Tian Xue-Hao, Zhang Hao, Wang Shen, Li Tong, Huang Xue-Mei, Yan Meng-Meng, Cao Xiao-Fei, Xu Bing, Wang Peng-Long, Lei Hai-Min
School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 100102, China.
School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100102, China.
Chin Herb Med. 2020 Apr 3;12(2):188-194. doi: 10.1016/j.chmed.2020.03.005. eCollection 2020 Apr.
There are some anthraquinones, anthraquinones and flavonones in which exhibited significant acidity, such as sennoside A/B and sennoside C/D. The current strategies used in separating these components are mainly based on conventional column chromatography which is time consuming, laborious and costly. This study is aimed at exploring a method of precipitation extraction of acid components in . Using alkaloid as a "hook", it is reasonable to use the principle of "acid-alkali complexation" to "fish" the acidic components in .
Isothermal titration calorimeter (ITC) was used to measure the extraction efficiency of different alkaloids. Then, alkaloid determined by ITC was mixed with extracting solution of to form complex. High performance liquid chromatography coupled with mass spectrometry (HPLC-MS) was used to investigate the ingredients "fished" by berberine (Ber). The mechanism of "fishing" process was explained by ITC, optical activity, fluorescence spectrometry and scanning electron microscope.
The ITC results proved that the choice of "hook" was particularly important in the process of "fishing". Among the hooks, the fishing efficiency of the isoquinoline alkaloids (Ber) was the highest, reaching 10.3%. Nine ingredients were detected and determined by HPLC-MS, and the main components were sennoside A/B and sennoside C/D. Based on ITC test of Ber and sennoside A, the combination mechanism of the two ingredients was a chemical reaction with a nearly binding ratio (2:1). Fluorescence and optical properties of the active ingredients were changed after complexation. By scanning electron microscope, we found that two types of components had obviously self-assembled behavior during the formation process.
Ber successfully "fished" the main acidic components, sennoside A/B and sennoside C/D, from . Combined with different characterizations, the "fishing" process was determined as a chemical association reaction induced by electrostatic interaction or π-π stacking. Therefore, with special identification ability, the "fishing" process had the potential of practical application.
存在一些蒽醌、蒽醌和黄酮酮，其中一些表现出显著的酸性，如番泻苷A/B和番泻苷C/D。目前用于分离这些成分的策略主要基于传统柱色谱法，该方法耗时、费力且成本高。本研究旨在探索一种沉淀萃取[具体物质]中酸性成分的方法。以生物碱作为“钩子”，利用“酸碱络合”原理“钓出”[具体物质]中的酸性成分是合理的。
使用等温滴定量热仪（ITC）测量不同生物碱的萃取效率。然后，将通过ITC确定的生物碱与[具体物质]的萃取溶液混合形成络合物。采用高效液相色谱-质谱联用（HPLC-MS）研究黄连素（Ber）“钓出”的成分。通过ITC、旋光性、荧光光谱和扫描电子显微镜解释“钓鱼”过程的机制。
ITC结果证明，在“钓鱼”过程中“钩子”的选择尤为重要。在这些钩子中，异喹啉生物碱（Ber）的钓鱼效率最高，达到10.3%。通过HPLC-MS检测并确定了9种成分，主要成分是番泻苷A/B和番泻苷C/D。基于Ber与番泻苷A的ITC测试，两种成分的结合机制是一种结合比接近（2:1）的化学反应。络合后活性成分的荧光和光学性质发生了变化。通过扫描电子显微镜，我们发现两种类型的成分在形成过程中具有明显的自组装行为。
Ber成功地从[具体物质]中“钓出”了主要酸性成分番泻苷A/B和番泻苷C/D。结合不同的表征，“钓鱼”过程被确定为由静电相互作用或π-π堆积诱导的化学缔合反应。因此，凭借特殊的识别能力，“钓鱼”过程具有实际应用潜力。
