DataSheet1_Chromatography Conditions Development by Design of Experiments for the Chemotype Differentiation of Four Bauhinia Species.pdf

The extensive use of medicinal herbs to traditionally treat disease persists for generations, and scientific evidence on plant-derived extracts has indicated their numerous biological activities. The Bauhinia, popular known as cow’s paw (“pata de vaca”), with more than 60 native species, are extensively used in Brazilian popular medicine for the control of diabetes. Therefore, in 2009, B. forficata, B. variegata and/or B. affinis were included in the Brazilian National List of Medicinal Plants of Interest to SUS (RENISUS - Brazil). In this context, this work reports the results of the chemical differentiation of B. forficata, B. variegata, B. longifolia, and B. affinis using liquid chromatography coupled to high-resolution mass spectrometry and unsupervised chemometric tools. Chromatographic conditions were optimized by using the design of experiments (DoE) and chromatographic knowledge. Furthermore, the chemical profile of the studied species was analyzed by principal component analysis (PCA) and hierarchical cluster analysis that differentiated the four species of Bauhinia, and 55 compounds were also inferred by MS2 experiments, some of them for the first time in B. affinis. In this manner, this work provides important information that could be used in quality control, development of new pharmaceuticals, and food products based on Bauhinia leaves, as well as to explain ethnomedicinal properties, pharmacological and toxicological actions.

传统上用于疾病治疗的药草历经多代仍被广泛使用，而针对植物源提取物的科学研究已证实其具备多种生物活性。羊蹄甲属（Bauhinia）俗称牛蹄草（“pata de vaca”），该属拥有超过60个本土物种，在巴西民间医学中被广泛用于糖尿病管控。因此，2009年，镰叶羊蹄甲（B. forficata）、洋紫荆（B. variegata）以及近缘羊蹄甲（B. affinis）被纳入巴西国家统一卫生系统（Sistema Único de Saúde，简称SUS）重点药用植物名录（RENISUS - 巴西）。在此背景下，本研究采用高效液相色谱-高分辨质谱联用法结合无监督化学计量学工具，对镰叶羊蹄甲、洋紫荆、长叶羊蹄甲（B. longifolia）以及近缘羊蹄甲进行化学区分并报告相关研究成果。本研究通过实验设计（Design of Experiments，DoE）结合色谱学知识对色谱条件进行了优化。此外，研究通过主成分分析（Principal Component Analysis，PCA）与系统聚类分析对受试物种的化学特征谱进行解析，成功区分了四种羊蹄甲属植物；同时通过二级质谱（MS2）实验推断出55种化合物，其中部分化合物为首次在近缘羊蹄甲中被发现。综上，本研究提供的重要信息可用于基于羊蹄甲属叶片的药品质量管控、新型药物与食品开发，同时有助于阐释其民族医药用途、药理与毒理作用。