LC-MS/MS同时测定不同产地雷公藤饮片中4种萜类成分的含量

Simultaneous Determination of Four Kinds of Terpenoids Composition in Prepared Slices of Radix et Rhizoma Tripterygii from Different Habitats by LC-MS/MS

目的建立简便、快速、灵敏的液相色谱-串联质谱法(LC-MS/MS)同时测定雷公藤饮片中雷公藤甲素、雷公藤红素、雷公藤内酯酮和雷公藤内酯甲的含量,为雷公藤饮片的质量控制与临床应用提供依据。方法采用液相色谱质谱联用技术。色谱条件:Agilent ZORBAX Eclipse Plus C18(2.1 mm×150 mm,3.5μm)色谱柱,乙腈-0.1%甲酸水为流动相进行梯度洗脱,流速为0.2 mL·min^(-1),柱温为35℃;采用电喷雾离子源(ESI),以正离子多反应监测模式进行检测,检测成分的离子对质荷比分别为:雷公藤甲素(361.1/145.1)、雷公藤内酯酮(358.9/143)、雷公藤红素(451.2/201.1)、雷公藤内酯甲(455.3/409.2)。结果雷公藤甲素、雷公藤内酯酮、雷公藤红素和雷公藤内酯甲分别在100~3000 ng·mL^(-1)、100~3000 ng·mL^(-1)、20~600 ng·mL^(-1)和50~1000 ng·mL^(-1)的浓度范围内,线性关系良好(r>0.999)。平均回收率为100.45%~102.74%,其RSD<3.15(n=6)。结论本研究所建立的LC-MS/MS色谱法稳定可靠、简便易行,可同时检测雷公藤饮片中雷公藤甲素、雷公藤红素、雷公藤内酯酮、雷公藤内酯甲4种萜类成分的含量,可为雷公藤饮片及雷公藤制剂的质量控制提供科学依据。不同地区的雷公藤饮片成分含量差异明显。

Objective To develop a LC- MS/MS method for simultaneous determination of triptolide, triptonide,celastrol and wilforlide A in prepared slices of Radix et Rhizoma Tripterygii from different habitats, thus to supply evidence for the quality control of the herbal medicinal material. Methods The assay was performed on a Zorbax Eclipse Plus C18（2.1 mm×150 mm,3.5 μm） and was the sample eluted with a gradient mobile phase which consisted of acetonitrile and 0.1 % formic acid water solution. The flow rate was 0.2 mL·min^（-1） and the temperature of the column was 35 ℃. The LC- MS/MS system was operated by using an electrospray ionization probe in the positive ion mode,and the scan mode was in multiple reaction ion monitoring（MRM） mode. The mass- to- charge ratio（m/z） was from 361.1 to 145.1 for triptolide,from 358.9 to 143 for triptonide,from 451.2 to 201.1 for celastrol,and from 455.3 to 409.2 for wilforlide A. Results The linear ranges of triptolide,triptonide,celastrol and wilforlide A were 100~3000ng·mL^（-1）,100~3000 ng·m L^（-1）,20~600 ng·m L^（-1） and 50~1000 ng·m L^（-1） respectively（r0.999）. The average recovery was from 100.45 % to 102.74 %, and RSD was lower than 3.15 %（n=6）. Conclusion A sensitive, accuracy and suitable LC- MS/MS method has been developed for the determination of triptolide,triptonide,celastrol and wilforlide A. The results will supply evidence for the quality control of prepared slices and compound preparation of Radix et Rhizoma Tripterygii. The results all indicated that the composition contents markedly vary in prepared slices of Radix et Rhizoma Tripterygii from different habitats.