基于双五氟苄基硫醚的气相色谱-质谱法测定大鼠脑组织中的H_(2)S

Determination of H_(2)S in rat brain tissues by GC-MS based on bispentafluorobenzyl sulfide

目的通过气相色谱-质谱(GC-MS)法测定硫化氢(H_(2)S)的衍生化产物双五氟苄基硫醚(bispentafluorobenzyl sulfide,C_(6)F_(5)CH_(2)SCH_(2)C_(6)F_(5)),建立大鼠脑组织中H_(2)S的测定方法。方法色谱条件:HP-5MS(30 m×250μm×0.25μm)色谱柱,色谱柱程序升温,进样口温度280℃。质谱条件:20 eV电子轰击离子源,离子源温度230℃,四极杆温度150℃,接口温度280℃。采用多反应检测扫描(MRM)模式,对C_(6)F_(5)CH_(2)SCH_(2)C_(6)F_(5)的离子对(m/z 394→181,m/z 181→161)进行定量、定性分析。结果脑组织样本中硫氢化钠(NaHS)在0.25~256μmol·L^(-1)范围内具有良好的线性关系,检测限为0.1μmol·L^(-1)。日内、日间精密度均小于15%,无明显的基质效应,回收率在90%以上。可选择性测定脑组织中的H_(2)S,测得大鼠脑皮质组织中基础H_(2)S浓度为(11.84±0.38)μmol·L^(-1),静脉注射NaHS后,脑组织中H_(2)S浓度呈剂量依赖性增高。结论C_(6)F_(5)CH_(2)SCH_(2)C_(6)F_(5)的GC-MS法是测定大鼠脑组织中H_(2)S的一种可靠、有效的方法,H_(2)S可透过血脑屏障进入脑组织。

Aim To establish a method for the determination of hydrogen sulfide(H_(2)S)in rat brain tissues by gas chromatography-mass spectrometry(GC-MS)on bispentafluorobenzyl sulfide(C_(6)F_(5)CH_(2)SCH_(2)C_(6)F_(5)).Methods Chromatographic conditions:The column was HP-5MS(30 m×250μm×0.25μm)and temperature programmed,the injection port temperature was 280℃.Mass spectrometry conditions:The electron bombardment ion source was 20 eV.The ion source,quadrupole and interface temperature was kept at 230,150 and 280℃,respectively.The MRM mode was used to quantitatively and qualitatively analyze the C_(6)F_(5)CH_(2)SCH_(2)C_(6)F_(5) ion pair(m/z 394→181,m/z 181→161).Results The concentration of sodium hydrosulfide(NaHS)in brain tissue samples had good linearity in the range of 0.25~256μmol·L^(-1).The limit of detection was 0.1μmol·L^(-1).The intra-day and inter-day precision were both less than 15%.There was no obvious matrix effect and the recovery rate was more than 90%.The H_(2)S concentration in brain tissues could be selectively determined.The basic H_(2)S concentration in rat brain cortex was measured to be(11.84±0.38)μmol·L^(-1).After intravenous injection of NaHS,the H_(2)S concentration in brain tissues increased significantly in a dose-dependent manner.Conclusions The GC-MS method based on C_(6)F_(5)CH_(2)SCH_(2)C_(6)F_(5) established here is reliable and effective to investigate H_(2)S in brain tissues,and H_(2)S could enter brain tissues through the blood-brain barrier.