高效液相色谱-串联质谱法测定雷公藤制剂中5种有效成分的含量

目的:建立高效液相色谱-串联质谱法测定雷公藤制剂中雷公藤甲素、雷公藤内酯酮、雷公藤晋碱、雷公藤次碱和雷公藤红素的含量。方法:采用电喷雾正电离模式和多反应监测模式。雷公藤甲素、雷公藤内酯酮、雷公藤晋碱和雷公藤次碱含量测定,以含5 mmol·L^-1碳酸氢铵的水溶液和纯甲醇作为流动相,采用梯度洗脱方式在ACE UltraCore SuperPhenylHexyl(2.1 mm×50.0 mm,5 μm)色谱柱上进行分离。雷公藤红素含量测定,以含0.5%甲酸的水溶液和含0.1%甲酸的乙腈作为流动相,采用等度洗脱在ACE Excel C₁₈-AR(2.1 mm×50.0 mm,5 μm)色谱柱上进行分离。结果:雷公藤甲素、雷公藤内酯酮、雷公藤晋碱、雷公藤次碱和雷公藤红素分别在2.4~1 200,2.0~1 000,80~40 000,80~40 000和10~5 000 ng·mL^-1范围内线性关系良好,平均加样回收率分别为 94.56%,102.36%,93.27%,95.67%和99.82%,RSD均低于5.13%。结论:本方法运行时间短,灵敏度高,精密度和重复性好,适用于雷公藤制剂中雷公藤甲素、雷公藤内酯酮、雷公藤晋碱、雷公藤次碱和雷公藤红素5种活性成分定量检测,可为雷公藤制剂多指标质量控制提供实验依据。

Determination of five active components in various Tripterygium wilfordii preparations by HPLC-MS/MS

OBJECTIVE To establish a method for the determination of triptolide, triptonide, wilforgine, wilforine and celastrol in various Tripterygium wilfordii preparations by high performance liquid chromatography-tandem mass spectrometry.METHODS ESI⁺ scan and multi-reaction monitoring mode were adopted. The determination of triptolide, triptonide, wilforgine and wilforine was performed on an ACE UltraCore SuperPhenylHexyl (2.1 mm×50.0 mm, 5 μm) chromatographic column with gradient elution by an aqueous solution containing 5 mmol·L^-1 ammonium bicarbonate and methanol as the mobile phase. For the analysis of celastrol, the chromatographic separation was conducted on an ACE Excel C₁₈-AR (2.1 mm×50.0 mm, 5 μm) column with isocratic elution by a mixture of buffer (0.5% formic acid in water) and acetonitrile containing 0.1% formic acid as the mobile phase.RESULTS The assay was linear within the range of 2.4-1 200, 2.0-1 000, 80-40 000, 80-40 000 and 10-5 000 ng·mL^-1 for triptolide, triptonide, wilforgine, wilforine and celastrol, respectively. The average recovery was 94.56%, 102.36%, 93.27%, 95.67% and 99.82%, respectively with the RSD below 5.13%.CONCLUSION The method has high sensitivity, good precision and repeatability with a short run time, which is suitable for the quantitative determination of triptolide, triptonide, wilforgine, wilforine and celastrol in various Tripterygium wilfordii preparations. It provides experimental basis for the quality control of multiple components in Tripterygium wilfordii preparations.