基于含量测定与药效活性相结合的川楝子质量评价
Quality Evaluation of Toosendan Fructus Based on Content Determination and Pharmacological Activity
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摘要: 目的 分析比较不同批次川楝子代表性成分川楝素、异川楝素含量及颜色值，分析比较不同批次川楝子提取物对TRAIL诱导非小细胞肺癌细胞凋亡的增效作用和对非小细胞肺癌细胞膜死亡受体DR5表达的影响，二者相结合共同评价川楝子的质量。方法 采用UHPLC-QqQ-MS/MS对其所含川楝素、异川楝素进行含量测定，色谱柱为 Poroshell 120 SB C18柱（2.1 mm×100 mm，1.8 μm），流动相为0.1%甲酸水溶液-乙腈，流速为0.3 mL/min，进样量为1 μL，柱温为35 ℃；同时采用分光测色仪测定川楝子饮片的颜色值。采用流式细胞仪检测非小细胞肺癌A549细胞凋亡率与细胞膜死亡受体DR5的表达。结果 10批川楝子中川楝素、异川楝素的含量分别为1.024~4.128，0.010~0.045 mg/g；平均颜色值（L*，a*，b*）为68.56，5.27，25.12。10批川楝子均能显著增加非小细胞肺癌细胞膜死亡受体DR5表达，增强TRAIL诱导非小细胞肺癌细胞凋亡。结论 建立川楝子饮片的含量测定与颜色检测方法，结合细胞药效活性，可准确用于川楝子的质量控制。
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关键词:
- 川楝子 /
- 川楝素 /
- 异川楝素 /
- UHPLC-QqQ-MS/MS /
- 细胞凋亡
Abstract: OBJECTIVE To compare the contents and color values of two representative components， toosendanin and isotoosendanin， in different batches of toosendan fructus， and to compare the effects of different batches of toosendan fructus extracts on the expression of DR5 and the apoptosis of NSCLC cells induced by TRAIL. METHEODS The contents of toosendanin and isotoosendanin in toosendan fructus were determined by UHPLC-QqQ-MS/MS. The analysis was performed on Poroshell 120 SB C18 (2.1 mm×100 mm，1.8 μm) column， mobile phase was 0.1% formic acid aqueous solution and acetonitrile with flow rate of 0.3 mL/min. The injection volume was 1 μL， and the column temperature was 35 ℃. Meanwhile， spectrophotometer was used to determine the color value of the samples. Flow cytometry analysis was used to detect apoptosis and the expressions of cell death receptor DR5 for TRAIL in non-small cell lung cancer cell line A549. RESULTS The contents of toosendanin and isotoosendanin in 10 batches of samples were 1.024~4.128 mg/g and 0.010~0.045 mg/g， respectively. The average of colour value (L*，a*，b*) was 68.56，5.27 and 25.12. The extracts from 10 batches of toosendan fructus could significantly increase the expression of DR5 and restore the apoptotic sensitivity of A549 cells towards TRAIL. CONCLUSION The content determination and color measurement， combined with pharmacological activity can be successfully applied to accurately evaluate the quality of toosendan fructus.-
Key words:
- toosendan fructus /
- toosendanin /
- isotoosendanin /
- UHPLC-QqQ-MS/MS /
- apoptosis
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