非诺贝特与代谢物的HPLC测定及在大鼠小肠淋巴转运中的应用  

Determination of Fenofibrate and Its Metabolite by HPLC and Its Application in Rat Intestinal Lymphatic Transport

作　　者：沈敏[1] 沈腾[2] 李中东[1] 弓艺涓[2] 施孝金[1] 钟明康[1] 

机构地区：[1]复旦大学附属华山医院临床药学研究室,上海200040 [2]复旦大学药学院药剂学教研室,上海201203

出　　处：《中国药学杂志》2011年第9期686-690,共5页Chinese Pharmaceutical Journal

基　　金：国家重大科技专项经费资助(2009ZX09310-006)

摘　　要：目的建立高效液相色谱法同时测定大鼠血浆和淋巴液中非诺贝特(fenofibrate,FEF)及代谢物非诺贝特酸(fenofibricacid,FEFA)的浓度,应用于大鼠在体内的FEF小肠淋巴转运研究。方法建立麻醉大鼠肠系膜淋巴插管和颈静脉插管模型,经十二指肠内给予FEF混悬液30 mg.kg-1后收集血液和淋巴液样品。血浆和淋巴液样品经含内标物萘普生的蛋白沉淀剂直接沉淀蛋白后,采用HPLC测定。色谱柱为Calesil ODS柱(4.6 mm×150 mm,5μm),流动相为甲醇-0.4%磷酸水溶液(pH3.2)(82∶18),流速为1.0 mL.min-1,检测波长为286 nm。结果 FEFA、FEF和IS的保留时间为4.25、9.15和3.3 min;血浆中FEFA线性范围为0.2～50μg.mL-1,淋巴液中FEFA、FEF线性范围均为0.1～25μg.mL-1(r>0.999);低、中、高3个质控浓度在血浆和淋巴液中的日内和日间精密度RSD均小于9%,准确度均在95%～110%,萃取回收率均大于84%。将HPLC应用于大鼠在体内的FEF小肠吸收研究,血浆中仅能检测FEFA,于(5.67±2.08)h达峰浓度为(13.29±2.17)μg.mL-1,淋巴液中能同时检测到FEFA和FEF,分别于(4.75±0.96),(3.75±1.26)h达峰浓度为(4.35±0.97),(0.51±0.13)μg.mL-1。结论本实验建立的RP-HPLC-UV法专属性及重现性好,准确可靠,可用于大鼠血浆和淋巴液中FEF和FEFA的同时测定。FEF经小肠吸收后部分经肠系膜淋巴转运进入全身血液循环。OBJECTIVE To develop a specific and accurate reversed-phase HPLC-UV method for simultaneous determination of fenofibrate（FEF） and its metabolite fenofibric acid（FEFA） in rats plasma and lymph fluid,then the method was applied in the study of intestinal lymphatic transport of fenofibate in rats.METHODS The intestinal mesenteric lymph duct and jugular vein of unconscious rats was cannulated for collecting blood and lymph fluid after intraduodenal adminstration of a single dose of 30 mg·kg-1 fenofibrate suspension.Plasma and lymph fluid samples were pretreated by protein precipitant containing internal standard naproxen and then analysed by the validated HPLC method.The analytical column was a Calesil ODS（4.6 mm×150 mm,5 μm）.The mobile phase was composed of methanol and 0.4% phosphoric acid solution（pH 3.2）（82∶18）.The flow rate was 1.0 mL·min-1.The wavelength of UV-detector was set at 286 nm.RESULTS The retention time of FEFA,FEF and IS were 4.25,9.15 and 3.3 min,respectively.The calibration curves showed good linear regression in the range of 0.2-50 and 0.1-25 μg·mL-1（r0.999） for plasma and lymph fluid samples.Intra-and inter-day variations were less than 9%,accuracy was 95%-110%,and extraction recovery was more than 84%.Both FEFA and FEF were detected in lymph fluid samples,while in plasma only FEFA was found.The values of ρmaxin plasma and lymph fluid samples were（13.29±2.17）,（4.35±0.97）,and（0.51±0.13） μg·mL-1,respectively and tmax were（5.67±2.08）,（4.75±0.96）,and（3.75±1.26） h,respectively.CONCLUSION The established and validated RP-HPLC-UV method is simple,sensitive and accurate for simultaneous determination FEF and FEFA in rats plasma and lymph fluid.The absorbed fenofibrate can be partially transported through mesenteric lymphatic routes into systemic circulation in rats.

关 键 词：非诺贝特 非诺贝特酸 高效液相色谱法 肠系膜淋巴管 插管 淋巴转运 

分 类 号：R917[医药卫生—药物分析学]