课件3‘液相色谱法.pptVIP

一、高效液相色谱中的速率方程 迁移的流动相 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 二、高效液相色谱仪 流程 检测器 Fixed Wavelength UV Detector 固定波长 Variable Wavelength UV Detector 可变波长 Diode Assay Detector (DAD) 二极管阵列 A. Fixed Wavelength UV Detector Typical flow-through cell B. Variable Wavelength UV Detector C. Diode Assay Detector Spectrum collection at any time Library search Purity check Quantitative analysis at several wavelengths 3-D plot of DAD 3. Features of UV detector Selective detection, such as aromatic species 选择性 High sensitivity 高灵敏度 Large linear range 线性宽 Good stability for the change of mobile phase and temperature 稳定 Suitable for gradient eluting.可做梯度 Workhorse detection method in HPLC 2. 示差折光检测器 Universal response，通用 Low sensitivity，灵敏度低 The baseline is seriously affected by temperature，温度敏感 Any changes in the eluent composition require the rebalancing of the detector. Not suitable for gradient eluting.不能做梯度 3 荧光检测器 Selective detection. Such as PAH, protein, or compounds made fluorescent by derivatization. 高选择性 High sensitivity. Up to 1000 times more sensitive than the UV detector. 高灵敏度 Its high specificity also results in less interferences from the background (i e from the solvent or from impurities). 干扰少 Suitable for gradient eluting.可做梯度 Detection at any excitation or emission wavelength. 4 ELSD Universal detection. Such as sugars, Fats, Surfactants, Polymers, Natural compounds. 通用 Better sensitivity than RI，灵敏度较高 Faster equilibration than RI，方便 Capable of detection in gradient HPLC systems.可梯度 Universal response gives a closer representation of sample mass than UV detectors, making quantification more accurate especially when standards are not available. 响应与质量有关 5 电化学检测器 Conductivity detection: measurement of electrical current carried by dissolved ions in an electric field.电导 DC Amperometric detection: measurement of current resulting from oxidation or reduction (electrolysis) of analyte molecules at the surface of an electrode.直流安培