HPLC法测定大鼠血液中H_2S的含量  

Development and validation of a HPLC method for determination of H_2S in rat blood

作　　者：李金芳[1] 马雪红[1] 宋百灵[1,2] 李新霞[1,3] 陈坚 

机构地区：[1]新疆医科大学药学院,乌鲁木齐830011 [2]新疆埃乐欣药业有限公司,乌鲁木齐830013 [3]新疆医科大学中心实验室,乌鲁木齐830011

出　　处：《新疆医科大学学报》2016年第1期56-58,63,共4页Journal of Xinjiang Medical University

基　　金：国家自然科学基金(81260486)

摘　　要：目的建立测定大鼠血液中H_2S含量的高效液相色谱法（HPLC法）。方法采用HPLC法,以乙腈-0.3%三乙胺=31：69（磷酸调pH=2.07）为流动相,检测波长为664 nm。将大蒜辣素（Allicin）加入大鼠血细胞中,37℃孵育10 min后,测定Allicin进入血细胞后释放的H_2S含量。结果HPLC法专属性符合要求,H_2S的线性范围为0.8~80μmol/L,定量限和检测限分别为0.8μmol/L和0.4μmol/L。将大蒜辣素加入大鼠血细胞中孵育后有H_2S产生,测得大蒜辣素产生的H_2S含量为3.23μmol/L。结论 HPLC法可用于生物样品中H_2S的含量测定,检测限满足内源性H2S测定要求。H2S是大蒜辣素在血细胞中孵育后的产物,大蒜辣素有望成为H_2S的供体。Objective To set up a HPLC method tor the determination of H_2S in rat blood.Methods Using a HPLC method,acetonitrile：0.3%trimethylamine（pH adjusted by phosphate to 2.07） =31：69 as mobile phase,and the detection wavelength was 664 nm.Allicin was added in rat blood cells and after incubation at 37°C for 10 min,measuring the concentration of H_2S,which was released by Allicin.Results HPLC method specificity met the determination of H_2S,a good linear relationship was obtained in the concentration range from 0.8 μmol/L to 80 μmol/L.The lowest limit of quantification（LLOQ） was 0.8 μmol/L,and the lowest limit of detection（LLOD） was 0.4μmol/L.H_2S can be detected after adding Allicin in rat blood cells and the concentration of H_2S was 3.23μmol/L.Conclusion This method was successfully applied to the determination of H_2S inbiological samples.The detection limits met the determination of endogenous H_2S.The results showed that H_2S was the product of allicin in the blood,which was expected to become the donor of H_2S.

关 键 词：大蒜辣素 HPLC 亚甲蓝法 H_2S 前药 

分 类 号：R914[医药卫生—药物化学]