CHROMATOGRAPHY

TLC – Thin Layer Chromatography

GC – Gas Chromatography known as (GLC) Gas Liquid

Chromatography

HPLC – High Performance / Highly Pressure Liquid

Chromatography

HPTLC - High Performance / Pressure Thin Layer Chromatography

 CHROMATOGRAPHY
 Technique by which mixture of components are separated
 This can be achieved by the properties of some components such as sizes
and how components reacts.
 Mainly used in toxicology and chemistry division for analysis of
pesticide, drugs and toxicant.
 Can be used as an analytical tool, feeding its output into a detector that
reads the contents of the mixture.
 And in particular mass spectrometry has long been used in criminal
investigations to help the police to determine the contents of a deceased
person’s stomach.
How is chromatography used to test for drugs?
 Chromatography uses a gas or liquid carrier medium to separate the urine
samples compounds by their molecular interactions with the carrier medium.

What is the main application of chromatography?

 Pharmaceuticals, clinical trials, environmental and chemical safety, food
and beverage, drug testing, forensic, petroleum creation, and molecular
biology are some of the most common uses of chromatography.
In which forensic cases chromatography can be used?
 Separation of complex mixture ( Known as chromatography ) is an essential tool
in forensic science. It is routinely used to identify and compare sample of drugs,
explosives, inks and biological sample such as saliva, urine. Blood and other.

How is chromatography used in crime scene investigation?

 Forensic scientists are able to use ink chromatography to solve crimes by
matching documents or stains found at a crime scene to the marker or pen that
belong to a suspect.
 Forensic scientists analyze the unknown ink and compare it to writing utensils
collected from possible suspects.
 History of Chromatography
Chromatography was first devised in Russia by the Italian-born scientist Mikhail Tsvet in
1900. He developed the technique and coined the term chromatography in the first decade of
the 20th century, primarily for the separation of plant pigments such as chlorophyll, 
carotenes, and xanthophylls. Since these components separate in bands of different colors
(green, orange, and yellow, respectively) they directly inspired the name of the technique.
New types of chromatography developed during the 1930s and 1940s made the technique
useful for many separation processes
 Principle of Chromatography
(how does chromatography work)

•Chromatography is based on the principle where molecules in mixture applied onto

the surface or into the solid, and fluid stationary phase (stable phase) is separating
from each other while moving with the aid of a mobile phase

•The factors effective on this separation process include molecular characteristics

related to adsorption (liquid-solid), partition (liquid-solid), and affinity or differences
among their molecular weights.

•Because of these differences, some components of the mixture stay longer in the
stationary phase, and they move slowly in the chromatography system, while others
pass rapidly into the mobile phase, and leave the system faster.
 CATEGORY OF CHROMATOGRAPHIC
TECHNIQUE

TLC – Thin Layer Chromatography

 Is an affinity-based methods used to separate compounds in a mixture.

 Is a highly versatile separation method that is widely used for both

qualitative and quantitative sample analysis.

 Can be used to analyze any substances class, including pesticides, steroids,

alkaloids, lipids, nucleotides, glycosides, carbohydrates, and fatty acids.
 GC – Gas Chromatography known as (GLC) Gas Liquid
Chromatography
 Is a common analytical technique used to separate and analyze volatile and semi
volatile compounds in a mixture.

 Is a popular analytical technique as it combines exceptional resolving power

with speed and sensitivity

 It is widely applied in many industries, including environmental, petroleum,

chemical, food and beverage, and pharmaceutical.

 In forensic investigations, gas chromatography is used in toxicology screening to

determine if a deceased person has ingested drugs or alcohol prior to death.

 It can also be used to tell if a victim of crime has been poisoned.

What type of chromatography is used for drug testing?

 Gas chromatography, The most sophisticated drug testing approach is gas

chromatography coupled with mass spectrum, which is regarded as a “gold
standard”.

 It is used in confirmatory testing, typically is preceded by a rapid immunoassay

method to eliminate the majority of the negative samples.

Which type of chromatography is used to test the hair and blood

samples at the crime scene?
 Gas chromatography ( also known as GC ) is one of the most common
methods used in forensic science.
HPLC – High Performance / Highly Pressure Liquid Chromatography

 Is a form of column chromatography that pumps a sample mixture or analyte in

a solvent (known as the mobile phase) at high pressure through a column with
chromatographic packing material.

 Has the ability to separate, and identify compounds that are present in any
sample that can be dissolved in a liquid in trace concentrations as low as parts
per trillion.
HPTLC - High Performance / Pressure Thin Layer Chromatography

 A number of enhancement can be made to the basic method of thin layer

chromatography to automate the different steps, to increase the resolution achieved,
and to allow more accurate quantitative measurements.

 Is one of the sophisticated instrumental techniques based on the full capabilities of

thin layer chromatography.

 Provide essential benefits, such as improved sample application, better and faster
separations, and less mobile phase usage.
What type of chromatography is used in forensic?
 Forensic scientists employ two types of planar chromatography. While they
are similar in that both have the mobile phase as a solvent, both have different
principles behind each method.

What are the possible applications for paper chromatography in

forensic?
 For investigation of crimes, paper chromatography is useful in the field of
forensic science, as this process can be successful carried out with even very
small quantities of material.
 Using this technique, sample from crime scene are collected to be analyzed
and identified. Used in DNA and RNA fingerprinting.
Paper chromatography
 Cases/crime cases related to the chromatography
The high profile courtroom drama unfolding around Casey Anthony–the Florida mother
accused of murdering her two-year-old daughter Caylee in 2008.

The smell of Anthony’s car has been central to the case against her, as various people who came in
contact with the car in the days following Caylee’s disappearance said it smelled like death. The
challenge for prosecutors is proving that a foul smell–a smell that defense attorneys say came from a
garbage bag left in the trunk–is actually the lingering odor of human decomposition.

To do so they brought in ORNL senior researcher Arpad Vass, whose work in odor detection had
been used to hunt cadavers before. Vass and a colleague were given a sealed can containing a scrap
of upholstery from the car, and using a syringe extracted some of the air inside.

They then ran it through a gas chromatography/mass spectrometry device to analyze it for
substances, and cross referenced those results against a database of more than 400 chemical traces of
decomposition that Vass has compiled. According to Vass, the air contained an “overwhelmingly
strong” scent of decomposition.
 Principle of Chromatography
PRINCIPLES OF CHROMATOGRAPHY
All chromatographic systems rely on the fact that a substance placed in contact with
two immiscible phases, one moving and one stationary, will equilibrate between them.

Chromatography result as evidence

In forensic investigations, gas chromatography is used in toxicology
screening to determine if a deceased person has ingested drugs or alcohol
prior to death. It can also be used to tell if a victim of crime has been
poisoned.