1. High Performance Liquid Chromatography (HPLC)
SMK Negeri 13 Bandung

2. CHROMATOGRAPHY
Separation procedure of dissolved substance by a dynamic differential migration process in chromatographic syem
Chromatographic system consists mobile phase and stationary phase‏
Usage
Purification of chemical substance
Qualitative and quantitative analysis of component in mixture
According to the mobile phasei:
-Liquid chromatography (mobile phase is liquid)
- Gas chromatography (mobile phase is gas)‏

3. High Performance Liquid Chromatography (HPLC) :
A method of liquid chromatography used to analyze organic compounds which are un volatile and thermolable (unstable on heat
Basic principle of HPLC
Separation of chemical compound from a mixture based on the physical properties difference

4. Based on the principle, HPLC divided in:
Isocratic HPLC
Used on analysis of chemical compound which are need no change of mobile phase composition, temperature ,pressure and flow during the injection and elution in column..
Gradient HPLC
Used on chemical compound which are need change of mobile phase composition, temperature ,pressure and flow during the elution in column.

5. Based on the polarity of mobile phase and stationary phase, HPLC divided in:
Normal phase chromatography
The stationary phase is more polar than the mobile phase
Reversed phase chromatography
The mobile phase is more polar than the stationary phase

6. Chromatography Equipment
1. Eluent bottle
2. Pump
3. Injector
4. Column
5. Detector
6. PC and printer

7. Main component of HPLC Eluant bottle‏ Pump‏
A bottle for mobile phase contain pure eluant
Pump‏
To pump injected sample or eluent. Much of pump have output pressure of 1.000–6.000 psi and produce flow up to 20 mL/ minute

8. Main component of HPLC Injector Column
A place to inject sample in to column
Column
- Very important part of HPLC
Made of plastic called cartridge, or made of stainless steel
Consist of stationary phase (octyl, octa decyl, phenyl, nitrile, amine) chemicaly bounded with porous silica particle in the inner column.

9. Main component of HPLC Detector‏ Computer and printer
UV-visible detector, fluorometer (especially for excitation and emission wavelength) and refraction detector
Computer and printer
Data processor sofware computer, moniter and printer to show chromatogram (graphic of sample absorbance to time)‏

10. High Performance Liquid Chromatography

11. THANK YOU