人工栽培桑黄HPLC指纹图谱及主成分含量测定

doi:10.3969/j.issn.1004-1524.2023.03.19

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (3): 658-665.DOI: 10.3969/j.issn.1004-1524.2023.03.19

人工栽培桑黄HPLC指纹图谱及主成分含量测定

钟石¹(), 曹英龙²^,³, 王文琼²^,³, 霍进喜¹, 孙雨晴¹, 宣利江²^,³, 李有贵¹^,^*()

1.浙江省农业科学院蚕桑与茶叶研究所,浙江杭州 310021
2.中国科学院上海药物研究所,原创新药研究重点实验室,上海 202009
3.中国科学院大学,北京 100049

收稿日期:2021-09-30 出版日期:2023-03-25 发布日期:2023-04-07
通讯作者: ^*李有贵,E-mail:liyougui3@126.com
作者简介:钟石(1977—),女,安徽合肥人,硕士,副研究员,主要从事天然药用资源相关研究。E-mail: 2196252@qq.com
基金资助:
浙江省自然科学基金(LQ21C150002);浙江省重点研发计划(2018C02003)

HPLC fingerprint of cultivated Sanghuangporus and content determination of main component

ZHONG Shi¹(), CAO Yinglong²^,³, WANG Wenqiong²^,³, HUO Jinxi¹, SUN Yuqing¹, XUAN Lijiang²^,³, LI Yougui¹^,^*()

1. Institute of Sericulture and Tea, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2. State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 202009, China
3. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2021-09-30 Online:2023-03-25 Published:2023-04-07

摘要/Abstract

摘要：

为建立人工栽培桑黄瓦宁纤孔菌的HPLC指纹图谱,采用化学计量学分析方法对其进行分析和筛选出桑黄的4个标志性成分,并以其中最主要成分Hypholomine B为指标建立含量测定方法。参照2020年版《中国药典》(四部)通则中高效液相色谱法,建立桑黄指纹图谱,利用“中药色谱指纹图谱相似度评价系统2004 A版”评价其相似度,确定共有峰,同时测定桑黄中苯乙烯吡喃酮类成分Hypholomine B的含量。结果表明,建立了10批桑黄的HPLC指纹图谱,共有12个共有峰,相似度均大于0.92。系统聚类分析(HCA)、主成分分析(PCA)和正交偏最小二乘判别分析(OPLS-DA)结果一致,将样品分为两类。在1.25~20.00 μg范围内,Hypholomine B的进样量与峰面积具有良好的线性关系(R²=0.999 9);精密度、稳定性和重复性试验的相对标准偏差(RSD)值均小于2.0%(n=6);加样回收率为95%~99%,其RSD值为1.48%;10批桑黄样品中Hypholomine B的含量范围为0.15%~0.95%。指纹图谱与HCA、PCA和OPLS-DA相结合可全面地评价桑黄药材质量,该方法稳定、可靠,可为桑黄质量评价提供参考。

关键词: 桑黄, 指纹图谱, 高效液相色谱法, 苯乙烯吡喃酮类, Hypholomine B

Abstract:

To establish HPLC fingerprint of cultivated Sanghuangporus(Sanghuang), screen out the signature component of Sanghuangporus through chemical pattern recognition, the content determination method of Sanghuangporus was established based on this index. According to the corresponding methods in the general rules of The Chinese Pharmacopoeia (four parts) in the 2020 edition, the fingerprint of Sanghuangporus was established, and the similarity was evaluated by using the similarity evaluation system of traditional Chinese Medicine Chromatographic Fingerprint 2004 Version A to determine the common peak. The content of Hypholomine B in Sanghuangporus was also determined. HPLC fingerprints of 10 batches of Sanghuangporus were established, with 12 common peaks, and the similarity was greater than 0.92. The results of HCA, PCA and OPLS-DA were consistent, and the samples were divided into two categories. The injection volume of Hypholomine B had a good linear range with its peak area in the range of 1.25-20.00 μg (R²=0.999 9). The RSD values of precision, stability and repeatability were all less than 2.0% (n=6). The recovery rate was 95%-99%, and the RSD was 1.48%. The content of Hypholomine B in 10 batches of mulberry yellow samples ranged from 0.15% to 0.95%. The fingerprint can be combined with HCA, PCA and OPLS-DA to comprehensively evaluate the quality of Sanghuangporus. The method is stable and reliable, providing reference for the quality evaluation of Sanghuangporus.

Key words: Sanghuangporus, fingerprints, high performance liquid chromatography, styrylpyranones, Hypholomine B

中图分类号:

S888

钟石, 曹英龙, 王文琼, 霍进喜, 孙雨晴, 宣利江, 李有贵. 人工栽培桑黄HPLC指纹图谱及主成分含量测定[J]. 浙江农业学报, 2023, 35(3): 658-665.

ZHONG Shi, CAO Yinglong, WANG Wenqiong, HUO Jinxi, SUN Yuqing, XUAN Lijiang, LI Yougui. HPLC fingerprint of cultivated Sanghuangporus and content determination of main component[J]. Acta Agriculturae Zhejiangensis, 2023, 35(3): 658-665.

图/表 12

参考文献 18

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编号 No.			栽培基质 Culture medium
S1	A1	浙江千济方医药科技公司Zhejiang Qianjifang Pharmaceutical Technology Co., Ltd.	椴木Basswood
S2	A2	浙江千济方医药科技公司Zhejiang Qianjifang Pharmaceutical Technology Co., Ltd.	椴木Basswood
S3	B1	海宁宏欣农业生物科技公司Haining Hongxin Agricultural Biotechnology Co., Ltd.	人工代料Manual substitution
S4	B2	海宁宏欣农业生物科技公司Haining Hongxin Agricultural Biotechnology Co., Ltd.	人工代料Manual substitution
S5	B3	海宁宏欣农业生物科技公司Haining Hongxin Agricultural Biotechnology Co., Ltd.	人工代料Manual substitution
S6	C1	淳安千岛湖桑都食用菌合作社Chun’an Qiandao Lake Sangdu Edible Fungus Cooperative	人工代料Manual substitution
S7	C2	淳安千岛湖桑都食用菌合作社Chun’an Qiandao Lake Sangdu Edible Fungus Cooperative	人工代料Manual substitution
S8	C3	淳安千岛湖桑都食用菌合作社Chun’an Qiandao Lake Sangdu Edible Fungus Cooperative	人工代料Manual substitution
S9	C4	淳安千岛湖桑都食用菌合作社Chun’an Qiandao Lake Sangdu Edible Fungus Cooperative	人工代料Manual substitution
S10	C5	淳安千岛湖桑都食用菌合作社Chun’an Qiandao Lake Sangdu Edible Fungus Cooperative	人工代料Manual substitution

编号 No.			栽培基质 Culture medium
S1	A1	浙江千济方医药科技公司Zhejiang Qianjifang Pharmaceutical Technology Co., Ltd.	椴木Basswood
S2	A2	浙江千济方医药科技公司Zhejiang Qianjifang Pharmaceutical Technology Co., Ltd.	椴木Basswood
S3	B1	海宁宏欣农业生物科技公司Haining Hongxin Agricultural Biotechnology Co., Ltd.	人工代料Manual substitution
S4	B2	海宁宏欣农业生物科技公司Haining Hongxin Agricultural Biotechnology Co., Ltd.	人工代料Manual substitution
S5	B3	海宁宏欣农业生物科技公司Haining Hongxin Agricultural Biotechnology Co., Ltd.	人工代料Manual substitution
S6	C1	淳安千岛湖桑都食用菌合作社Chun’an Qiandao Lake Sangdu Edible Fungus Cooperative	人工代料Manual substitution
S7	C2	淳安千岛湖桑都食用菌合作社Chun’an Qiandao Lake Sangdu Edible Fungus Cooperative	人工代料Manual substitution
S8	C3	淳安千岛湖桑都食用菌合作社Chun’an Qiandao Lake Sangdu Edible Fungus Cooperative	人工代料Manual substitution
S9	C4	淳安千岛湖桑都食用菌合作社Chun’an Qiandao Lake Sangdu Edible Fungus Cooperative	人工代料Manual substitution
S10	C5	淳安千岛湖桑都食用菌合作社Chun’an Qiandao Lake Sangdu Edible Fungus Cooperative	人工代料Manual substitution

编号No.	相似度Similarity
S1	0.966
S2	0.985
S3	0.989
S4	0.975
S5	0.942
S6	0.984
S7	0.924
S8	0.955
S9	0.968
S10	0.973

编号No.	相似度Similarity
S1	0.966
S2	0.985
S3	0.989
S4	0.975
S5	0.942
S6	0.984
S7	0.924
S8	0.955
S9	0.968
S10	0.973

成分 Component	特征值 Eigenvalue	方差的贡献率 Contribution rate of variance/%	累计的方差贡献率 Cumulative variance contribution rate/%
1	5.070	42.248	42.248
2	2.584	21.532	63.780
3	1.727	14.392	78.172
4	1.221	10.172	88.343

人工栽培桑黄HPLC指纹图谱及主成分含量测定

HPLC fingerprint of cultivated Sanghuangporus and content determination of main component

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编号 No.	批号 Batch No.	峰面积 Peak area	含量 Content/%
S1	A1	2 082.7	0.21
S2	A2	4 129.5	0.45
S3	B1	5 992.5	0.66
S4	B2	8 471.5	0.95
S5	B3	1 564.4	0.15
S6	C1	2 138.3	0.21
S7	C2	3 107.8	0.33
S8	C3	2 169.5	0.22
S9	C4	3 696.9	0.39
S10	Y1	5 086.7	0.56

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