论文部分内容阅读
目的建立一种测定血液中虫草素含量的反相高效液相色谱法。方法大鼠全血0.4 ml,加入甲醇1.6 ml,4℃12000 r/min离心10 min,取上清液50℃氮气吹干,用0.15 ml甲醇复溶,12000 r/min离心15 min,取上清液10μl进样;色谱条件为ODS-SP C18色谱柱(4.6 mm×250 mm,5μm),以甲醇∶水=27∶73为流动相;流速0.8 ml/min;柱温35℃;紫外检测波长259 nm;检测血中虫草素含量。结果虫草素浓度在0.1~5.0μg/ml范围内线性关系良好,相关系数为0.9998;浓度为0.2、1.0、5.0μg/ml时日内RSD(%)分别为5.38、0.63、2.95,日间RSD(%)分别为2.20、4.41、2.57;检出限(S/N=3)为0.05μg/ml;全血中浓度为0.1、0.8、1.5μg/ml时回收率为85%~102%。结论该反相高效液相色谱法灵敏度高,重复性好,能有效分离检测血中虫草素含量。
Objective To establish a method for the determination of cordycepin in blood by reversed-phase high performance liquid chromatography. Methods Rat whole blood 0.4 ml, adding methanol 1.6 ml, centrifuged at 12000 r / min at 4 ℃ for 10 min, the supernatant was dried under nitrogen at 50 ℃, reconstituted with 0.15 ml methanol, centrifuged at 12000 r / min for 15 min, 10 μl of supernatant was injected into the column. The chromatographic conditions were ODS-SP C18 column (4.6 mm × 250 mm, 5 μm) with methanol: water = 27:73 as mobile phase, flow rate of 0.8 ml / min, Wavelength 259 nm; detection of cordycepin in blood. Results Cordycepin had a good linearity in the range of 0.1-5.0 μg / ml, with a correlation coefficient of 0.9998. The intraday RSD (%) at the concentrations of 0.2, 1.0 and 5.0 μg / ml were 5.38, 0.63 and 2.95, respectively. %) Were 2.20,4.41,2.57 respectively. The detection limit (S / N = 3) was 0.05μg / ml. The recoveries were 85% -102% at 0.1,0.8,1.5μg / ml in whole blood. Conclusion The reversed-phase high-performance liquid chromatography with high sensitivity, good reproducibility, can effectively separate and detect cordycepin in blood.