-6	m/z (MS/MS)	References
1¹⁾	Chlorogenic acid	C₁₆H₁₈O₉	354.095 08	[M-H]^-	-0.8	353[M-H]^-;268[M-H-CHO-COOH]^-;162[M-H-C₇H₁₁O₆]^-	[15]
2	7-Hydroxycoumarin	C₉H₆O₃	162.031 69	[M+H]⁺	-0.3	135[M+H-CO]⁺;107[M+H-2CO]⁺	[16]
3¹⁾	Caffeic acid	C₉H₈O₄	180.042 26	[M-H]^-	0.7	179[M-H]^-;135[M-H-COOH]^-;90[M-H-C₆HO]^-	[17]
4	Rutin	C₂₇H₃₀O₁₆	610.153 39	[M-H]^-	-3.2	301[M-H-C₁₂H₂₀O₉]^-;273[M-H-C₁₃H₂₀O₁₀]^-;255[M-H- C₁₃H₂₂O₁₁]^-;151[M-H-C₂₀H₂₆O₁₂]^-	[18]
5¹⁾	Ferulic acid	C₁₀H₁₀O₄	194.057 91	[M-H]^-	-0.9	193[M-H]^-;133[M-H-CH₃-COOH]^-	[19]
6	Isochlorogenic acid C	C₂₅H₂₄O₁₂	516.126 78	[M-H]^-	-2.6	515[M-H]^-;353[M-H-C₉H₇O₃]^-;324[M-H-C₁₀H₇O₄]^-	[20]
7	Isochlorogenic acid A	C₂₅H₂₄O₁₂	516.126 78	[M-H]^-	-2.6	515[M-H]^-;380[M-H-C₈H₇O₂]^-;336[M-H-C₉H₇O₄]^-	[21]
8	Quercitrin	C₂₁H₂₀O₁₁	448.100 56	[M-H]^-	-2.8	447[M-H]^-;301[M-H-Rha]^-	[22]
9	Astin A	C₂₅H₃₃C₁₂N₅O₇	585.175 7	[M-H]^-	-3.4	584[M-H]^-;512[M-H-Cl₂]^-;362[M-H-C₈H₁₃N₂OCl₂]^-; 352[M-H-C₁₃H₁₄NO₃]^-	[23]
10¹⁾	Quercetin	C₁₅H₁₀O₇	302.042 65	[M-H]^-	-0.7	301[M-H]^-;151[M-H-C₈H₆O₃]^-;122[M-H-C₉H₇O₄]^-; 107[M-H-C₉H₆O₅]^-	[24]
11	Asterin	C₂₅H₃₃C₁₂N₅O₆	569.180 79	[M+H]⁺	0.1	570[M+H]⁺;485[M+H-C₄H₆NO]⁺;464[M+H-C₇H₈N]⁺; 338[M+H-C₁₃H₁₅N₂O₂]⁺
12¹⁾	Kaempferol	C₁₅H₁₀O₆	286.047 74	[M-H]^-	0.9	285[M-H]^-;153[M-H-C₇HO₃]^-;152[M-H-C₈H₅O₂]^-	[25]
13	Apigenin	C₁₅H₁₀O₅	270.052 82	[M-H]^-	-2.2	239[M-H-CH₃O]^-;153[M-H-C₈H₅O]^-;115[M-H- C₇H₅O₂-2OH]^-
14	Perillen	C₁₈H₃₀O₂	278.224 58	[M+H]⁺	-0.7	277[M-H]^-;196[M-H-C₅H₅O]^-;210[M-H-C₄H₃O]^-
15	Cholesteryl formate	C₂₈H₄₆O₂	414.349 78	[M+H]⁺	-2.7	415[M+H]⁺;327[M+H-C₆H₁₆]⁺	[26]
16	Poriferasterol monoglucoside	C₃₅H₅₈O₆	574.423 34	[M-H]^-	-3.4	573[M-H]^-;411[M-H-C₆H₁₀O₅]^-;440[M-H-C₅H₉O₄]^-
17¹⁾	Shionone	C₃₀H₅₀O	426.386 17	[M+H]⁺	0	427[M+H]⁺;372[M+H-C₄H₇]⁺;318[M+H-C₈H₁₃]⁺; 304[M+H-C₈H₁₁O]⁺;236[M+H-C₁₃H₁₉O]⁺;140 [M+H-C₂₀H₃₁O]⁺	[27]

Peak No.	Predicted compounds	Formula	m/z (Observe)	Molecular ion peak	Error /×10^-6	m/z (MS/MS)	References
1¹⁾	Chlorogenic acid	C₁₆H₁₈O₉	354.095 08	[M-H]^-	-0.8	353[M-H]^-;268[M-H-CHO-COOH]^-;162[M-H-C₇H₁₁O₆]^-	[15]
2	7-Hydroxycoumarin	C₉H₆O₃	162.031 69	[M+H]⁺	-0.3	135[M+H-CO]⁺;107[M+H-2CO]⁺	[16]
3¹⁾	Caffeic acid	C₉H₈O₄	180.042 26	[M-H]^-	0.7	179[M-H]^-;135[M-H-COOH]^-;90[M-H-C₆HO]^-	[17]
4	Rutin	C₂₇H₃₀O₁₆	610.153 39	[M-H]^-	-3.2	301[M-H-C₁₂H₂₀O₉]^-;273[M-H-C₁₃H₂₀O₁₀]^-;255[M-H- C₁₃H₂₂O₁₁]^-;151[M-H-C₂₀H₂₆O₁₂]^-	[18]
5¹⁾	Ferulic acid	C₁₀H₁₀O₄	194.057 91	[M-H]^-	-0.9	193[M-H]^-;133[M-H-CH₃-COOH]^-	[19]
6	Isochlorogenic acid C	C₂₅H₂₄O₁₂	516.126 78	[M-H]^-	-2.6	515[M-H]^-;353[M-H-C₉H₇O₃]^-;324[M-H-C₁₀H₇O₄]^-	[20]
7	Isochlorogenic acid A	C₂₅H₂₄O₁₂	516.126 78	[M-H]^-	-2.6	515[M-H]^-;380[M-H-C₈H₇O₂]^-;336[M-H-C₉H₇O₄]^-	[21]
8	Quercitrin	C₂₁H₂₀O₁₁	448.100 56	[M-H]^-	-2.8	447[M-H]^-;301[M-H-Rha]^-	[22]
9	Astin A	C₂₅H₃₃C₁₂N₅O₇	585.175 7	[M-H]^-	-3.4	584[M-H]^-;512[M-H-Cl₂]^-;362[M-H-C₈H₁₃N₂OCl₂]^-; 352[M-H-C₁₃H₁₄NO₃]^-	[23]
10¹⁾	Quercetin	C₁₅H₁₀O₇	302.042 65	[M-H]^-	-0.7	301[M-H]^-;151[M-H-C₈H₆O₃]^-;122[M-H-C₉H₇O₄]^-; 107[M-H-C₉H₆O₅]^-	[24]
11	Asterin	C₂₅H₃₃C₁₂N₅O₆	569.180 79	[M+H]⁺	0.1	570[M+H]⁺;485[M+H-C₄H₆NO]⁺;464[M+H-C₇H₈N]⁺; 338[M+H-C₁₃H₁₅N₂O₂]⁺
12¹⁾	Kaempferol	C₁₅H₁₀O₆	286.047 74	[M-H]^-	0.9	285[M-H]^-;153[M-H-C₇HO₃]^-;152[M-H-C₈H₅O₂]^-	[25]
13	Apigenin	C₁₅H₁₀O₅	270.052 82	[M-H]^-	-2.2	239[M-H-CH₃O]^-;153[M-H-C₈H₅O]^-;115[M-H- C₇H₅O₂-2OH]^-
14	Perillen	C₁₈H₃₀O₂	278.224 58	[M+H]⁺	-0.7	277[M-H]^-;196[M-H-C₅H₅O]^-;210[M-H-C₄H₃O]^-
15	Cholesteryl formate	C₂₈H₄₆O₂	414.349 78	[M+H]⁺	-2.7	415[M+H]⁺;327[M+H-C₆H₁₆]⁺	[26]
16	Poriferasterol monoglucoside	C₃₅H₅₈O₆	574.423 34	[M-H]^-	-3.4	573[M-H]^-;411[M-H-C₆H₁₀O₅]^-;440[M-H-C₅H₉O₄]^-
17¹⁾	Shionone	C₃₀H₅₀O	426.386 17	[M+H]⁺	0	427[M+H]⁺;372[M+H-C₄H₇]⁺;318[M+H-C₈H₁₃]⁺; 304[M+H-C₈H₁₁O]⁺;236[M+H-C₁₃H₁₉O]⁺;140 [M+H-C₂₀H₃₁O]⁺	[27]

DPPH	ABTS
S1	32.97	S12	43.57	S1	43.04	S12	47.52
S2	34.00	S13	38.15	S2	46.29	S13	46.29
S3	53.57	S14	35.26	S3	46.05	S14	45.63
S4	40.09	S15	35.56	S4	60.32	S15	51.92
S5	46.10	S16	29.98	S5	65.71	S16	57.65
S6	49.46	S17	38.15	S6	67.47	S17	47.28
S7	34.70	S18	34.62	S7	59.20	S18	52.16
S8	44.49	S19	30.07	S8	50.21	S19	46.43
S9	34.62	S20	31.87	S9	50.99	S20	46.19
S10	30.54	S21	31.22	S10	51.20	S21	44.91
S11	35.58	S22	35.67	S11	50.53	S22	51.92

DPPH	ABTS
S1	32.97	S12	43.57	S1	43.04	S12	47.52
S2	34.00	S13	38.15	S2	46.29	S13	46.29
S3	53.57	S14	35.26	S3	46.05	S14	45.63
S4	40.09	S15	35.56	S4	60.32	S15	51.92
S5	46.10	S16	29.98	S5	65.71	S16	57.65
S6	49.46	S17	38.15	S6	67.47	S17	47.28
S7	34.70	S18	34.62	S7	59.20	S18	52.16
S8	44.49	S19	30.07	S8	50.21	S19	46.43
S9	34.62	S20	31.87	S9	50.99	S20	46.19
S10	30.54	S21	31.22	S10	51.20	S21	44.91
S11	35.58	S22	35.67	S11	50.53	S22	51.92

DPPH correlation results	ABTS correlation results
17	0.907	1	17	0.873	1
9	0.868	2	9	0.866	2
4	0.863	3	5	0.859	3
14	0.857	4	4	0.847	4
13	0.848	5	12	0.845	5
2	0.846	6	2	0.835	6
7	0.844	7	6	0.833	7
5	0.838	8	13	0.827	8
12	0.832	9	7	0.82	9
10	0.817	10	15	0.814	10
11	0.816	11	1	0.805	11
6	0.81	12	14	0.795	12
15	0.808	13	11	0.786	13
1	0.782	14	8	0.763	14
8	0.776	15	3	0.759	15
3	0.738	16	10	0.758	16
16	0.589	17	16	0.617	17

DPPH correlation results	ABTS correlation results
17	0.907	1	17	0.873	1
9	0.868	2	9	0.866	2
4	0.863	3	5	0.859	3
14	0.857	4	4	0.847	4
13	0.848	5	12	0.845	5
2	0.846	6	2	0.835	6
7	0.844	7	6	0.833	7
5	0.838	8	13	0.827	8
12	0.832	9	7	0.82	9
10	0.817	10	15	0.814	10
11	0.816	11	1	0.805	11
6	0.81	12	14	0.795	12
15	0.808	13	11	0.786	13
1	0.782	14	8	0.763	14
8	0.776	15	3	0.759	15
3	0.738	16	10	0.758	16
16	0.589	17	16	0.617	17

祁紫菀UPLC指纹图谱的建立及其抗氧化谱效关系研究

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李蕊 , 徐荣荣 , 张楚涵 , 李葆林 , 甄亚钦 , 田宇柔 , 高乐 , 梁策 , 牛丽颖 , 王迎春 ^* , 冯薇 ^*

中国药学杂志 | 论著 2025,60(8): 809-817

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中国药学杂志 | 论著 2025, 60(8): 809-817

祁紫菀UPLC指纹图谱的建立及其抗氧化谱效关系研究

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李蕊, 徐荣荣, 张楚涵, 李葆林, 甄亚钦, 田宇柔, 高乐, 梁策, 牛丽颖, 王迎春^*, 冯薇^*

作者信息

河北中医药大学药学院, 中药材品质评价与标准化河北省工程研究中心, 石家庄 050091

李蕊,女,硕士研究方向:中药药效物质基础

通讯作者:

^*王迎春,女,硕士,高级实验师研究方向:中药药效物质基础Tel:(0311)85216828;

冯薇,女,博士,教授,博士生导师研究方向:中药药效物质基础 Tel:(0311)85216828

Establishment of UPLC Fingerprint and Effects of Antioxidant Related to Its Spectrum Study of Aster tataricus cv.Qiziwan

Rui LI, Rongrong XU, Chuhan ZHANG, Baolin LI, Yaqin ZHEN, Yurou TIAN, Le GAO, Ce LIANG, Liying NIU, Yingchun WANG^*, Wei FENG^*

Affiliations

School of Pharmaceutical Sciences, Quality Evaluation and Standardization Hebei Province Engineering Research Center of Traditional Chinese Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050091, China

出版时间: 2025-04-15 doi: 10.11669/cpj.2025.08.005

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摘要

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目的建立祁紫菀超高效液相色谱(UPLC)指纹图谱,采用超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF-MS)对共有峰进行成分鉴定,测定各批次祁紫菀抗氧化活性,并探讨祁紫菀的化学成分与抗氧化活性的谱效关系。方法采用《中药色谱指纹图谱相似度评价系统(2012版)》绘制22批祁紫菀药材的指纹图谱并进行相似度评价,标定共有峰。以1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除率和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除率为抗氧化指标,考察祁紫菀的抗氧化活性。采用灰色关联度和偏最小二乘回归分析祁紫菀化学成分与抗氧化活性的谱效关系。结果建立了22批不同批次祁紫菀药材的指纹图谱,其相似度为0.892~0.995。聚类分析可将祁紫菀母根、纯根和根茎区分开。利用对照品对比和UPLC-MS解析指认出17个共有峰。通过DPPH自由基和ABTS自由基清除实验发现,22批祁紫菀均有抗氧化能力,灰色关联分析表明,紫菀酮的关联度最高。结合偏最小二乘回归分析,初步证实芦丁、阿魏酸、异绿原酸A、astin A、槲皮素、紫苏烯和紫菀酮是祁紫菀发挥抗氧化活性的主要成分。结论本实验成功建立了祁紫菀的指纹图谱及其抗氧化活性谱效关系,可为祁紫菀的药效物质基础研究与质量控制提供参考。

关键词

祁紫菀 / 超高效液相色谱-四极杆-飞行时间质谱 / 指纹图谱 / 抗氧化 / 谱效关系

Abstract

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OBJECTIVE To establish UPLC fingerprint of Aster tataricus cv.Qiziwan, identify the compounds of common peaks by UPLC-Q-TOF-MS technology, determine antioxidant activity of each batch and study the relationship between effects of antioxidant and spectrum of Aster tataricus cv.Qiziwan. METHODS The fingerprints of 22 batches of Aster tataricus cv.Qiziwan were drawn and evaluated by the the Similarity Evaluation System of TCM Chromatographic Fingerprint (2012 edition), and the common peaks were calibrated. The antioxidant activity of Aster tataricus cv.Qiziwan was investigated by using the free radical scavenging rate of 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) and 2, 2'-azinobis- (3-ethylbenzthiazoline-6-sulphonate) (ABTS) free radical scavenging rate as the antioxidant index. The spectral effect relationship between chemical constituents and antioxidant activity of Aster tataricus cv.Qiziwan was analyzed by grey correlation degree and partial least squares regression. RESULTS The fingerprints of 22 batches of Aster tataricus cv.Qiziwan from different batchs were established, and the similarity was 0.892-0.995. Cluster analysis can distinguish rhizome with mother root, root and rhizome. Seventeen common peaks were identified by comparison of reference materials and UPLC-MS analysis. The DPPH free radical and ABTS free radical scavenging experiments showed that 22 batches of Aster tataricus cv.Qiziwan had antioxidant capacity, and the grey correlation analysis showed that shionone had the highest correlation degree. Combined with partial least squares regression analysis, it was preliminarily confirmed that rutin, ferulic acid, isochlorogenic acid A, astin A, quercetin, perillen and shionone were the main components of Aster tataricus cv.Qiziwan with antioxidant activity. CONCLUSION In this study, the fingerprint and antioxidant activity spectrum effect relationship of Aster tataricus cv. Qiziwan are successfully established, which can provide reference for the pharmacodynamic material basis research and quality control of Aster tataricus cv.Qiziwan.

Key words

Aster tataricus cv.Qiziwan / UPLC-Q-TOF-MS / fingerprint / antioxidant / spectrum-effect relationship

引用本文

李蕊, 徐荣荣, 张楚涵, 李葆林, 甄亚钦, 田宇柔, 高乐, 梁策, 牛丽颖, 王迎春, 冯薇. 祁紫菀UPLC指纹图谱的建立及其抗氧化谱效关系研究. 中国药学杂志, 2025 , 60 (8) : 809 -817 . DOI: 10.11669/cpj.2025.08.005

Rui LI, Rongrong XU, Chuhan ZHANG, Baolin LI, Yaqin ZHEN, Yurou TIAN, Le GAO, Ce LIANG, Liying NIU, Yingchun WANG, Wei FENG. Establishment of UPLC Fingerprint and Effects of Antioxidant Related to Its Spectrum Study of Aster tataricus cv.Qiziwan[J]. Chinese Pharmaceutical Journal, 2025 , 60 (8) : 809 -817 . DOI: 10.11669/cpj.2025.08.005

正文

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祁紫菀为菊科植物紫菀(Aster tataricus L. f.)的干燥根和根茎。春、秋二季采挖,除去有节的根茎(习称“母根”)和泥沙,编成辫状晒干,或直接晒干^[1]。现代研究证明母根的有效成分含量同样符合《中国药典》2020年版标准^[2],但现代紫菀均采用无性繁殖的方式人工栽培,母根被用作繁殖材料,故除去母根只保留无节的根茎^[3]。河北省安国市古称祁州,以传统的“八大祁药”闻名海外。目前安国所产紫菀,根粗且长,质柔韧。因其质地纯正,药效良好,畅销全国各地,故名“祁紫菀”^[4]。本品润肺下气,消痰止咳,常用于痰多喘咳、新久咳嗽、劳嗽咳血^[5],具有镇咳祛痰、抗癌、抗衰老、抗病毒等药理作用^[6]。有研究表明,紫菀抗癌、抗衰老的作用可能与抗氧化活性相关^[7]。目前,祁紫菀的研究主要集中在总黄酮化学成分含量测定^[8]、挥发油化学组分的研究^[9]、对紫菀中紫菀酮、槲皮素、芹菜素的含量测定^[10]以及基于一测多评法对紫菀多成分含量测定及质量评价^[11]等方面,针对祁紫菀超高效液相色谱(UPLC)指纹图谱及抗氧化谱效关系方面的研究未见报道。

中药指纹图谱具有信息量大、特征性强、整体性强的特点,能够直观地展现制剂中各药味的化学组成与分布,为全面控制质量和深入药效研究提供了重要依据^[12]。中药谱效关系则可以将中药指纹图谱与中药药效通过各种数据处理方法进行分析,筛选出与药效密切相关的中药成分^[13]。本实验采用UPLC法建立了祁紫菀的指纹图谱,并结合聚类分析对祁紫菀进行质量评价,同时采用超高效液相色谱-质谱联用技术(UPLC-MS)和对照品进行比对,对共有峰进行鉴定。利用1,1-二苯基-2-三硝基苯肼(DPPH)法和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)法检测体外抗氧化能力,通过谱效关系,初步揭示祁紫菀抗氧化能力的活性物质,为祁紫菀的质量控制及开发研究提供理论支持。

1 仪器与试药

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1.1 仪器

H-Class型超高效液相色谱仪(美国Waters公司);Triple-TOF^TM6600⁺型四极杆串联飞行时间高分辨质谱仪(美国AB SCIEX公司,Analyst TF 1.8.1数据采集软件、PeakView 1.2数据处理软件);YB-150型多功能粉碎机(永康市速锋工贸有限公司);CPA225D型电子天平、BSA224S-CW型电子天平(德国赛多利斯有限公司);KQ-250E型超声波清洗器(昆山市超声仪器有限公司);H2050R高速离心机(湖南赛特湘仪离心机仪器有限公司);ELx800型酶标仪(美国Bio-Tek公司);总抗氧化能力(DPPH法)试剂盒(苏州科铭生物技术有限公司);总抗氧化能力(ABTS法)试剂盒(苏州科铭生物技术有限公司)。

1.2 药品与试剂

对照品:绿原酸(批号BWC9013-2016,纯度≥98%)、阿魏酸(批号BWJ4346-2016,纯度≥99%)、槲皮素(批号BWC9014-2016,纯度≥98%)、山柰酚(批号BWC9039-2016,纯度≥98%)(北京北方伟业计量研究院);咖啡酸(DSTDK001301,纯度≥98%)、紫菀酮(DST221008-033,纯度≥98%)(成都乐美天医药科技有限公司);甲酸、乙腈为色谱纯,其他试剂均为分析纯;实验用水为超纯水(Millipore超纯水系统制备)。

22批祁紫菀药材自河北安国收集,经侯芳洁副教授鉴定为菊科植物紫菀(Aster tataricus L.f.)的干燥根和根茎。其中,S1~S12为不带母根的茎条(根茎),S13、S14为母根,S15~S22为纯根。22批祁紫菀药材的信息见表1。

2 方法与结果

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2.1 祁紫菀指纹图谱的建立

2.1.1 色谱条件

菲罗门 Titank C₁₈色谱柱(2.1 mm×150 mm,1.8 μm),流动相为0.2%甲酸水溶液(A)-乙腈(B)梯度洗脱(0~3 min,5%~9%B;3~13 min,9%~10%B;13~15 min,10%~16%B;15~20 min,16%B;20~48 min,16%~28%B;48~54 min,28%~51%B;54~66 min,51%~100%B;66~74 min,100%B;74~76 min,100%~5%B),流速为0.3 mL·min^-1,柱温为40 ℃,进样量为2 μL。分时段波长如下:325 nm(0~48 min)、260 nm(48~70 min)、203 nm(70~76 min)。

2.1.2 对照品溶液制备

分别精密称取绿原酸、咖啡酸、槲皮素、阿魏酸、山柰酚、紫菀酮对照品适量,加甲醇溶解,制成单一对照品储备液。精密量取上述各对照品储备液适量,加甲醇稀释,制成质量浓度分别为117.7、7.92、12.84、37.49、64.82和347.28 μg·mL^-1的混合对照品溶液。

2.1.3 供试品溶液制备

取紫菀粉末(过60目筛)2.0 g,精密称定,置50 mL锥形瓶中,加入20 mL甲醇,称定质量,超声处理(功率250 W,频率40 kHz)30 min,静置至室温,再次称定质量,用甲醇补足减失的质量,摇匀,用0.22 μm微孔滤膜滤过,取续滤液,即得。

2.2 指纹图谱方法学考察

2.2.1 精密度实验

取S1号紫菀粉末,按“2.1.3”项下方法制备供试品溶液,按“2.1.1”项下色谱条件连续进样6次,以1号峰(绿原酸)为参照峰,计算各共有峰的相对保留时间相对标准偏差(RSD)为0.05%~0.30%,相对峰面积的RSD分别为2.00%、2.89%、2.65%、2.09%、1.66%、1.62%、1.57%、1.80%、1.43%、2.90%、1.67%、2.16%、1.58%、1.34%、1.50%和1.92%,均小于2.91%,结果表明,仪器精密度良好。

2.2.2 重复性实验

取S1号紫菀粉末,按“2.1.3”项下方法平行制备供试品溶液6份,按“2.1.1”项下色谱条件测定,以1号峰(绿原酸)为参照峰,计算各共有峰的相对保留时间RSD为0.07%~1.60%,相对峰面积的RSD分别为0.11%、0.98%、0.29%、0.17%、0.12%、0.09%、0.09%、0.19%、0.16%、0.42%、0.11%、0.19%、0.20%、0.14%、0.16%和0.10%,均小于0.99%,表明该方法重复性良好。

2.2.3 稳定性实验

取S1号紫菀粉末,按“2.1.3”项下方法制备供试品溶液,按“2.1.1”项下色谱条件在24 h内连续进样6次,以1号峰(绿原酸)为参照峰,计算各共有峰的相对保留时间RSD为0.13%~1.48%,相对峰面积的RSD分别为0.87%、1.61%、0.69%、0.75%、0.71%、0.75%、0.71%、1.00%、1.00%、0.88%、0.71%、0.78%、0.75%、0.54%、0.68%和0.76%,均小于1.62%,结果表明,样品溶液在24 h内相对稳定。

2.3 对照指纹图谱的生成与相似度分析

取22批祁紫菀样品适量按“2.1.3”项下方法制备供试品溶液,按“2.1.1”项下色谱条件依次进样检测,记录色谱图。将22批祁紫菀药材样品的UPLC图谱导入《中药色谱指纹图谱相似度评价系统》(2012 版)软件,设定S1为参照色谱图,选取时间窗格宽度0.1 min,通过中位数生成对照指纹图谱,运用多点校正对标记(Mark)色谱峰进行匹配,计算相似度。结果表明,除S13、S14的相似度为0.892、0.898外,其余批次样品与对照指纹图谱的相似度均大于0.919,见表2。其中,S13、S14为祁紫菀母根,其余样品为祁紫菀根茎与纯根。纯根与根茎类样品相似度较高,化学成分相似,质量较稳定。共标定17个共有峰,见图1。鉴于紫菀母根非《中国药典》2020年版正式收录的药用部位,因此,本实验为确保实验结果的科学性与合规性,将含有紫菀母根的样品批次S13与S14从中剔除后再生成对照图谱。对照指纹图谱共有峰模式见图2。

2.4 聚类分析

将22批祁紫菀样品的17个共有峰的峰面积导入SPSS 26.0,采用中位数联接系统聚类法,在平方欧氏距离为10时,22批祁紫菀样品聚为两大类,S1~S12、S15~S22聚为第一类,S13、S14聚为第二类。其中,S13、S14为祁紫菀母根,其余批次为纯根与根茎,表明祁紫菀纯根与根茎相似度较高,与母根差异较大,与相似度结果一致。22批祁紫菀的聚类分析谱系图见图3。

2.5 指纹图谱中共有峰的鉴定

2.5.1 色谱条件离子模式:分别在正、负离子模式下分析检测样品。

仪器参数优化后设置如下:离子源(正离子模式):电喷雾离子源(ESI);雾化电压(ISVF):+5 500 V;雾化温度(TEM):550 ℃;解簇电压(DP):+80 V;碰撞能量(CE):+45 eV;雾化气压力(Gas 1,N₂)、辅助气压力(Gas 2,N₂)、气帘气压力(Cur,N₂)分别为:344 737.85、379 211.63、241 316.49 Pa;高分辨质谱扫描质量范围设置为m/z 100~1 200,累积时间:250 ms;碎片离子扫描质量范围设置为50~1 200,累积时间:100 ms;碰撞能量扩展(CES):20 eV,信息依赖获取(IDA)方式开启。

离子源(负离子模式):ESI;雾化电压(ISVF):-4 500 V;TEM:550 ℃;DP:-80 V;CE:-45 eV;Gas 1、Gas 2、Cur分别为:344 737.85、379 211.63、241 316.49 Pa;其余各时间等参数同正离子模式保持一致。

2.5.2 共有峰鉴定

按“2.1.3”制备供试品溶液,“2.1.1”项下液相条件和“2.5.1”项下质谱条件下获得紫菀的正、负离子流图,通过采用Peak View 1.2软件中“XIC Manager”功能进行数据分析,通过比对精确相对分子质量和特征碎片离子^[14],鉴定出祁紫菀指纹图谱中共有峰的结构,结果见表3。祁紫菀的共有峰鉴定以峰10(槲皮素)为例,在负离子模式下,其准分子离子峰为m/z 301 [M-H]^-,母离子通过黄酮类化合物的经典裂解途径——逆狄尔斯-阿尔德反应(RDA)裂解(途径Ⅰ和Ⅱ),得到碎片离子m/z 151[M-H-C₈H₆O₃]^-和m/z 107[M-H-C₉H₆O₅]^-,其均为黄酮类化合物的特征碎片离子。准分子离子峰m/z 301[M-H]^-再丢失1分子-C₈H₃O₅得到碎片离子m/z 122[M-H-C₈H₃O₅]^-。其二级质谱和裂解过程见图4。

2.6 抗氧化活性考察

2.6.1 DPPH自由基清除率

取祁紫菀供试品溶液5 μL至EP管中,加15 μL甲醇稀释,加入380 μL的DPPH试剂,充分混匀,室温避光孵育20 min,取200 μL置96孔板中,即得样品溶液。平行实验重复3次。取20 μL甲醇至EP管中,加入380 μL DPPH试剂,充分混匀,室温避光孵育20 min,取200 μL置96孔板中,即得空白溶液。酶标仪515 nm下测定吸光度值。

祁紫菀总抗氧化能力以自由基清除率表示,见公式1。

公式(1)DPPH自由基清除率(%)=[(A_空白-A_测定)/A_空白]×100%

2.6.2 ABTS自由基清除率

按试剂盒说明书制备ABTS工作液,将上述“2.1.3”项下供试品溶液稀释40倍后,分别精密吸取10 μL至EP管中,加入ABTS工作液190 μL,充分混匀,取200 μL置96孔板中,即得样品溶液。平行实验重复3次。取体积分数70%甲醇10 μL至EP管中,加入ABTS工作液190 μL,充分混匀,取200 μL置96孔板中,即得空白溶液。酶标仪734 nm下测定吸光度值。祁紫菀的总抗氧化能力以ABTS自由基清除率表示(公式2):

公式(2)ABTS自由基清除率(%)=[(A_空白-A_测定)/A_空白]×100%

2.6.3 自由基清除活性测定结果

本实验采用DPPH、ABTS法评价祁紫菀的抗氧化能力,抗氧化活性结果见表4。结果表明,22批祁紫菀均具有清除自由基的能力,且祁紫菀对ABTS自由基具有更强的清除力。DPPH自由基清除率均在30%以上,ABTS自由基清除率均在43%以上,祁紫菀母根与根茎、纯根的抗氧化活性相差不大,说明药用部位对祁紫菀抗氧化活性无显著影响。

2.7 灰色关联度分析

参考Liu^[28]的灰色关联分析方法,对原始数据进行无量纲化处理,采用初值法,以各批祁紫菀药材(剔除母根)UPLC指纹图谱共有峰峰面积数据为比较序列,抗氧化活性为参考序列,计算灰色关联系数。

关联度计算结果见表5。通过关联度可初步确定共有峰所对应的化学成分与抗氧化活性之间的联系。关联度越大,说明化学成分与抗氧化作用的关联越大,反之则表示关联越小^[29]。实验结果可知,除16号峰外,祁紫菀17个共有峰清除DPPH和ABTS 2种自由基活性的关联度均大于0.7^[30],有较高的关联度,表明祁紫菀中的多个组分可能对抗氧化性具有协同增效作用。各共有峰清除DPPH自由基的关联序为17>9>4>>14>13>2>7>5>12>10>11>6>15>1>8>3>16,清除ABTS自由基的关联序为17>9>5>4>12>2>6>13>7>15>1>14>11>8>3>10>16,其中与抗氧化活性关联度最大的是17号峰(紫菀酮)。

2.8 偏最小二乘回归分析(PLS)

2.8.1 PLS回归方程的建立

灰色关联度较为直观地反映不同峰与抗氧化作用的关联度大小,但无法呈现两者的正负相关性,故需结合PLS综合分析。以各批祁紫菀药材(剔除母根)UPLC指纹图谱共有峰峰面积数据为自变量x,以祁紫菀提取物抗氧化能力为因变量y,导入SIMCA 14.1软件作PLS模型拟合,得到回归方程y₁=-0.054 011 1x₁+0.328 721x₂+0.058 301 3x₃+0.118 432x₄+0.106 862x₅+0.033 875 2x₆+0.116 79x₇-0.062 372 1x₈+0.124 063x₉+0.055 667 9x₁₀+0.036 719 3x₁₁+0.076 471 5x₁₂+0.041 831 5x₁₃+0.122 915x₁₄+0.018 005 5x₁₅+0.008 410 73x₁₆+0.153 66x₁₇,y₂=-0.020 731 8x₁+0.037 154 1x₂+0.060 098 8x₃+0.099 948 4x₄+0.114 109x₅+0.028 446 5x₆+0.093 399 6x₇-0.007 859 91x₈+0.096 463 1x₉+0.056 051 8x₁₀+0.038 816 9x₁₁+0.076 226 4x₁₂+0.371 925x₁₃+0.094 348 9x₁₄+0.041 200 8x₁₅+0.067 71x₁₆+0.117 33x₁₇。回归方程中y₁、y₂分别对应DPPH、ABTS的自由基清除率,回归系数为正值说明其与自由基清除率呈正相关,负数则表示与药效呈负相关。结果见图5、6。结果表明,峰2、3、4、5、6、7、9、10、11、12、13、14、15、16、17与DPPH自由基清除能力呈正相关,贡献度依次为17>9>14>4>7>5>12>3>10>13>11>6>2>15>6;峰2、3、4、5、6、7、9、12、13、14、15、16、17与ABTS自由基清除能力呈正相关,贡献度依次为17>5>4>9>14>7>12>16>3>10>15>11>13>2>6。

2.8.2 变量投影重要性(variable importance in projection, VIP)分析

VIP是反映自变量对因变量解释能力的重要指标,其值越大说明该自变量对因变量的解释能力越强。一般认为当VIP>1时,自变量在解释因变量时具有显著重要性。将自变量X(各共有峰峰面积)与因变量Y(自由基清除率)导入SIMCA 14.1 软件进行变量投影重要性分析,生成VIP,见图7、8。由图7、8可知,在以DPPH为指标时,VIP值由大到小的峰号顺序为17>9>14>4>7>5>10,以ABTS自由基清除能力为指标时,VIP值由大到小的峰号顺序为17>9>14>5>4>10>12>7>6>13>2。

3 讨论

收起

紫菀中富含多种药效成分^[31],本实验以《中国药典》2020年版一部规定的紫菀甲醇提取物为对象,建立了指纹图谱,并对其相似度进行评价。纯根与根茎类样品相似度较高,化学成分相似,采用中位数联接系统聚类法,可以清晰地将祁紫菀母根分开。以DPPH自由基和ABTS自由基的清除率为抗氧化活性的药效学指标,分析了22批祁紫菀的抗氧化活性。结果显示,22批祁紫菀均有抗氧化能力,且药用部位对其抗氧化活性无显著影响。以灰色关联度>0.7,PLS分析回归系数为正数(呈现正相关)且VIP值>1为标准^[30],筛选出祁紫菀产生抗氧化作用的关键成分。可综合得出17(紫菀酮)、9(astin A)、14(紫苏烯)、4(芦丁)、7(异绿原酸A)、5(阿魏酸)和10(槲皮素)号峰是与紫菀抗氧化作用关联度较大的色谱峰,说明这7个峰是反映其药效的重要特征峰,对抗氧化活性贡献较大。

在深入探究祁紫菀药材的母根、纯根及其根茎的化学成分特征时,相较于纯根与根茎,母根中紫菀酮的峰面积显著降低,降幅约为30%,这一发现为药典不将母根作为药用部位的规定提供了实验依据。然而,母根中绿原酸(标记为1号峰)的峰面积相较于纯根与根茎显著高出近1倍,而咖啡酸(标记为3号峰)的峰面积则高出约1.5倍,这一发现提示母根在特定化学成分富集方面具有独特优势。鉴于上述差异,建议在针对紫菀中某一类特定化学成分进行深入研究时,应优先考虑选取该成分含量较高的部位作为分析对象,以优化实验设计与资源利用率。后续研究计划对祁紫菀母根、纯根及根茎中的有效成分进行更为全面与深入的定量分析,旨在明确这些成分在不同药用部位的具体含量及其变化规律。为紫菀药材的质量控制标准建立、药效物质基础阐明以及资源的合理利用提供重要参考,进一步推动中药材现代化研究与产业发展。

基金

收起

河北省省级科技计划项目资助(21372503D)

参考文献

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文献

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参考文献引证文献

排序方式：

[1]

Ch.P (2020) Vol Ⅰ(中国药典2020年版.一部)[S]. 2020:177-178.

[2]

GUO

W N

, CHENG

, FANG

C W

. Studies on structure, accumulation of chief chemical constituents and content of Aster tataricus L. f. ' maternal root[J]. Lishizhen Med Mater Med Res(时珍国医国药), 2016, 27(11):2614-2616.

[3]

H Y

, LI

W J

, DING

X Y

, et al. Herbal textual research on Asteris Radix et Rhizoma in famous classical formulas[J]. Chin J Exp Tradit Med Form(中国实验方剂学杂志), 2024, 30(7):20-30.

[4]

LIANG

C Y

. Accumulation characteristics, speciationanalysis, and risk assessment of heavymetals in Aster tataricusL.f.[D]. Baoding: Hebei University, 2024.

[5]

CHENG

C G

, CHENG

Z F

, LIU

X H

. Microwave-assisted and pressurized solvent extraction of shionone from Tatarian aster root[J]. Chin Pharm J(中国药学杂志), 2006, 41(7):502-505.

[6]

YAO

P B

, LIU

Y L

, ZHU

Z G

, et al. Effects and aster water extract on proliferation and invasion of human lung cancer A549 cells, and tumorigenesis ability of nude mice[J]. Chin J Pharm Anal(药物分析杂志), 2022, 42(3):380-386.

[7]

PENG

W J

, XIN

R H

, REN

L H

, et al. Progress on chemical constituents and pharmacological effects of Aster tataricus[J]. Prog Vet Med(动物医学进展), 2015, 36(3):102-107.

[8]

Y Y

, LI

H J

, HOU

G G

, et al. Purification process for total flavonoids in Asteris Radix et Rhizoma with macroporous absorption resin[J]. Chin Tradit Pat Med(中成药), 2019, 41(3):501-505.

[9]

YANG

, XIAO

Y Q

, LIANG

R X

, et al. Studies on expectorant compounds in volatile oil from root and rhizome of Aster tataricus[J]. China J Chin Mater Med(中国中药杂志), 2008, 33(3):281-283.

[10]

SHI

X L

, ZHU

. Determination of apigenin, quercetin and Asterone in aster from different places by HPLC wavelength switching[J]. Chem Eng Des Common(化工设计通讯), 2022, 48(7):184-186.

[11]

X Y

, ZHANG

Y X

, ZHAO

R Z

, et al. Quality evaluation of Aster souliei based on quantitative analysis of multi-components by single marker[J]. Acta Pharm Sin(药学学报), 2022, 57(3):775-782.

[12]

LIN

, WANG

, GAO

S S

, et al. Quality evaluation of Aster souliei based on quantitative analysis of multi-components by single marker[J]. Res Pract Chin Med(现代中药研究与实践), 2022, 36(5):54-59.

[13]

Y R

, ZHOU

W W

, WANG

Z Y

, et al. Relationship between antioxidant activity and spectrum-effect of Hawthorn leaf extracts[J]. Chin Pharm J(中国药学杂志), 2020, 55(20):1673-1679.

[14]

, LI

. The effects of shionone on NF-κB pathway and oxidative stress in ovalbumin-induced asthmatic immature rats[J]. Curr Immunol(现代免疫学), 2023, 43(3):226-232.

[15]

YANG

L F

, TIAN

, ZHENG

, et al. Analysis of the flavonoids in Ziziphi Spinosae Semen using UPLC/ESI-Q TRAP-MS/MS[J]. Spec Wild Econ Anim Plant Res(特产研究), 2022, 44(3):91-100.

[16]

ZHENG

, HUANG

, ZHANG

J L

, et al. Study on the chemical composition of Jiangxi specialty Chinese medicine Chaxiong based on UPLC-Q-TOF-MS/MS[J]. Nat Prod Res Dev(天然产物研究与开发), 2024, 36(1):52-62.

[17]

XIAO

G L

, ZHONG

H X

, HUANG

H J

, et al. Identification of chemical constituents in Xiao'er Qingyan Granules by UPLC-Q-TOF-MS/MS[J]. Chin J Mod Appl Pharm(中国现代应用药学), 2022, 39(12):1627-1636.

[18]

GAO

Y T

, SONG

X P

, LI

, et al. Fingerprint and identification of chemical constituents of the extract of Fructus Ligustri Lucidi-Ecliptae Herba[J]. J Nanjing Univ Tradit Chin Med(南京中医药大学学报), 2024, 40(4):399-412.

[19]

CUI

W H

, DENG

X L

, CHEN

C Y

. Rapid identification of chemical constituents in Taraxaci Herba Roots based on UPLC-Q-Exactive-Orbitrap MS[J]. Henan Sci(河南科学), 2023, 41(7):978-984.

[20]

CHEN

F C

, GAO

S H

, WANG

Z P

, et al. Identification of chemical components in Actinidia chinensis root by UHPLC-Q-TOF-MS/MS[J]. J China Pharm(中国药房), 2020, 31(14):1725-1731.

[21]

LIAO

, CHEN

, LI

Y R

, et al. Qualitative analysis of Zhuang Medicine Tongfeng Li'an Capsule by Q-Exactive Orbitrap HRMS mass spectrometry[J]. Chin Pharm J(中国药学杂志), 2024, 59(14):1339-1346.

[22]

R N

, LUO

W Z

, WEI

R R

, et al. Rapid identification of constituents of Urtica hyperborea using UPLC-ESI-Q-TOF-MS/MS method[J]. China J Chin Mater Med(中国中药杂志), 2019, 44(8):1607-1614.

[23]

CHEN

S L

, LIU

, FENG

M H

, et al. Identification of peptides in Aster tataricus by the strategy of UHPLC-Q-TOF-MS/MS combined with molecular network[J]. J Chin Mass Spectrom Soc(质谱学报), 2023, 44(3):397-411.

[24]

SHI

, LI

, ZHOU

, et al. Analysis of chemical constituents and components absorbed into plasma of Ardisia crenata based on UPLC-QE-HF-MS/MS[J]. J China Pharm(中国药房), 2024, 35(3):316-321.

[25]

LUO

Y T

, ZENG

Y J

, PENG

J Y

, et al. Chemical composition analysis of Tibetan medicine Ji-ni-de-xie based on UPLC-Q-Executive Orbitrap MS technology[J]. Chin J New Drugs(中国新药杂志), 2022, 31(17):1727-1735.

[26]

YANG

R R

. Quality control of Caulis Spatholobi and its antithrombotic effect on zebrafish[D]. Beijing: Beijing University of Chinese Medicine, 2018.

[27]

CHEN

Y J

, WU

, WEI

Z Y

, et al. Identification of chemical constituents in Aster tataricus by UHPLC-Q-TOF-MS[J]. Acta Pharm Sin(药学学报), 2019, 54(9):1645-1654.

[28]

SHU

Y H

, JIAN

Q P

, ZHENG

Y K

, et al. Research on the spectrum-effect relationship of antimicrobial activity of coptidis rhizoma based on grey relational analysis and orthogonal partial least squares method[J]. Lishizhen Med Mater Med Res(时珍国医国药), 2023, 34(9):2298-2302.

[29]

HUANG

Y F

, CAI

, RAN

, et al. Preliminary discovery of quality marker of herb Meconopsis integrifolia (non flower parts) based on spectrum-effect relationship[J]. Chin Tradit Herb Drugs(中草药), 2020, 51(17):4521-4530.

[30]

LIU

T H

, MA

, WANG

B C

, et al. Effective components and mechanism of Cistanches Herba aqueous extract against diabetic nephropathy based on the spectral-effect relationship[J]. Chin J Mod Appl Pharm(中国现代应用药学), 2024, 41(10):1313-1323.

[31]

CAI

Y J

, SHI

X L

, LIU

H Y

, et al. Effective components and pharmacological effects of Asteris Radix[J]. J Tradit Chin Vet Med(中兽医医药杂志), 2023, 42(2):39-42.

2025年第60卷第8期

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doi: 10.11669/cpj.2025.08.005

接收时间：2024-10-23
首发时间：2025-11-12
出版时间：2025-04-15

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收稿日期：2024-10-23

基金

河北省省级科技计划项目资助(21372503D)

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河北中医药大学药学院, 中药材品质评价与标准化河北省工程研究中心, 石家庄 050091

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^*王迎春,女,硕士,高级实验师研究方向:中药药效物质基础Tel:(0311)85216828;

冯薇,女,博士,教授,博士生导师研究方向:中药药效物质基础 Tel:(0311)85216828

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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No.	Medicinal part	Regions(in Chinese)	No.	Medicinal part	Regions(in Chinese)
S1	Rhizome	Wangqizhuang,Anguo,Hebei(河北安国王奇庄)	S12	Rhizome	Fucun,Anguo,Hebei(河北安国伏村)
S2	Rhizome	Wangqizhuang,Anguo,Hebei(河北安国王奇庄)	S13	Mother root	Mingguandian,Anguo,Hebei(河北安国明官店)
S3	Rhizome	Wangqizhuang,Anguo,Hebei(河北安国王奇庄)	S14	Mother root	Huozhuang,Anguo,Hebei(河北安国霍庄)
S4	Rhizome	Duancun,Anguo,Hebei(河北安国段村)	S15	Root	Dawunü,Anguo,Hebei(河北安国大五女)
S5	Rhizome	Duancun,Anguo,Hebei(河北安国段村)	S16	Root	Cuizhuang,Anguo,Hebei(河北安国崔庄)
S6	Rhizome	Huozhuang,Anguo,Hebei(河北安国霍庄)	S17	Root	Nanliu,Anguo,Hebei(河北安国南流)
S7	Rhizome	Huozhuang,Anguo,Hebei(河北安国霍庄)	S18	Root	Xihe,Anguo,Hebei(河北安国西河)
S8	Rhizome	Xishizhuang,Anguo,Hebei(河北安国西化庄)	S19	Root	Jiaozhuang,Anguo,Hebei(河北安国焦庄)
S9	Rhizome	Cuizhuang,Anguo,Hebei(河北安国崔庄)	S20	Root	Jiaozhuang,Anguo,Hebei(河北安国焦庄)
S10	Rhizome	Nanliu,Anguo,Hebei(河北安国南流)	S21	Root	Mingguandian,Anguo,Hebei(河北安国明官店)
S11	Rhizome	Mingguandian,Anguo,Hebei(河北安国明官店)	S22	Root	Mingguandian,Anguo,Hebei(河北安国明官店)

No.

Medicinal part

Regions(in Chinese)

No.

Medicinal part

Regions(in Chinese)

Rhizome

Wangqizhuang,Anguo,Hebei(河北安国王奇庄)

S12

Rhizome

Fucun,Anguo,Hebei(河北安国伏村)

Rhizome

Wangqizhuang,Anguo,Hebei(河北安国王奇庄)

S13

Mother root

Mingguandian,Anguo,Hebei(河北安国明官店)

Rhizome

Wangqizhuang,Anguo,Hebei(河北安国王奇庄)

S14

Mother root

Huozhuang,Anguo,Hebei(河北安国霍庄)

Rhizome

Duancun,Anguo,Hebei(河北安国段村)

S15

Root

Dawunü,Anguo,Hebei(河北安国大五女)

Rhizome

Duancun,Anguo,Hebei(河北安国段村)

S16

Root

Cuizhuang,Anguo,Hebei(河北安国崔庄)

Rhizome

Huozhuang,Anguo,Hebei(河北安国霍庄)

S17

Root

Nanliu,Anguo,Hebei(河北安国南流)

Rhizome

Huozhuang,Anguo,Hebei(河北安国霍庄)

S18

Root

Xihe,Anguo,Hebei(河北安国西河)

Rhizome

Xishizhuang,Anguo,Hebei(河北安国西化庄)

S19

Root

Jiaozhuang,Anguo,Hebei(河北安国焦庄)

Rhizome

Cuizhuang,Anguo,Hebei(河北安国崔庄)

S20

Root

Jiaozhuang,Anguo,Hebei(河北安国焦庄)

S10

Rhizome

Nanliu,Anguo,Hebei(河北安国南流)

S21

Root

Mingguandian,Anguo,Hebei(河北安国明官店)

S11

Rhizome

Mingguandian,Anguo,Hebei(河北安国明官店)

S22

Root

Mingguandian,Anguo,Hebei(河北安国明官店)

Sample No.	Similarity	Sample No.	Similarity	Sample No.	Similarity
S1	0.926	S9	0.995	S17	0.975
S2	0.936	S10	0.976	S18	0.957
S3	0.954	S11	0.969	S19	0.920
S4	0.947	S12	0.951	S20	0.919
S5	0.929	S13	0.892	S21	0.955
S6	0.981	S14	0.898	S22	0.931
S7	0.985	S15	0.924
S8	0.978	S16	0.982

Sample No.

Similarity

Sample No.

Similarity

Sample No.

Similarity

0.926

0.995

S17

0.975

0.936

S10

0.976

S18

0.957

0.954

S11

0.969

S19

0.920

0.947

S12

0.951

S20

0.919

0.929

S13

0.892

S21

0.955

0.981

S14

0.898

S22

0.931

0.985

S15

0.924

0.978

S16

0.982

Peak No.	Predicted compounds	Formula	m/z (Observe)	Molecular ion peak	Error /×10^-6	m/z (MS/MS)	References
1¹⁾	Chlorogenic acid	C₁₆H₁₈O₉	354.095 08	[M-H]^-	-0.8	353[M-H]^-;268[M-H-CHO-COOH]^-;162[M-H-C₇H₁₁O₆]^-	[15]
2	7-Hydroxycoumarin	C₉H₆O₃	162.031 69	[M+H]⁺	-0.3	135[M+H-CO]⁺;107[M+H-2CO]⁺	[16]
3¹⁾	Caffeic acid	C₉H₈O₄	180.042 26	[M-H]^-	0.7	179[M-H]^-;135[M-H-COOH]^-;90[M-H-C₆HO]^-	[17]
4	Rutin	C₂₇H₃₀O₁₆	610.153 39	[M-H]^-	-3.2	301[M-H-C₁₂H₂₀O₉]^-;273[M-H-C₁₃H₂₀O₁₀]^-;255[M-H- C₁₃H₂₂O₁₁]^-;151[M-H-C₂₀H₂₆O₁₂]^-	[18]
5¹⁾	Ferulic acid	C₁₀H₁₀O₄	194.057 91	[M-H]^-	-0.9	193[M-H]^-;133[M-H-CH₃-COOH]^-	[19]
6	Isochlorogenic acid C	C₂₅H₂₄O₁₂	516.126 78	[M-H]^-	-2.6	515[M-H]^-;353[M-H-C₉H₇O₃]^-;324[M-H-C₁₀H₇O₄]^-	[20]
7	Isochlorogenic acid A	C₂₅H₂₄O₁₂	516.126 78	[M-H]^-	-2.6	515[M-H]^-;380[M-H-C₈H₇O₂]^-;336[M-H-C₉H₇O₄]^-	[21]
8	Quercitrin	C₂₁H₂₀O₁₁	448.100 56	[M-H]^-	-2.8	447[M-H]^-;301[M-H-Rha]^-	[22]
9	Astin A	C₂₅H₃₃C₁₂N₅O₇	585.175 7	[M-H]^-	-3.4	584[M-H]^-;512[M-H-Cl₂]^-;362[M-H-C₈H₁₃N₂OCl₂]^-; 352[M-H-C₁₃H₁₄NO₃]^-	[23]
10¹⁾	Quercetin	C₁₅H₁₀O₇	302.042 65	[M-H]^-	-0.7	301[M-H]^-;151[M-H-C₈H₆O₃]^-;122[M-H-C₉H₇O₄]^-; 107[M-H-C₉H₆O₅]^-	[24]
11	Asterin	C₂₅H₃₃C₁₂N₅O₆	569.180 79	[M+H]⁺	0.1	570[M+H]⁺;485[M+H-C₄H₆NO]⁺;464[M+H-C₇H₈N]⁺; 338[M+H-C₁₃H₁₅N₂O₂]⁺
12¹⁾	Kaempferol	C₁₅H₁₀O₆	286.047 74	[M-H]^-	0.9	285[M-H]^-;153[M-H-C₇HO₃]^-;152[M-H-C₈H₅O₂]^-	[25]
13	Apigenin	C₁₅H₁₀O₅	270.052 82	[M-H]^-	-2.2	239[M-H-CH₃O]^-;153[M-H-C₈H₅O]^-;115[M-H- C₇H₅O₂-2OH]^-
14	Perillen	C₁₈H₃₀O₂	278.224 58	[M+H]⁺	-0.7	277[M-H]^-;196[M-H-C₅H₅O]^-;210[M-H-C₄H₃O]^-
15	Cholesteryl formate	C₂₈H₄₆O₂	414.349 78	[M+H]⁺	-2.7	415[M+H]⁺;327[M+H-C₆H₁₆]⁺	[26]
16	Poriferasterol monoglucoside	C₃₅H₅₈O₆	574.423 34	[M-H]^-	-3.4	573[M-H]^-;411[M-H-C₆H₁₀O₅]^-;440[M-H-C₅H₉O₄]^-
17¹⁾	Shionone	C₃₀H₅₀O	426.386 17	[M+H]⁺	0	427[M+H]⁺;372[M+H-C₄H₇]⁺;318[M+H-C₈H₁₃]⁺; 304[M+H-C₈H₁₁O]⁺;236[M+H-C₁₃H₁₉O]⁺;140 [M+H-C₂₀H₃₁O]⁺	[27]

Peak
No.

Predicted
compounds

Formula

m/z
(Observe)

Molecular
ion peak

Error
/×10^-6

m/z
(MS/MS)

References

1¹⁾

Chlorogenic acid

C₁₆H₁₈O₉

354.095 08

[M-H]^-

-0.8

353[M-H]^-;268[M-H-CHO-COOH]^-;162[M-H-C₇H₁₁O₆]^-

[15]

7-Hydroxycoumarin

C₉H₆O₃

162.031 69

[M+H]⁺

-0.3

135[M+H-CO]⁺;107[M+H-2CO]⁺

[16]

3¹⁾

Caffeic acid

C₉H₈O₄

180.042 26

[M-H]^-

0.7

179[M-H]^-;135[M-H-COOH]^-;90[M-H-C₆HO]^-

[17]

Rutin

C₂₇H₃₀O₁₆

610.153 39

[M-H]^-

-3.2

301[M-H-C₁₂H₂₀O₉]^-;273[M-H-C₁₃H₂₀O₁₀]^-;255[M-H-
C₁₃H₂₂O₁₁]^-;151[M-H-C₂₀H₂₆O₁₂]^-

[18]

5¹⁾

Ferulic acid

C₁₀H₁₀O₄

194.057 91

[M-H]^-

-0.9

193[M-H]^-;133[M-H-CH₃-COOH]^-

[19]

Isochlorogenic acid C

C₂₅H₂₄O₁₂

516.126 78

[M-H]^-

-2.6

515[M-H]^-;353[M-H-C₉H₇O₃]^-;324[M-H-C₁₀H₇O₄]^-

[20]

Isochlorogenic acid A

C₂₅H₂₄O₁₂

516.126 78

[M-H]^-

-2.6

515[M-H]^-;380[M-H-C₈H₇O₂]^-;336[M-H-C₉H₇O₄]^-

[21]

Quercitrin

C₂₁H₂₀O₁₁

448.100 56

[M-H]^-

-2.8

447[M-H]^-;301[M-H-Rha]^-

[22]

Astin A

C₂₅H₃₃C₁₂N₅O₇

585.175 7

[M-H]^-

-3.4

584[M-H]^-;512[M-H-Cl₂]^-;362[M-H-C₈H₁₃N₂OCl₂]^-;
352[M-H-C₁₃H₁₄NO₃]^-

[23]

10¹⁾

Quercetin

C₁₅H₁₀O₇

302.042 65

[M-H]^-

-0.7

301[M-H]^-;151[M-H-C₈H₆O₃]^-;122[M-H-C₉H₇O₄]^-;
107[M-H-C₉H₆O₅]^-

[24]

Asterin

C₂₅H₃₃C₁₂N₅O₆

569.180 79

[M+H]⁺

0.1

570[M+H]⁺;485[M+H-C₄H₆NO]⁺;464[M+H-C₇H₈N]⁺;
338[M+H-C₁₃H₁₅N₂O₂]⁺

12¹⁾

Kaempferol

C₁₅H₁₀O₆

286.047 74

[M-H]^-

0.9

285[M-H]^-;153[M-H-C₇HO₃]^-;152[M-H-C₈H₅O₂]^-

[25]

Apigenin

C₁₅H₁₀O₅

270.052 82

[M-H]^-

-2.2

239[M-H-CH₃O]^-;153[M-H-C₈H₅O]^-;115[M-H-
C₇H₅O₂-2OH]^-

Perillen

C₁₈H₃₀O₂

278.224 58

[M+H]⁺

-0.7

277[M-H]^-;196[M-H-C₅H₅O]^-;210[M-H-C₄H₃O]^-

Cholesteryl formate

C₂₈H₄₆O₂

414.349 78

[M+H]⁺

-2.7

415[M+H]⁺;327[M+H-C₆H₁₆]⁺

[26]

Poriferasterol monoglucoside

C₃₅H₅₈O₆

574.423 34

[M-H]^-

-3.4

573[M-H]^-;411[M-H-C₆H₁₀O₅]^-;440[M-H-C₅H₉O₄]^-

17¹⁾

Shionone

C₃₀H₅₀O

426.386 17

[M+H]⁺

427[M+H]⁺;372[M+H-C₄H₇]⁺;318[M+H-C₈H₁₃]⁺;
304[M+H-C₈H₁₁O]⁺;236[M+H-C₁₃H₁₉O]⁺;140
[M+H-C₂₀H₃₁O]⁺

[27]

DPPH	ABTS
S1	32.97	S12	43.57	S1	43.04	S12	47.52
S2	34.00	S13	38.15	S2	46.29	S13	46.29
S3	53.57	S14	35.26	S3	46.05	S14	45.63
S4	40.09	S15	35.56	S4	60.32	S15	51.92
S5	46.10	S16	29.98	S5	65.71	S16	57.65
S6	49.46	S17	38.15	S6	67.47	S17	47.28
S7	34.70	S18	34.62	S7	59.20	S18	52.16
S8	44.49	S19	30.07	S8	50.21	S19	46.43
S9	34.62	S20	31.87	S9	50.99	S20	46.19
S10	30.54	S21	31.22	S10	51.20	S21	44.91
S11	35.58	S22	35.67	S11	50.53	S22	51.92

DPPH

ABTS

Batch No.

Clearance rate/%

Batch No.

Clearance rate/%

Batch No.

Clearance rate/%

Batch No.

Clearance rate/%

32.97

S12

43.57

43.04

S12

47.52

34.00

S13

38.15

46.29

S13

46.29

53.57

S14

35.26

46.05

S14

45.63

40.09

S15

35.56

60.32

S15

51.92

46.10

S16

29.98

65.71

S16

57.65

49.46

S17

38.15

67.47

S17

47.28

34.70

S18

34.62

59.20

S18

52.16

44.49

S19

30.07

50.21

S19

46.43

34.62

S20

31.87

50.99

S20

46.19

S10

30.54

S21

31.22

S10

51.20

S21

44.91

S11

35.58

S22

35.67

S11

50.53

S22

51.92

DPPH correlation results	ABTS correlation results
17	0.907	1	17	0.873	1
9	0.868	2	9	0.866	2
4	0.863	3	5	0.859	3
14	0.857	4	4	0.847	4
13	0.848	5	12	0.845	5
2	0.846	6	2	0.835	6
7	0.844	7	6	0.833	7
5	0.838	8	13	0.827	8
12	0.832	9	7	0.82	9
10	0.817	10	15	0.814	10
11	0.816	11	1	0.805	11
6	0.81	12	14	0.795	12
15	0.808	13	11	0.786	13
1	0.782	14	8	0.763	14
8	0.776	15	3	0.759	15
3	0.738	16	10	0.758	16
16	0.589	17	16	0.617	17

DPPH correlation results

ABTS correlation results

Peak No.

Order

Peak No.

Order

0.907

0.873

0.868

0.866

0.863

0.859

0.857

0.847

0.848

0.845

0.846

0.835

0.844

0.833

0.838

0.827

0.832

0.82

0.817

0.814

0.816

0.805

0.81

0.795

0.808

0.786

0.782

0.763

0.776

0.759

0.738

0.758

0.589

0.617