[关键词]
[摘要]
研究赤灵芝总三萜最佳提取工艺条件,评价三萜类化合物对脂肪氧化酶的抑制效果。采取单因素实验和响应面法分析提取时间、提取次数、液料比和乙醇浓度对赤灵芝三萜得率的影响,筛选最佳工艺参数;利用高效液相色谱-电喷雾质谱联用技术(HPLC-ESI-MS)对赤灵芝中三萜类提取物化学成分进行分析鉴定,以脂肪氧化酶为生物靶分子,采用超滤质谱技术筛选酶抑制剂,探究三萜类化合物对脂肪氧化酶的抑制效果。结果显示:最佳工艺条件为料液比1:30、乙醇浓度75%、提取次数3次和提取时间1.5 h;在这些条件下,总三萜得率为1.04%;从赤灵芝中筛选出5种脂肪氧化酶抑制剂,5种化合物对1.0 U/mL脂肪氧化酶活性有较强抑制作用,抑制效果的强弱顺序为灵芝酸B(52.64%)>灵芝酸C2(40.21%)>灵芝酸D2(31.25%)>灵芝酸F(12.69%)>灵芝酸A(10.82%)。实验结果表明,响应面法筛选赤灵芝三萜的提取工艺参数准确可靠,可用以赤灵芝三萜的生产制备;赤灵芝中三萜类活性成分可抑制脂肪氧化酶,表明有缓解炎症的功效,研究结果可为赤灵芝中抗炎成分的进一步研究提供一定理论基础。
[Key word]
[Abstract]
In this study, the optimal extraction conditions of the total triterpenes from Ganoderma lucidum were investigated, and the inhibitory effect of triterpenoids on lipoxygenase was evaluated. Single factor experiments and response surface method were used to analyze the effects of extraction time, extraction times, liquid-to-material ratio, and ethanol concentration on the yield of triterpenes from Ganoderma lucidum, and screen the optimal process parameters. High performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS) was used to analyze and identify the chemical constituents of the triterpenoid extracts from Ganoderma lucidum. Lipoxygenase was used as the biological target molecule, and ultrafiltration mass spectrometry was used to screen enzyme inhibitors and examine the inhibitory effect of the triterpenoids on lipoxygenase. The results revealed that the optimal process conditions were material-to-liquid ratio of 1:30, ethanol concentration of 75%, three extraction times, and extraction time of 1.5 h. Under these conditions, the extraction rate of total triterpenes was 1.04%; five types of lipoxygenase inhibitors were screened out from Ganoderma lucidum, which demonstrated a strong inhibitory effect on the activity of lipoxygenase at 1.0 U/mL. The order of the inhibitory effect was as follows: ganoderic acid B (52.64%) >ganoderic acid C2 (40.21%) >ganoderic acid D2 (31.25%) >ganoderic acid F (12.69%) >ganoderic acid A (10.82%). The experimental results showed that the extraction process parameters of red Ganoderma lucidum triterpenes screened by the response surface methodology are accurate and reliable, and can be used for the processing and preparation of Ganoderma lucidum triterpenes. The active triterpenoid components in Ganoderma lucidum can inhibit lipoxygenase, indicating their abilities to relieve inflammation. The research results can provide a theoretical basis for further studies on the anti-inflammatory components in Ganoderma lucidum.
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[基金项目]
国家自然科学基金项目(31870336);吉林省科技发展计划项目(20200201114JC);长师大自科合字(2020)第011号