| The extraction, purification and analysis method of the biologically active composition chlorogenic acid in ramie leaf and paclitaxel in Taxus were discussed.(1) Technological extraction process of chlorogenic and paclitaxel with organic solvent was experimented.The solvent of 70% ethanol aqueous was used to extract from ramie leaf three times for a duration of two hours, ratio of feed-stock weight to the volume of solvent is 1:10. Temperature is 70℃ and pH is 4.0, the extraction ratio was 83%.The extraction ratio of paclitaxel was 90.24% when extracted with 1:10 of 85% ethanol aqueous, twice at 75 ℃ and 2 hours.(2) Effects of mobile phase, acid and velocity on the analysis of chlorogenic acid and paclitaxel were discussed as well.The content of chlorogenic acid in ramie leaf were determined by HPLC and the analysis condition of chlorogenic acid was as follows: ODS column (200x4.6 mm, 5 um), 18% methanol aqueous and 1.0% acetic acid as mobile phase, UV detector at 326 nm, injection volume was 6 U L, the experimental separating time was 14 min. The calibration curve was Y=1.618 X 106X-23081.3 (r=0.9997) and the linear determination range is 20mg/L-160mg/L, the recovery was 97% -101 %. The method could meet the requirement of quickly analysis and various results; the content of chlorogenic acid is 0.52%.The analysis condition of paclitaxel was as follows: methanol: watenacetic acid=55:44:1 (v/v), UV detector at 221 nm, the experimental separating was 28min and the calibration cure was y = 13021.7 + 1.01 X 106x (r=0.9990) which was linear in the concentration range of 0.05mg/mL-0.5mg/mL and the recovery was 96% -104%.(3) A method for the colorimetry of chlorogenic acid was studied. The optimal coloration and examining conditions were as follows: appropriative sample was added in the colorimetric tube, then evaporated methanol, added 1.00mL H2O, 0.100mL acetic acid aqueous, 2.00mL urea aqueous and 0.250mL sodium nitrite aqueous, blendedthem evenly, 0.500mL sodium hydroxide aqueous tree minutes later. The adsorption value was measured at 510nm within one hour with water as consultation. The calibration curve was A=0.037+0.1602X (r=0.9998) and the linear is in the range of 0.05mg-0.6mg, the recovery was 95% - 107%, the RSD of the content of chlorogenic acid was 2.4%. (4) Technological process of purification for chlorogenic acid and paclitaxel were completed by chromatographic column.Employed with 15X400 mm column, the installation was simple and the adsorption and desorption was easy to operate. In this experiment, the ethanol was used which is no pollution and no poison. Chlorogenic acid was absorbed well by AB-8 resin. The condition obtained was as follows: The content of chlorogenic acid in injection sample is 1.4mg/mL and pH is 4.0 (4% Nacl was added, w/v) at the flow rate 1.5BV/h, the injection sample must be added again and the adsorption time is 30min, then eluted by 30% ethanol aqueous which pH is 9.0 at the rate 1.0BV/h, the desorption ratio was 95% and the optimum adsorption amount by chlorogenic acid adsorption-desorption experiment was 23.8mg/mL. The purity of chlorogenic acid reached 41.31% from 3.53%.The Al2O3 chromatographic process established the conversion and separation of paclitaxel from Taxus, the column is 15 X 250 mm while the reaction time is 30min, the balancing liquid is 2% CH3OH and 98% CHCl3 and the eluent compositions is 4% CH3OH and 96% CHCl3. The purity of paclitaxel reached 16.59% from 1.87% with the recovery of 170% after the one chromatographic step.
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