摘要
目的:建立液相微萃取光化学荧光高效液相色谱法测定生物样品中枸橼酸氯米芬顺反异构体的含量。方法:应用自制的液相微萃取装置,对生物样品中的枸橼酸氯米芬顺反异构体萃取后,分离采用 Sinochrom ODS—BP C_(18)色谱柱(200 mm×4.6mm,5μm);流动相为甲醇-水(70∶30,每1L 水含0.25 mL 磷酸、50μL三乙胺);流速为0.8 mL·min^(-1);荧光检测器的激发波长和发射波长分别为255 nm 和378 nm;柱温20℃。结果:利用该方法可以将血浆、尿液和肝脏组织匀浆中的枸橼酸氯米芬顺反异构体同时提取分离,浓缩倍数可达5～7倍;枸橼酸氯米芬在血浆、尿液和肝脏中的线性范围分别为0.05～20μg·mL^(-1),0.02～20μg·mL^(-1),0.04～40μg·g^(-1);检出限分别为20 ng·mL^(-1),10μg·mL^(-1),20 ng·g^(-1);精密度试验的 RSD 小于11.7%。结论:本文将液相微萃取技术应用于生物样品中枸橼酸氯米芬顺反异构体的提取分离,利用高效液相色谱法荧光检测,获得了较好的效果。
Objective：To establish a method for determining the concentration of the E and Z isomers of clomifene in biological matrices by liquid - phase microextraction combined with photochemical fluorescence high performance liquid chromatography. Methods： By liquid - phase microextraction apparatus self - made, the E and Z isomers of clomifene citrate were extracted simultaneously, separated on a Sinochrom ODS - BP C18 column （ 200 mm × 4. 6 mm, 5 μm） ;The mobile phase consisted of methanol - water（ 70： 30, with 0. 25 mL of phosphoric acid and 50μL of triethylamine per litre） at a flow rate of 0. 8 mL · min^- 1 ; determined with fluorescence detector with excitation wavelength at 255 nm and emission wavelength at 378 nm. Column temperature was controlled about 20 ℃. Replasma, urine and liver respectively ; The detection limit （ S/N = 3 ） were 20 ng · mL^- 1,10 ng 2 mL^- 1 and 20 ng · g^-1 respectively for this method; The relative standard deviation of precision was below 11.7%. Conclusion： The liquid -phase microextraction is first applied in abstraction of clomifene in biological sample, combined with photochemical fluorescence high performance liquid chromatography, more sensitivity are achieved. The method can be applied for analysis of the E and Z isomers of clomifene in biological matrices.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2008年第1期49-53,共5页
Chinese Journal of Pharmaceutical Analysis
基金
山西省自然科学基金资助项目(20031104)
关键词
枸橼酸氯米芬
液相微萃取
光化学荧光
高效液相色谱
clomifene citrate
liquid - phase microextraction
photochemical fluorescence
high performance liquid chromatography
