Professional Documents
Culture Documents
CHROMATOGRAPHY
NAUREEN SHEHZADI
M.PHIL. (PHARMACEUTICAL CHEMISTRY)
UNIVERSITY COLLEGE OF PHARMACY,
UNIVERSITY OF THE PUNJAB, LAHORE, PAKISTAN
OUTLINE
• History • How GC works?
• Definition • Factors affecting sample
• Principle of separation separation
• Requirements for GC • Chromatogram analysis
• Types • Derivatization of samples
• Instrumentation • Compound identification
• The carrier gas • Applications
• The flow regulators or flow
meters
• The injection port
• The column
• Stationary phases
• Supports
• Temperature control devices
• Detectors
• Recorders and integrators
HISTORY
• German graduate student “Fritz Prior” developed solid state
gas chromatography in 1947.
Stationary phase:
1. Solid
2. Liquid
Mobile phase:
Gas
PRINCIPLE OF SEPARATION
Principle of separation
in gas
chromatography
Partition Adsorption
REQUIREMENTS FOR GC
• Low molecular weight
• Thermally stable compound
• High vapor pressure
• Low boiling point
TYPES
Types of gas
chromatography
Gas-liquid Gas-solid
chromatography chromatography
1- Gas-liquid chromatography
REQUIREMENTS OF A CARRIER:
Flow meters
§ Glass
§ Stainless steel
§ Copper tube
Glass columns are inert and highly fragile. These are used for
biological samples or for compounds which undergo reaction
with copper or stainless steel.
Types of columns (based on content)
Porous packing
Packed column
Non-porous
packing Liquid coating
Column
Wall-coated open
tubular column
Support-coated open
tubular column
(i) Packed column
These columns are not packed but inner walls are coated with a
liquid layer. These are used for partition chromatography.
§ WCOT
§ PLOT
§ SCOT
5- Stationary phases
The phase chosen must meet following requirements;
Adiponitrile Hydrocarbons
Apiezon L Alcohols, Aldehydes, Ketones, Fatty acids,
Asphalt Pesticides
Aromatics
AN IDEAL DETECTOR:
Detector
Detector
Concentration dependent
Flow dependent detectors detectors
Mass spectrometer
9- Recorders and integrators
• RECORDERS record the baseline and all the peaks obtained
Or
EXAMPLES:
• Low vapor pressure organic acids can be converted to low
boiling acid chlorides
COMPOUND IDENTIFICATION
There are two techniques that are used to identify compounds
eluting from the column.
Qualitative analysis
Gc applications
Quantitative
analysis
Miscellaneous
Applications
Headspace GC
analysis
1- Qualitative analysis
Qualitative identification by GC is achieved by two ways;
Pharmaceutical analysis
Drug detections
Petrochemical industry
Forensic analysis
Environmental sciences
Agrochemical industry
Paint industry
Biomedical field
Food industry
For example;
• Lipids, Proteins, Carbohydrates & Vitamins etc.
• Steroids
• Preservatives
• Colorants & Flavors
• Texture modifiers
• Drugs
• Pesticide residues
• Trace elements
Food industry
Dairy products:
• Boranes
• Silanes
• Germanes
• Organotin
• Lead compounds
Forensic analysis
Examples are;