CHROMATOGR
APHY
Samson, Silvano,
Sedillo
CHROMATOGR
• Chromatography is APHY
a technique for separating the
components, or solutes, of a mixture based on the
relative amounts of each solute distributed between a
moving fluid stream, called the mobile phase, and a
contiguous stationary phase.
CHROMATOGR
APHY
• Chromatography is usually based on the principle of
partition of solute between two phases. It usually
consists of a Mobile Phase and a Stationary Phase.
-The Mobile Phase usually refers to the mixture of the
substances to be separated and dissolved in a liquid or a
gas
-The Stationary Phase is a porous solid matrix through
TYPES OF
CHROMATOGRAP
HY
• Gas chromatography - the
mixture of interest is vaporized
and carried through a stationary
phase (usually a metal or glass
separation column) with an inert
glass, usually nitrogen or
helium. Larger molecules in the
mixture take longer to pass
through the column and reach
the detector at the far end.
• Thin-layer Chromatography
(TLC) - the stationary phase is a
thin layer of solid material,
usually silica-based, and the
mobile phase is a liquid in
which the mixture of interest is
dissolved. Thin-layer
chromatography comes with the
advantage of photographing
well, making its output easy to
• Liquid Chromatography - the
mixture of interest is dissolved in a
liquid and passed through a solid
stationary phase, which is often
made of a silica material. Several
varieties of liquid chromatography
exist, depending on the relative
polarities of the mobile and
stationary phases (normal-phase
versus reverse-phase) and whether
the mobile phase is pressurized
(high-performance).
• Paper Chromatography - is a
technique used for the
separation of compounds
based on the differential
solubility in the stationary
phase and mobile phase
(solvent). In the amino acid
separation by paper
chromatography, the solvent
travels across the paper, it
carries the amino acids with it.
PROCEDURE
1. Sample Application: The sample is applied to the stationary
phase, usually in liquid or gas form. This can be done by
spotting a small volume of the sample onto the chromatography
paper or injecting it into the chromatography column.
2. Mobile Phase Migration: The mobile phase (solvent or gas)
is allowed to move through the stationary phase. As the mobile
phase moves, it carries the components of the sample along
with it.
PROCEDURE
3. Separation: The components of the sample interact differently
with the stationary phase based on their chemical properties. This
results in differential migration rates, causing the components to
separate from each other as they move through the stationary
phase.
4. Detection: As the separated components elute from the
chromatography system, they can be detected using various
methods such as UV-Vis spectroscopy, fluorescence, or mass
spectrometry. Detection allows for the identification and
PROCEDURE
5. Analysis: Finally, the data obtained from the chromatogram
can be analyzed to determine the composition, purity, or
concentration of the components in the sample. Comparing
retention times or peak areas can aid in qualitative and
quantitative analysis.
VIDEO
REAL-LIFE APPLICATION
How can Chromatography be used in real life?
• Pharmaceutical and Clinical Testing
-Chromatography plays an important role in the safety of
pharmaceuticals. Pharmaceutical companies use
chromatography to quantify and analyze compounds for
contaminants.
-Vaccination creation is also an application of chromatography.
Chromatography can be used to determine which antibodies are
the best for fighting and neutralizing certain diseases.
REAL-LIFE APPLICATION
• Food and Beverage
- Chromatography is used to separate and analyze additives,
vitamins, proteins, amino acids, and other nutritional compounds
in food items. Chromatography can also be used to determine
expiration dates by distinguishing the number of organic acids
present as well as to detect any harmful toxins that may have
been added to the food item.
-Chromatography can be used to make sure every bottle of a
drink prepared is consistent.
REAL-LIFE APPLICATION
• Drug Testing
-Chromatography can be very useful in drug testing and clinical
toxicology reports. Chromatography can separate and analyze
substances found in urine samples.
• Forensics
-Gas chromatography can be used for more in-depth forensics
procedures, for example, crime scene analysis to test evidence
such as blood, hair, and fabric samples to further understand
what may have happened at the scene
REAL-LIFE APPLICATION
• Molecular Biology Studies
-Chromatography in nucleic acid research plays a role in
accelerating the identification process of nucleobases,
nucleotides, and nucleosides, as well as identifying their
oxidization process.
• Petroleum
-Gas chromatography is used to analyze finished gas products
and refining processes. Chromatography is most notably used in
the analysis of natural and refinery gas for BTU content and
hydrocarbon composition.
REFERENCE:
(https:// (https://
www.britannica.com)
(https:// www.orgchemboulder.com)
(https://
chem.libretexts.org)
(https:// www.lcservicesltd.co.uk)
(https://
blog.perkinelmer.com)
(https:// (www.tutorialspoint.com)
www.careerpower.in)
(https:// https://youtu.be/PvHvx7k7U
www.shimadzu.com)
(https:// PU?si=nWvtWgQocKsL6Iv
G
byjus.com)
(http://
)
www.rnlkwc.ac.in)
(https://
www.sciencedirect.com)
Thank
You
