加压溶剂提取高效液相色谱法测定天然和人工虫草中的麦角甾醇、核苷及其碱基  

Simultaneous determination of ergosterol, nucleosides and their bases from natural and cultured Cordyceps by pressurized solvent extraction and high performance liquid chromatography

作　　者：李鹏[1] 李绍平[1] 龚元香[1] 王一涛[1] 

机构地区：[1]澳门大学中华医药研究所

出　　处：《药学学报》2004年第11期917-920,共4页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目 (3 0 2 715 85 ) ;澳门大学研究基金资助项目 (RG0 45 0 2 -0 3S LSP ICMS FST) .

摘　　要：目的　建立同时测定虫草中麦角甾醇、核苷及其碱基含量的简便方法。方法　正交法优化加压溶剂提取条件 ;HPLC法同时测定天然和人工虫草中上述成分的含量。结果　以甲醇为提取溶剂 ,提取温度 16 0℃ ,提取时间 5min ,提取压力 10MPa ,循环 1次 ,提取 1次。采用ZorbaxNH2 分析柱 (2 5 0mm× 4 6mmID ,5 μm) ,流动相为A(乙腈 ) B(10mmol·L- 1 醋酸铵溶液 )二元梯度洗脱 :0 - 5 0min ,B %为 0→ 15 % ;5 0 - 2 5 0min ,B %为 15 %→ 2 0 % ;2 5 0 - 35 0min ,B %为 2 0 %→ 4 0 % ;35 0 - 4 5 0min ,B %为 4 0 %→ 80 % ;4 5 0 - 5 0 0min ,B %为 80 %。结论　本法能够快速、简便地测定虫草中麦角甾醇、核苷及其碱基的含量。Aim To establish a simple method for simultaneous determination of ergosterol, nucleosides and their bases in Cordyceps by HPLC coupled with pressurized solvent extraction (PSE). Methods The extraction was performed by using PSE and the PSE condition was optimized by using orthogonal test, the contents of ergosterol, nucleosides and their bases in Cordyceps were determined by HPLC. Results The optimized condition of PSE extraction for ergosterol, nucleosides and their bases in Cordyceps was obtained as follow: solvent, methanol; temperature, 160 ℃; static extraction time, 5 min; pressure, 10 MPa and one extraction cycle and one time. A ZORBAX NH 2 column (250 mm×4.6 mm ID, 5 μm) and a ZORBAX NH 2 guard column (12.5 mm×4.6 mm ID, 5 μm) were used for simultaneous determination of ergosterol, nucleosides and their bases. Solvents that constituted the mobile phase were A (acetonitrile) and B (10 mmol·L -1 ammonium acetate in water). The elution conditions applied were: 0-5.0 min, linear gradient 0→15% B; 5.0-25.0 min, linear gradient 15%→20% B; 25.0-35.0 min, linear gradient 20%→40% B; 35.0-45.0 min, linear gradient 40%→80% B; 45.0-50.0 min, 80% B isocratic. The flow-rate was 0.6 mL·min -1 and the injection volume was 20 μL. The system operated at 25 ℃. Ergosterol was monitored and quantified at 275 nm, nucleosides and their bases at 254 nm.Conclusion High performance liquid chromatography coupled with pressurized solvent extraction is a rapid and simple method for simultaneous determination of ergosterol, nucleosides and their bases in Cordyceps.

关 键 词：加压溶剂提取 高效液相色谱法 虫草 麦角甾醇 核苷 碱基 

分 类 号：R917[医药卫生—药物分析学]