RP-HPLC法测定人血浆中的替勃龙

Method for detection of tibllone in human plasma by reversed-phase high performance liquid chromatography

采用反相高效液相色谱法检测人体血浆中替勃龙的含量.以5倍样品体积的乙腈萃取2次,经氮气吹干,甲醇溶解后,上机分析.采用Promosil-C18色谱柱(4.6mm×250mm,5μm),流动相:甲醇-乙腈-水,流速:1.0mL/min,进样量:10μL,检测波长:204nm,柱温:25℃.在优化的色谱条件下,替勃龙在0.5～250.0μg/mL内线性关系良好(r=0.999 9),最低检测限为0.05μg/mL,平均回收率(n=3)为92.7%,RSDs小于1.6%.本方法简单,灵敏,准确,适合于体内替勃龙含量的检测与分析.

A method of reversed-phase high performance liquid chromatography（RF-HPLC） was developed for the determination of tibolone in the plasma of people.The samples were extracted twice with 5 times voloum of acetonitrile.After the solvent was removed with nitrogen,the residues were dissolved in methanol.The separation column was Promosil-C18（250 mm×4.6 mm,5 μm） and the mobile phase was V（methanol）∶V（acetonitrile）∶V（water）（50∶30∶20）,The flow rate was 1.0 mL/min,The injection volume was 10 μL and the detection wavelength was set at 204 nm,The column temperature was 25 ℃.Under the optimized separation conditions,the calibration curves showed good linearity within the concentrations of 0.5-250.0 μg/mL（r=0.999 9）,The limit of detection was 0.05 μg/mL,The average recovery（n=3） was 92.7%,and the RSDs were less than 1.6%.This method is simple,sensitive and accurate,and suitable for the determination of tibolone in biological samples.