中文摘要：

类固醇激素对脑功能的调节越来越受到人们的关注,但其在脑组织中极其微量而且脑组织样品量相对少,这对检测方法提出了更高的挑战。本研究优化了脑组织中提取类固醇激素的预处理方法,发现乙醇的提取效果最好。经过固相萃取柱进一步纯化的样品,基质干扰明显减少。采用75μm内径的C18捕集柱和分析柱,通过阀切换连接,构建了在线微柱富集纳升液相色谱分析系统,结合纳喷离子源串联质谱法实现高灵敏检测脑组织中类固醇激素。系统考察了方法的精密度、定量曲线、回收率等分析特性。日内和日间精密度分别在4.5%～12.8%和8.3%～20.3%之间。除了孕酮略有离子抑制外,其它激素基质效应不明显,而且低、中、高3个浓度下的回收率基本都在60%～115%之间。不同类固醇激素的检出限差异较大,在0.02～10μg/L之间。与常规尺寸液相色谱-质谱系统相比,相同进样量下,该方法的灵敏度显著提高了57～714倍,同时流动相消耗量降低了几百倍。将本方法用于大鼠脑组织中类固醇激素的检测,检测到4种类固醇激素,分别为睾酮（Testosterone）0.47 ng/g,肾上腺酮（Corticosterone）26.2 ng/g,醛甾酮（Aldosterone）0.49 ng/g和皮质醇（Hydrocortisone）0.06 ng/g。本方法具有灵敏度高、样品需要量少和溶剂消耗少等优势,可用于生物组织样品中类固醇激素的测定。

英文摘要：

The modulation of steroid hormones on brain is attracting more and more attention.However,the detection of steroid hormones is faced with great challenge owning to the very low content in brain tissue.In this work,the preparation conditions of steroid hormones extracted from brain tissue were optimized.Ethanol was the best extraction solvent for brain homogenization as it readily penetrated cell membranes and was a good solvent for steroid hormones.After purified by Oasis HLB SPE,the matrix effect was reduced significantly.An on-line capillary solid phase extraction-nano liquid chromatographic system was built based on valve-switching 75 μm i.d.trap column and analytical column.Tandem mass spectrometer with NanoESI source was used as the detector for the determination of ten brain steroid hormones.The method showed good precision（intraday RSD 4.5%-12.8% and inter day RSD 8.3%-20.3%） and linearity（r0.995）.The limits of detection in solution were from 20 ng/L to 10 μg/L for 10 steroid hormones.The sensitivity of the method was gained in two orders of magnitude compared with that of conventional HPLC/MS when the equal amounts of the steroid hormones were injected.The recoveries were above 60% at three concentrations except for androsterone while the matrix effect was inapparent except for progesterone.Brain tissue samples from mouse were analyzed using the new method.Four steroids were identified and quantified including testosterone 0.47 ng/g,corticosterone 26.2 ng/g,aldosterone 0.49 ng/g and hydrocortisone 0.06 ng/g.The results showed the method has many advantages such as high sensitivity,low sample amount and solvent consumption which is suitable for the determination of steroid hormones in biological tissue.