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首页> 外文期刊>Journal of mass spectrometry: JMS >Study of the fragmentation of arginine isobutyl ester applied to arginine quantification in Aedes aegypti mosquito excreta
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Study of the fragmentation of arginine isobutyl ester applied to arginine quantification in Aedes aegypti mosquito excreta

机译：精氨酸异丁酯片段化技术在埃及伊蚊蚊精液中精氨酸定量研究中的应用

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摘要

It has been demonstrated that argininolysis and uricolysis are involved in the synthesis and excretion of urea in Aedes aegypti female mosquitoes. To further investigate the metabolic regulation of urea in female mosquitoes, it is desirable to have a rapid and efficient method to monitor arginine (Arg) concentration in mosquito excreta. Thus, a procedure currently used for the identification of Arg in urea cycle disorders in newborn babies was adapted to analyze Arg in A. aegypti excreta. The fragmentation patterns of the isobutyl esters of Arg and ~(15)N _2-Arg (labeled at the guanidino group) were explored by electrospray ionization (ESI)-tandem mass spectrometry and fragmentation pathways not described before were characterized. In addition, Arg, ~(18)O _2-Arg, ~(15)N _2-Arg and ~(15)N _2- ~(18)O _2-Arg were also analyzed to elucidate some of the minor fragments in greater detail. Mosquito excreta from individual females were collected before and at different times after feeding a blood meal, mixed with ~(15)N _2-Arg, an internal standard, and then derivatized as isobutyl esters. Based on the fragmentation mechanisms of Arg standards, studied by MS ~2 and MS ~3, Arg in the mosquito excreta was successfully analyzed by ESI-multiple reaction monitoring in a triple-quadrupole mass spectrometer. Arg excretion was monitored over a 120 h window before and after feeding female mosquitoes with a blood meal, with the maximum level of Arg excretion observed at 36-48 h post blood feeding. This method provides an efficient and rapid tool to quantify Arg in individual blood-fed mosquitoes, and can be applied to other organisms, whose small size severally limits the use of conventional biochemical analysis.
机译：已经证明精氨酸分解和尿液分解参与埃及伊蚊雌蚊的合成和排泄。为了进一步研究雌性蚊子中尿素的代谢调节，希望有一种快速有效的方法来监测蚊子排泄物中精氨酸（Arg）的浓度。因此，目前用于鉴定新生儿尿素循环异常中的Arg的方法适用于分析埃及伊曲菌排泄物中的Arg。通过电喷雾电离（ESI）-串联质谱研究了Arg和〜（15）N _2-Arg（标记在胍基上）的异丁酸酯的碎裂模式，并表征了之前未描述的碎裂路径。此外，还对Arg，〜（18）O _2-Arg，〜（15）N _2-Arg和〜（15）N _2-〜（18）O _2-Arg进行了分析，以阐明一些较小的碎片。详情。在喂食血粉之前和之后的不同时间收集来自各个雌性的蚊子排泄物，将其与内标物〜（15）N _2-Arg混合，然后衍生为异丁酯。基于MS〜2和MS〜3研究的Arg标样的断裂机理，在三重四极杆质谱仪中通过ESI-多重反应监测成功地分析了蚊虫排泄物中的Arg。在给雌性蚊子加血粉之前和之后，在120小时内监测Arg的排泄，在喂血后36-48 h观察到最大的Arg排泄水平。该方法提供了一种有效，快速的工具，可以定量检测单个血源性蚊子中的Arg，并且可以应用于其他生物，这些生物的小尺寸分别限制了常规生化分析的使用。

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