1、HPLC,How Does HPLC Separate Mixtures,speaker:Li changwei,Construction,There are five part in the Efficient HPLC.,precompressor(填装器),infusion apparatus （输液器）,separator（分离器）,detector 检测器,data processor（数据处理器）,HPLC,Efficient HPLC,Efficient HPLC,precompresso,inject needle tubing,ventricle with high pres
2、sure,Generally choose,or,To be beneficial to the repeatability of analytic sample,infusion apparatus,high-pressure pump,the pressure is l.474.4X107Pa,flow velocity can be adjusted smoothly,When liquid flow through chromato bar,diffusion effect may be fallen, and accelerate the velocityof liquid whic
3、h can be beneficial to heighten resolution,reclaiming materials and keeping bioactivity.,mobile phase container,gradient maker,mobile phase vary with the character of sample,infusion apparatus,infusion apparatus,include,character,the polarity of eluant,ionic strength,PH,As a result,all kinds of subs
4、tances can be separated effectively.,Separator,chromatographic column,length l=1050cm,inside diameter r=25mm,raytrace:titanium alloyand some stationary phases,stationary phase which are composed of base material and stationary liquid,Detector,a.Ultraviolet-detector,Generally speaking,there are three
5、 different kinds of detector:,This detector is used for the sample which can absorb ultraviolet and visible light.,There are some specialtys in it.,First of all,this detector can be used in a vast range,it can be used for detecting Pr,amino acid ,amino acid,hormone and polypeptide.,Secondly,it is se
6、nsitive.,Detector,Detector,Thirdly,it has wide linearity region,Lastly,this detector can be used for sample which is washed out by grads-solution.,Detector,b.differential-refractometer detector,If the refractive indices of mobile phase are different,the sample may be detected by it.,But it can not b
7、e used in trace bound and the sample which is washed out by grads-solution.,Detector,c.fluorescence-detector,Under certain conditions,the light intensity of fluorescent material gathers head with its concentration.,So,this detector can be used in a narrow range,it can be only used for detecting some
8、 Pr,amino acid ,amino acid,hormone and polypeptide.,Data processor,this system is used for,data collection,storage,exhibition,mimeograph,deal with data,Data processor,Data processor,Y:voltage X:time,Theory,mobile phasein reservoir,is injected into,high-pressure pump,system,A.,When sample-liquid flow
9、 into mobile phase,mobile phase drag liquid into chromatographic column(solid).,B.,Theory,Because of the different partition coefficients,when sample-liquid flow through mobile phase and solid,and sample going from place to place oppositely,by adsorption-desorption time after time.,Theory,As a resul
10、t,sample-liquid is broken down into many single components,and also they issue from chromatographic column respectively.,C.,Theory,Concentration of single components will be converted into electrical signal which can be recorded by grapher , and also these datum wil be printed in the way of atlas.,D.,Theory,Theory,THATS ALL Thank you,
