学校：哈尔滨商业大学
教师：王宝庆
学生：刘彪
班级：生物制药 2012级药学二班
A selective pretreatment method for determination of endogenous active brassinosteroids in plant tissues: double layered solid phase extraction combined with boronate affinity polymer monolith microextraction 选择性预处理方法测定内源活跃:油菜甾醇内酯类植物生长调节剂在植物组织:双分层的固相萃取结合整体boronate关联聚合微萃取 Abstract(文摘)
Background: Brassinosteriods (BRs), a group of important phytohormones, have various effects on plant growth and
development. However, their physiological functions in plants have not been fully understood to date. Endogenous BRs in plant tissue are extremely low and the elucidation of BRs functions relies on sensitive detection method. Reported methods for the
determination of BRs required large amount of plant tissue, tedious pretreatment process, and were lack of selectivity. Therefore, development of a simple and selective method for the sensitive quantification of BRs is highly needed.
背景:Brassinosteriods(BRs),一群重要的植物激素,对植物的生长和发育有不同的影响。然而到目前为止,它们在植物中的生理功能还没有被完全理解。内源性BRS在植物组织极低并且BRS的函数说明依赖于敏感的检测方法。报告所需的BRS方法测定大量的植物组织,
繁琐的预处理过程,缺乏选择性。因此,开发一个简单的和有选择性的方法敏感的量化管理是非常必要的。
Results: We established a pretreatment method of BRs in plant tissues by employing double layered solid phase extraction (DL/SPE) combined with boronate affinity polymer monolith
microextraction (BA/PMME). After the initial depigmentation with DL/SPE cartridge, BA/PMME was employed to selectively extract BRs from sample matrix. Uniquely, most sample matrix was successfully removed by BA monolith purification. Using this method, BRs was determined by liquid chromatography-mass spectrometry (LC-MS). Endogenous active BRs could be detected in only 1 g fresh weigh (FW) leaves or 0.5 g FW flower tissues. 结果:我们建立了BRS的预处理方法，在植物组织中采用双分层的固相萃取(DL / SPE)结合boronate整体关联聚合微萃取(BA / PMME)。在最初的脱色与DL / SPE墨盒,英航/ PMME来选择性地从样本中提取的BRS矩阵。独特的,大多数样本矩阵被英航庞然大物净化成功移除。使用这种方法,黄铜是由液相色谱-光谱法(质)。内源性活性BRS只能检测到1克重量(FW)鲜叶或0.5 g弗兰克-威廉姆斯花组织。 Conclusion: A DL/SPE-BA/PMME pretreatment method for the determination of endogenous brassinosteroids in plant tissues was developed and validated. The proposed method was sensitive and
selective. Besides, it may be further developed for the
determination of other BRs including their precursors and conjugates.
结论:DL / SPE-BA / PMME预处理方法的确定对于内生油菜甾醇内酯类植物生长调节剂在植物组织是一种开发和验证。该方法是敏感性和选择性。除此之外,这可能进一步发展其他的BRS包括他们的前体和轭合物。
Keyword: Brassinosteroid, Boronate affinity chromatography, Polymer monolith microextraction, Double layered solid phase extraction, HPLC-ESI-MS/MS.
关键字:Brassinosteroid Boronate亲和色谱法、聚合物庞然大物微萃取,双分层的固相萃取,HPLC-ESI-MS /MS
Background Brassinosterods (BRs), confirmed as the sixth plant hormone, are a group of naturally occurring polyhydroxy steroids
[1]. Since the first BR was discovered in 1970 [2], around 60 natural occurrence of BRs have been reported with wide occurrence in the plant kingdom [1]. BRs have multiple functions on various physiological and metabolic processes and normally occur in
extremely low concentration. Many biologists have been dedicated to the researches on signal transduction, biosynthesis, degradation and metabolic pathway of BRs [3-5].
背景 Brassinosterods(BRS),确认为第六植物激素,是一群天然聚羟基类固醇[1]。自1970年发现了第一个BR[2],大约60个自然产生的BR已报告发生在植物王国[1]。BRs在极低的浓度下的不同的生理和代谢过程中可以有多个功能。许多生物学家一直致力于研究RBS的信号转导、生物合成、降解和代谢途径 [3 - 5]。