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Method Article
Traditional Chinese medicine (TCM) represents a rich treasure trove of natural treatment options and plays an important role in addressing diverse health needs worldwide. This study presents a comprehensive experimental investigation on the extraction, separation, and physicochemical properties of rutin, the key active component of Flos sophorae immaturus.
Traditional Chinese medicine (TCM) has long served as a rich repository of natural therapeutic solutions, playing a pivotal role in addressing diverse healthcare needs worldwide. Among the various herbs used in TCM, Sophora flavescens is known for its wide range of medicinal properties. Its application dates back centuries and continues to be highly valued in modern medicine, particularly for its therapeutic benefits in treating cardiovascular diseases. Rutin, a bioactive flavonoid compound, is widely distributed across the plant kingdom, with the highest concentration found in Flos sophorae immaturus. Rutin has been reported to reduce abnormal capillary permeability and fragility and acts as a protective agent for cardiovascular and cerebrovascular vessels. This study presents a detailed experimental protocol for the extraction, separation, and characterization of the physicochemical properties of rutin derived from Flos sophorae immaturus. Additionally, the protocol for the extraction of flavonoids from TCM using the alkali extraction and acid precipitation method is introduced and described.
Flos sophorae immaturus (FSI), the dried flower bud of Sophora japonica L., has been a cornerstone of traditional Chinese medicine (TCM) for centuries. Its main active components include rutin, isoquercitrin, Sophorae Flos, and other flavonoids and saponins. These constituents collectively confer a variety of pharmacological effects, such as clearing heat and detoxifying, cooling blood and hemostasis, and lowering blood pressure. In clinical practice, FSI is primarily valued for its ability to protect capillary permeability1, maintain cardiovascular system function2, cool blood, arrest bleeding....
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Rutin is a flavonoid containing multiple phenolic hydroxyl groups and is soluble in hot alkaline solutions, followed by acidification and precipitation. It can be extracted using the alkali extraction-acid precipitation process. The reagents and equipment used in this study are listed in the Table of Materials.
1. Experimental preparation
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In the physicochemical identification experiment of rutin, the solution turned red after the addition of magnesium powder and a small amount of concentrated hydrochloric acid, followed by shaking, indicating the presence of a flavonoid parent nucleus. In the Molisch reaction, a purple ring formed at the interface, indicating a positive result and the presence of sugar moieties in the molecular structure. The sample solution turned bright yellow upon the addition of 2% chloro-oxidation reagent in methanol, suggesting the .......
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The extraction of rutin is primarily based on the alkali extraction-acid precipitation (AEAP) method6, which offers the advantages of simple equipment requirements, ease of operation, high efficiency, and low cost. This technique is often supplemented by microwave-assisted extraction (MAE)7 and ultrasound-assisted extraction (UAE)8, which reduce reagent consumption, accelerate extraction time, and improve overall yield.
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The authors of this paper hereby declare that they have disclosed any and all conflicts of interest that may have arisen in the course of conducting this research and writing this article. We confirm that we have no financial, personal, or professional relationships that could potentially bias our interpretation of the results or conclusions presented in this work. Specifically, we have no ties to any pharmaceutical companies, research institutions, or other entities that could benefit from the findings reported here. Finally, we affirm that our findings and conclusions are solely based on the data and analysis presented in this paper and are not influenced by any external factors. We remain committed to the highest standards of academic integrity and ethical conduct in all our research activities.
We would like to express our profound gratitude to the Pharmaceutical Chemistry Laboratory at the Beijing University of Chinese Medicine for providing the experimental platform. Their support and resources have been invaluable in the conduct of our research and experiments. We deeply appreciate their dedication to scientific excellence and their commitment to fostering the development of young researchers.
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| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| 1-Naphthol | MACKLIN | C16245764 | |
| Acetic acid | MACKLIN | C12662811 | |
| Acetonitrile | MACKLIN | C16129374 | |
| Aluminium Chloride | MACKLIN | C16485922 | |
| Citric acid | MACKLIN | C16429480 | |
| DMSO | FUCHEN (Tianjin) Chemical Reagent Co., LTD | 20230306 | |
| Ethanol | FUCHEN (Tianjin) Chemical Reagent Co., LTD | 20250102 | |
| Ethyl acetate | FUCHEN (Tianjin) Chemical Reagent Co., LTD | 20240318 | |
| Flos sophorae immaturus | Beijing Tongrentang | 10092026329782 | |
| Formic acid | FUCHEN (Tianjin) Chemical Reagent Co., LTD | 20150402 | |
| MestReNova | Mestrelab Research | / | |
| Methanol | innochem | C16365684 | |
| Silica gel 60F254 | STEEMA | 20210222 | |
| Xcalibur | Thermo Fisher Scientific | / | |
| Zirconyl chloride octahydrate | MACKLIN | C15350627 |
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