Department of Pharmacognosy, Faculty of Life Sciences, University of Vienna, Althanstraße 14, 1090 Vienna, Austria; Innovation Centre of the Faculty of Chemistry Ltd., Studentski trg 12-16, 11000 Belgrade, Serbia.
Department of Pharmacognosy, Faculty of Life Sciences, University of Vienna, Althanstraße 14, 1090 Vienna, Austria.
J Chromatogr A. 2019 Jun 7;1594:190-198. doi: 10.1016/j.chroma.2019.02.006. Epub 2019 Feb 4.
Quality control is a crucial step in the production of effective and safe herbal remedies. The aim of this study was to develop a new, simple, and high throughput procedure for the quality assessment of herbal drugs using a high-performance thin-layer chromatography (HPTLC)/bioautography and UPLC-MS/MS approach combined with chemometrics. This was exemplarily shown for Morus alba L. root bark (sāng bái pí; SBP). Bioautography assays were developed for the identification of constituents with radical scavenging (DPPH assay) and antimicrobial activities (Bacillus subtilis, Escherichia coli) of 18 different M. alba samples, which was supported by UPLC-MS/MS analysis. Further, the combination of bioautography and chemometrics identified those samples with the most bioactive constituents. Plant materials collected from Serbia (11 samples) showed higher both radical scavenging and antimicrobial activities compared to samples provided from China (7 samples). Principal component analysis (PCA) confirmed the discrimination of geographically different samples and recognized their main markers responsible for differences between Serbian and Chinese samples. Most importantly for quality assessment, the combined HPTLC/bioautography and UPLC-MS/MS approach not only allowed for a fast chemical profiling of the investigated samples and their unambiguous identification, but also for the disclosure of major and minor bioactive constituents present in SBP.
质量控制是生产有效和安全草药疗法的关键步骤。本研究旨在开发一种新的、简单的高通量方法，用于使用高效薄层色谱（HPTLC）/生物自显影和 UPLC-MS/MS 方法结合化学计量学评估草药药物的质量。这通过桑白皮（SBP）的实例得到了证明。为了鉴定具有自由基清除（DPPH 测定）和抗菌活性（枯草芽孢杆菌、大肠杆菌）的成分，开发了生物自显影测定法，对 18 种不同的桑白皮样品进行了研究，这得到了 UPLC-MS/MS 分析的支持。此外，生物自显影和化学计量学的结合确定了具有最生物活性成分的那些样品。与来自中国（7 个样品）的样品相比，从塞尔维亚（11 个样品）收集的植物材料显示出更高的自由基清除和抗菌活性。主成分分析（PCA）证实了对地理上不同的样品的区分，并确定了它们的主要标记物，这些标记物负责区分塞尔维亚和中国样品之间的差异。最重要的是，用于质量评估，HPTLC/生物自显影和 UPLC-MS/MS 方法的组合不仅允许快速分析研究样品的化学特征，并对其进行明确的鉴定，而且还可以揭示 SBP 中存在的主要和次要生物活性成分。
