目的:建立同时测定雷公藤饮片中雷公藤甲素、雷公藤内酯酮、雷酚内酯、雷公藤次碱、雷公藤内酯甲、雷公藤红素6个活性成分RP-UPLC-PDA分析方法。方法:雷公藤饮片采用乙酸乙酯超声提取,甲醇溶解分离,利用RP-UPLC-PDA法对其6个成分进行定量分析,采用AcquityTM UPLC BEH C18 (100 mm×2.1 mm, 1.7 μm)色谱柱,柱温30 ℃,以乙腈(A)-0.1%甲酸水(B)为流动相,进行梯度洗脱,流速0.4 mL·min-1,进样量2 μL。结果:6个活性成分在测定浓度范围内线性关系良好(0.999 2≤r≤0.999 7);加样回收试验中平均回收率为99.2%~103.1%,RSD为1.2%~2.9%。对10批不同产地雷公藤饮片进行含量测定,结果显示不同产地饮片中成分含量具有差异性,雷公藤甲素含量最高为140.2 μg·g-1,最低为103.2 μg·g-1;雷公藤内酯酮含量最高为224.7 μg·g-1,最低为112.2 μg·g-1;雷酚内酯含量最高为306.7 μg·g-1,最低为189.6 μg·g-1;雷公藤次碱含量最高为283.2 μg·g-1,最低为211.2 μg·g-1;雷公藤内酯甲含量最高为31.2 μg·g-1,最低为16.8 μg·g-1;雷公藤红素含量最高为87.6 μg·g-1,最低为52.1 μg·g-1。结论:建立RP-UPLC-PDA法能够同时测定雷公藤饮片中6个成分,方法稳定,结果可靠,适用于雷公藤饮片定量分析测定。
Objective: To establish the RP-UPLC-PDA method for simultaneous determination of triptolide, triptonide, triptophenolide, wilforine, wilforlide A and celastrol in Tripterygii Radix. Methods: Tripterygii Radix were extracted with ethyl acetate and the extracts were dissolved and separated by methanol. The six components were determined by RP-UPLC-PDA method. The chromatographic column was AcquityTM UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm), the column temperature was 30 ℃, the injection volumn was 2 μL, the flow rate was 0.4 mL·min-1 and the mobile phase was acetonitrile (A)-0.1% formic acid (B). Results: The linear relationship of six components was good (0.999 2≤r≤0.999 7) in the concentration ranges. The average recoveries were 99.2%-103.1% and RSDs were 1.2%-2.9%. The contents in 10 batches of Tripterygii Radix from different habitat were determined. The results showed that the contents of Tripterygii Radix in prepared pieces from different producing areas were different. The highest content of triptolide was 140.2 μg·g-1, and the lowest content was 103.2 μg·g-1. The highest content of triptonide was 224.7 μg·g-1 and the lowest content was 112.2 μg·g-1. The highest content of triptophenolide was 306.7 μg·g-1 and the lowest content was 189.6 μg·g-1. The highest and lowest contents of wilforine were 283.2 μg·g-1 and 211.2 μg·g-1. The highest content of wilfhabitat was 31.2 μg·g-1 and the lowest content was 16.8 μg·g-1. The highest content of celastrol was 87.6 μg·g-1, and the lowest content was 52.1 μg·g-1. Conclusion: The RP-UPLC-PDA method can simultaneously determine six components in Tripterygii Radix. The method is reliable and stable, which is suitable for quantitative analysis and determination of Tripterygii Radix.
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