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| 桑树桑黄总三萜的提取纯化及其生物活性研究 |
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刘 坤1,2, 付鑫垚1, 郭 彤1, 张婉月1, 赵庆生3, 孔泽娟4, 程 华4*, 宋 恒5
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1. 河北经贸大学 生物科学与工程学院,石家庄 050061；2. 河北经贸大学数学与统计学院,石家庄 050061；3. 中国科学院过程工程研究所,北京 100190；4. 河北省科学院生物研究所,石家庄 050081；5. 太安谷(浙江）中药饮片有限公司桑树桑黄研究室,浙江 嘉兴 314000
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| 摘要: |
| 为了优化桑树桑黄总三萜的提取工艺，分析其化学成分及其抗肿瘤和抗氧化活性。该研究采用响应面法优化桑树桑黄总三萜的提取工艺，经D101型大孔树脂和Sephadex LH-20层析联用纯化总三萜，结合UHPLC-ESI-MS技术分析三萜成分、并通过CCK8法、DPPH法和ABTS法对其进行抗肿瘤和抗氧化活性测定。结果表明：（1）最佳提取工艺为提取时间55 min、提取温度45 ℃、乙醇浓度75%、液料比35∶1（mL·g-1），此条件下总三萜含量为10.34 mg·g-1。（2）纯化后的Fraction a（Fa）组分纯度为47.98%，提高了3.8倍；从Fa中检测出8种三萜成分，即甘草次酸、黄柏酮、熊果酸、大戟醇、24，25-二氢羊毛甾醇、大豆皂苷B、羊毛甾醇、山楂酸。（3）200 μg·mL-1时Fa对肿瘤细胞PC3的抑制率为68.65%，高于5-氟尿嘧啶66.3%；Fa对DPPH和ABTS自由基均有较高的清除能力，半数清除率分别为42.76、66.24 μg·mL-1。该研究为桑树桑黄总三萜的进一步研究和利用提供实验数据支持。 |
| 关键词: 桑树桑黄，三萜，响应面优化，纯化，抗肿瘤，抗氧化 |
| DOI：10.11931/guihaia.gxzw202404018 |
| 分类号:R283.6 |
| Fund project:河北省重点研发计划项目(21327109D)；河北省科学院基本科研业务费制度试点项目（2024PF04） |
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| Extraction, purification and biological activity of total triterpenoids from Sanghuangporus sanghuang |
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LIU Kun1,2, FU Xinyao1, GUO Tong1, ZHANG Wanyue1, ZHAO Qingsheng3, KONG Zejuan4, CHENG Hua4*, SONG Heng5
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1.College of Bioscience and Bioengineering,Hebei University of Economics and Business;2.Institute of Process Engineering Chinese Academy of Sciences;3.Institute of biology,Hebei academy of science;4.Institute of Sanghuangporus sanghuang,Peace Angel Zhejiang Traditional Chinese Medicine Slices Co,Ltd
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| Abstract: |
| Aim to optimize the extraction process of total triterpenes from Sanghuangporus sanghuang, purify the total triterpenes, analyze its chemical composition, as well as its antitumor and antioxidant activities. This study first used the response surface method to optimize the extraction process of total triterpenes from S. sanghuang. Secondly, D101 macroporous resin and Sephadex LH-20 chromatography were used to purify the total triterpenes. Meanwhile UHPLC-ESI-MS technology was used to analyze the composition of triterpenes, Finally, the anti-tumor and antioxidant activities of total triterpenes were studied by the methods of CCK8, DPPH and ABTS. The results were as follows: (1) The optimal conditions were extraction time 55, ultrasonic temperature 45 ℃, ethanol concentration 75% and a liquid-solid ratio of 35:1 (mL·g-1). Under these conditions, the total triterpene content was 10.34 mg·g-1. (2) The purity of Fraction a (Fa) was increased by 3.8 times to 47.98%. Eight triterpene components were identified from Fa: glycyrrhetinic acid, xibutanone, ursolic acid, euphorbiol, 24, 25-dihydrolanosterol, soya saponin B, lanosterol and cratonic acid. (3) The inhibition rate of Fa on PC3 was 68.65% at 200 μg·mL-1, which was higher than that of 5-fluorouracil (66.3%). Fa exhibited high DPPH and ABTS radical-scavenging activities with IC50 values of 42.76 and 66.24 μg·mL-1, respectively. This study provides a theoretical foundation for further research and utilization of total triterpenes from S. sanghuang. |
| Key words: Sanghuangporus sanghuang, triterpene, response surface optimization, purification, antitumor, antioxidant |
