《Introduction to High Performance Liquid Chromatography.ppt》由会员分享，可在线阅读，更多相关《Introduction to High Performance Liquid Chromatography.ppt（31页珍藏版）》请在三一办公上搜索。
1、Introduction to High Performance Liquid Chromatography,2,In This Section,We Will Discuss:,The differences between High Performance Liquid Chromatography and Gas Chromatography.The components of the high performance liquid chromatograph(HPLC).The separation process.The chromatogram.The most common mo
2、des of HPLC.,3,Youve Got a Problem to Solve,4,Separation Techniques,I have two separation techniques in my lab,High Performance Liquid Chromatographyand Gas Chromatography.Which should I use?,5,Comparison of HPLC and GC,Sample Volatility,Sample Polarity,HPLC,No volatility requirement Sample must be
3、solublein mobile phase,GC,Sample must be volatile,HPLC,GC,Separates both polar andnon polar compounds PAH-inorganic ions,Samples are nonpolarand polar,6,Comparison of HPLC and GC,7,Comparison of HPLC and GC,Sample Thermal Lability,Sample Molecular Weight,HPLC,Analysis can take placeat or below roomt
4、emperature,GC,Sample must be able to survive high temperature injection port and column,HPLC,GC,No theoretical upper limitIn practicality,solubility islimit.,Typically 500 amu,8,Comparison of HPLC and GC,Sample Preparation,Sample Size,HPLC,Sample must be filteredSample should be insame solvent as mo
5、bilephase,GC,Solvent must be volatileand generally lower boiling than analytes,HPLC,GC,Sample size based uponcolumn i.d.,Typically 1-5 L,9,Comparison of HPLC and GC,10,How can We Analyze the Sample?,Carbohydrates1.fructose2.Glucose3.Saccharose4.Palatinose5.Trehalulose6.isomaltose,Zorbax NH2(4.6 x 25
6、0 mm)70/30 Acetonitrile/Water1 mL/min Detect=Refractive Index,11,Separations,Separation in based upon differential migration between the stationary and mobile phases.,Stationary Phase-the phase which remains fixed in the column,e.g.C18,Silica,Mobile Phase-carries the sample through the stationary ph
7、ase as it moves through the column.,12,Separations,13,Separations,14,Separations,15,Separations,16,Separations,17,Separations,18,Separations,19,Separations,20,Separations,21,Separations,22,Separations,23,Separations,24,Separations,25,Separations,26,Separations,27,Separations,28,The Chromatogram,29,HPLC Analysis Parameters,Mobile Phases,Flow RateComposition,Injection Volume,Column Oven Temperature,WavelengthTime Constant,30,Modes of High Performance Liquid Chromatography,31,HPLC Applications,
