应用离子对反相高效液相色谱法测定动物肝脏中APS和PAPS的含量

Determination of APS and PAPS by Ion-pair Reversed-Phase phase High-Performance Liquid Chromatography in Liver of Animals

[目的]建立适于动物肝脏中APS和PAPS含量测定的离子对反相高效液相色谱法,测定几种动物肝脏中APS和PAPS的含量.[方法]色谱条件：Ultimate AQ-C18反相色谱柱;流动相100mM KH2PO4,1mM四丁基氢氧化铵和甲醇,用4M的氢氧化钾调节pH至7.4;流速1 mL/min,上样量为10 μL,柱温25℃,检测波长254 nm.[结果]动物肝脏中APS含量（nmol/g）：羊：2.4-21.3;牛：0.9-1.1;马：1.4 -3.5;猪：1.4 -6.7;大鼠：9.5 -21.1.动物肝脏中PAPS含量（nmol/g）：羊：7.2-15.7;牛：4.7 -28.5;马：14.1-20.8;猪：7.7 -30.1;大鼠：23.5～74.5.[结论]该方法具有样品处理简单、待测成分分离度高（APS 27.7 min,PAPS 32.4min）、灵敏度高（APS 0.45 μg/mL,PAPS 0.48 μg/mL）的特点;测定的几种动物肝脏中APS和PAPS的含量具有种间差异性,牛、羊、猪、马等四种家畜中,猪肝脏中APS和PAPS的含量均最高.

[Objective] To develop a rapid ion-pair reversed-phase high-performance liquid chromatography （RP-HPLC） method for the quantitative determination of APS and PAPS in liver of animals.[Method] The chromatographic conditions were：Ultimatewere：ultimate AQ-C 18 reversed-phase column; mobile phase 100mM KH2PO4,1mM terabutylammonium hydroxide,and methanol.,brought to pH PH 7.4 with potassium hydroxide （4M） was used to be,filtered through a 0.45 μm Millipore filter and degassed prior to use.The flow-rate was 1.0 mL/min.The injection volume was 10 μL.Detection was performed at a wavelength of 254 nm under a constant temperature （25 ℃）.[Result] The contents of APS in animal livers （nmol/g） were：sheep：2.4 ～ 21.3 ; cattle：0.9 ～ 1.1 ; househorse：1.4 ～ 3.5 ; pig：1.4 ～ 6.7 ; rat：9.5 ～ 21.1.The contents of PAPS in animal livers （nmol/g） were：sheep：7.2 ～ 15.7 ; cattle：4.7 ～ 28.5 ; house：14.1 ～ 20.8 ; pig：7.7 ～ 30.1 ;rat：23.5 ～ 74.5.[Conclusion] The method has characteristics of simple sample processing,higher resolution （APS 27.7min,PAPS 32.4min） and sensitivity （APS 0.45 μg/mL,PAPS 0.48 μg/mL）.Results showed that there were differences among species in APS and PAPS concentrations of livers,with pigs having the highest contents.