1st Quarter SY 2016-2017

Preliminary Test and Solubility Test: Prospects for Forensic Application

ABSTRACT

During a forensics investigation, unknown samples are often sent to laboratories for identification. With
technological advancements at hand, most of the basic laboratory procedures are to be forgotten. In this
experiment, the researchers employed Preliminary Testing and Solubility Testing in determining the
identity of the unknown in the hopes that the published results can parallel that of the machines
identification capacity. In the Preliminary Tests, physical properties of the samples were determined such
as: a) Physical State at Room Temperature; b) Odor of the Compound; c) Color of the Compound. Under
the preliminary test, ignition tests were also employed to determine: d) the kind of flame the sample will
produce; e) presence of gas evolution; f) presence of residues of total ignition. In the Solubility Tests,
samples were exposed to different solvents; the solubility of the compound to a specific solvent elicits its
possible identity. The unknown samples that were subjected to solubility tests tested B and S respectively.
As results have shown, it is possible to use the different tests stated in this experiment for forensic
analysis.

Keywords: Preliminary Testing, Solubility Testing, Ignition Test, Solubility Class

INTRODUCTION groups added, the solubility of these substances

become less and less. Even the smallest amount of
Pathology is a branch of medical science that trace crystals left adhering on the sides of the
involves the close examination of how a disease container is enough to determine the identity of the
spreads, how is it caused, and how it behaves in said compound.
nature. While forensic pathology centers on the cause
of death to the organism especially when if the death In this experiment, the researchers were tasked to
is not caused by natural means. Forensic pathology identify the possible identity of the compound using
also involves the in depth compression of the preliminary testing and solubility testing. As an
pathological findings that employ procedures in additional objective to this experiment, we are to
morphology, biochemistry, and molecular biology. determine if preliminary and solubility testing can be
Biochemical findings during the autopsy play an used in forensics.
important part in investigating the pathophysiological
changes after death which morphological data cannot METHODOLOGY
normally distinguish. Moreover, molecular biology
2.1 Materials and Chemicals Used
plays an important role in detecting dynamic
functional changes complexed with Table 2.1.1 List of Reagents Used
pathophysiological reactions. TESTS REAGENTS
PRELIMINARY COPPER NITRATE
Since time immemorial, humans used poisonous TEST
substances to eliminate competitions, shape ACETIC ACID
civilizations, and direct progress as they see fit. With
time, the compounds used for making poison became SOLUBILITY TESTS BUTYL BROMIDE
more complicated as more functional groups and ETHANOL
SUCROSE
various geometries were incorporated to them.
BUTYRALDEHYDE
However, with larger geometries and functional ETHANOIC ACID
1st Quarter SY 2016-2017

METHYLAMINE 2.2 Preliminary Tests

NITROPHENOL
DIETHYL ETHER For this part of the experiment, Copper Sulfate and
BENZOIC ACID Acetic Acid were the reagents used as representative
PROPANONE compounds. Unknown samples 6a and 6b were also
5% HCl
used for this part of the experiment to narrow down
5% NaOH
5% NaHCO3 the identity of the unknown.
CONC H2SO4
15% HCl The experiment began by carefully observing the
PHENOLPHTHALEI physical properties of the representative compounds
N and the unknowns. All observations were noted on
their designated tables (See Tables 3.1.1, and 3.1.3).
The physical properties that were observed are: a)
All chemical reagents listed on Table 2.1.1 were all Physical State; b) Color; c) Odor;
used in their respective tests. The reagents were
prepared and sorted by the laboratory assistant prior For the ignition tests, small amount of the
to the experiment. All reagent bottles were shaken representative compounds and unknowns were placed
before introducing the reagent to the test samples to in their individual crucibles. The set-up in Figure 2.2
ensure that gathered results would not be false was copied and assembled as seen.
positives. Additionally, all the apparatus listed on
Table 2.1.2 were retrieved and returned in good
condition at the Stock Room. No damages and
additional fees were given to the students as there
were no breakages or lost items on the list.

Fig
ure 2.2 Schematic Diagram for the Ignition Test

After the set-up had been replicated, the crucible was

then exposed to the open of the Bunsen burner. The
subsequent reactions were observed and described.
Additionally, some important parameters were also
Table 2.1.2 List of Borrowed
Materials taken into consideration like: a) Flame color; b) The
BORROWED manner of how the sample would melt; c) Presence of
MATERIALS precipitates
MICRO TEST
TUBES In the case of precipitate formation, further tests were
PIPETS conducted such as: a) rehydration and acid-base
TEST TUBE
RACKS property determination using the litmus paper; b) Gas
CRUCIBL evolution by the addition of HCl; c) determination of
E the metal present by placing an ample amount of
CLAY
TRIANGLE
residue in the nichrome wire then exposing it to open
HOT flame.
PLATE
2.3 Solubility Test
1st Quarter SY 2016-2017

Figure 2.3 Cascade of Reaction Leading to the

Appropriate Solubility Class Figure 2.4 Preliminary Identity of the Compound
According to its Solubility Class
The experiment started by dissolving a pinch (solid),
or a drop (liquid), of sample to 1 milliliter of water in The solubility class was determined by referring to
a micro test tube. Afterwards, the test tube was Figure 2.3, and in order to obtain the preliminary
shaken and the solubility of the compound was identity of the sample refer to Figure 2.4
observed. As suggested by the laboratory manual, if
the sample didnt readily dissolve, wait for a while RESULTS AND DISCUSSIONS
and agitate. If there were no observable changes, then
3.1 Preliminary Test
the sample is insoluble to water.
Table 3.1.1 Physical Properties of Acetic Acid and
For test samples soluble to water, a freshly pipetted Copper Nitrate
sample was introduced to 1 milliliter of diethyl ether COMPOUNDS COPPER ACETIC ACID
(just a drop). If the sample was soluble, test the NITRATE
solution against a litmus paper to determine its pH. PHYSICAL LIQUID LIQUID
STATE
For test samples insoluble to water, a freshly pipetted COLOR BLUE COLORLESS
sample was introduced to 1 milliliter of 5% HCl. If ODOR ODORLESS VINEGAR/SOU
the sample remained in soluble, a fresh drop of the R
sample was introduced to 5% NaOH.

For test samples soluble to NaOH, a fresh of the test
sample was then introduced to 1 milliliter of 5%
NaHCO3. The sample procedure was done for those
insoluble to NaOH, however, instead to NaHCO3, the
fresh drop of the sample was introduced to
concentrated H2SO4.
2.4 Assessing the Identity of the Solubility Class
Copper Nitrate and Acetic Acid. Copper Nitrate and
Acetic Acid were observed to be in their liquid state
at room temperature which was approximately at
250C. Copper Nitrate was observed to be blue in
colour and was odourless while Acetic Acid was
observed to be colourless and had a sour/vinegar-like
smell (accounting that Acetic Acid and Vinegar are
the same compound). [All information stated were
tabulated and seen in Table 3.1.1]

Table 3.1.2 Ignition Test Results for Copper Nitrate

and Acetic Acid
COMPOUND COPPER ACETIC ACID
1st Quarter SY 2016-2017

NITRATE Unknown Samples 6a and 6b. Unknown sample 6a

IGNITION FLAME FLAME was observed to be in its liquid state while unknown
TEST COLOR: COLOR: sample 6b was observed to be in its solid state at
BLUE GREEN PURPLE (NO room temperature which was approximately at 250C.
COLOR Unknown samples 6a and 6b were both observed to
DARK GREEN CHANGE
be odourless, however, unknown sample 6a was
RESIDUE OCCURRED)
WAS observed to colourless while unknown sample 6b was
FORMED; NO RESIDUE observed to be white in colour. [All information
ACIDIC WAS FOUND; stated were tabulated and seen in Table 3.1.3]
WHITE
VAPOR Table 3.1.4 Ignition Test Results for Unknown
EVOLUTION Samples 6a and 6b
WAS COMPOUND UNKNOWN UNKNOWN 6b
OBSERVED 6a
IGNITION FLAME SLOW MELTING
TEST COLOUR: (CARAMELIZING
During the ignition test for Copper Nitrate, a blue GREEN )
green flame was observed and dark green residues FLAME
were formed. The observed blue-green flame was COLORLESS ->
accounted to the presence of copper in the sample. WHITE BRIGHT YELLOW
RESIDUE -> CARAMEL
The residue was hydrated, by adding a drop water, to
WAS COLOUR
test the acid-base property of the residue using a blue FORMED;
litmus paper; the Copper Nitrate residue tested
positive for the acidity test because the blue litmus NEUTRAL
paper turned red after placing it to the hydrated pH
residue. For the ignition test of the Acetic Acid, no
colour change was observed in the flame of the
Bunsen burner. The absence of any colour change in For the ignition test of Unknown Sample 6a, a green
the flame signified the absence of any metal in the flame was observed and white residues were formed.
Acetic Acid. There were no residue formation during The observed green flame signified the presence of
the ignition test for Acetic Acid but formations of barium in the sample. The formed residue tested
white vapour were visible during and after the total neutral for both blue and red litmus paper.
ignition of Acetic Acid. [All information stated were Additionally, during and after the total ignition of the
tabulated and seen in Table 3.1.2] sample, a rattling sound was emitted by the sample.
For the ignition test of Unknown Sample 6b, no
colour change to the flame was observed. The
absence of colour change signified that the absence
of any metal in the structure of the sample. Unknown
sample 6b was observed to caramelized; a slow
Table 3.1.3 Physical Properties of Unknowns 6a and transition from colourless to a caramel-colour. As the
6b
unknown sample melted, a caramelizing smell
COMPOUNDS UNKNOWN UNKNOWN 6b
6a became abundant. In this early stage, the identity of
PHYSICAL LIQUID SOLID the sample was narrowed down to a sugar compound.
STATE [All information stated were tabulated and seen in
COLOR COLOURLES WHITE Table 3.1.4]
S
ODOR ODORLESS ODORLESS 3.2 Solubility Test
1st Quarter SY 2016-2017

There are three independent rules that govern the

solubility of organic compounds namely: a) Organic
molecules with a smaller surface area are more
soluble in water than those with larger organic
molecules; b) Organic molecules that are polar and
capable of hydrogen bonding are more soluble than
organic molecules that are non-polar; c) Molecules
that assume their ionic configuration are more soluble
in water than their neutral configuration; d)
Molecules with more electronegative atoms are more
stable and acidic compared to molecules with less
electronegative atoms.

Table 3.2.1 Water and Ether Soluble at Neutral pH

COMPOUNDS ETHANO PROPANONE
L
WATER Figu
ETHER re 3.2.1 Pathway for Compounds Positive for SN
HCl
NaOH Table 3.2.2 Water and Ether Soluble at Acid and
NaHCO3 Base Ph
H2SO4 COMPOUNDS ETHANOI METHYL
REACTION TO NEUTRAL NEUTRAL C ACID AMINE
LITMUS
WATER
REACTION TO No reaction No reaction
PHENOLPHTHALEIN
ETHER
SOLUBILITY CLASS SN SN HCl
NaOH
signifies that solute dissolves on the said solvent
NaHCO3
Water-Ether Soluble Compounds at Neutral pH. H2SO4
REACTION TO ACID BASE
Water can readily dissolve small organic compounds LITMUS
such as alcohols, ketones, amines, carboxylic acids, REACTION TO Colourless Pink (base)
and a few phenols. In the experiment, ethanol and PHENOLPHTHALEIN (acid)
propanone are both water soluble compounds. Their SOLUBILITY CLASS SA SB
dissolution to diethyl ether was attributed to the signifies that solute dissolves on the said solvent
principal rule of like dissolves like wherein polar
Water-Ether Soluble Compounds at Acid and Base
solvents dissolve polar solutes. When tested against
pH. The principles used for Water-Ether Soluble
blue and red litmus paper, both molecules tested
Compounds at Neutral pH govern this solubility test
neutral; hence, the solubility class for both ethanol
as well. The main difference is that Ethanoic Acid
and propanone is SN (Seen in Figure 3.2.1). Some of
tested acidic for both litmus and phenolphthalein
the possible compounds under SN are mono
while Methyl Amine tested basic for both pH tests;
functional alcohols and ketones which are the nature
hence, the solubility class for Ethanoic Acid is S A and
of ethanol and propanone respectively.
for Methyl Amine SB (Seen in Figure 3.2.2). Ethanoic
Acid represents the carboxylic acid family, under S A,
while Methyl Amine represents the aliphatic amine
family, under SB.
1st Quarter SY 2016-2017

Figu
re 3.2.2 Pathway for Compounds Positive for S A and Figure 3.2.3 Pathway for Compounds Positive for S
SB
Table 3.2.4 NaHCO3 Soluble
Table 3.2.3 Water Soluble, Ether COMPOUNDS BENZOIC
Insoluble ACID
COMPOUNDS SUCROSE WATER X
WATER ETHER
ETHER X HCl X
HCl NaOH
NaOH NaHCO3
NaHCO3 H2SO4
REACTION TO No reaction
H2SO4
LITMUS
REACTION TO NEUTRAL
REACTION TO No reaction
LITMUS
PHENOLPHTHALEIN
REACTION TO NO
SOLUBILITY CLASS A1
PHENOLPHTHALEIN REACTION
signifies that solute dissolves on the said solvent; X
SOLUBILITY CLASS S
signifies that solute did not dissolve on the said solvent.
signifies that solute dissolves on the said solvent; X
signifies that solute did not dissolve on the said solvent.
NaHCO3 Soluble Compounds. Acids that readily
Water-Soluble, Ether Insoluble Compound. As dissolve in sodium hydroxide and sodium
stated earlier, water can readily dissolve small, bicarbonate are classified as strong acids. Being
organic compounds. Sucrose is a polar molecule, and soluble with both sodium hydroxide and sodium
as a principal rule sucrose will readily dissolve in bicarbonate, Benzoic Acid is classified as a strong
water. However, even if sucrose is a polar molecule, acid; hence, belonging to solubility class A 1. A
it cannot readily dissolve in diethyl ether due to the sample of the compounds under this class includes
large size. Since sucrose is insoluble in ether, sucrose carboxylic acids that contain more than 6 carbons in
belongs to solubility class S (Seen in Figure 3.2.3). A its chain (Seen in Figure 3.2.4).
possible identity of a molecule belonging to this class
is sugars which sucrose is.
1st Quarter SY 2016-2017

Figure 3.2.4 Pathway for Compounds Positive for A1 Figure 3.2.5 Pathway for Compounds Positive for N

Table 3.2.5 H2SO4 Soluble Table 3.2.6 H2SO4 Insoluble

COMPOUNDS BUTYRALDEHYDE DIETHYL COMPOUNDS BUTYL BROMIDE NITROPHENOL
ETHER
WATER X X WATER X X
ETHER ETHER
HCl X X HCl X X
NaOH X X NaOH X X
NaHCO3 X X NaHCO3 X X
H2SO4 H2SO4 X X
REACTION TO NEUTRAL NEUTRAL
REACTION TO ACIDIC NEUTRAL LITMUS
LITMUS REACTION TO
REACTION TO COLOURLESS NO NO REACTION NO
PHENOLPHTHALEIN
PHENOLPHTHALEIN
REACTION REACTION
SOLUBILITY X X
SOLUBILITY N N CLASS
CLASS

H2SO4 Soluble Compounds. Organic molecules that
readily dissolve in concentrated sulphuric acid
contain the following functional groups: a) alkene
functional group; b) oxygen; c) nitrogen.
Buteraldehyde and diethyl ether are both soluble in
concentrated sulphuric acid, thereby, classifying both
buteraldehyde and diethyl ether under solubility class
N (Seen in Figure 3.2.5)
H2SO4 Insoluble Compounds. Organic compounds
that does not dissolve in concentrated sulphuric acid
belongs to the last solubility class, X. Compounds
such haloalkane and deactivated aromatic compounds
like butyl bromide and nitrophenol, respectively.
1st Quarter SY 2016-2017

Table 3.2.7 Unknown Samples For the solubility test, very much all the known
COMPOUNDS UNKNOWN 6a UNKNOWN 6b samples were classified as they were with sucrose as
WATER X the only exception. For sucrose, the factors that
ETHER affected its solubility are: a) its polarity on the
HCl hydroxide; Sucrose was soluble for both water and
NaOH
ether, a description for Class S. Class S includes
NaHCO3
H2SO4 carbohydrates as one of its representative families.
REACTION TO NEUTRAL NEUTRAL
LITMUS For the unknown samples, unknown sample 6a was
REACTION TO CLOUDY NO found to have a Barium metal latched on its structure
PHENOLPHTHALEIN
COLOUR but none for unknown sample 6b. Unknown sample
CHANGE 6a produced precipitates while for unknown 6b it
SOLUBILITY B S
CLASS caramelized. The caramelization signifies that sample
6b is a heavy sugar. It was assumed to be a bulky
compound because it did not dissolve in ether.
Unknown Samples 6a and 6b. Unknown Sample 6a
was found to be insoluble in water but soluble in REFERENCES
hydrochloric acid. It tested neutral for litmus paper
test but produced a cloudy mixture upon the addition 1. [ CITATION Mae10 \l 13321 ]
of phenolphthalein. The results from the solubility
2. Signature Lab Series ANAL 0727 and
tests signified that Unknown Sample 6a belonged to 0728. Cengage Learning.
the solubility class B. Compounds that belong to this 3. De Castro, K. A., and Baluyut, J. Y. (n.d.)
class include aliphatic amines, ethers, and anilines Organic Chemistry Laboratory Manual For
with one phenyl group that is attached to nitrogen. Chemistry Students Part 2
For Unknown Sample 6b, it was found soluble to
both water and ether. Since it was soluble in ether, the
solubility class for this compound should be SN, SA,
or SB; however, it was declared as S because of the
manner it melted, caramelization. Furthermore, it
paralleled the results for sucrose signifying that the
compound is a simple sugar which is represented by
class S.

CONCLUSION

The experiment simulated the importance of the

Preliminary Test and Solubility Test as a precursor
step in qualifying and determining the identity of the
unknown sample.

For the preliminary test, copper nitrate produced a

signature flame profile which is not present on Acetic
Acid and either of the researchers unknown samples.
The signature flame was produced by the copper
metal present in the structure of copper nitrate. For
acetic acid, a total ignition was observed in less than
a minute without any formation of residues.