Toxicology

Laboratory
Merry Cris B. Mallari, RPh
 Jens Martensson
Environmental
Toxicants
2
Air Pollution
Vapors, Mist, Smoke

• Environmental quality
• Wildlife

Jens Martensson
• Public health

Smog > most chemically complex

dimension of air pollution.
> mixture of smoke and fog
> eye-irritating photochemical
reaction products of auto exhaust

3
Other Oxidants
Peroxyacetyl nitrate (PAN)

 responsible for much of the eye-stinging

activity of smog

Jens Martensson
 it is more soluble and reactive than O3,
and hence rapidly decomposes in mucous
membranes before it can penetrate into
the respiratory tract.
 the cornea is a sensitive target and is
prominent in the burning/stinging
discomfort often associated with oxidant
smog’s.

4
Formaldehyde
Aka. Formalin, Formol
 Primary sensory irritant
 Because it is very soluble in water, it is absorbed in
mucous membranes in the nose, upper respiratory
tract, and eyes.
 The dose–response curve for formaldehyde is steep:
 0.5–1 ppm yields a detectable odor
 2–3 ppm produces mild irritation
 4–5 ppm is intolerable to most people.
 Formaldehyde is thought to act via sensory C-fibers
that signal locally as well as through the trigeminal
nerve to reflexively induce broncheoconstriction
Jens Martensson
through the vagus nerve.
 Respiratory frequency and minute volume also
decreased, but these changes were not statistically
significant until >10 ppm. The no observed effect level
(NOEL) using these lung function criteria is about
0.05 ppm.
5
Formaldehyde
 The general pattern of the irritant response and its rapid recovery is similar to that produced
by higher concentrations of SO2.

 Like SO2, breathing through a tracheal cannula to bypass nasal scrubbing greatly augments the

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irritant response, indicating that deep lung irritant receptors can also be activated by this
vapor.

 The irritancy of inhaled formaldehyde vapor, again like SO2, has been shown to be potentiated
by water-soluble salt aerosols.

 Irritancy appears to be augmented in proportion to the aerosol concentration, but the

potentiation could not be accounted for by a simple aerosol “carrier” effect

6
Formaldehyde
Two aspects of formaldehyde toxicology
 One is its near ubiquitous
presence in indoor
atmospheres as an off-gassed
product of construction
materials such as plywood,
furniture, or improperly
polymerized urea-
formaldehyde foam insulation
 Complaints of formaldehyde
irritation in industry have
been reported at 50 ppb
 In studies relating household
Jens Martensson
formaldehyde to chronic effects, children
were found to have significantly lower
peak expiratory flow rates (about 22% in
homes with 60 ppb) than did unexposed
children, and asthmatic children were
affected below 50 ppb. Thus, this irritant
vapor can cause respiratory effects, and
perhaps act as an allergen, at commonly
experienced exposure levels
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Formaldehyde
 Second, a longtime concern regarding formaldehyde has been its potential carcinogenicity.

 Formaldehyde causes nasopharyngeal cancer, leukemia, sinonasal cancer and nasal cancer in

Jens Martensson
animals.

 Nasal cancer had been induced empirically with formaldehyde vapor in a 2-year study where
rats were exposed to 2, 6, or 14 ppm 6 hours per day, 5 days per week. The incidence of nasal
squamous cell carcinomas was zero in the control and 2-ppm groups, 1% in the 6-ppm group,
and 44% in the 14-ppm group.

 Exposure-related induction of squamous metaplasia occurred in the respiratory epithelium of

the anterior nasal passages in all exposed groups. A 20-fold increase in cell proliferation in the
nasal epithelium occurred after 5 days to 14 ppm. Mice were much less sensitive; only one
carcinoma was seen at 14 ppm. The detection of DNA adducts in the two species paralleled
the difference in the incidence of tumors as well as regional dosimetry.
8
Thank
You
 ACTIVITY 5:
FORMALDEHYDE
OBJECTIVES:
AT THE END OF THE ACTIVITY, THE STUDENTS ARE EXPECTED TO;

 STATE THE POSITIVE RESULTS OF THE GENERAL ALDEHYDE REACTIONS

 LIST DOWN THE HEALTH EFFECTS OF FORMALDEHYDE
 DETERMINE THE DIFFERENT OTHER ROUTES OF EXPOSURE TO
FORMALDEHYDE
 DISCUSS THE APOMORPHINE TEST, PHLOROGLUCINOL TEST, RENIN’S
PHENYLHYDRAZINE TEST, AND HEXAMETHYLENE TETERAAMINE TEST.
HANDLING FORMALDEHYDE

 Formaldehyde should always be used with adequate ventilation,

preferably in a fume hood, to minimize inhalation of vapor
formaldehyde.
 Always use chemical goggles or a face shield when handling
formaldehyde to minimize the risk of even a small splash or vapor
exposure to the corneas.
FORMALDEHYDE
SYNONYMS: FORMALIN, FORMIC ALDEHYDE, METHANAL, METHYL ALDEHYDE

PHYSICAL PROPERTIES:
 Flammable
 Colorless gas at room temperature
 Has a pungent and suffocating odor

CHEMICAL PROPERTIES:
 Molecular formula: CH2O
 Molecular weight: 30.03
 Boiling point: –19.5°C
 Melting point:–92°C
 Density: 0.8153 g/𝑐𝑚3
 Specific gravity: 1.067 with respect to air
 Solubility: Very soluble in water; soluble in alcohol and ether
USES OF FORMALDEHYDE:

 Used as a preservative in medical laboratories, mortuaries, and consumer products

such as hair smoothening and straightening agents.
 Embalming agent.
 It is used in the industrial field in the construction of plywood, chipboard, insulation
materials, paint and plastic materials, textile industry, carpets, furniture, wall
coverings, and household cleaning products.
 Preservative in soaps, deodorants, shampoos, detergents, cleaning
agents, etc. against microbial contamination
 Used as disinfectant
TOXICITY:
Lethal dose: 30 mL
Toxic effect: Metabolic acidosis
Treatment: Folic acid, Sodium Bicarbonate (NaHCO3)
Mechanism of Toxicity:
Formaldehyde causes precipitation of proteins and will cause coagulation necrosis of
exposed tissue leading cell death. When inhaled, it produces immediate local irritation of
the upper respiratory tract and has been reported to cause spasm and edema of the larynx
as it is rapidly absorbed. When ingested, formaldehyde is rapidly absorbed but may be
delayed with the presence of food. Once absorbed, formaldehyde is metabolized to formic
acid by aldehyde dehydrogenase. Formic acid causes acid-base imbalance leading to
metabolic acidosis.
FORMALDEHYDE CONTAINING PRODUCTS:

 Resins used in the manufacture of composite wood products (i.e.,

hardwood plywood, particleboard and medium-
density fiberboard);
 Building materials and insulation;
 Household products such as glues, permanent press fabrics,
paints and coatings, lacquers and finishes, and paper products;
 Preservatives used in some medicines, cosmetics and other
consumer products such as dishwashing liquids and fabric
softeners; and
 Fertilizers and pesticides.
 PROCEDURE: GENERAL ALDEHYDE REACTION

• SILVER NITRATE TEST • FEHLING’S TEST

2 mL of 5 drops of
1-2 mL of Heat to boil. Fehling reagent
Formaldehyde
Formaldehyde

Positive result: Dark gray precipitate or silver

mirror at the bottom of test tube. Positive result: Reddish brown
precipitate
• NESSLER’S TEST
5 drops of
Nessler’s reagent
Positive result: Gray precipitate 2 mL of
Formaldehyde
Heat to boil.
 • SCHIFF’S TEST • RESORCINOL TEST

1 mL of 5%
resorcinol
1 mL of 40%
5 drops of NaOH sol’n
Schiff’s reagent
1 mL of Formaldehyde
2 mL of Formaldehyde Heat to
Warm the
boiling.
Solution.

Positive result: Bright Pink solution Positive result: Red coloured solution
 DATA
TEST OBSERVATION/RESULT ILLUSTRATION/PICTURES

SILVER NITRATE Dark gray precipitate or silver mirror

TEST

FEHLING’S TEST Reddish brown precipitate

NESSLER’S TEST Gray precipitate

SCHIFF’S TEST Bright pink or magenta coloured

solution

RESORCINOL TEST Red coloured Solution

1. List down the health effects of formaldehyde.

When formaldehyde is present in the air at levels exceeding 0.1

ppm, some individuals may experience effects such as watery eyes,
burning sensations in the nose, eyes, and throat as well as
coughing, wheezing, and skin irritation. Long term exposure to
formaldehyde has been shown to be associated with an increased
risk of cancer of the nose and accessory sinuses, nasopharyngeal
and oropharyngeal cancer and lung cancer in humans.
2. Aside from the Gas-phase inhalation, What are the other routes of
exposure to fomaldehyde?

Formaldehyde exposure is most common through gas-phase inhalation.

However it can also occur through liquid-phase skin absorption.
Workers maybe exposed during direct production treatment of
materials, and production of resins. Moreover, the possible routes of
exposure to formaldehyde are ingestion, inhalation, dermal absorption,
and rarely, blood exchange as in dialysis.
3. Discuss
the other laboratory test for detection of
formaldehyde including positive results.
A. APOMORPHINE TEST
Opium alkaloids, particularly morphine and apomorphine, have been used
considerably in the identification of formaldehyde. In the presence of strong acid, the
reaction is that of a phenol with an aldehyde is producing a color.
B. PHLOROGLUCINOL TEST
Add unequal quantity of 15% HCL -> Sprinkle a pinch of phloroglucinol. Positive result
would produce red in color.
C. RIMINI’S PHENYLHYDRALAZINE TEST
Add 10 drops of 0.5% phenylhydralazine HCL -> add 2 drops of 5% Na
nitroprusside -> add 10 drops 10% NaOh. Positive result would produce
deep blue in color.
D. HEXANEMETHYLENE TETRA AMINE TEST
It is used for the detection of formaldehyde. It uses ammonia for the test. It
will then be evaporated slowly to produce white precipitate as the positive
result.
CONCLUSION:

Therefore, Silver nitrate reacts with formaldehyde to produce silver mirror. With
fehling’s test, reddish brown precipitate is produced with the presence of
formaldehyde. Nessler’s reagent interacts with formaldehyde to produce gray
precipitate. Schiff’s test produces magenta coloured solution, and resorcinol test
gives off red colored complex. Formaldehyde can cause watery eyes, burning
sensations in the nose, eyes, and throat as well as coughing, wheezing, and skin
irritation. For long term exposure, it increases the risk of cancer of the nose and
accessory sinuses, nasopharyngeal and oropharyngeal cancer and lung cancer in
humans. Aside from Gas-phase inhalation, exposure to formaldehyde can also occur
through liquid-phase skin absorption, ingestion, inhalation, dermal absorption, and
rarely, blood exchange as in dialysis. Lastly, we have discuss the Apomorphine test,
phloroglucinol test, renin’s phenylhydrazine test, and hexamethylene teteraamine
test in question number 3.
PRESENTED BY GROUP 4:

ALIMPOLO
BENITEZ
BRIZO
CORO
DACUMOS
ELLORIMO
TANGA-AN
ACTIVITY 6:
ACETONE
GROUP 5:

BANGKAYLAN
CUBELO
MAWALI
SANTIAGO
 OBJECTIVES:
AT THE END OF THE ACTIVITY , THE STUDENTS ARE EXPECTED TO:
1. DETERMINE THE POSITIVE RESULT OF THESE DIFFERENT
TESTS.
2. DIFFERENTIATE LIEBEN’S IODOFORM TEST FROM LEGAL TEST.
3. EXPLAIN THE LONG-TERM HEALTH EFFECTS WHEN EXPOSED
TO ACETONE.
4. EXPLAIN WHAT IS REYNOLD’S TEST.
✖ Acetone is a clear or colorless liquid that smells like nail polish remover.
When exposed to the air, it quickly evaporates and remains highly
flammable. Acetone is dangerous to use around an open flame. It is
completely miscible in water and is soluble in benzene and ethanol.
✖ Chemical name: 2-Propanone
✖ Molecular formula: C3H6O
✖ Molar mass: 58.08 g.mol-2
✖ Melting point – (-95.4 degree Celsius)
✖ Other names - Propanone, propanone di-methyl ketone
✖ Mechanism of toxicity: Ketones contain acetone and travel through the
bloodstream until they are broken down in the liver. Acetone poisoning
can occur when an abnormally high amount of ketones is present.
✖ Toxicity rating: Acute toxicity
✖ LETHAL DOSE
✖ the minimum lethal dose for a 150-lb man is estimated to be 100ml of
acetone LD:50ml
✖ 200ml of acetone cause red and swollen throat, erosions in the soft palate
and esophagus, elevated blood glucose levels, and became comatose for
12hrs.
TREATMENT
- Induce emesis
- Neutralization by milk and water
Use:
Nail polish remover, solvent for fats
✖ Sympotoms
-respiratory depression
Hypothermia
bradychardia
 APPARATUS & CHEMICAL REAGENTS

APPARATUS/EQUIPMENTS
✖ Test tubes. Graduated cylinder, medicine droppers. Litmus
paper, hot plate. Water bath

CHEMICAL REAGENTS
✖ Acetone
✖ Lugol’s iodine
✖ Potassium hydroxide
✖ Sodium nitroprusside
✖ Acetic acid
PROCEDURE
Lieben’s iodoform test
Legal’s test
 RESULTS

Lugol’s reagent mixed with

acetone and KOH will result
into yellowish white precipitate
A reddish-yellow colour will appear
when sodium nitroprusside is added to
acetone and KOH solution. The
solution will turn into a green colour
when heated.
 ACETONE
USES
solvent in paint, inks, adhesives, thinners,
degreasers, varnishes, lacquers
-chemical intermediate in the production of
lubricating oils, chloroform and various
pharmaceuticals and pesticides.
Mechanism of toxicity
Ketones contain acetone and travel through the
bloodstream until they are broken down in the
liver. Acetone poisoning can occur when an
abnormally high amount of ketones is present.
EFFECTS
Acetone acts primarily as a depressant of the
central nervous system. Signs of toxicity are
ataxia, sedation, coma, respiratory
depression, gastrointestinal disorders,
hyperglycemia, ketonemia, acidosis, hepatic
and renal damage.
Acetone is excreted mainly through the lungs and
urine. Excretion is rapid following a single oral
dose but may not be complete in 24 hours; 40-70%
may be excreted through the lungs, 15-30% in the
urine, and 10% through the skin. Injection of 5
g/hour into humans resulted in a slow rate of
excretion.
Products containing acetone
 -furniture polish
 -rubbing alcohol
 -nail polish remover
 -paint remover
 -glue
 -cleaning agents

Laboratory measure in handling acetone

 make sure the area in which the acetone is used is well-ventilated
 -wear proper PPE (gloves, goggles, mask)
 -if you work with acetone on a surface, make sure the surface it’s a
the surface that doesn’t soak up the liquid
 -if you use acetone in your business, make sure you have got the
proper licensing in place to do so
QUESTIONS
What are the long-term health effects of exposure to
acetone?
● Breathing acetone can cause nose, throat, lung, and eye
irritation; headaches; light-headedness; confusion;
increased pulse rate; effects on blood; nausea; vomiting;
unconsciousness and possibly coma. There are no
related studies regarding the long-term health effects of
acetone on humans. However, in animal studies, Kidney,
liver, and nerve damage, increased birth defects and
lowered ability to reproduce (males only) occurred in
animals exposed long-term.
Will acetone acts synergistically with other material?

● Acetone has increased the liver toxicity of chemicals

such as carbon tetrachloride, chloroform,
trichloroethylene, bromodichloromethane,
dibromochloromethane, N-nitrosodimethylamine and
1,1,2-trichloroethane, the lung toxicity of styrene and the
toxicity of acetonitrile and 2,5-hexanedione in laboratory
animals. It also appears to inhibit the metabolism and
elimination of ethyl alcohol that increases its toxicity.
What is Reynold’s Test? Discuss the procedure and give the
positive result.

● Reynold’s test is used to detect acetone. The positive

result is black color develops when freshly prepared
mercuric oxide is added to acetone and shaken and
then ammonium sulfide is added.
 CONCLUSION
In conclusion acetone or otherwise known as propanone is considered to be the simplest
and smallest ketone. It is an organic compound with the formula (CH₃)₂CO. Acetone is
medically used as an antiseptic; it is also used in producing pills and liquid medicines in
order to modify their density. When using acetone one must be properly equipped with
the proper protective equipment, avoid physical contact with acetone and if contaminated
wash the contaminated area thoroughly. Lieben’s iodoform test is intended to show if a
solution has the presence of methyl ketones, legal’s test, on the other hand is attest to
known if a certain compound has the presence of acetone.
THANK YOU!
Name ______________________________ Points earned _________________
Course and Year ___________________ Date Performed _____________

ACETONE

OBJECTIVES:

At the end of this activity, the students are expected to

1.
2.
3.
4.

APPARATUS/EQUIPMENTS

Test tubes. Graduated cylinder, medicine droppers. Litmus paper, hot plate. Water bath

CHEMICAL REAGENTS

Acetone Lugol’s iodine Potassium hydroxide

Sodium nitroprusside Acetic acid

PROCEDURE

1. Lieben’s iodoform test

Add 1 ml of Lugol’s to 1 ml of acetone then KOH drop by drop until the color is yellow
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2. Legal’s test

Add a few drops of freshly prepared sodium nitroprusside solution to 2 ml acetone and then
KOH solution. Observe the color change. Add an excess acetic acid to acidify the solution.
Observe the result and heat the solution gently
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______________________________________________________________________________
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ILLUSTRATIONS/PICTURES

LIEBEN’S IODOFORM TEST LEGAL’S TEST

CONCLUSION:
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
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______________________________________________________________________________
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_______________________

QUESTIONS
1. What are the long-term health effects of exposure to acetone?
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
________________________________________________
2. Will acetone act synergistically with other material?
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________
3. What is Reynold’s test? Discuss the procedure and positive result
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
___________________________________________________
Name ______________________________ Points earned _________________
Course and Year ___________________ Date Performed _____________

FORMALDEHYDE
OBJECTIVES:

At the end of this activity, the students are expected to

1.
2.
3.
4.

APPARATUS/EQUIPMENT

Test tubes, medicine dropper, test tube holder, hot plate, water bath, graduated cylinder

CHEMICAL REAGENTS

Formaldehyde Silver nitrate Fehling’s reagent

Nessler’s reagent Schiff‘s reagent 5% resorcinol
40% NaOH 25% HCl

PROCEDURES

General aldehyde reaction

A. Silver nitrate test

Add 1 ml to 2 ml of formaldehyde. Heat the solution to boiling.

___________________________________________________________________________
___________________________________________________________________________

B. Fehling’s test

Heat 2 ml of formaldehyde with 5 drops of Fehling’s reagent

___________________________________________________________________________
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_________________________________________________

C. Nessler’s test

Add 5 drops of Nessler’s reagent to 2 ml of formaldehyde. Heat the solution to boiling

___________________________________________________________________________
___________________________________________________________________________
________________________________________________
 D. Schiff’s test

Add 5 drops of Schiff’s reagent to 2 ml of formaldehyde. Warm the solution.

___________________________________________________________________________
___________________________________________________________________________
______________________________________________________________________

E. Resorcinol test

To 1 ml of solution add a mixture of an equal volume of 5% resorcinol and 40% NaOH

solution. Heat to boiling.
___________________________________________________________________________
___________________________________________________________________________
______________________________________________

DATA

TEST OBSERVATION/RESULTS
Silver Nitrate testy
Fehling’s test
Nessler’s test
Schiff’s test
Resorcinol test

ILLUSTRATIONS/PICTURES

SILVER NITRATE FEHLING’S TEST NESSLER’S TEST

 SCHIFF’S TEST RESORCINOL TEST

CONCLUSION:
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______________________________________________________________________________
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______________________________________________________________________________
_______________________

QUESTIONS:

1. List down the health effects of formaldehyde

______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
________________________________________________

2. Aside from the gas phase inhalation, what are the other routes of exposure to formaldehyde?
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
________________________________________________________

3. Discuss the other laboratory test for detection of formaldehyde including positive result
A. APOMORPHINE TEST
___________________________________________________________________________
___________________________________________________________________________
_____________________________________________________
B. PHLOROGLUCINOL TEST
___________________________________________________________________________
___________________________________________________________________________
___________________________________________________________

C. RIMINI’S PHENYLHYDRALAZINE TEST

___________________________________________________________________________
___________________________________________________________________________
_______________________________________________________________

D. HEXANEMETHYLENE TETRA AMINE TEST

___________________________________________________________________________
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