| 引用本文:牛俐燃1，2，李冠聪1，2，杨光明1，2，潘扬1，3，蔡宝昌1，2.聚酰胺色谱法联合中压液相制备色谱法提取分离镰形棘豆中活性黄酮类成分[J].世界中医药,2019,(02):. |
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| 聚酰胺色谱法联合中压液相制备色谱法提取分离镰形棘豆中活性黄酮类成分 |
| Extraction and Separation of Bioactive Flavonoids from Oxytropis Falcata by Polyamide Chromatography Combined with Medium-pressure Liquid Chromatography |
| 投稿时间：2019-01-10 |
| DOI：10.3969/j.issn.1673-7202.2019.02.005 |
| 中文关键词: 镰形棘豆；黄酮化合物；聚酰胺色谱法；中压制备色谱法；A549细胞 质谱法 |
| English Keywords:Oxytropis falcata Flavonoids Polyamide column chromatography Medium-pressure liquid chromatography A549 cells Mass spectrometry |
| 基金项目:国家自然科学基金项目（81473420，30902012）；江苏高校优势学科建设工程资助项目（PAPD）；江苏高校品牌专业建设工程资助项目（PPZY2015A070） |
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| 中文摘要: |
| 目的:对藏药镰形棘豆中黄酮类成分进行提取分离，并将得到的黄酮化合物进行抗人肺腺癌A549细胞的活性筛选。方法:镰形棘豆药材浸提液经乙酸乙酯萃取，该萃取部位通过聚酰胺柱色谱梯度洗脱，结合中压液相制备色谱法进一步纯化分离，获得单一黄酮化合物，通过高效液相色谱法（HPLC）、质谱法（MS）及核磁共振波谱法（NMR）对化合物进行分析、鉴定；采用CCK-8法，检测黄酮化合物对人肺腺癌A549细胞的抑制作用。结果:从镰形棘豆的乙酸乙酯部位分离得到2个黄酮类单体，鉴定为7-羟基二氢黄酮及2'，4'-二羟基二氢查尔酮，HPLC分析两者纯度均大于98%；CCK-8法结果显示，7-羟基二氢黄酮及2'，4'-二羟基二氢查尔酮均能显著抑制A549细胞活力，并呈量效关系，48 h的IC50值分别为137.6 μg/mL和63.83 μg/mL。结论:分离得到的2个黄酮成分对A549细胞均有良好的抑制作用，为后期抗肺癌机制研究提供了实验基础。聚酰胺柱色谱法具有富集黄酮的作用，适用于分离黄酮类成分，聚酰胺色谱法联合中压制备液相色谱法，提取分离镰形棘豆中黄酮类成分，简便易行，为分离黄酮类成分提供参考。 |
| English Summary: |
| To extract and isolate the flavonoids from Oxytropis falcata (Of)，and screen bioactive flavonoids on human lung adenocarcinoma A549 cells in vitro.Methods:Crude extraction was extracted from Of by ethyl acetate.Ethyl acetate extraction was further separated by polyamide column chromatography with gradient elution and purified by medium-pressure liquid chromatography to obtain the flavonoids; High performance liquid chromatography (HPLC)，mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR) were used to analyze and elucidate the structure of the obtained flavonoids.CCK-8 analysis was used to detect the inhibited viability of flavonoids on human lung adenocarcinoma A549 cells in vitro.Results:There were 2 flavonoids isolated from Of ethyl acetate fraction，and were identified as 7-hydroxy dihydroflavone and 2'，4'-dihydroxy dihydrochalcone.Purity of them were both greater than 98% by HPLC analyses; The results of CCK-8 showed that 7-hydroxy dihydroflavone and 2'，4'-dihydroxy dihydrochalcone could significantly inhibit the viability of A549 cells in a dose dependent manner.The IC50 values were 137.6 μg/mL and 63.83 μg/mL at 48 h，respectively.Conclusion:The 2 flavonoids have effective bioactivities on inhibiting proliferation of human lung adenocarcinoma A549 cells，which could provide the experimental basis for the further studies on anti-lung cancer mechanism.Polyamide column chromatography is suitable for enrichment and isolation of flavonoids.Polyamide column chromatography combined with medium pressure liquid chromatography is simple and easy to prepare various flavonoids.Thus，the method could provide a reference for obtaining flavonoids from Chinese medicines. |
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