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高效液相色谱法测定大鼠子宫组织中氟尿嘧啶的浓度 被引量:1

The determination of fluorouracil in rat uterus tissue by high performance liquid chromatography
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摘要 目的建立大鼠子宫组织中氟尿嘧啶(fluorouracil,5FU)的高效液相色谱(high performance liquid chromatography,HPLC)测定方法,为研究输注FUDR后5FU在体内的组织分布及代谢转换提供方法学基础。方法色谱柱:Shim-pack CLC-ODS柱(4.6mm×250mm,5μm);流动相:含2.8×10-4mol·L-1四丁基硫酸铵和0.05mol·L-1磷酸二氢钠(pH=4)的水溶液;流速:0.6ml·min-1;检测波长:254nm。将大鼠子宫组织样本精密称重并预先剪碎后,定量加入0.9%氯化钠注射液作为匀浆介质,匀浆仪下匀浆均匀后,取匀浆液200μl,以溴尿嘧啶为内标,采用正丙醇/乙醚(20:80)为提取液,进行两次提取后,合并提取液,常温氮气下吹干,用100μl流动相复溶后进样分析。结果本方法在0.050~5.000mg·L-1范围内线性良好,相关系数r=0.9996(n=4)。低、中、高3个浓度质控样本的批内变异(RSD)均在1.59%~5.24%之间,批间变异(RSD)在3.76%~5.26%之间,方法学回收率在101.60%~103.91%之间,检测限为0.050mg·L-1(相当于子宫组织FUDR浓度0.100μg·g-1)。结论本方法灵敏度高,重现性好,为下一步研究大鼠体内输注FUDR后FUDR的代谢物5FU在体内的组织分布转运提供方法学基础。 Objective To improve a HPLC method for the determination of fluorouracil in rat uterus tissue. Methods 5FU was quantitated by the reversed-phase HPLC using a Shim-pack CLC-ODS column (4.6mm×250mm,5μm). The mobile phase was consisted of 2.8×10^-4mol·L^-1 tetrabutyl-ammonium hydrogen sulfate in 0.05 mol·L^-1 sodium dihydrogen phosphate (pH=4) solution at a flow rate of 0.6 ml·min^-1. The detection wavelength was at 254nm. After identical volume of saline was added, each rat uterus tissue was homogenated using a homogenated apparatus. With 5-Bromouracil as the internal standard, 200μl rat uterus homogenated solution was extracted by n-propyl alcohol/aether. The organic layer was evaporated under nitrogen stream and the residual was reconstituted with the mobile phase. Results The linearity was obtained over the range of 0.050 to 5.000 mg·L^-1 of fluorouracil in saline with a good correlation coefficient (r=0.9996, n=4). The coefficients of intra -run and inter-run variation were 1.59%-5.24% and 3.76%-5.26% respectively. The mean recoveries were 103.91%, 101.60% and 102.90% for the low, middle and high concentration of check samples, respectively. The limit of detection was 0.050 mg·L^-1 (about 0.100μg·g^-1 FUDR in rat tissue). Conclusion The method was sensitive, specific and simple. It is suitable for the distribution research of 5FU in vivo after i.v FUDR in rats.
出处 《中国现代医药杂志》 2007年第2期32-34,共3页 Modern Medicine Journal of China
关键词 氟尿嘧啶 高效液相 大鼠子宫组织 Fluorouracil High performance liquid chromatography Rat uterus tissue
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  • 1冯婉玉,蔡爽,申建伟.高效液相色谱法测定人血浆中氟尿嘧啶浓度[J].中国药学杂志,2003,38(4):289-290. 被引量:10
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