摘要
采用高效液相电化学检测的方法测定了大鼠下丘脑内组胺和5_羟色胺。色谱柱为NUCLEOSILRC18(250mm×4mm ,10μm) ,流动相 :磷酸二氢钠溶液 (25mmol/L ,内含乙二胺四乙酸二钠0.5mmol/L ,辛烷磺酸钠3mmol/L,pH4.8) -乙腈=100∶14(体积比 ) ,流速0.8mL/min,检测电位0.6V。大鼠下丘脑匀浆液经低温高速离心 ,上清液直接进样测定5_羟色胺 ,另取上清液衍生后进样测定组胺。组胺在0.075～1.8μg/mL ,5_羟色胺在0.0625～1.25μg/mL浓度范围内 ,线性关系良好。组胺测定的日内和日间相对标准偏差分别小于1.0%和11.4 %,5_羟色胺测定的日内和日间相对标准偏差分别小于3.4 %和6.3 %,两者回收率分别为94.1 %～102.3%和72.8 %～89.1 %。正常雄性SD大鼠下丘脑内组胺和5_羟色胺的平均含量分别为0.280±0.029μg/mL和0.224±0.068μg/mL。本法简便、准确、快捷 。
Ahighperformance liquid chromatograph(HPLC)withanelectrochemical detector(ECD)and a NUˉCLEOSIL R C 18 column(250mm×4mm,i.d.10μm)was used for the determination of histamine(HA)and5-hydroxytryptamine(5-HT)in rat′s hypothalamus.The mobile phase was a mixture of NaH 2 PO 4 (25mmol/L,containing0.5mmol/LdisodiumEDTAand3mmol/Lsodiuml-octanesulfonate at pH4.8)and acetonitrile in a ratio of100∶14(by volume).The flowrate was0.8mL/min,and the detector voltage was set at0.6V.The homogenate of rat′s hypothalamus was centrifuged at4℃.A part of the supernatant fraction was injected for the direct determination of5-HT,and another part of the supernatant was allowed to react with a derivatizaˉtion reagent before the analysis of HA.Good linearity was found in the range of0.075～1.8μg/mL for HA and0.0625～1.25μg/mL for5-HT.The within-day and intra-day RSDs of the method were≤1.0%and11.4%for HA,and≤3.4%and6.3%for5-HT,respectively.The recoveries of HA and5-HT were94.1%～102.3%and72.8%～89.1%,respectively.The average contents of HA and5-HTin hypothalaˉmus of normal male Sprague-Dawley rats were0.280±0.029μg/mL and0.224±0.068μg/mL,respecˉtively.The method is simple,accurate and rapid,and can be used for the determination of HA and5-HT in rat′s hypothalamus.
出处
《分析测试学报》
CAS
CSCD
北大核心
2004年第6期36-39,共4页
Journal of Instrumental Analysis
基金
广东省自然科学基金资助项目 (980644)
