《食品化学与分析 Food Chemistry and Analysis》课程PPT教学课件（英文版）Liquid Chromatography

CHROMATOGRAPHY Chromatography basically involves the separation of mixtures due to differences in the distribution coefficient (equilibrium distribution) of sample components between 2 different phases. One of these phases is a mobile phase and the other is a stationary phase

CHROMATOGRAPHY Chromatography basically involves the separation of mixtures due to differences in the distribution coefficient (equilibrium distribution) of sample components between 2 different phases. One of these phases is a mobile phase and the other is a stationary phase

Definition: Different affinity of these 2 components to stationary phase causes the separation. Concentration of component A in stationary phase Concentration of component A in mobile phase Distribution Coefficient (Equilibrium Distribution )

Definition: Different affinity of these 2 components to stationary phase causes the separation. Concentration of component A in stationary phase Concentration of component A in mobile phase Distribution Coefficient (Equilibrium Distribution )

Kinds of Chromatography 1. Liquid Column Chromatography 2. Gas Liquid Chromatography 3. Thin-layer Chromatography

Kinds of Chromatography 1. Liquid Column Chromatography 2. Gas Liquid Chromatography 3. Thin-layer Chromatography

LIQUID COLUMN CHROMATOGRAPHY A sample mixture is passed through a column packed with solid particles which may or may not be coated with another liquid. With the proper solvents, packing conditions, some components in the sample will travel the column more slowly than others resulting in the desired separation

LIQUID COLUMN CHROMATOGRAPHY A sample mixture is passed through a column packed with solid particles which may or may not be coated with another liquid. With the proper solvents, packing conditions, some components in the sample will travel the column more slowly than others resulting in the desired separation

A + B + C OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOO OOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOO OOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOO OOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO Sample (A+B+C) Column Solid Particles (packing material￾stationary phase) Eluant (eluate) D I A G R A M O F S I M P L E L I Q U I D C O L U M N C H R O M A T O G R A P H Y A B C Solvent(mobile or moving phase)

A + B + C OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOO OOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOO OOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOO OOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO OOOOOOOOOO OOOOOOOOOOO OOOOOOOOOOO Sample (A+B+C) Column Solid Particles (packing material￾stationary phase) Eluant (eluate) D I A G R A M O F S I M P L E L I Q U I D C O L U M N C H R O M A T O G R A P H Y A B C Solvent(mobile or moving phase)

The 4 basic liquid chromatography modes are named according to the mechanism involved: 1. Liquid/Solid Chromatography (adsorption chromatography) A. Normal Phase LSC B. Reverse Phase LSC 2. Liquid/Liquid Chromatography (partition chromatography) A. Normal Phase LLC B. Reverse Phase LLC 3. Ion Exchange Chromatography 4. Gel Permeation Chromatography (exclusion chromatography) FOUR BASIC LIQUID CHROMATOGRAPHY

The 4 basic liquid chromatography modes are named according to the mechanism involved: 1. Liquid/Solid Chromatography (adsorption chromatography) A. Normal Phase LSC B. Reverse Phase LSC 2. Liquid/Liquid Chromatography (partition chromatography) A. Normal Phase LLC B. Reverse Phase LLC 3. Ion Exchange Chromatography 4. Gel Permeation Chromatography (exclusion chromatography) FOUR BASIC LIQUID CHROMATOGRAPHY

LIQUID SOLID CHROMATOGRAPHY   Si - O - H   Normal phase LS Reverse phase LS Silica Gel The separation mechanism in LSC is based on the competition of the components of the mixture sample for the active sites on an absorbent such as Silica Gel

LIQUID SOLID CHROMATOGRAPHY   Si - O - H   Normal phase LS Reverse phase LS Silica Gel The separation mechanism in LSC is based on the competition of the components of the mixture sample for the active sites on an absorbent such as Silica Gel

LIQUID SOLID CHROMATOGRAPHY Si - OH HEXANE OH C-CH3 CH3 CH3- C CH3 CH3 OH OH CH3 CH3

LIQUID SOLID CHROMATOGRAPHY Si - OH HEXANE OH C-CH3 CH3 CH3- C CH3 CH3 OH OH CH3 CH3

WATER-SOLUBLE VITAMINS 1. Niacinamide 2. Pyridoxine N CONH2 N CH2OH CH2OH HO H3C 3. Riboflavin N N NH N CH2 HOCH HOCH HOCH CH2OH O H3C O H3C Cl N S N H3C N CH2 NH2 CH3 CH2CH2OH 4. Thiamin

WATER-SOLUBLE VITAMINS 1. Niacinamide 2. Pyridoxine N CONH2 N CH2OH CH2OH HO H3C 3. Riboflavin N N NH N CH2 HOCH HOCH HOCH CH2OH O H3C O H3C Cl N S N H3C N CH2 NH2 CH3 CH2CH2OH 4. Thiamin

WATER-SOLUBLE VITAMINS 0 5 10 15 20 Column: u Bondapak C18 Solvent: MeOH Sample: Water-Soluble Vitamins Inject 1 2 3 4

WATER-SOLUBLE VITAMINS 0 5 10 15 20 Column: u Bondapak C18 Solvent: MeOH Sample: Water-Soluble Vitamins Inject 1 2 3 4