在线SPE-HPLC系统在补骨脂二氢黄酮甲醚药代动力学研究中的应用

目的: 探讨在线固相萃取(SPE)-高效液相色谱(HPLC)系统在小鼠血浆中补骨脂二氢黄酮甲醚药代动力学研究的应用。 方法: 基于在线SPE-HPLC技术的Ultimate 3000系统测定补骨脂二氢黄酮甲醚血药浓度。色谱条件:采用Venusil MP C₁₈(150 mm×4.6 mm,5 μm)色谱柱,流动相为乙腈-5 mmol·L^-1磷酸二氢钾缓冲液(pH 3.5),流速1.0 mL·min^-1,梯度洗脱;使用Capcell MF Ph-1在线SPE柱(10 mm×4 mm,5 μm),洗脱剂为5 mmol·L^-1磷酸二氢钾缓冲液(pH 3.5);检测波长238 nm。WinNonlin 5.2软件计算药动学参数。 结果: 补骨脂二氢黄酮甲醚线性范围为20.0~4000 ng·mL^-1,最低定量限为20.0 ng·mL^-1,提取回收率为90.5%~94.6%,日内、日间精密度试验的RSD均小于10%,短期稳定性、冻融稳定性及长期稳定性准确度为92.38%~101.7%。 结论: 该方法工作流程简化,样品分析快速,提高了工作效能,其结果可信且重复性好。

Objective: To study the application of online solid phase extraction-high performance liquid chromatography(SPE-HPLC) systems in the plasma pharmacokinetic study of bavachinin in mouse. Methods: Online SPE-HPLC systems were applied on determination of bavachinin in mouse plasma.The chromatographic conditions were as follows:a C₁₈ column(Venusil MP,150 mm×4.6 mm,5μm) was adopted with a linear gradient elution,and the mobile phase consisted of acetonitrile-5 mmol·L^-1 monopotassium phosphate buffer at a flow rate of 1.0 mL·min^-1;a MF Ph-1 online SPE column(10 mm×4 mm,5 μm) was adopted with isocratic elution,and the washing solvent was 5 mmol·L^-1 monopotassium phosphate buffer;the detected wavelength was set at 238 nm.The pharmacokinetic parameters were calculated by WinNonlin 5.2 software. Results: The linear range of the calibration curve was 20.0-4000 ng·mL^-1,and the lower limit of qualification was 20 ng·mL^-1.The extraction recovery was 90.5%-94.6%.The RSD of intra-day and inter-day precision was less than 10%.The accuracy of short-term stability,freeze-thaw stability and long-term stability was from 92.38% to 101.7%. Conclusion: The method is proved to be simple,rapid and high efficiency,and the result is reliable and reproducible.

[1] ZHAO Lu-hua(赵陆华),HUANG Chao-yu (黄朝瑜),TU Ying (屠颖),et al.Determination of six constituents in Fructus Psoraleae by HPLC(HPLC法同时测定补骨脂药材中6种成分的含量).Chin J Nat Med(中国天然药物),2005,3(4):242
[2] YANG Rong-ping(杨荣平),WANG Bin-hao(王宾豪),TU Yong-qin(涂永勤),et al.Determination of coumarin and flavonoid in Psoralen corylifolia L.in cornpound Buguzhi sustained-release tablet by HPLC(HPLC测定复方补骨脂缓释片中补骨脂香豆素类和黄酮类成分的含量).Chin Tradit Pat Med(中成药),2007,29(1):68
[3] Marchi I,Viette V,Badoud F,et al.Characterization and classification of matrix effects in biological samples analyses.J Chromatogr A,2010,1217(25):4071
[4] Ye Guan,Li Yin-zeng,Li Yan-yan,et al.SPE-HPLC method for the determination and pharmacokinetic studies on paeoniflorin in rat serum after oral administration of traditional Chinese medicinal preparation Guan-Xin-Er-Hao decoction.J Pharm Biomed Anal,2003,33:521
[5] Sheng Yuxin,Li Lie,Wang Chuanshe,et al.Solid-phase extraction-liquid chromatographic method for the determination and pharmacokinetic studies of albiflorin and paeoniflorin in rat serum after oral administration of Si-Wu decoction.J Chromatogr B,2004,806:127
[6] Rao RN,Vali RM,Shinde DD,et al.On-line 2D-LC-ESI/MS/MS determination of rifaximin in rat serum.Biomed Chromatogr,2009,23(11):1145
[7] Chen LG,Yu AM,Zhuang XD,et al.Determination of andrographolide and dehydroandrographolide in rabbit plasma by on-line solid phase extraction of high-performance liquid chromatography.Talanta,2007,74(1):146
[8] Chiuminatto U,Gosetti F,Dossetto P,et al.Automated online solid phase extraction ultra high performance liquid chromatography method coupled with tandem mass spectrometry for determination of forty-two therapeutic drugs and drugs of abuse in human urine.Anal Chem,2010,82(13):5636