高效液相的色谱法.ppt

?

第三章高效液相色谱法

HighPerformanceLiquidChromatography

(HPLC)

;一、高效液相色谱中的速率方程;

滞留的流动相;

涡流扩散项纵向扩散流动相传质滞留区传质固定相内传质

;5;二、高效液相色谱仪;检测器;FixedWavelengthUVDetector固定波长

VariableWavelengthUVDetector可变波长

DiodeAssayDetector(DAD)二极管阵列

;A.FixedWavelengthUVDetector;Typicalflow-throughcell;B.VariableWavelengthUVDetector;C.DiodeAssayDetector;Spectrumcollectionatanytime

Librarysearch

Puritycheck

Quantitativeanalysisatseveralwavelengths

;3-DplotofDAD;IsoabsorbanceplotofDAD;3.FeaturesofUVdetector;17;2.示差折光检测器;Universalresponse，通用

Lowsensitivity，灵敏度低

Thebaselineisseriouslyaffectedbytemperature，温度敏感

Anychangesintheeluentcompositionrequiretherebalancingofthedetector.

Notsuitableforgradienteluting.不能做梯度

;3荧光检测器;Selectivedetection.SuchasPAH,protein,orcompoundsmadefluorescentbyderivatization.高选择性

Highsensitivity.Upto1000timesmoresensitivethantheUVdetector.高灵敏度

Itshighspecificityalsoresultsinlessinterferencesfromthebackground(iefromthesolventorfromimpurities).

干扰少

Suitableforgradienteluting.可做梯度

Detectionatanyexcitationoremissionwavelength.;22;4ELSD;Universaldetection.Suchassugars,Fats,Surfactants,Polymers,Naturalcompounds.通用

BettersensitivitythanRI，灵敏度较高

FasterequilibrationthanRI，方便

CapableofdetectioningradientHPLCsystems.可梯度

UniversalresponsegivesacloserrepresentationofsamplemassthanUVdetectors,makingquantificationmoreaccurateespeciallywhenstandardsarenotavailable.响应与质量有关

;25;26;5电化学检测器;Selectivedetection:suchascarboxylic,alcohols,carbohydrates;amines;sulfates,inorganicionsetc.

Highsensitivity.

Conductivity:theworkhorsedetectionmethodinionchromatography.

DCamperometry:neurochemicalanalyses.

Pulsedamperometry:carbohydrates.;附件;梯度洗提;31;装置

低压梯度（内梯度）：溶剂在常压下混合，再用高压泵输送至柱系统。简单、经济，只需一个泵，所用溶剂的元数没有限制。

高压梯度（外梯度）：一般由两台高压泵构成，每台泵输送一种溶剂。溶剂在混合室混合后，在输???柱系统。溶剂的可压缩性和热力学体积的变化可能影响输入柱子中溶剂的组成。;溶剂A;