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Compilation

of
Experiments
in Forensic
Chemistry
Laboratory Manual

Compiled by: Honeylynn E.


Edles
TABLE OF CONTENTS
Laboratory Safety Rules and
Regulations............................................................................. 3

Globally Harmonized
System...........................................................................................
.... 4

Exercise # 1
Laboratory Apparatus and Equipment used in Forensic
Chemistry...................................... 5
Experiment # 1
The Optical
Microscope.....................................................................................
................. 7
Experiment # 2
Dry and Wet
Mounting........................................................................................
.............. 9
Experiment # 3
Fingerprinting.................................................................................
................................... 11
Experiment # 4
Detection of Blood Kastle-Meyer Test
(Phenolphthalein).................................................18
Experiment # 5
Blood Spatter
Analysis..........................................................................................
............. 21
Experiment # 6
Blood Pattern
Analysis..........................................................................................
............. 25
Experiment # 7
Blood
Typing............................................................................................
.......................... 29
Experiment # 8
Identification of
Semen............................................................................................
.......... 31
Experiment # 9
Chemical Test for
Explosives ......................................................................................
........ 33
Compilation of Experiments in Forensic Chemistry [Type text] Page 2
Experiment # 10
Hair
Analysis..........................................................................................
............................. 35
Experiment # 11
Fiber
Analysis..........................................................................................
........................... 38
Experiment # 12
Physical Properties of
Glass..............................................................................................
. 41
Experiment # 13
Restoration of Erased Serial Numbers by Suitable Etching
Solutions................................... 44
Experiment # 14
Casting and
Moulding........................................................................................
................. 46
Experiment # 15
Identification of Inks using Paper
Chromatography............................................................. 48
Experiment # 16
Soil
Analysis..........................................................................................
.............................. 51
Experiment # 17
Determination of Alkaloids using Paper
Chromatography................................................... 52
Appendix A
Munsell Color
Chart..............................................................................................
..............

Laboratory Safety Rules and Regulations


1. The chemistry laboratory is a place for serious work. Do not perform
activities without your teachers permission. Never work alone in the
laboratory. Work only when your teacher is present.
2. Study your lab activity before you come to the lab. If you are in doubt
about any procedures, ask your teacher for help.
3. Safety goggles and a laboratory apron must be worn whenever you work
in the lab. Gloves should be worn whenever you use chemicals that cause
irritations or can be absorbed through the skin.
4. Contact lenses should not be worn in the lab, even if goggles are worn.
Lenses can absorb vapors and are difficult to remove in an emergency.

Compilation of Experiments in Forensic Chemistry [Type text] Page 3


5. Long hair should be tied back to reduce the possibility of it catching fire.
6. Avoid wearing dangling jewelry or loose, draping clothing. The loose
clothing may catch fire and either the clothing or jewelry could catch on
chemical apparatus.
7. Wear shoes that cover the feet at all times. Bare feet or sandals are not
permitted in the lab.
8. Know the location of the fire extinguisher, safety shower, eyewash, fire
blanket, and first aid kit. Know how to use the safety equipment provided for
you.
9. Report any accident, injury, incorrect procedure, or damaged equipment
immediately to your teacher.
10. Handle chemicals carefully. Check the labels of all bottles before
removing the contents. Read the labels three times: before you pick up the
container, when the container is in your hand, and when you put the bottle
back.
11. Do not return unused chemicals to reagent bottles.
12. Do not take reagent bottles to your work area unless specifically
instructed to do so. Use test tubes, paper, or beakers to obtain your
chemicals. Take only small amounts. It is easier to get more than to dispose
of excess.
13. Do not insert droppers into reagent bottles. Pour a small amount of the
chemical into a beaker.
14. Never taste any chemical substance. Never draw any chemicals into a
pipette with your mouth. Eating, drinking, chewing gum, and smoking are
prohibited in the laboratory.
15. If chemicals come into contact with your eyes or skin, flush the area
immediately with large quantities of water. Immediately inform your teacher
of the nature of the spill.
16. Keep combustible materials away from open flames. (Alcohol and
acetone are combustible.

Globally Harmonized System


As of June 1, 2015, the Hazard Communication Standard requires pictograms
on labels to alert users of the chemical hazards to which they may be
exposed. Each pictogram consist of a symbol o a white background framed
within a red border and represents a distinct hazard(s). The pictogram on the
label is determined by the chemical hazard classification .

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Exercise # 1
Laboratory Apparatus and Equipment used in Forensic
Chemistry

Compilation of Experiments in Forensic Chemistry [Type text] Page 5


Name:______________________________ Date Submitted:_______________
Yr and Sec: __________________________ Date Performed:_______________
Group No. ____________________________ Score:________________________

I. Objectives:

a.

b.

c.

II. Directions: Complete the table below.

Apparatus Name Function

Name Function
Apparatus

Compilation of Experiments in Forensic Chemistry [Type text] Page 6


`

Experiment # 1
The Optical Microscope
Name:______________________________ Date Submitted:_______________

Compilation of Experiments in Forensic Chemistry [Type text] Page 7


Yr and Sec: __________________________ Date Performed:_______________
Group No. ____________________________ Score:________________________

Introduction

The purpose of a microscope is to magnify a small object or to magnify the fine


details of a larger object in order to examine minute specimens that cannot be seen
by the naked eye

I. Objectives:
a.

b..

c.

II. Materials: Optical Microscope

II. Parts of the Optical Microscope

Compilation of Experiments in Forensic Chemistry [Type text] Page 8


No Part Function

10

11

12

13

Guide Question:
1. Briefly explain how the optical microscope magnifies objects.

Experiment # 2
Dry and Wet Mounting

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Introduction
Wet mount slides use water to suspend once living things for better viewing. Dry
mount slides can be used for many different objects.

II. Materials:
Microscope Toothpick
Slides and cover slips Tweezers
Newspaper Clear Nail Polish
Onion skin

III. Procedure:
Wet mounting: The letter
1. Obtain a clean, dry microscope slide and cover slip. Place the slide in front
of you.
2. Carefully cut a lowercase e from the newspaper.
3. Use the medicine dropper to place 1 or 2 drops of water in the middle of
the microscope slide.
4. Use tweezers to place the e right side up on the drop of water.
5. Hold the cover slip very carefully by its edges, at an angle of about 45 to
the surface of the slide. Gently lower the cover slip over the sample. If any
air bubbles get trapped, carefully move the cover slip with your finger to
free them.
6. View the sample under the microscope. Start with low power and then
move to medium power.
Dry Mounting: Letter e
1. Using a crucible tong in holding the microscope, gently heat the slide.
2. Drop a small amount of colourless nail polish on the center of the slide.
3. Gently put the letter e on the nail polish and heat again.
4. After heating, add another 2 drops of colorless nail polish on the mounted
letter e. Gently heat the slide.
5. Gently heat the cover slip. Slightly bend the cover slip and slowly cover the
letter e on the slide.
6. If bubbles occur, repeat the whole procedure. If there are no bubbles, the
specimen is ready to be observed under the microscope.
Wet Mounting: Onion Cells and Cheek cells
1. Obtain a very thin piece of onion tissue. Place the onion tissue on the slide.
2. Add a drop of methylene blue or iodine solution on the onion tissue. Remove
excess liquid.
3. Gently place the cover slip on top of the onion tissue. If bubbles occur, repeat
the whole procedure.
4. If there are no bubbles, you may observed the specimen under a microscope,
5. Repeat the procedure using the cheek cells. In obtaining the specimen, use a
toothpick and gently scrape your inside cheek.

REPORT SHEET
Experiment # 2

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Dry and Wet Mounting
Name:______________________________ Date Submitted:_______________
Yr and Sec: __________________________ Date Performed:_______________
Group No. ____________________________ Score:________________________

Observation:

Wet mount: letter Dry mount: letter Onion Tissue/Skin Cheek Cells
e e
Observation: Observation: Observation: Observation:

Guide Questions:

1. What is the purpose of wet or dry mounting?

2. What is the difference between wet and dry mounting?

Name:______________________________ Date Submitted:_______________

Yr and Sec: __________________________ Date Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 3
Fingerprinting
Compilation of Experiments in Forensic Chemistry [Type text] Page 11
Introduction
Fingerprints are some of the most important and useful pieces of forensic
evidence collected by the forensic scientist. Individual prints are unique. Fingerprint
types are inherited, but exact patterns are not. Compare your prints to those of your
parents and siblings. No two persons fingerprints are alike, not even identical twins.
This fact allows a persons fingerprints to be considered as direct evidence for
identification.
Three main fingerprint patterns are found in the human populationarch,
loop, and whorl. Approximately 69% of all prints are loops. The rarest type of
fingerprint is the arch, making up approximately 6% of the total population. That
leaves about 25% of the population as whorls. Each of these groups is subdivided.
Look at the following examples.

Arches
The plain arch consists of simple ridges that flow from left to
right, with a rise or hill in the center. The tented arch appears
to have a ridge that supports the arches like the center post in
a tent.

Loops
There are three parts of a loop pattern: deltas, typelines, and Figure 1. Arches
looping. Loop patterns always develop ridges that separate to form a delta. All loops
must have at least one delta. The ridges that separate or diverge at the delta are
typelines. The third part of a loop pattern is the looping.
Loops must have at least one curving and returning ridge. They may have as many
as 20.
In a loop pattern, ridges flow from one side with a rise. Then, they curve and return
to the
same side from which they started.

Whorls
Unlike arches and loops, whorls are often unique. Whorls are ridge patterns that
have a minimum of two deltas. Plain whorls are target-shaped patterns with two
deltas. An imaginary line connecting the two deltas must cross at least one circle.

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Central pocket loop whorls must
have at least one circle and two deltas.
An imaginary line connecting the two
deltas must not cross the circle
between the deltas.

Double loop whorls must have two separate loop formations inside two delta
formations.

Accidental whorls are unusual patterns with three deltas or a combination of a loop
and a tented arch.

Compilation of Experiments in Forensic Chemistry [Type text] Page 13


I. Objectivies:

a.

b.

c.

A. Dusting and Lifting Latent Fingerprints


Materials:
1. Dusting powder
Talc
Starch
Charcoal
Iron fillings
Zinc carbonate
2. Beakers
3. Brush
4. Tape
Procedure:
1. Place 3 g of charcoal powder, 9 g zinc carbonate and 01g of talcum
powder in a mortar and pestle. Grind the mixture to a find powder
2. Place the object on a clean, flat surface with the suspected location of the
latent prints accessible. (Dont forget to wear gloves.)

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3. Choose the dark or light fingerprint dusting
powder, according to which will provide better
contrast with the color of the surface.
4. Transfer a small amount of the dusting powder
into the lid or work directly from the jar that
contains the powder. Dip just the tips of the
bristles of the brush into the powder so that a small amount of powder is
retained by the bristles. Tap the brush gently to return excess powder to
the container.
5. Under a strong light, use a circular, twirling motion to sweep the brush
gently over the area, allowing the bristles to just barely contact the
surface. Continue depositing powder lightly until the latent fingerprint
begins to develop, concentrating on that area as it becomes clearer where
the latent prints are on the surface. When the ridges begin to appear,
change the direction of motion to follow the direction of the ridges. Once
the fingerprint is developed clearly, stop dusting immediately. Beginners
tend to overdevelop prints, which almost invariably causes loss of detail if
not loss of the entire print.

Lifting Developed Prints


1. Choose one of your better developed prints. (If possible, shoot an image of a
print before attempting to lift it. Accidents happen.)
2. Wearing gloves, lift the free end from the roll of lifting tape and smoothly pull
out about 6 to 7.5 cm (2.5 to 3") of tape from the roll. Dont touch the sticky
surface of the tape, and do not cut the tape from the roll.
3. Press the free end of the tape into contact with the surface, starting 5 to 6 cm
from the nearest part of the dusted print. Make sure the tape adheres firmly
to the surface.
4. Beginning at the free end, use your fingers to
carefully press the tape down onto the surface,
making sure that no air bubbles are trapped.
5. Continue pressing tape onto the surface, unrolling
more as necessary, until you have covered the
entire print with tape and continued for a couple
of centimeters past the print.
6. Using the roll as a handle, peel the tape from the
surface using one smooth motion, as shown in the picture. It helps to put one
finger on the free end of the tape to make sure the tape doesnt curl back on
itself.
7. Stick the free end of the tape near one edge of a transfer card of a color that
contrasts with powder you used to dust the print. Make sure the free end
adheres tightly to the transfer card, and then carefully press the tape into
contact with the transfer card, making sure to avoid air bubbles.
8. Cut the used tape from the roll and press the free end into contact with the
transfer card. Label the transfer card with your initials, the date and time,
and the object from which the print was lifted.

B. Revealing Latent Fingerprints Using Iodine Fuming


Materials:
1. Iodine crystals 3. Starch water (rice)
2. Round Container 4. Ziplock bags

Compilation of Experiments in Forensic Chemistry [Type text] Page 15


5. Jar 6. spray battle
Procedure:
1. Open the zip-lock bag and transfer one or two tiny iodine crystals to it. An
amount the size of a pinhead is sufficient.
2. Place your first specimen inside the bag, expand the bag so that it contains
some air space, and then zip the bag closed.
3. The crystals begin sublimating immediately, filling the bag with iodine vapor.
Depending on the size of the bag and specimen, the amount of iodine you
use, and how you vaporize it, latent prints should start becoming visible
within anywhere from a few seconds to a few minutes as faint orange
smudges on the specimen.
4. Allow the fuming to continue until the ridge detail is evident in the developing
prints or until no further change occurs. Depending on the specimen, the
amount of iodine vapour present, and other factors, this may require
anything from just a minute or so to several hours or more.
5. When development appears to be complete, quicklyremove the first
specimen, insert the second specimen, and re-zip the bag. (Be careful not to
inhale the iodine vapor, which is irritating and has a strong chlorine-like odor.)
You can use the iodine vapor already present in the chamber to develop
additional specimens, adding a few more iodine crystals as required to keep
the chamber filled with iodine vapor.
6. Place the specimen on a clean, flat surface and examine it carefully under
strong light with the magnifier. Depending on the specimen, you may see
only faint orange smudges with little or no ridge detail, or you may see well-
developed orange prints with considerable ridge detail visible.
7. Regardless of how much or how little ridge detail is visible, forensic
technicians always make an image of the specimen immediately after fuming
it
8. If you have a camera capture an image of the fumed fingerprints for your
records. Record the pertinent details for the specimen in your lab notebook.
9. After you have a good image of the original fumed specimen, use a starch
solution to develop the iodine stains. You can spray the specimen with the
starch solution. or gently swab the solution onto the fumed prints using a
cotton ball, cotton swab, or the corner of a paper towel. You neednt drench
the specimen; just dampening it is sufficient. The iodine reacts with the
starch instantly, changing the color of the fumed prints to blue-black.
10.Record your observations in your lab notebook and tape the developed
specimen into your lab notebook

Revealing Latent Fingerprints Using Ninhydrin


Materials:
1. Ninhydrin solution
2. Zinc chloride solution
3. Acetone
4. Spray bottle
5. Alcohol Lamp
6. Tripod
Method 1:

Compilation of Experiments in Forensic Chemistry [Type text] Page 16


1. If you are spraying or swabbing, place the specimen print side up on paper
towels or old newspaper to protect the work surface.
1. Hold the bottle about 15 cm away from where you think the print might be,
and spray a light coat on the area. Dont drench it, but make sure the entire
surface is dampened with ninhydrin solution
2. Wait a few moments until much of the ethanol evaporates, then spray again.
3. Place one of the treated specimens aside for a day or two to allow
development to occur at room temperature.
4. Allow the area to dry completely. The print will appear only when the area is
completely dry. Use the hair dryer to help dry the area.
Method 2:
1. Repeat procedure 1 and 2 in Method 1
2. If you are spraying or swabbing, place the specimen print side up on paper
towels or old newspaper to protect the work surface.
3. Apply ninhydrin solution sufficient to dampen the surface of the specimen,
Dont drench it, but make sure the entire surface is dampened with ninhydrin
solution.
4. Allow the specimens to air dry for a few minutes. Some bluish or purplish
ninhydrin stains may be faintly visible at this point, but do not be concerned
if no stains are evident.
5. Place one of the treated specimens aside for a day or two to allow
development to occur at room temperature.
6. Make a sandwich with two thicknesses of paper towels, followed by the
specimen (print side up), and then two more layers of paper towels.
7. Place the specimen on top of GI sheet. Make sure to move it from time to
time and cover with another GI sheet.
8. Occasionally, spray water on the paper sandwich to increase humidity
9. When development appears to be complete, place the specimen on a clean,
flat surface and examine it carefully under strong light with the magnifier. You
should see fingerprints, which may vary from featureless smudges to prints
showing considerable ridge detail.
Ninhydrin After-Treatments
1. Develop the prints with ninhydrin, as described in the preceding procedure,
and then cut the sheet into four pieces, each of which contains approximately
the same number of similar-appearing prints.
2. Label one of these pieces Control and keep it for comparison to the other
three pieces, each of which will receive a different after-treatment.
3. Label the first test specimen Ninhydrin, and subject it to the ninhydrin
development process, including heat/humidity development, described in the
preceding procedure.
4. Label the second test specimen Blank.and instead of using ninhydrin use
acetone.
5. Label the third test specimen Zinc. Spray this specimen with the diluted
zinc chloride solution to dampen the surface, allow it to air dry, and then
dampen the specimen again with zinc chloride solution and allow it to air dry.
Finally, use the heat/humidity development process described in the
preceding procedure to develop the zinc treated specimen.

Data and Analysis:

Compilation of Experiments in Forensic Chemistry [Type text] Page 17


Material Used Fingerprint Description

White Powder

Black Powder

Iodine Fuming

Ninhydrin

Review Questions:
Dusting
1. Should dusting be the first or last method attempted to raise latent fingerprints?
Why?

2. Is dusting better suited for porous or nonporous surfaces?

Iodine Fuming
1. For what type(s) of specimen is iodine fuming best suited?

2. Is iodine fuming normally the first method attempted for revealing latent
prints? Why?

3. What other types of forensically significant prints might iodine fuming be


used to develop?

Compilation of Experiments in Forensic Chemistry [Type text] Page 18


Ninhydrin
1. Which two common fingerprint development methods must be used before
ninhydrin, if they are to be used at all?

2. What four after-treatment methods are commonly used to enhance


fingerprints developed with ninhydrin?

3. Which, if any, of the after-treatment methods increased the detail visible in


your ninhydrin treated prints

References:
Thompson R., Thompson B. 2012.Illustrated Guide to Home Forensic
Sciences. OReiley Media Inc, Sebastopol, CA
Chemistry Forensics Student Edition , Columbus, OH

Name:______________________________ Date Submitted:_______________

Yr and Sec: __________________________ Date Performed:_______________

Compilation of Experiments in Forensic Chemistry [Type text] Page 19


Group No. ____________________________ Score:________________________

Experiment # 4
Detection of Blood Kastle-Meyer Test (Phenolphthalein)

Introduction
In this activity, you will be comparing the results of the catalase test using
hydrogen peroxide with the phenolphthalein test, to see how each reacts with
blood and other substances.

I. Objective
a.

b.

c.

II. Materials
Substances to be Blood stains 3% hydrogen proxide
identified Cotton swab/ Filter Kastle-Meyer Reagent
Blood Samples (all paper 95% ethanol
animal blood) Spot plate

III. Procedure

A. Using Cotton Swab


1. Sample stain with clean moist cotton swab
2. Add a drop of hydrogen peroxide.
3. Repeat the procedure 1. This time add 2 drops of ethanol
4. Add a drop of Kastle-Meyer Reagent and a drop of hydrogen peroxide.

B. Using the spot plate (fiber)

1. Put one drop of the samples on a cloth


2. Collect stain on a thread and place it on the spot plate.
3. Add a drop of hydrogen peroxide.
5. Repeat the procedure 1 and 2 This time add 2 drops of ethanol
6. Add a drop of Kastle-Meyer Reagent and a drop of hydrogen peroxide.

C. Using filter paper

1. Using a piece of filter paper or paper towel, rub the stain suspected to be
blood to collect a sample. If the stain is dry, you may moisten the paper

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slightly with distilled water. You will not see a visible blood stain on your filter
paper.
2. Lay the paper out so that the spot that you rubbed is exposed. Add 1 drop of
ethanol to the center of the paper.
3. Follow this with 1 drop of reduced phenolphthalein and 1 drop of 3%
hydrogen peroxide.
4. If a pink color appears within 5 s, a positive test should be recorded. No pink
color within 5 s is a negative test result and should be recorded as such.

NOTE: False readings can occur. It is typical for tested filter paper to turn pink after
an extended amount of time. Do not consider this in your test results. A pink color
after phenolphthalein has been applied but before hydrogen peroxide has been
applied normally indicates a false positive due to an oxidant being present.

Data and Analysis

A. Hydrogen Peroxide ONLY

Observations:
Substance Cotton Swab Spot Plate Filter paper

Tomato Sauce
Rust Extract
Red Food Coloring
Cow Blood
Pig Blood
Rabbit Blood
Chicken Blood
Frog Blood
Human Blood
Unknown A

Unknown B

B. Kastle-Meyer Test or Phenolphthalein Test

Observations:
Substance Cotton Swab Spot Plate Filter paper

Compilation of Experiments in Forensic Chemistry [Type text] Page 21


Tomato Sauce
Rust Extract
Red Food Coloring
Cow Blood
Pig Blood
Rabbit Blood
Chicken Blood
Frog Blood
Human Blood
Unknown A

Unknown B

Guide Questions:

1. Which of the method is easier to use?

2. Explain the mechanism behind the Kastle-Meyer Test.

Name:______________________________ Date Submitted:_______________

Yr and Sec: __________________________ Date Performed:_______________

Group No. ____________________________ Score:________________________

Compilation of Experiments in Forensic Chemistry [Type text] Page 22


Experiment # 5
Blood Spatter Analysis
Introduction:
Crime scenes that involve bloodshed often contain a wealth of information in
the form of bloodstains. The pattern, size, shape, and the location of such stains
may be very useful in the reconstruction of the events that occurred
The experiment will let the students learn how to reconstruct the crime scene by
merely analyzing the bloodstain pattern. Different categories of bloodstain pattern
will be figured out and calculation of the angle of impact will be determined

I. Objectives
a.

b.

c.

II. Materials
Hammer Artificial blood
Knife/ Cutter Sponge
Manila paper Syringe
Newspaper Medicine Dropper

III. Procedure
A. Making the Artificial Blood
Make your own blood using cornstarch and dye. Mix approx. 40 g of
cornstarch with 80 mL of water. Add 160 mL of corn syrup (optional) and mix. Add
2-3 teaspoons of red dye or until the color of the mixture is that of blood. Dont get
it on your clothes.
B. Types of Bloodstain Pattern
A. Passive bloodstain
1. Drop By using a medicine dropper, drop 3 drops of blood in three
different places, 25 cm away from the paper. Repeat the same procedure
on different heights (50cm, 75 cm and 100cm). Repeat the procedure using
the knife.
2. Drip Drop 8 drops of blood on the paper on the same spot, 25 cm away
from the paper. Repeat the same procedure on different heights (50cm, 75
cm and 100cm). Repeat the procedure using the knife
3. Pool Using 1 mL of blood, place the blood 25 cm away from the paper and
note the pattern. Repeat the same procedure on different heights (50cm, 75
cm and 100cm)
B. Transfer Bloodstain
1. Contact Bleeding From a bloodied object, transfer the blood unto the
unstained paper producing a replica of the mark
Compilation of Experiments in Forensic Chemistry [Type text] Page 23
2. Swipe From a bloodied object, transfer the blood to the paper. Swipe the
mark, using your hands several times to erase the original mark of the object.
C. Projected Bloodstain
1. Arterial Spurt- Using a syringe, draw 5mL of blood and push the plunger
acroos a vertical paper producing a vertical stain characteristic of an arterial
spurt.
2. Splash onto floor- Start off walking at slow walking rate along the paper
from one to end to another. Repeat the procedure using a normal walking
rate and a fast walking rate. Repeat the procedure using the knife
D. Impact Spatter
1. Stamping- Place 10 mL of blood on paper the using a shoe heel, stamp on
the pool of blood.
1. Horizontal surface - Pour two tablespoons of blood onto a damp
sponge. The sponge should not be dripping wet but rather just moist.
Strike the sponge with a hammer. Repeat the same procedure using a
knife.
2. Vertical surface
a. Tape large sheets of paper on a wall. Make sure the paper
reaches the floor and goes pretty high up.
b. Lay newspaper on the floor underneath the papered wall.
c. Pour two tablespoons of blood onto a damp sponge. The sponge
should not be dripping wet but rather just moist.
d. Lay the sponge on the floor about 2 feet from the wall.
e. Strike the sponge with the hammer.
f. Repeat the procedure from a-d, strike the sponge in swishing
manner.
Data and Analysis
1. Drop
Height Picture or Image Description

2. Drip
Height Picture or Image Description

Compilation of Experiments in Forensic Chemistry [Type text] Page 24


3. Pool
Height Picture or Image Description

B. Transfer Bloodstain
Type Picture or Image Description

Contact
Bleeding

Swipe

C. Projected Spatter
Type Picture or Image Description

Arterial
Spurt

Splash

D. Impact Spatter
Type Picture or Image Description

Stamping

Compilation of Experiments in Forensic Chemistry [Type text] Page 25


Vertical
Surfaces

Horizontal
Surfaces

Guide Questions
1. Which tool made a low-velocity spatter? Medium-velocity? High
velocity?

2. Are the computations and measurements same or precise with each


other?

3. What were the factors that affected your exact measurements of blood
spatters?

4. What could be some reasons for some errors in measurement?

Name:______________________________ Date
Submitted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Compilation of Experiments in Forensic Chemistry [Type text] Page 26


Group No. ____________________________ Score:________________________

Experiment # 6
Blood Pattern Analysis
Introduction
Blood Pattern Analysis the examination of the shapes, locations, and
distribution patterns of bloodstains, in order to provide an interpretation of the
physical events which gave rise to their origin. The experiment considers
bloodstains on a single target surface such as a floor. If multiple surfaces are
introduced, complexity significantly increases.

Materials
Artificial Blood (Experiment # 4) Tape
Hammer Graphing paper
Sponge (2 in. thick) Meter stick
Manila paper Protractor
Newspaper Ring stand
String (yarn)

Procedure:
A. The Blood Stain (You may use the specimen in Experiment # 4)
1. Tape large sheets of paper on a wall. Make sure the paper reaches the floor
and goes pretty high up.
2. Lay newspaper on the floor underneath the papered wall.
3. Pour two tablespoons of blood onto a damp sponge. The sponge should not
be dripping wet but rather just moist.
4. Lay the sponge on the floor about 2 feet from the wall.
5. Strike the sponge with the hammer.

B. Angle of Impact
1. Accurately measure the width
and length of a given bloodstain.
This must be measured to the
nearest millimetre.
2. Divide the width of the stain by
the length of the stain in order to
obtain the width to length ratio.
3. Calculate the inverse sine of this
ratio.
4. This value is the angle of impact.

C. Defining the Area of Origin

I. Triangulation Method
1. Place the ring stand on the Area of Convergence
Compilation of Experiments in Forensic Chemistry [Type text] Page 27
2. Write the calculated Angle of Impact next to each stain.
3. Using string, masking tape and a protractor, raise the string to the calculated
angle and attach it to the ring stand.
4. Do the same for a minimum of 3 stains.
5. The place on the ring stand where the string from each stain meets is the
Area of Origin.
6. Measure the height of the Area of Origin.

II. Tangent Method


Using the same data from Part B, the following formula is used to calculate the Area
of Origin. You need to use a scientific calculator for this activity.
TAN i = H/D
o i = angle of impact
o D = distance from stain to area of convergence
o H = unknown distance above target surface

III. Graphing Method


The scales of both axes used in the graph must be the same. The X-axis represents
the target plane and graphs the distance from the back-edge of the stain to the
AOC. The target plane is the floor. Make sure the axis of the graph is long enough.
The Z-axis represents the height above the target plane in this example the target
plane is the floor. This is an unknown value it is what you are trying to find out.
Make this scale the same as the X axis. Use a separate graphing paper for this
method.
1. Mark on the X-axis of the graph the AOC distance (cm) for each different
stain. You need to graph a minimum of 3 stains.
2. Using a protractor, draw a line from the mark on the X-axis, at the calculated
angle of impact, to the Z-axis. See the example below.
3. Repeat this procedure for each stain.
4. The point at which the lines from the X-axis converge on the Z-axis
establishes the probable height of the Area of Origin.

DATA AND ANALYSIS

A. Angle of Impact
STAIN WIDTH LENGTH ANGLE OF
IMPACT
1
2
3
4
5
Calculation:

B. Determining the Point of Origin by using Triangulation Method


Compilation of Experiments in Forensic Chemistry [Type text] Page 28
STAIN ANGLE OF IMPACT HEIGHT OF THE ORIGIN
1
2
3
4
5

C. Determining the Point of Origin using Tangent Method

DISTANCE FROM HEIGHT OF THE


STAIN ANGLE OF IMPACT
POC ORIGIN
1
2
3
4
5
Calculation:

D. Determining Point of Origin by using Graphical Method

DISTANCE FROM HEIGHT OF THE


STAIN ANGLE OF IMPACT
POC ORIGIN
1
2
3
4
5

Summary:

TRIANGULAR GRAPHICAL
STAIN TANGENT METHOD
METHOD METHOD
1
2
3
4
5

Questions

Which method do you think is the best method?

Compilation of Experiments in Forensic Chemistry [Type text] Page 29


What is your reason for this selecting this method?

Which method do you think is the worst method?

Why?

REFERENCES

Forensic investigations: Bloodstain analysis experiments (teacher activity


information)
FSB07 | revised June 2013 | The University of Western Australia

Compilation of Experiments in Forensic Chemistry [Type text] Page 30

D. Summary: Point of Origin


Name:______________________________ Date
Submitted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 7
Blood Typing

Introduction
One way in which blood may be characterized is by the presence or absence of
molecules located on the surfaces of the red blood cells. In the ABO blood typing
group, there are two types of chemical molecules or antigens that are found on the
red blood cells: A and B.
If a red blood cell has only A antigens on it, it is type A.
If the red blood cell has only B antigens on it, it is type B.
If the red blood cell has both A and B antigens on it, it is type AB.
If neither A nor B antigens are on the surface, it is type O

Objectives:
a.

b.

c.

Materials
simulated A, B, AB, and O blood toothpicks
simulated anti-A and anti-B sera unknown blood samples (prepared by
spot plate teacher)

Procedure

Part 1: Observing the Reactions of Known Blood Samples with Antiserum


1. Gently swirl or shake the container with blood type A to re-suspend the cells.
2. Using a clean spot plate, place three drops of blood type A into two of the
wells.
3. Add three drops of anti-serum A to one well and three drops of anti-B serum to
the other.
4. Gently swirl the plate or use a clean toothpick to stir the solution (30 to 60
seconds).
5. Observe carefully, looking for clumping (agglutination) around the edges.
6. Record your results in a data table.
7. Repeat Steps 1 to 6 for blood types B, AB, and O, making sure all equipment is
clean prior to use.

Compilation of Experiments in Forensic Chemistry [Type text] Page 31


Part 2: Identifying the Blood Types of Unknown Samples (Practice)
1. Obtain two unknown blood samples from the teacher.
2. Repeat the procedure from Part 1 with the unknowns.
3. Record your results in a data table.
4. Compare your results with those from Part 1 to identify the unknowns.
Data and Analysis:

Observing the Reactions of Known Blood Samples with Antiserum


Type Anti-A Sera Anti-B Sera Conclusion
A
B
AB
O

Identifying the Blood Types of Unknown Samples (Practice)


Unknown Anti-A Sera Anti-B Sera Blood Type
A
B
C

Guide Questions:
1. Explain how blood collected at the scene of a crime could be used in a criminal
investigation.

2. Identify and describe other situations, in addition to criminal investigations,


where a working
knowledge of blood types and the procedure to determine blood type might be
used.

Compilation of Experiments in Forensic Chemistry [Type text] Page 32


Name:______________________________ Date
Submitted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 8
Identification of Semen
Introduction
The examination of semen and other seminal stains is an important part in
the routine investigation of sexual offenses like cases of rape, adultery, sodomy and
bestiality.

Objectives:
a.

2b.

c.

Materials:
Microscope slide
Cover slip
Microscope
Medicinal dropper

Reagent

Florence Reagent
Methylene Blue
pH paper

Procedure

A. Physical Examination
a. Examine and describe the characteristics of each stain (one month,
one week, one day and fresh stain.
B. Chemical Examination
a. Florence Test
1. Divide the cloth with stains into small bits.
2. Soak in 0.5% saline solution.
Compilation of Experiments in Forensic Chemistry [Type text] Page 33
3. Transfer a piece of cloth to the slide and carefully tease
until the fluid evaporates.
4. Add a drop of Florence reagent and cover with a cover
slip.
5. Observe under a microscope.
C. Microscopical Examination
a. Fresh semen sample
1. Put a small amount of fresh semen on the microscope
slide.
2. Cover with the cover slip.
3. Observe under the microscope
b. Staining
1. Put a small sample of the material on the slide,( if dry add
a drop of distilled water).
2. Allow to dry and stain with methylene blue.
3. Wash with water and dry
4. Observe under a microscope.

Data and Analysis

Physical Examination
Sample Color Smell Appearance pH
One Month
One week
One day
Fresh Semen
Analysis:

Chemical Examination
Sample Observation Result
(+/-)
One Month
One week
One day
Fresh Semen
Analysis:

Microscopical Examination
Sample Observation
One Month
One week
One day
Fresh Semen
Analysis

Compilation of Experiments in Forensic Chemistry [Type text] Page 34


Name:______________________________ Date
Submitted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 9
Chemical Test for Explosives

Introduction
Salts of Nitrates such as potassium nitrate (KNO 3) and ammonium
nitrate(NH4NO3) are used as ingredients in fireworks, gunpowder and explosives. It
is also used in pickling meat (KNO3) and fertilizers (NH4NO3). Addition of
Diphenylamine Reagent (Lunges Reagent) to nitrates results in the production of
blue color. Nesslers Reagent is used to identify ammonium samples. Positive result
of this reagent is brown-orange precipitate.

Materials
Spot plate 1N NaOH
Dropper Potassium Nitrate
Test tube Ammonium Nitrate
Litmus paper Gunpowder (if available)
Diphenylamine reagent Fire cracker
Nesslers reagent Fertilizer

Procedure
A. Test for Nitrates
1. To a spot plate, place 2mg of the sample
2. Add 3 drops of DPA reagent
B. Test for Ammonium Nitrate
1. Transfer 10 mg of sample to a test tube.
2. Add 5 drops of 1N NaOH to make it alakaline. Heat the solution and place a
red litmus paper on the tip of the test tube. (Alkaline-Red litmus paper turns
to blue)
3. Add 2 drops of water.
4. Add 5 drops of Nesslers reagent

Compilation of Experiments in Forensic Chemistry [Type text] Page 35


Data and Analysis
A. Test for Nitrates
Powder Observation Nitrate(+/-)

Potassium Nitrate
Ammonium Nitrate
Firecracker
Gunpowder
Fertilizer
B. Test for Ammonium Nitrates
Powder Observation Nitrate(+/-)

Potassium Nitrate
Ammonium Nitrate
Firecracker
Gunpowder
Fertilizer

Guide Questions:
1. Explain briefly the rationale of the DPA test and Nesslers Reagent.
Write the chemical reactin for each test.

References

Dacil-Caete, AM, Sangil, MC. 2014. Forensic Chemistry and Toxicology,


Revised Edition. Wisemans Books Trading Inc. Quezon City, NCR
Thompson R., Thompson B. 2012.Illustrated Guide to Home Forensic
Sciences. OReiley Media Inc, Sebastopol, CA

Compilation of Experiments in Forensic Chemistry [Type text] Page 36


Name:______________________________ Date
Submitted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 10
Hair Analysis

Introduction
The examination of hair and fiber is an important phase in criminal
investigation. Almost all of the violent crimes include the possibility of physical
contact between the criminal and the victim. The successful investigation of crimes
of violence such as rape, murder, assault, kidnapping, etc are frequently materially
assisted by the examination of hair and fiber

Materials
Hair
Microscope slide
Coverslips
Microscope
Water
Nail polish-clear

Procedure
A. Microscopic Examination of Hair
1. Examine the hair with your eyes, the hand lens, and microscope. Record your
observations. Note characteristics such as colour, thickness, texture, and any
unique characteristics such as split ends.
2. Remember to observe both ends of the hair.
3. Make a wet mount slide of a hair.
4. Observe the hair with the compound microscope and record its
characteristics, including colour, thickness, texture, and any unique
characteristics such as split ends. Remember to observe both ends of the
hair. Look for medulla, cuticle, and scales. Look for the distribution of pigment
granules.
5. Record and make drawings of your observations.
6. Repeat this procedure for the other hair samples. Note and record any
differences
7. Examine and record your observations

B. Make and Observe Scale Casts of Human Hair


1. Brush a thin layer of clear nail polish onto the middle third of a microscope
slide.
2. Carefully press the hair specimen into the tacky nail polish until it adheres.
3. Allow the nail polish to dry.
4. Using the forceps, carefully pull the hair specimen away from the slide in one
smooth motio Using forceps to remove the hair specimen from the dried nail
polish

Compilation of Experiments in Forensic Chemistry [Type text] Page 37


5. Examine the scale cast under high magnification by both transmitted and
incident light, adjusting the brightness, angle, and contrast of the lighting to
optimize visibility of the scale pattern. Use your ocular micrometer to
measure the size of the scales, and note any variations in the pattern or size
of the scales over the length of the shaft. Record your observations in your
lab notebook.
6. Repeat steps 1 through 5 for each specimen.
7. If you have the necessary equipment, shoot an image of a representative
scale pattern,Make a sketch and paste it in your laboratory sheet.

Data and Analysis

A. Microscopic Examination (Wet Mount)


Hair Image or Sketch Observation

Head

Pubic Hair (Male)

Pubic Hair (Female)

Armpit

Legs

Dog

Cat

Compilation of Experiments in Forensic Chemistry [Type text] Page 38


B. Scales of Hair

Hair Image or Sketch Observation

Head

Pubic Hair (Male)

Pubic Hair (Female)

Armpit

Legs

Dog

Cat

Guide Question:

1. Which two somatic regions yield hairs that are most significant forensically?
Why?

2. What is the primary forensic value of determining the scale pattern on a hair
specimen?

Reference:

Compilation of Experiments in Forensic Chemistry [Type text] Page 39


Thompson R., Thompson B. 2012.Illustrated Guide to Home Forensic
Sciences. OReiley Media Inc, Sebastopol, CA
Name:______________________________ Date
Submtted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 11
Fiber Analysis

Introduction
Fibers are trace evidences that are transferred through personal contact, struggle or
impact. In crimes such as rape, homicide and hit and run, the transfer of fiber or thread may
occur between the victim and the assailant. These evidences may be in small amounts but
their value is a great import in the crime investigation.

Materials
Compound light microscope Watch glass
Slide 50 ml beaker
Cover slip 10% NaOH
Forceps 5% Oxalic acid
White paper Millons Reagent
Alcohol lamp Conc. Sulfuric acid
Match Conc and Dil Ammonium Hydroxide
Samples of wool, rayon, silk, Iodine Solution
polyester, and cotton Different Types of Fibe

Procedure
Microscopic Examination
1. Prepare a wet-mount slide of the fi ber by placing it on the slide, adding a
drop of water, and covering the fiber and water with a cover slip.
2. Examine the fiber under low, medium and high magnifi cation with your
microscope. Sketch what you see. Note any pits or striations on the fi ber.
Place the sketch and notes in your data table.
3. Repeat this procedure with the samples of wool, rayon, silk, polyester, and
cotton. Sketch each of these samples at low, medium and high power. Place
your sketch and notes about the fi bers in your data table.
B. Flame Test
1. Holding the fiber in the forceps, bring it close to, but not touching, the flame.
Describe the fibers behavior as it approaches a flame: does it begin to melt,
ignite, or curl?
2. Holding the fiber in forceps, touch the fi ber to a fl ame. Does it ignite quickly
or slowly? Does it sputter, drip, or melt?
3. Remove the fi ber from the fl ame and describe how it behaves. Does it self-
extinguish, continue to burn, or continue to glow?
Compilation of Experiments in Forensic Chemistry [Type text] Page 40
4. Note any odor associated with the fiber in the flame. Does it smell like
vinegar or hair?
5. What kind of residue is left after the fiber is removed from the fl ame? Does
the fiber leave a white, fluff y ash, a hard bead, or a melted?
C. Chemical Analysis

1. Place sample in a small beaker and cover with 10% NaOH solution. Cover with
watch glass and boil gently over a burner for 10 15 minutes
Sample dissolves wool or silk
Sample does not dissolve - Cotton, linen, wild silk or cellulose silk
o Wash sample with water, place in beaker with 5% oxalic acid and boil
for 15-20 mins
o Remove and boil in several changes of distilled water or until sample
gives no test for starch (blue color in iodine). Dry thoroughly between
pieces of filter paper and divide into several portions
Put one portion in a watch glass add one or two drops of
Millons reagent and warm slightly
Pink or rose red silk
Not collored Cotton or Linen
Immerse another portion for exactly two minutes in conc
sulfuric acid, rinse well in distilled water and then dilute in
ammonium hydroxide. Remove and dry
Sample gelatinized; soluble in water cotton
Sample little affected; not soluble linen
In a sample add few drops of iodine sol, allow to remain
for one minute, wash in water and place in a watch glass
of distilled water.
Sample remains blue or black mercerized cotton
Loses color in few minutes, fading to a yellow or
brown cotton

Data and Analysis

A. Microscopic Examination
Type of Fiber Image or Sketch Description

Compilation of Experiments in Forensic Chemistry [Type text] Page 41


B. Flame Test
Type of Fiber Image or Sketch Description

C. Chemical Analysis

Sample 10% NaOH Millons Conc. Sulfuric Iodine


Reagent Acid Solution

Guide Questions

1. Among the various test for fibers, which is the most reliable? Why?
Compilation of Experiments in Forensic Chemistry [Type text] Page 42
Reference:
Garcia, M.A.2014.Forensic Chemistry and Toxicology, Purely Books & Trading
Publishing Corp,
Manila, Philippines
Name:______________________________ Date
Submtted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 12
Physical Properties of Glass

Introduction
Glass is encountered as evidence in burglaries, homicides, assaults and hit
and run offenses. The fragments are easily embedded in shoes and clothing during
the enactment of the crime. Physical properties such as density and refractive index
will help in determining if the glass pieces from the crime scene matches the
fragments that found in the suspect.

Materials
Different types of glass fragments Microscope
Graduated cylinder, 10 mL Isopropyl alcohol or acetone
Forceps Glycerin
Test tube(s) Olive oil
Test tube rack Cassia oil
Beaker, 100 mL Slided (with deep cavity as well)

Procedure:
A. Determining the Density of Glass
1. Weigh about 10 grams of one of the glass specimen
2. Transfer 4 mL of water to the graduated cylinder. Record the initial volume.
3. Carefully add the glass fragments to the graduated cylinder. Record the final
volume.
4. Calculate the volume by subtractive the final volume to the inital volume
(water discplacement method: V=Vf- - Vi.)
5. Calculate the density by dividing the mass of the glass fragments over the
volume you have obtained from Step 4.
6. Compare your results on the table below and make a tentative identification
of the type of glass.

Table 1: Densities of common glass(g/cm3

Glass Type Density


Automobile headlight glass 2.20-2.29
Automobile window glass 2.53-2.75
Compilation of Experiments in Forensic Chemistry [Type text] Page 43
Fused quartz/ silica 2.18-2.21
Labware Borosilicate glass (Kimax, 2.23-2.72
Pyrex, Duran)
Lead glass 2.65-6.00
Soda-lime-slicate (flat glass, bottles, 2.40-2.63
window glass)
Glass ceramic 2.53
Glass type

B. Refractive Indices of Glass


Put a deep cavity on the slides. This will serves as the well four your refractive index
liquid and glass fragment.
Place a small piece of the glass fragment on the middle range of the cavity. Make
sure that the glass fragment is completely submerged in the liquid
Place the slide on the microscope stage and observe it at 40x and 100x
Use the Bech line method in determining the refractive index (RI). If the RI of the
liquid matches the glass, the glass will be invisible. If the RI of the glass is closely
matched to the RI liquid, the edges of the glass will be poorly defined. If the RI of
the glass and the liquid differ significantly, the edges of the glass will standout
against the liquid. You may use the table below for the proportion of olive cassia
oil or glycerine.

Table 2. Refractive Indices of some Liquid


Table
Cassia Refractive
Olive Oil
Oil Index
All None -1.467
28 2 -1.477
26 4 -1.486
24 6 -1.496
22 8 -1.506
20 10 -1.515
18 12 -1.525
16 14 -1.535
14 16 -1.544
12 18 -1.554
10 20 -1.564
8 22 -1.573
6 24 -1.583
4 26 -1.593
2 28 -1.602
None 2 -1.60
Water 1.33
Acetone 1.36
Glycerine 1.47

Data and Analysis


A. Density of Glass

Compilation of Experiments in Forensic Chemistry [Type text] Page 44


Glass Mas Initial Final Volume Density Densit Perce
s Volume Volu (V=Vf- - (D=M/V) y nt
me Vi) (Table Error
1)
Glass
(drinking)
Window
(house)
Headlights
(automobile
)
Labware
Bottle
B. Refractive Indices of Glass
Glass Observation RI Liquid Refractive
Used Index
Glass (drinking)

Window (house)

Headlights
(automobile)
Labware

Bottle

Guide Questions
1. Why is the density of the liquid used to determine the density of a specimen by
displacement immaterial?

2. You are presented with one questioned glass specimen and a dozen known
specimens. Propose a method for doing a quick preliminary screening to determine
if the RI of the questioned specimen matches one or more of the known specimens.

Compilation of Experiments in Forensic Chemistry [Type text] Page 45


Reference:
Thompson R., Thompson B. 2012.Illustrated Guide to Home Forensic
Sciences. OReiley Media Inc, Sebastopol, CA

Name:______________________________ Date
Submitted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 13
Restoration of Erased Serial Numbers by Suitable Etching
Solutions
Introduction
Macro-etching is the application of chemical solution on the metal surface
where the serial number is normally located. Serial number refers to the series of
number that is punched or pressed into a particular item to distinguish it from
another of common type. It is a series of alphanumeric digits that is placed in items
to distinguish each.

Materials
Coins
Any metal material that has corroded serial numbers
Bond paper
50 % v/v Nitric acid
Copper (II) chloride solution (See Appendix A)
Hydrochloric acid

Procedure
A. For clean visible serial numbers
1. Remove the grease from the specimen using alcohol or gasoline
2. Place the bondpaper over the serial number
3. Stencil it with a pencil until the serial number appears
B. Number Restoration Hard Metal
1. Clean the surface of the specimen with cotton soaked in the reagents listed
below depending on the type of material.
a. Hard metal Nitric acid solution, Cupric chloride solution
b. Soft Metal Hydrogen peroxide in acetic acid
2. Wipe the suface with clean cotton until it is dry.

Compilation of Experiments in Forensic Chemistry [Type text] Page 46


3. Apply black powder (you may use fingerprint powder) until the letter are
revealed.
4. Lift the number with a lifting tape.
5. Place the lifted number on the bond paper

Data and Analysis

A. For clean visible serial numbers (Paste your results)

Observation

B. Number Restoration

Lifted Serial Numbers Lifted Serial Numbers

Guide Questions
1. How will you know if the serial number is tampered or not?

Compilation of Experiments in Forensic Chemistry [Type text] Page 47


References

Dacil-Caete, AM, Sangil, MC. 2014. Forensic Chemistry and Toxicology,


Revised Edition. Wisemans Books Trading Inc. Quezon City, NCR
Thompson R., Thompson B. 2012.Illustrated Guide to Home Forensic
Sciences. OReiley Media Inc, Sebastopol, CA
Name:______________________________ Date Submtted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________


Score:________________________

Experiment # 14
Casting and Moulding

Introduction
In many cases, footwear evidences can lead to positive identification of a
particular known shoe that made the print. Footwear evidence can provide
investigators with certain information that can assist them in locating a suspect.
Casting provides lifelike and actual-size molding of the original impression including
uneven surfaces and depths. It also back-ups photographs from the crime scene.

Materials
Plaster of Paris/ dental stone
Zip-lock bags
Mixing bowl
Stirring rod
Talcum Powder
form

Procedure
A. Preparation of the Plaster of Paris
1. Place 5 pounds of Plaster of Paris in a mixing bowl and add 15 oz of water.
2. Mix for 5-7 minutes or until pancake batter consistency.
B. Pouring
1. Put the form around the impression.
2. Sprinkle the impression with talcum powder.
3. Pour the mixture starting on side of the impression, using the stirring rod as
the guide.
4. You may use the Ziplock bag by tearing one edge of the plastic.
5. After pouring about half of the mixture, add the reinforcement material
(sticks) and then complete the plaster pouring
Compilation of Experiments in Forensic Chemistry [Type text] Page 48
6. Allow the cast the cast to set for an hour. When the cast is firm and soft,
scratch the date, case number, initials of the investigator and other pertinent
information.

Data and Analysis

A. Image of the Casted Impression

B. Observation

Guide Questions:
Why do you need a spoon or stick to guide the mixture to the impression?

Give some techniques in storing and transporting the cast.

What is the chemical name of Plaster of Paris? Write chemical formula.

Compilation of Experiments in Forensic Chemistry [Type text] Page 49


References
Dacil-Caete, AM, Sangil, MC. 2014. Forensic Chemistry and Toxicology,
Revised Edition. Wisemans Books Trading Inc. Quezon City, NCR

Name:______________________________ Date
Submtted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 15
Identification of Inks using Paper Chromatography
Introduction

Most samples of matter are impure mixtures of two or more substances.


Chromatography is a widely used experimental technique for the separation of a
mixture of compounds into its individual components. The word chromatography
means "separation of colors" but today chromatography iused for both colored and
colorless substances.
The separation process is based on the fact that porous solids adsorbs
different substances to different extremes depending upon their polarity. The term
Adsorption refers to the adhesion or stickiness of a substance to the surface of
another substance, as opposed to the term absorption which refers to a substance
penetrating into the inner structure of another substance. A mixture to be
separated is first applied to an immovable porous solid (like paper, or alumina, or
fine silica sand) called the stationary phase. The components of the mixture then
get washed along the porous solid by the flow of a solvent called the mobile
phase. The mobile phase can be liquid (as in column, paper, or thin-layer
chromatography) or it can be a gas (as in gas chromatography).
Substances can be identified by the heights they reach on the completed
chromatogram by calculating Rf (rate of flow or retention factor) values. The Rf
value is a constant for a given substance under the same experimental conditions.
The Rf value may be calculated from the following equation. The Rf value itself is
unitless.

Distance of t h e center of t h e sample spot


Distance of t h e solvent
Rf = t h e origin t h e origin

Compilation of Experiments in Forensic Chemistry [Type text] Page 50


Materials
Chromatography paper (or Whattman Filter paper)
Beaker
Watch glass (big enough to cover the mouth of the beaker)
Color pens or markers
Suspected document
0.1% NaCl solution
Solvent (i.e. ethanol, hexane, acetone)

Procedure
1. Obtain a 9 x 14 cm piece of chromatography paper.
2. Use a pencil to draw the origin line about 1.5 cm from the bottom of the long
edge of the paper.
3. Use a pencil to make 8 small evenly spaced vertical marks every 1.5 cm
along the length of the origin line, starting about 1.5 cm from the edge of the
paper. Number these marks 1 through 8 just below each with a pencil.
4. Select a water soluble black felt pen or marker and touch it briefly. Make a
black spot about 1 or 2 mm in diameter. With pencil write the name "black"
below the origin line.
5. Do the same procedure with other inks.
6. Lightly brush the cotton swab with a solvent (can absorb the ink from the
paper) on the suspected document. Wet the swab with the same solvent that
is used in the previous procedure. Extract the ink by squeezing the swab. Use
a capillary tube in making a spot to the chromatography paper. Label the
spot.
7. Roll the chromatography paper into a cylinder with the
origin line at the bottom and the dye spots on the
outside of the cylinder. Use two fingers of one hand to
hold the ends of the paper close together, about 2 mm
apart. Staple the top ends and then staple the bottom
ends. The ends of the paper should not touch.
8. Add about 10 mL of 0.1% salt (NaCl) solution to a 400-
mL beaker. The solution should be about 0.5 cm deep
9. Place the beaker on your work bench where it will not be bumped or
disturbed
10. Being careful that the paper does not touch the sides of the beaker, carefully
place the paper cylinder into the beaker containing the salt solution. Make
sure the origin line is at the bottom. The origin line must not be below the
surface of the eluent.
11.Cover the beaker with a watch glass. Do not disturb the beaker while the
eluent rises up the paper.
12.Watch the eluent as it moves up the paper and see what happens as it comes
into contact with the ink and food colors. Leave the paper in the beaker until
the eluent front is about 1.5 cm from the top of the paper.
13. When the eluent front nears the top, remove the chromatogram and open
the cylinder by tearing the paper at the staples. Set the chromatogram on an
empty beaker or a paper towel to dry.
14.After 2 or 3 minutes, or when the paper appears to be drying out, mark the
final position of the eluent front with a pencil line.
15.Outline the main color spots and calculate the Rf values of each dye.

Compilation of Experiments in Forensic Chemistry [Type text] Page 51


Data and Analysis
A. Chromatogram: (Recreate as accurately as possible.)

B. Distance Travelled by Solvent Front

Color Color of Spot Rf


Distance
(List all if more
Travelled
than one)

Guide Questions:

What is the advantage of allowing the eluent front to rise to near the top of
the TLC sheet rather than stopping when only half-way up?

Compilation of Experiments in Forensic Chemistry [Type text] Page 52


References
CH104 Paper and TLC.pdf
Name:______________________________ Date
Submitted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 16
Soil Analysis

Introduction
Soil is one of the most common forms of physical evidence found at crime
scenes. For example, a vehicle suspected of having been used in an armed robbery
may later be found to have soil from the crime scene adhering to its tires or wheel
wells, thereby establishing that that vehicle was present at the crime scene.
Similarly, a suspected rapist or mugger may have soil adhering to his clothes or
shoes. (Soil specimens are often particularly easy to obtain from shoes or boots with
deep tread. If soil found on a suspects shoes or clothing is consistent with soil
present at the crime scene, it establishes that the suspect was probably present at
that scene.

Materials
Beaker Munsell color chart pH paper with pH paper
Test tubes Magnifying glass color chart
Pipette or droppers Microscope Sample of soil from
Test tube stopper Scoop different location

Procedure:
A. Determination of Color and Texture of Soil
Color
1. Use a plastic spoon to scoop a small amount from the crime scene.
2. Observe the color of the soil using a magnifying glass and microscope.

Compilation of Experiments in Forensic Chemistry [Type text] Page 53


3. Using the Munsell chart, assign the color grade of the samples being
analyzed.
Texture
1. Use your touch, naked eyes, the aid of magnifying glass and microscope to
determine the texture of your sample.
2. Classify your soil sample wheter it is sandy, silt, clay or combination
Foreign Materials
1. Using a magnifying glass, note for the inclusion of other materials such as
vegetation, roots, hair, fibers, etc.

B. Determination of pH in Soil Samples


1. Scoop a small amount of soil and place in a test tube.
2. Add 6 mL of water to the test tube.
3. Stopper the test tube and turn it upside down for 2-3 minutes. DO NOT
SHAKE THE TEST TUBE.
4. Allow the solution to settle for about 1 minute.
5. Dip the tip of the pH paper and observe the color change.
6. Compare the color on the pH paper with pH indicator.

Data and Analysis


A. Determination of Color and Texture of Soil
Sample Color Grade Texture Foreign Materials

B. Determination of pH in Soil Samples


Sample pH

Guide Questions:
Are the types and color grades of your sample the same? Can color grade and
texture be used as a differentiating tool in crime scene investigation?

From the foreign materials present in your sample, what do these tell you
about the type of soil?

Compilation of Experiments in Forensic Chemistry [Type text] Page 54


From the results of the pH test, what conclusions can be drawn as to the
relative acidity and basicity of soil from different location.

Reference:
Garcia, M.A.2014.Forensic Chemistry and Toxicology, Purely Books & Trading
Publishing Corp,
Manila, Philippines

Name:______________________________ Date
Submitted:_______________

Yr and Sec: __________________________ Date


Performed:_______________

Group No. ____________________________ Score:________________________

Experiment # 17
Determination of Alkaloids using Thin-Layer
Chromatography
Introduction
Before the use of spectroscopy and other instrumental analysis techniques
became common in forensic toxicology, analysis was done strictly by wet-chemistry
techniques. Because many different plant alkaloidssuch as nicotine, strychnine,
aconitine, and coniineare fatal in small doses, forensic chemists during the 19th
and early 20th centuries devoted considerable effort to developing reagents that
could detect alkaloids at very low concentrations. Many of these reagents are
general alkaloid reagents, providing positive results with many or most alkaloids.
Others were used to differentiate specific alkaloids.

Materials
Centrifuge tubes Mortar and Pestle
Beaker TLC Plate
Test tube (small) 6M Hydrochloric acid
Test tube, 2.5 cm x 15 cm Distilled water
Erlenmeyer flask, 250 mL Dragendorff Reagent
Graduated cylinder, 10 mL Spinach leaves
Capillary tube Cactus
Stirring rod Black pepper seeds
Spot plate
Compilation of Experiments in Forensic Chemistry [Type text] Page 55
Procedure
A. Preparation of Plant Samples
1. Separately crush the plant samples. Make sure to clean the mortar and pestle
before using it.
2. Transfer the crushed material to separate test tubes and add 3 mL distilled
water and 0.25 mL 6M HCl using a pipette.
3. Swirl to mix the contents
4. Stand the test tubes in a test tuibe rack and swirl occasionally, then allow the
dilute acid to extract any alkaloids present in the specimen.
B. Determination of Alkaloids using Thin-layer Chromatography
1. Draw a pencil line (light) across the 2.5cm x 10cm TLC plate about 1.5 cm
from the end of the strip. Put a small dot near the center of each line.
2. Draw a pencil line (light) about 1 cm from the other end of the strip.
3. Place 2 mL of hot water to the 2.5 cm x 15 cm test tube. Stopper the test
tube. Put the tube inside the erlenmeyer flask,
4. Using a toothpick or capillary tube, spot each strips with the corresponding
alkaloid solution.
5. When you have spotted all the strips with the samples, place a new spot on
the dried spot. Repeat the procedure until you have spotted it at least five
times. Allow the strips to dry completely.
6. Transfer three drops of each plant sample in the same well in a spot plate.
Mix the solution using toothpick. Repeat steps 1-4 for the sample.
7. Using one of the strips, check the water level inside the test tube. The water
must not be high enough to reach the spot on the strip. As quickly as
possible, remove the stopper from the test tube, slide the strip (2cm mark
first) and stopper it again.
8. Keep a close eye on the tubes. The TLC plate draws up water from the bottom
tube by capillary action. When the solvent reaches up on the 1 cm mark,
remove the strip.
9. Place the strips on a paper towel and let it dry.
10.Once the strips are dried, place one strip on another paper towel.
11.Transfer a couple of drops of Dragendorffs reagent to a cotton swab.
12.Put the tip of the cotton swab to the top center of the strip and wipe
downward, dampening the center of the strip along its length until you reach
the spot. If necessary, add more reagent to the cotton swab. The goal is not
to soak the entire strip, but to dampen the area of the strip where the
alkaloid traces are present. Depending on the particular alkaloid and the
amount present in the trace, a yellow-orange patch may appear immediately,
or it may take a few minutes to become visible.
13.Repeat the procedure for each plant sample. Use fresh swabs for each
sample.
14.Get the Rf (as explained in Experiment # 16)

Data and Analysis:


Sample Observation Dragendo Distance Distance of the Retention
(Preparation) rffs from the Travelled by Factor
Reagent origin to the Solvent (Rf)
(+/-) the spot
Guide Questions:
If the Rf value of a questioned specimen is the same as that of one of your
known specimens, can you identify the questioned alkaloid specimen as
matching the known specimen?

REFERENCES:
Thompson R., Thompson B. 2012.Illustrated Guide to Home Forensic
Sciences. OReiley Media Inc, Sebastopol, CA
Chemistry Forensics Student Edition , Columbus, OH
Appendix A
Munsell Color Chart