补骨脂酚超临界CO2萃取工艺优化及萃取物抗肿瘤活性研究OA
Optimization of supercritical CO2 extraction process for bakuchiol and antitumor activity of the extract
目的 优化补骨脂酚的超临界CO2萃取工艺,并评价在该最适宜条件下萃取物的体外抗肿瘤活性.方法 采用单因素实验考察萃取压力(15 MPa、20 MPa、25 MPa)以及乙醇浓度(30％、50％、70％)对补骨脂萃取物得率和萃取物质量损失率的影响.采用高效液相色谱法(HPLC)测定萃取物中补骨脂酚的含量;通过CCK-8法检测在最适宜条件下萃取物对多种肿瘤细胞的增殖抑制作用,同时评估其对正常细胞的细胞毒性.结果 在考察萃取压力单因素实验中,15 MPa、20 MPa、25 MPa条件下萃取物得率分别为7.90％、17.30％、17.15％,萃取物质量损失率分别为4.00％、1.00％、10.00％.因此,最适宜萃取压力为20 MPa.在萃取压力为20 MPa时,30％、50％、70％乙醇条件下萃取物得率分别为12.23％、7.77％、14.17％,失重率分别为47.3％、26.0％、23.3％,其中70％乙醇组萃取物得率最高,30％乙醇组次之.HPLC分析结果显示20 MPa 30％乙醇条件下萃取釜Ⅱ中补骨脂超临界CO2萃取物的补骨脂酚含量为47.4％,20 MPa 70％乙醇条件下萃取釜Ⅱ中补骨脂酚含量为26.19％,且色谱图显示20 MPa 70％乙醇条件下的补骨脂超临界萃取物有更多的杂峰,说明30％乙醇夹带剂条件为最适宜浓度.体外抗肿瘤细胞活性结果显示,最适宜条件下(20 MPa 30％乙醇)萃取物对人肝癌细胞HepG2、顺铂耐药肝癌细胞BEL-7402/DDP、人乳腺癌细胞MCF-7、阿霉素耐药乳腺癌细胞MCF-7/ADR均具有显著的增殖抑制作用,且呈剂量依赖性(5～40 μg/mL);对正常肝细胞LX-2毒性较低,安全性较好.结论 成功构建了补骨脂酚的绿色高效超临界CO2萃取工艺.体外活性实验证实,该工艺所得萃取物能显著抑制耐药肿瘤细胞的增殖,并对正常细胞表现出低毒性.
Objective To optimize the supercritical CO2 extraction process for bakuchiol and to evaluate the in vitro antitumor activity of the extract obtained under the optimal conditions.Methods Single-factor experiments were conducted to investigate the effects of extraction pressure(15,20,25 MPa)and ethanol concentration(30％,50％,70％)on the yield and mass loss rate of the Buguzhi(Psoraleae Fructus)extract.The bakuchiol content in the extract was determined by high-performance liquid chromatography(HPLC).The CCK-8 assay was used to assess the inhibitory effects of the extract obtained under optimal conditions on the proliferation of various tumor cell lines,as well as its cytotoxicity against normal cells.Results In the single-factor pressure experiments,the extract yields at 15,20,and 25 MPa were 7.90％,17.30％,and 17.15％,respectively,while the corresponding mass loss rates were 4.00％,1.00％,and 10.00％.Therefore,the optimal extraction pressure was determined to be 20 MPa.At 20 MPa,with ethanol concentrations of 30％,50％,and 70％,the extract yields were 12.23％,7.77％,and 14.17％,and the mass loss rates were 47.3％,26.0％,and 23.3％,respectively.The 70％ethanol group gave the highest yield,followed by the 30％ethanol group.HPLC analysis showed that under 20 MPa and 30％ethanol,the bakuchiol content in the extract from extraction vessel Ⅱ was 47.4％,whereas under 20 MPa and 70％ethanol it was 26.19％.Moreover,the chromatogram of the 70％ethanol extract exhibited more impurity peaks,indicating that 30％ethanol as the entrainer was the optimal concentration.In vitro antitumor activity tests demonstrated that the extract obtained under the optimal conditions(20 MPa,30％ethanol)significantly inhibited the proliferation of human hepatoma HepG2 cells,cisplatin-resistant hepatoma BEL-7402/DDP cells,human breast cancer MCF-7 cells,and doxorubicin-resistant breast cancer MCF-7/ADR cells in a dose-dependent manner(5～40 μg/mL).Meanwhile,it showed low cytotoxicity against normal liver LX-2 cells,indicating good safety.Conclusion A green and efficient supercritical CO2 extraction process for bakuchiol was successfully established.In vitro bioactivity assays confirmed that the extract obtained by this process significantly suppresses the proliferation of drug-resistant tumor cells while exhibiting low toxicity to normal cells.
陈鑫;戴宏芳;向贞虹;李晨煜;贺卫和;黄伟荣;覃艳
湖南中医药大学药学院,湖南长沙 410208湖南中医药大学药学院,湖南长沙 410208湖南中医药大学药学院,湖南长沙 410208湖南中医药大学药学院,湖南长沙 410208湖南中医药大学药学院,湖南长沙 410208长沙理工大学物理与电子科学学院,湖南长沙 410114||云南衡金药业有限公司,云南楚雄 675000湖南中医药大学药学院,湖南长沙 410208
医药卫生
补骨脂酚超临界CO2萃取工艺优化高效液相色谱抗肿瘤活性
bakuchiolsupercritical CO2 extractionprocess optimizationhigh-performance liquid chromatography(HPLC)antitumor activity
《药食同源杂志》 2026 (6)
516-524,9
湖南中医药大学中药粉体与创新药物省部共建国家重点实验室培育基地开放基金资助项目(24PTKF1017).
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