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机构地区:[1]广东省物料实验检测中心,广州市东风东路751号510080 [2]广州开发区环境监测站,广州开发区友谊路105号510730
出 处:《光谱实验室》2011年第4期2074-2078,共5页Chinese Journal of Spectroscopy Laboratory
摘 要:运用高效液相色谱-电感耦合等离子体-质谱法(HPLC-ICP-MS)研究了硒代胱氨酸(SeCys2)在目前广泛应用的水浴振荡提取和方便快速的超声提取过程中的稳定性。在水溶性SeCys2的提取过程中,不管是37℃水浴振荡过程还是超声提取过程对SeCys2的稳定性影响都不大。在酶溶性SeCys2的提取过程中,由于SeCys2分子很容易在Se—Se间断裂,而在水浴振荡或超声作用下酶容易发生多肽链断裂或空间构型变化,游离出来的硒代半胱氨酸(SeCys)和酶分子中断裂出来的小分子结合形成新的存在形式,从而导致了SeCys2在C18柱的洗脱时间发生迁移。The stability of selenocystine(SeCys2) in the most widely used extraction procedures of Se species(traditional water bath shaking and fast ultrasonic extraction)was studied by high performance liquid chromatography(HPLC)-inductively coupled plasma-mass spectrometry(ICP-MS).The results showed that SeCys2 was stable by both water bath(37℃) shaking and ultrasonic extraction for water leaching of Se species.During the enzymatic hydrolysis,the Se—Se bond of SeCys2 fractured easily,SeCys was free from SeCys2,and rupture of polypeptide chain or space configuration changes of pronase E caused by external forces(water bath shaking or sonication).The effects of water bath shaking and ultrasonic extraction to the enzyme were different,and SeCys were easily combined with the fragments of enzyme,which caused the transfer of retention time of SeCys2 on C18 column.
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