Quantificational Analyses Of Rice Endogenous Constituents By High Performance Liquid Chromatography Coupled With Electrospray Ionization Tandem Mass Spertrometry

Plant endogenous coustituents are important physiolygical compounds in plant life cycle.It is essential to quatify and quantify the endogenous constituents in fresh plant tissues.This paper arms to develope some rapid and efficient methods for quatification analyses of some endogenous constistuents,such as nicotianamine, zeatin,glucose,fructose and sucrose.The new methods for separation and determination are significant of plant life activities.It includes three parts as follow:In the first chapter,a new method for separation and determination of nicotianamine was developed by HPLC-ESI/MS.The fress rice tissues was extracted with ddH₂O at 80℃for 30min and then to centrifigate.The supernatant was separated directed by a reverse phase column,and then the effusion was analyzed by ESI-MS.The linear range is 0.127-65 ng/20μL,and the limite of detection at three times signal to noise(LOD,3*S/N)is 3.175 ng/mL.In the sencond chapter,the zeatin in rice was analyzed for the first time by 2D-HPLC and using "Heart-cutting" mode.The rice sample extracting with 80% methanol-H₂O was separated primarily in the first dimension,then the aimed part of first dimension run is introduced to separate futher into the second dimension by switching a six port valve.The linear range is 0.0537-27.5 ng,and the LOD(3*S/N) is 0.026 ng.The aavantages of 2D-HPLC are high purification potential, reproducibility,and unattended operation.The third chapter,a useful HPLC-ELSD method for quatification analyses of sugars-glucose,fructose and sucrose was estabilished.Before analyzed by HPLC-ELSD,the sample first was extracted by 80%ethanol-H₂O at 90℃for 40min and purified by chloroform-Isoamylol(24:1).The linear range of glucose,fructose and sucrose is 0.44-5.275,0.228-5.5 and 0.422-10.15μg/μL,and the LOD(3*S/N)is 0.11,0.114 and 0.212μg/μL.