m Thin layer chromatography is the simplest

of all the chromatographies.

m It is still being used widely for qualitative
analysis of samples in various dimensions
of forensics due to its low cost and
simplicity.
m 2 TLC plate is a sheet of glass, metal, or
plastic which is coated with a thin layer of a
solid adsorbent (usually silica or alumina).

m 2 small amount of the mixture to be

analyzed is spotted near the bottom of this
plate. The TLC plate is then placed in a
shallow pool of a solvent in a developing
chamber so that only the very bottom of
the plate is in the liquid. This liquid, or the
eluent, is the mobile phase, and it slowly
rises up the TLC plate by capillary action.
m 2s the solvent moves past the spot that was
applied, an equilibrium is established for
each component of the mixture between
the molecules of that component which
are adsorbed on the solid and the
molecules which are in solution.
m In principle, the components will differ in
solubility and in the strength of their
adsorption to the adsorbent and some
components will be carried farther up the
plate than others.
m !hen the solvent has reached the top of
the plate, the plate is removed from the
developing chamber, dried, and the
separated components of the mixture
are visualized. If the compounds are
colored, visualization is straightforward.
Usually the compounds are not colored,
so a UV lamp is used to visualize the
plates.
m Take a Tlc plate made up of Silica ²G
and cut it appropriately according to
our requirements.
m From the bottom mark a line at a
distance of about 2 cm and another line
at a distance of 10 cm from this 2 cm line
m Mark points equidistantly on the line 2
cm above the bottom.
m The sample needs to be concentrated in
appropriate solvent which in this case is
petroleum.
m Once a concentrated sample is
obtained it should be covered to avoid
evaporation.
m ¢ow at the marked points apply the
sample and allow them to dry off.
m 2gain spotting is done and is repeated
3-4 times to concentrate the sample.
m Then the tlc plate is placed either
vertically or at an angle of 20 degrees
inside the solvent chamber that contains
appropriate solvent that can be varied
and chosen as per our need.
m Solvent rises in tlc plate due to capillary
action and thereby causes the
separation of various components of
ganja due to difference in their
adsorption.
m Plate is removed once solvent reaches
the 10 cm line and then is dried.
m Spots can be visualized by using various
reagents like dragondroff reagent, Fast
blue etc.
m Spots can also be visualized using Ultra
visible.
m 2long with the sample we also apply the
standard of the suspected drug and
thereby compare their spots position.
m This not only help us to identify that
whether suspected drug is present but
also help us to know about whether any
impurity is present or not.
m The solvent choice is the key to Tlc.
m Solvent choice depends on
temperation,envoirnment,polarity etc.
m Before carrying out tlc we carry out
various color tests that give us an idea
about which of the narcotic drug may
be present and thus we can carry out tlc
for the suspected drugs.
m The compound runs as a streak rather than
a spot =>The sample was overloaded.
m The sample runs as a smear or a upward
crescent.=> Compounds which possess
strongly acidic or basic groups (amines or
carboxylic acids) sometimes show up on a
TLC plate with this behaviour. 2dd a few
drops of ammonium hydroxide (amines) or
acetic acid (carboxylic acids) to the eluting
solvent to obtain clearer plates.
m The sample runs as a downward crescent.=>Likely, the
adsorbent was disturbed during the spotting, causing
the crescent shape.

m ¢o spots are seen on the plate. You might not have

spotted enough compound, perhaps because the
solution of the compound is too dilute. Try
concentrating the solution, or, spot it several times in
one place, allowing the solvent to dry between
applications. Some compounds do not show up under
UV light; try another method of visualizing the plate. Or,
perhaps you do not have any compound because your
experiment did not go as well as planned.
m If the solvent level in the developing jar is
deeper than the origin (spotting line) of the
TLC plate, the solvent will dissolve the
compounds into the solvent reservoir
instead of allowing them to move up the
plate by capillary action. Thus, you will not
see spots after the plate is developed.
m There are 3 gas cylinders, each for
different gas namely
hydrogen,nitrogen,oxygen.
m 2ll these 3 gas cylinders are now
connected to the column or detectors
according to their usage.
m 2 column contains coiled coils which
contains stationary phase(usually
containing silica gel).This column is
present in a oven to provide appropriate
temp.
m Column is so coiled so that a large
length to the column can be provided
for better separation of compounds.
m The separated compounds goes to
detector which can of varied type.
m Mainly used detectors are FID·s-flame
ionization detector.
m These FID·s give out their results to
computers that provides
chromatograms.
m It is estimated that 10-20% of the known
compounds can be analyzed by GC. To be
suitable for GC analysis, a compound must
have sufficient volatility and thermal
stability. If all or some of a compound or
molecules are in the gas or vapour phase
at 400-450°C or below, and they do not
decompose at these temperatures, the
compound can probably be analyzed by
GC.
m One or more high purity gases are supplied to
the GC. One of the gases (called the carrier
gas) flows into the injector, through the column
and then into the detector. 2 sample is
introduced into the injector usually with a
syringe or an exterior sampling device. The
injector is usually heated to 150-250°C which
causes the volatile sample solutes to vaporize.
The vaporized solutes are transported into the
column by the carrier gas. The column is
maintained in a temperature controlled oven.
m The solutes travel through the column at
a rate primarily determined by their
physical properties, and the temperature
and composition of the column. The
various solutes travel through the column
at different rates. The fastest moving
solute exits (elutes) the column first then
is followed by the remaining solutes in
corresponding order.
m 2s each solute elutes from the column, it
enters the heated detector. 2n
electronic signal is generated upon
interaction of the solute with the
detector. The size of the signal is
recorded by a data system and is
plotted against elapsed time to produce
a chromatogram.
m The ideal chromatogram has closely
spaced peaks with no overlap of the
peaks. 2ny peaks that overlap are
called co eluting. The time and size of a
peak are important in that they are used
to identify and measure the amount of
the compound in the sample. The size of
the resulting peak corresponds to the
amount of the compound in the sample.
m 2 larger peak is obtained as the
concentration of the corresponding
compound increases. If the column and all
of operating conditions are kept the same,
a given compound always travels through
the column at the same rate. Thus, a
compound can be identified by the time
required for it to travel through the column
(called the retention time).
m The identity of a compound cannot be
determined solely by its retention time. 2
known amount of an authentic, pure
sample of the compound has to be
analyzed and its retention time and peak
size determined. This value can be
compared to the results from an unknown
sample to determine whether the target
compound is present (by comparing
retention times) and its amount (by
comparing peak sizes).
m The detection of organic compounds is
most effectively done with flame ionization
m However, typically the biochemical
compounds have a greater amount of
carbon present than other elements. This
means that a particular compound may be
more easily detected using flame ionization
over the other methods because of higher
carbon concentration and also flame
ionization's sensitivity
m In order to detect these ions, two
electrodes are used to provide a
potential difference. The positive
electrode doubles as the nozzle head
where the flame is produced. The other,
negative electrode is positioned above
the flame
m the design has been modified into a tubular
electrode, commonly referred to as a collector
plate. The ions thus are attracted to the
collector plate and upon hitting the plate,
induce a current. This current is measured with
a high-impedance picoammeter and fed into
an integrator. How the final data is displayed is
based on the computer and software. In
general, a graph is displayed that has time on
the x-axis and total ion on the y-axis.
m The current measured corresponds roughly to
the proportion of reduced carbon atoms in the
flame.
m 2n Ê
Ê
Ê , also called an
Ê
Ê, is a substance that contains a
great amount of stored energy that can
produce an explosion, a sudden
expansion of the material after initiation,
usually accompanied by the production
of light, heat, sound, and pressure.
m The energy stored in an explosive
material may be
m chemical energy, such as nitro-glycerine
or grain dust
m pressurized compressed gas, such as a
gas cylinder or aerosol can
m nuclear, such as fissile isotopes of
uranium-235 and plutonium-239
m [xplosive materials may be categorized by
the speed at which they expand. Materials
that detonate (explode faster than the
speed of sound) are said to be MM
Ê
Ê and materials that deflagrate
are said to be  Ê
Ê. [xplosives
may also be categorized by their sensitivity.
Sensitive materials that can be initiated by
a relatively small amount of heat or pressure
are primary explosives and materials that
are relatively insensitive are secondary
explosives.
m 2 chemical explosive may consist of
either a chemically pure compound,
such as nitro-glycerine, or a mixture of an
fuel and a oxidizer, such as black
powder or grain dust and air.
m ¢
Êʉ 2 highly unstable and
sensitive liquid.
m 2Ê
Ê
Ê ʉ 2 very unstable white
organic peroxide.
m 6¢6‰ Yellow insensitive crystals that can be
melted and cast without detonation.
m ¢
Êʉ 2 nitrated polymer which
can be a high or low explosive depending
on nitration level and conditions.
m  
, K 6¢, 
‰ Very powerful explosives
which can be used pure or in plastic
explosives
m FT-IR is one of important techniques to
identify the presence or absence of an
explosives and its quantity.
ëÊ The clear liquid that
can be separated from
clotted blood. Serum differs
from plasma, the liquid
portion of normal unclotted
blood containing the red and
white cells and platelets. It is
the clot that makes the
difference between serum
and plasma

ëÊThe examination and

analysis of body fluids
including saliva, semen, urine,
and blood.
 2 complex mixture of cells, enzymes,
proteins & inorganic substances
 Fluid portion of blood is called the
plasma (55% of blood content)
 primarily water
 red cells (erythrocytes)
 white cells (leukocytes)
 platelets
m Red blood cells (erythrocytes), white
blood cells (leukocytes), and platelets
are the solid materials suspended in
plasma.
m 2ntigens, usually proteins, are located
on the surface of red blood cells and
are responsible for blood-type
characteristics.
m 2
ü based on having 2, B,
both, or no antigens on red blood cells
m  M 
ü may be present on red blood cells;
positive if present and negative if not
m 2
Êü a substance that can stimulate the
body to make antibodies. Certain antigens
(proteins) found in the plasma of the red blood
cell·s membrane account for blood type.
m 2
ü a substance that reacts with an
antigen
m 2

ü clumping of red blood cells; will
result if blood types with different antigens are
mixed.
Blood typing requires two antiserums‰
anti-2 and anti-B. By inserting a droplet of
these antiserums in samples of blood, one
can determine which samples maintain a
normal appearance and which samples
become clotted, or agglutinated, under
microscopic examination. Type-2 blood will
be agglutinated by anti-2 serum; Type-B
blood will be agglutinated by anti-B serum;
Type-2B blood by both; and Type-O blood by
neither.
m To determine whether or not
blood is present at a crime
scene, forensic investigators
use color or crystalline tests.

m The _

  was widely

implemented until it was
discovered to be a known
carcinogen.(.0.25gm
benzidine+acetic acid 25ml
then 3%Hhydrogen peroxide)

m The current * 

  ,
which uses the chemical,
phenolphthalein, operates on
the fact that when
phenolphthalein comes in
contact with hemoglobin, it
releases peroxidase enzymes
that cause a bright pink color
to appear.
`
This chemical is used by crime scene investigators to locate
traces of blood, even if it has been cleaned or removed.
Investigators spray a luminol solution is throughout the area
under investigation and look for reactions with the iron
present in blood, which causes a Ê ÊÊÊ.
One problem is that other substances also react, such some
metals, paints, cleaning products, and plant materials.
2nother problem is that the chemical reaction can destroy `
other evidence in the crime scene.  Ê 


ÊÊ
This chemical is also capable of detecting latent or old
blood, similar to luminol. It is ideal for fine stains or smears
found throughout a crime scene. 2fter the solution has been
sprayed onto the substance or area suspected to contain ÊÊ
blood, a UV light and goggles are used to detect any  Ê 

illuminated areas, which appear greenish-white if blood is `M

present. It may also react to many of the same things as

luminol (copper and bleach).
6Ê
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Blood is injected into a rabbit; antibodies are
formed; the rabbit·s blood is extracted as an
antiserum; the antiserum is placed on sample blood.
The sample will react with human proteins if human
blood is present. This test is very sensitive and requires
only a small amount of blood.
6MÊKÊ


Ê

Blood can also be categorized using the Rh

(Rhesus disease) factor. If an individual
has a positive Rh factor, this means that
his/her blood contains a protein that is
also found in Rhesus monkeys. The Rh
factor, like other antigens, can be found on
the surface of red blood cells.
 General points
m !et blood has more value than dried blood
because more tests can be run.
X For example, alcohol and drug content can be
determined from wet blood only. Blood begins to
dry after 3-5 minutes of exposure to air. 2s it dries,
it changes color towards brown and black.

m _    

 
  

    .
m 2 number of immunological assay techniques
are commercially available for detecting drugs
through antigen-antibody reaction.
m One such technique, the enzyme-multiplied
immunoassay technique ([MIT), is used by
toxicologists because of its speed and high
sensitivity for detecting drugs in urine.
m In a typical [MIT analysis, antibodies that will
bind to a specific drug are added to the
subject·s urine.
m Other immunoassay procedures are also
available, such as radioimmunoassay (RI2)
which uses drugs labeled with radioactive tags.

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m The rape victim must undergo a medical
examination as soon as possible after the
assault.
m 2t that time the appropriate items of physical
evidence including clothing, hairs, and vaginal
and rectal swabs can be collected for
subsequent laboratory examination.
m 2ll outer and undergarments should be
carefully removed and packaged separately in
paper (not plastic) bags.
m Bedding, or the object upon which the assault
took place, may also be carefully collected.
m Pubic combings m Oral swabs & smear
m Pubic hair m Head hairs
standard/reference
m Blood sample
sample
m Fingernail scrapings
m [xternal genital dry-skin
areas m 2ll clothing
m Vaginal swabs and m Urine specimen
smear
m Cervix swab
m Rectal swabs and
smear
m Sculptures created with
modeling clay and other
materials
m High-resolution three-
dimensional computer images.
m Usually require both artist and
forensic anthropologist.
m Computer programs are used
to maneuver scanned
photographs of remains, facial
features and other available
reconstructions.
m Usually most effective in victim
identification
m Superimposing
photograph of
individual believed to
be the owner of skeletal
remains over X-ray of
the unidentified skull.

m If X-ray and
photograph are the
same individual, the
anatomical features of
face should align
accurately.
m Sculptors need to know depth of skin
that overlays skull for life-like faces.
2round mouth and between eyes
are heavily concentrated depths.
Depth measurements are available
for people of all age, race and size.
m Small pegs are used as facial depth
indicators and are fixed into the skull
or cast. Strips of clay that match
height of pegs are placed between
pegs and then clay is used to fill the
gaps.
m [ffectiveness of clay reconstruction
depends wholly on the skill of the
sculptor.
m The sculptor then work on aspects of face that
give the most character and parts that expire
fastest as the body decays.
m There are certain rules during reconstruction, ex‰
ears are around the same length as nose though
elderly people usually have longer ears. !hen
facial features are complete, sculptor makes
mold from clay head using plaster of Paris silicone
rubber.
m The facial features of victim present the most
information on identification.
m Reconstruction includes
building muscles. Sculptors
estimate muscle structures
by noting shape and size of
certain facial bones that
affect shape of the muscles
previously attached to them.
Using their experience,
sculptors can construct face
by shaping each muscle and
fixing them on the skull. Final
step is covering clay muscles
with layer of clay skin
because it resembles real
skin when smoothed over.
 Skull and Teeth Gender
Determinations

ë 
† Male skulls are larger than female
skulls
†Male orbits are not circular
Picture‰
†Male skulls are not smooth
http‰//medstat.med.utah.edu/kw/osteo/forensics/sex/bo
nesksex.html † Square chins indicate male
† Round chins indicate female

6ÊÊ
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† Males have much larger teeth
† Males have a larger mandible
Picture‰
www.sav-
ondrugs.com/shop/templates/surgeries/graphics/Chinaugmentation_1.j
pg
 2ge Determinations
†Skull

† The cranium of a baby is not solid, but divided into four pieces.

† Studying a skull that is 18-23 years of age demonstrates‰

- 2 relatively larger skull size.
† - Closer (almost meeting) brain sutures.
2ge Determination‰ Jaws & Teeth
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The Forensic 2rtist
2 forensic artist is called in when a sketch of a person is
needed to notify the public that he/she is missing, identify
a suspect, or eliminate a suspect.

Usually, the forensic artist goes about his/her job by

carefully listening to an eyewitness or victim s description
of the missing person, then accurately translating his/her
depiction into a composite drawing.

This sketch is then put on flyers, and given to the police,

television news stations, etc. in order to help with the
search.
2-D Facial Reconstruction
KÊÊ
†The unidentified skull is placed on a stand,
in the Frankfort
† Horizontal position.
†Tissue markers are placed on the skull.
†2 ruler is placed beside the skull.
†Photos are taken of the skull·s frontal and
profile views.
†Following the skull·s natural contours, and
using tissue markers as guidelines, the
forensic artist creates a sketch of the
unidentified person.
†2pproximate measurements for the mouth,
nose, and eyes are made.
†Hair type, and style are determined by
other evidence collected from the crime
scene that give clues as to the kind of life
the unidentified person led.
3-D Facial Reconstruction

KÊÊ

2 skull is placed on a workable stand in the

Horizontal
position.
Referring to tissue depth data previously taken,
artificial eyes are centered in the skull s eye
sockets.
Tissue markers are glued right onto the skull.
Following the skull s contours, and restrictions set
by the tissue
markers, clay is placed directly on the skull.
Measurements are made to determine mouth
and nose thickness, length, and width.
3D scanner
 3D Reconstruction‰
Finishing Touches
2ny information compiled from forensic anthropologists
about the deceased such as the geographical
locations of where he/she lived, or his/her occupation is
used to complete construction.

Clay or wig is put on the skull to create hair.

The completed face is photographed.

The forensic artist now takes over and sketches the

constructed face.
m There are certain accuracy limits that arise
during reconstruction despite sculptor
experience.
m Sculptors can guess hairstyles but cannot
create expressions on a person·s face to make
sculpture completely life like.
m But sculpture can be successful when it assist in
nudging someone's memory or help narrow
down search by exempting anyone who does
not look like the reconstructed face.
m Insufficient tissue thickness data

m Most pressing issue is data used to average facial

tissue thickness. Data available to forensic artists
are still very limited in ranges of ages, sexes, and
body builds