利用代谢组学方法探索一种商业进口药用树(Prunus africana (Hook.f.) Kalkman)的化学地理变异

IF 3.8 2区 农林科学 Q1 PLANT SCIENCES
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引用次数: 0

摘要

非洲樱桃(Prunus africana (Hook.f.) Kalkman)的茎皮传统上用于治疗胃痛、胸痛、疟疾、发烧、糖尿病和高血压。已从茎皮中分离出大量成分,包括β-谷甾醇和熊果酸,据报道它们具有抗炎活性。这项研究旨在确定从喀麦隆、刚果民主共和国(DRC)和津巴布韦这三个非洲国家采集的非洲李样本之间的化学分型差异。共从 19 个种群中采集了 95 份茎皮样本,其中喀麦隆 11 份(n = 55),刚果民主共和国 4 份(n = 20),津巴布韦 4 份(n = 20)。样品用乙酸乙酯、二氯甲烷和甲醇提取(分别为 10 mL 中提取 1 g、10 mL 中提取 1 g 和 5 µL 中提取 2 mg)。使用 MetaboAnalyst 4.0 软件对高效薄层色谱(HPTLC)、超高效液相色谱-质谱联用(UPLC-MS)、一维气相色谱-飞行时间-质谱联用(1D GC-ToF-MS)获得的色谱数据和质子核磁共振(1H NMR)获得的光谱数据进行化学计量分析,以探索化学型变异。通过二维气相色谱-飞行时间质谱法(2D GC-ToF-MS)对每个国家的样品进行分析,以分辨在一维气相色谱分析中共同沉降的化合物。采用经过验证的超高效液相色谱-光电二极管阵列检测(UPLC-PDA)方法对所选化合物(β-谷甾醇和熊果酸)进行定量。通过这四种技术获得的化学图谱对每个国家的样品来说都非常相似。津巴布韦样品的 HPTLC 图谱与其他两个国家的明显不同,在 Rf 值为 0.80 时有一个独特的条带。通过对 HPTLC 数据(使用 rTLC 软件)、UPLC-MS、1D GC-ToF-MS 和 1H-NMR 数据进行多变量分析,发现了三个国家特有的聚类。通过构建 PCA 和 PLS-DA 分数图进行了化学计量分析,随后构建了投影图中的重要变量,从而选出并初步确定了对三个群组的分离有重大贡献的标记化合物。定量分析显示,与刚果民主共和国和喀麦隆相比,津巴布韦样品中的β-谷甾醇含量更高(29.2 - 119 µg/克干重(DW)提取物),而喀麦隆样品中的熊果酸含量最高(1.80 - 141 µg/克干重提取物)。HPTLC, UPLC-MS, 1D and 2D GC-ToF-MS and 1H-NMR 是对从三个国家采集的样本进行质量控制的有效工具。喀麦隆样本中熊果酸含量较高,而津巴布韦样本中的β-谷甾醇含量较高。与二维气相色谱-串联质谱(2D GC-ToF-MS)相比,一维气相色谱-串联质谱(1D GC-ToF-MS)在三个国家检测到的化合物数量较少,从而提高了分离度和分辨率。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Exploring the chemogeographical variation of a commercially important medicinal tree (Prunus africana (Hook.f.) Kalkman) using a metabolomics approach

The stem-bark of Prunus africana (Hook.f.) Kalkman (African cherry) is traditionally used for the treatment of stomach ache, chest pain, malaria, fever, diabetes and high blood pressure. A large number of constituents have been isolated from the stem-bark including β-sitosterol and ursolic acid, which were reported to have anti-inflammatory activity. The aim of the study was to establish chemotypic variation among Prunus africana samples collected from three African countries, Cameroon, Democratic Republic of Congo (DRC) and Zimbabwe. Ninety-five stem-bark samples were collected from a total of nineteen populations, 11 in Cameroon (n = 55), 4 in Democratic Republic of Congo (n = 20) and 4 in Zimbabwe (n = 20). The samples were extracted with ethyl acetate, dichloromethane and methanol (1 g in 10 mL, 1 g in 10 mL and 2 mg in 5 µL, respectively). Chemometric analysis of chromatographic data obtained from high-performance thin layer chromatography (HPTLC), ultra performance liquid chromatography coupled with mass spectrometry (UPLC–MS), one-dimensional gas chromatography-time-of-flight-mass spectrometry (1D GC–ToF–MS) and spectroscopic data obtained from proton nuclear magnetic resonance (1H NMR) were done using MetaboAnalyst 4.0 software to explore chemotypic variation. A sample from each country was analysed by two-dimensional gas chromatography-time-of-flight-mass spectrometry (2D GC–ToF–MS) to resolve compounds that co-eluted in the 1D GC analysis. Quantification of selected compounds (β-sitosterol and ursolic acid) were done using a validated ultra performance liquid chromatography-photodiode-array detection (UPLC–PDA) method. The chemical profiles obtained from each of the four techniques were very similar for the samples from each country. The HPTLC profiles for Zimbabwe samples were distinctly different from the other two countries with a unique band at Rf value of 0.80. Multivariate analysis of the HPTLC data (using rTLC software), UPLC–MS, 1D GC–ToF–MS and 1H–NMR data revealed three clusters that were country specific. Chemometric analysis through the construction of PCA and a PLS–DA scores plots was performed, followed by the construction of variable important in projection plots, whereby marker compounds contributing significantly to the separation of the three clusters were selected and tentatively identified. Quantitative analysis revealed that samples from Zimbabwe contained higher levels of β-sitosterol (29.2 – 119 µg/g dry weight (DW) of the extract) compared to DRC and Cameroon, while samples from Cameroon were found to contain the highest levels of ursolic acid (1.80 – 141 µg/g DW of the extract). HPTLC, UPLC–MS, 1D and 2D GC–ToF–MS and 1H–NMR are valid tools for quality control of P. africana plant materials after revealing three clusters of samples collected from the three countries. Cameroon samples had high content of ursolic acid, while Zimbabwe samples showed high levels of β-sitosterol. The number of compounds detected across the three countries by 1D GC–ToF–MS was less compared to 2D GC–ToF–MS, which enhanced separation and resolution.

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来源期刊
Journal of Applied Research on Medicinal and Aromatic Plants
Journal of Applied Research on Medicinal and Aromatic Plants Pharmacology, Toxicology and Pharmaceutics-Drug Discovery
CiteScore
6.40
自引率
7.70%
发文量
80
审稿时长
41 days
期刊介绍: JARMAP is a peer reviewed and multidisciplinary communication platform, covering all aspects of the raw material supply chain of medicinal and aromatic plants. JARMAP aims to improve production of tailor made commodities by addressing the various requirements of manufacturers of herbal medicines, herbal teas, seasoning herbs, food and feed supplements and cosmetics. JARMAP covers research on genetic resources, breeding, wild-collection, domestication, propagation, cultivation, phytopathology and plant protection, mechanization, conservation, processing, quality assurance, analytics and economics. JARMAP publishes reviews, original research articles and short communications related to research.
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