High performance(pressure
Upcoming SlideShare
Loading in...5
×
 

High performance(pressure

on

  • 1,280 views

 

Statistics

Views

Total Views
1,280
Views on SlideShare
1,280
Embed Views
0

Actions

Likes
0
Downloads
54
Comments
0

0 Embeds 0

No embeds

Accessibility

Categories

Upload Details

Uploaded via as Microsoft PowerPoint

Usage Rights

© All Rights Reserved

Report content

Flagged as inappropriate Flag as inappropriate
Flag as inappropriate

Select your reason for flagging this presentation as inappropriate.

Cancel
  • Full Name Full Name Comment goes here.
    Are you sure you want to
    Your message goes here
    Processing…
Post Comment
Edit your comment

High performance(pressure High performance(pressure Presentation Transcript

  • HIGH PERFORMANCE(PRESSURE/PRICED) LIQUID CHROMATOGRAPHY ALTHOUGH GAS CHROMATOGRAPHY IS MOST WIDELY USED TECHNIQUE IN THE ANALYSIS OF OILS & FATS,BUT THE DESTRUCTION OF SAMPLE UNDER ANALYSIS & OTHER LIMITATION LIKE LOW MOL WT,HIGH VOLATILITY & HIGH STABILITY IN GAS PHASE NECESSITATED IN THE DEVELOPMENT OF NEWER TECHNIQUE AND HPLC IS THE RESULT OF THAT ONLY.
  • HPLC LIQUID CHROMATOGRAPHY INVOLVES SEPARATION OF COMPONENTS DUE TO DIFFERENCE IN EQUILIBRIUM DISTRIBUTION OF SAMPLE COMPONENTS BETWEEN A LIQUID MOBILE PHASE & STATIONARY PHASE (ONE WHICH DISTINGUISHES THIS FROM OTHER CHROMATOGRAPHIC TECHNIQUE).
  • INSTRUMENTATIONGradientController • Pump Column Detecor Injector Mobile Phases
  • MOBILE PHASE THERE ARE TWO TYPES OF MOBILE PHASES WIDELY USED BY ANALYST: ISOCRATIC ELUTION:BOTH MOBILE PHASE COMPOSITION & FLOW RATE IS FIXED. GRADIENT ELUTION: BOTH MOBILE PHSE COMPOSITION AND FLOW RATE IS PROGRAMMED TO GET BETTER ELUTION AND RESOLUTION
  • Types of HPLC Reverse-phase (polar mobile phase/non- polar stationary phase/somewhat polar analytes) Normal Phase (non-polar mobile phase/polar stationary phase/non-polar analytes) Adsorption (non-polar mobile phase/polar stationary phase/non-polar analytes); isomer separation Ion-Exchange (salts/ionic stationary phase) Size-exclusion (aqueous/gel for large MW solutes, >104)
  • STATIONARY PHASE IN THIS WE WILL BE TALKING ABOUT THE STATIONARY PHASES OF NORMAL PHASE AND REVERSE PHASE HPLC NORMAL PHASE:
  • STATIONARY PHASE REVERSE PHASE:
  • NORMAL PHASE NONPOLAR COMPONENTS ARE RETAINED WHILE NON POLAR COMPONENTS ARE ELUTED FIRST. INCREASE IN POLARITY OF MOBILE PHASE DECREASES RETENTION TIME & VICE-VERSA. FOR ANALYSIS OF OILS AND FATS THESE COLUMN IS GENERALLY NOT USED.
  • NORMAL PHASE
  • REVERSE PHASE HYDROPHILIC COMPOUNDS ELUTE MORE QUICLKY THAN HYDROPHOBIC COMPOUNDS. RETENTION TIME CAN BE INCREASED BY ADDITION OF NONPOLAR SOLVENTS. REVERSED PHASE TLC IS USED IN SEPERATION OF COMPOUNDS ACCORDING TO NO OF C ATOMS. RP GAVES BETTER RESOLUTUION BECAUSE NONPOLAR- NONPOLAR INTERACTION IS MUCH LARGER THAN POLAR-POLAR INTERACTION
  • REVERSE PHASE .