Supplementary file 1_E-Nose and HS-SPME-GC-MS unveiling the scent signature of Ligusticum chuanxiong and its medicinal relatives.pdf

IntroductionTo explore the origin and evolution of Ligusticum Chuanxiong, we conducted a component analysis of Ligusticum Chuanxiong and its medicinal relatives.
MethodsThis study encompassed seven species from various origins, including Chuanxiong (Ligusticum chuanxiong Hort.), Gansu Chuanxiong (Ligusticum chuanxiong cv. Gansu), Yunnan Chuanxiong (Ligusticum chuanxiong cv. Yunnan), Japanese Chuanxiong (Cnidium officinale Makino), Fuxiong (Ligusticum sinense ‘Fuxiong’), Gaoben (Ligusticum sinense), and Liaogaoben (Ligusticum jeholense), comprising 27 distinct materials. We employed headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) to identify various odor profiles from these species using electronic nose technology (E-nose). The method effectively identified volatile constituents in the leaves of these seven species.
ResultsResults indicated that odor differences between L. chuanxiong and its medicinal relatives were predominantly observed in sensors W1W and W1S. Linear discriminant factor analysis (LDA) successfully distinguished five of the relatives; however, L. chuanxiong and L. sinense exhibited high odor similarity, limiting complete differentiation in some samples. HS-SPME-GC-MS identified a total of 118 volatile constituents, with eight differential volatiles identified: trans-Neocnidilide, β-Caryophyllene, β-Selinene, 5-Pentylcyclohexa-1,3-diene, (E)-Ligustilide, Butylphthalide, Neophytadiene, and Senkyunolide. Hierarchical cluster analysis (HCA) grouped L. chuanxiong, L. sinense, L. jeholense, and L. chuanxiong cv. Gansu together, highlighting the close relationship between L. chuanxiong and L. sinense. Joint analysis revealed a significant positive correlation between sensor W1W and the differential volatile component β-Caryophyllene, suggesting its potential for distinguishing closely related species.
DiscussionThis study provides a foundational understanding of volatile components in the leaves of L. chuanxiong and its medicinal relatives using E-nose combined with HS-SPME-GC-MS, contributing to the discussion on their interspecific odor characteristics and relationships.

引言 为探究川芎（Ligusticum Chuanxiong）的起源与演化，本研究对川芎及其药用近缘物种开展了组分分析。
方法 本研究纳入了7个不同来源的物种，包括川芎（Ligusticum chuanxiong Hort.）、甘肃川芎（Ligusticum chuanxiong cv. Gansu）、云南川芎（Ligusticum chuanxiong cv. Yunnan）、日本川芎（Cnidium officinale Makino）、抚芎（Ligusticum sinense ‘Fuxiong’）、藁本（Ligusticum sinense）、辽藁本（Ligusticum jeholense），共计27份独立材料。本研究采用顶空固相微萃取-气相色谱-质谱联用法（headspace solid-phase microextraction-gas chromatography-mass spectrometry，HS-SPME-GC-MS）结合电子鼻技术（electronic nose technology，E-nose）对上述物种的气味特征进行分析。该方法可有效鉴定这7个物种叶片中的挥发性成分。
结果 结果显示，川芎与其药用近缘物种的气味差异主要体现在传感器W1W和W1S上。线性判别因子分析（linear discriminant factor analysis，LDA）可成功区分其中5个近缘物种；但川芎与藁本（Ligusticum sinense）气味相似度极高，导致部分样本无法完全区分。通过HS-SPME-GC-MS共鉴定出118种挥发性成分，其中8种为差异挥发性成分：反式新蛇床内酯（trans-Neocnidilide）、β-石竹烯（β-Caryophyllene）、β-芹子烯（β-Selinene）、5-戊基-1,3-环己二烯（5-Pentylcyclohexa-1,3-diene）、(E)-藁本内酯（(E)-Ligustilide）、丁基苯酞（Butylphthalide）、新植二烯（Neophytadiene）以及洋川芎内酯（Senkyunolide）。系统聚类分析（hierarchical cluster analysis，HCA）将川芎、藁本、辽藁本以及甘肃川芎聚为一类，凸显了川芎与藁本之间的近缘关系。联合分析显示，传感器W1W与差异挥发性成分β-石竹烯存在显著正相关，表明该成分可用于区分亲缘关系较近的物种。
讨论 本研究通过电子鼻结合顶空固相微萃取-气相色谱-质谱联用法，为川芎及其药用近缘物种叶片的挥发性成分研究提供了基础认知，有助于深入探讨其种间气味特征与亲缘关系。