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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
引用本文:李鹏,李绍平,龚元香,王一涛.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[J].药学学报,2004,39(11):917-920.
作者姓名:李鹏  李绍平  龚元香  王一涛
作者单位:1. 澳门大学,中华医药研究所,澳门;成都中医药大学,四川,成都,610075
2. 澳门大学,中华医药研究所,澳门;南京中医药大学,国家规范化中药药理实验室,江苏,南京,210029
3. 澳门大学,中华医药研究所,澳门
基金项目: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160 ℃,提取时间5 min,提取压力10 MPa,循环1次,提取1次。采用Zorbax NH2分析柱(250 mm×4.6 mm ID, 5 μm),流动相为A(乙腈)-B(10 mmol·L-1醋酸铵溶液)二元梯度洗脱:0-5.0 min,B%为0→15%;5.0-25.0 min,B%为15%→20%;25.0-35.0 min,B%为20%→ 40%;35.0-45.0 min,B%为40%→ 80%;45.0-50.0 min,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。

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
收稿时间:2004-02-09

Simultaneous determination of ergosterol, nucleosides and their bases from natural and cultured Cordyceps by pressurized solvent extraction and high performance liquid chromatography
LI PengLI Shao-ping,GONG Yuan-xiang,WANG Yi-tao.Simultaneous determination of ergosterol, nucleosides and their bases from natural and cultured Cordyceps by pressurized solvent extraction and high performance liquid chromatography[J].Acta Pharmaceutica Sinica,2004,39(11):917-920.
Authors:LI PengLI Shao-ping  GONG Yuan-xiang  WANG Yi-tao
Institution:Institute of Chinese Medical Sciences, University of Macau, Macau, China.
Abstract: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 degrees C; static extraction time, 5 min; pressure, 10 MPa and one extraction cycle and one time. A ZORBAX NH2 column (250 mm x 4.6 mm ID, 5 microm) and a ZORBAX NH2 guard column (12.5 mm x 4.6 mm ID, 5 microm) were used for simultaneous determination of ergosterol, nucleosides and their bases. Solvents that constituted the mobile phase were A (acetonitrile) and B (10 mmol x 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 x min(-1) and the injection volume was 20 microL. The system operated at 25 degrees C. 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.
Keywords:pressurized solvent extraction  high performance liquid chromatography  Cordyceps  ergosterol  nucleoside  base
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