Kimberly Ann A.
Vipinosa
Expt 1: Ethanol/Ethyl Alcohol/Grain
Alcohol/Wine, Neutral Or Rectified
Spirit
Ethanol properties
 Molecular formula c2h6o
 Molar mass 46.07 g·mol −1
 Density 0.789 g/cm 3 (at 25°c)
 Melting point −114 °c (−173 °f; 159
k)
 Boiling point 78.37 °c (173.07 °f;
351.52 k)
 VD: 0.5-0.7l
 Lethal dose: 300-400 pure ethanol;
1 ½ pint of whiskey
 Use: wine,disinfectant
 Effect: cns depression
 Antidote: fomepizole, disulfiram
Tests
 Berthelot’s test sx -> + few
drops benzyl cl -> + excess
10% naoh (+) aromatic odor
 Chromic acid test sx -> 2 drops
k dichromate -> + 4 drops
h2so4 (+) green solution with
chico odor
 Ethyl acetate test sx-> + 3ml
conc. H2so4 -> + few na
acetate crystals-> warm (+)
plastic balloon odor
 Lieben’s iodoform test sx-> 1 ml
lugol’s solution, then warm -> +
koh solution-> cool (+) yellowish
white precipitate
 vitali’s test sx in evap dish -> +
3 drops cs2, evaporate-> + koh
pellet (+) red solution
Clinical Toxicology Laboratory
Expt 2: methanol/ methyl alcohol/
denatured alcohol/ wood alcohol
Methanol properties
• Molecular formula ch4o
• Molar mass 32.04 g·mol −1
• Appearance colorless liquid
• Density 0.7918 g·cm −3 0.7925
g·cm −3 @20°c [2]
• Melting point −97.6 °c (−143.7
°f; 175.6 k)
• Boiling point 64.7 °c (148.5 °f;
337.8 k)
• vd: 0.6-0.77 l/kg
• fatal dose: 20-240 ml
• use: denaturant, anti freeze,
formalin synthesis
• effect: blindness
• antidote: fomepizole, ethanol
• mechanism of toxicity: alcohols
are metabolised via alcohol
dehydrogenase to their toxic
metabolites. Methanol is slowly
metabolised to formaldehyde
which is rapidly metabolised to
formate, the metabolite mainly
responsible for methanol
toxicity. Formate metabolism
depends upon the folate pool
which is small in primates
compared with other animals.
Therefore, formate accumulates
in primates during methanol
intoxication and is mainly
responsible for the metabolic
acidosis in the early stage of
intoxication. In late stages
lactate may also accumulate,
mainly due to formate inhibition
of the respiratory chain.
Tests
a. Oxidation tests
• glowing copper wire -> sx
in porcelain dish ->
plunge glowing cu spiral
Kimberly Ann A. Vipinosa
into solution -> heat (+)
bluish green with
formalin odor
• with kmno4 sx -> + 10
drops conc. H2so4 (+)
decolorized
• with k2cr2o7 sx-> +2
drops k2cr2o7 -> + 10
drops h2so4 (+) green
b. Phloroglucinol test mix 2 ml
0.1% phloroglucinol in 1 ml koh
solution -> + sx (+) pink to red
c. Rimini’s phenylhydrazine test
sx-> +10 drops of
penylhydrazine hcl -> + 2 drops
0.5% na nitroprusside -> +10
drops of 20%/6n naoh (+) violet
Expt 3: acetone/ b-keto propane/ 2-
propanone/ dimethylketone
Acetone properties
• Molecular formula c3h6o
• molar mass 58.08 g·mol −1
• Appearance colorless liquid odor
pungent, irritating, floral
• Density 0.791 g cm −3
• Melting point −95 °c; −139 °f; 178
k • effect: metabolic acidosis, eye
• Boiling point 56 °c; 133 °f; 329 k
• lethal dose: 50 ml
• uses: solvent, nail polish remover
• effect: cns depression
• no antidote ( neutralization with
milk or water)
• mot: 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.
Tests:
 legal’s test sx-> 5 drops of 0.5%
na nitroprusside -> +6n naoh
solution -> observe color
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change -> acidify with acetic
acid (+) red to orange; when
heated: green
 lieben’s iodoform test sx-> +1
ml lugol’s solution -> warm
gently -> + koh (+) yellowish
white precipitate
 reynold’s test sx-> +2 ml
mercuric cl-> +1 ml of alcoholic
koh -> shake and filter-> 5
drops nh4s to clear filtrate ->
observe (don’t shake) (+) black
zone
Expt 4: formaldehyde/ formalin/
formol/ methanal/ methyl aldehyde
Formaldehyde properties
• Molecular formula ch2o
• Molar mass 30.03 g·mol −1
• Appearance colorless gas
• Density 0.8153 g/cm³ (−20 °c)
• Melting point −92 °c (−134 °f;
181 k)
• Boiling point −19 °c (−2 °f; 254
k)
• lethal dose: 30 ml
• use: embalming agent
irritation
• treatment: folic acid, nahco3
• mot: the exact mechanism of
action of formaldehyde toxicity
is not clear, but it is known that
it can interact with molecules on
cell membranes and in body
tissues and fluids (e.g., proteins
and dna) and disrupt cellular
functions. High concentrations
cause precipitation of proteins,
which results in cell death.
Absorption from the respiratory
tract is very rapid; absorption
from the gastrointestinal tract is
also rapid, but may be delayed
Kimberly Ann A. Vipinosa
by ingestion with food. Once
absorbed, formaldehyde is
metabolized to formic acid,
which may cause acid-base
imbalance and a number of
other systemic effects.
Tests:
a. General aldehyde rxn
 silver nitrate test sx-> + few
drops ammoniacal agno3->
heat to boiling (+) silver mirror
 fehling’s test sx -> + 5 drops
fehling’s reagent (+) brick red
precipitate
 nessler’s test sx-> + few drops
nessler’s reagent-> heat to
boiling (+) gray precipitate
b. Hehner’s test sx-> +2 ml fresh
unboiled milk & 7 ml 25% hcl->
+ few drops fecl3-> boil gently
(+) purple ring at junction
c. Hexamethylene tetraamine test
sx-> + nh3-> evaporate slowly
(+) white precipitate
d. Phloroglucinol test sx-> +
unequal qtty of 15% hcl ->
sprinkle a pinch of
phloroglucinol (+) red
e. Resorcinol test sx-> + mixture
of equal vol. Of 5% resorcinol
&40% naoh (+) red
f. Rimini’s phenylhydrazine test
sx-> +10 drops of 0.5%
phenylhydrazine hcl -> +2 drops
of 5% na nitroprusside )-> +10
drops 10% naoh (+) deep blue
Clinical Toxicology Laboratory
Expt 5: phenol/ carbolic acid/
hydroxybenzene
Phenol properties
• Molecular formula c6h6o
• Molar mass 94.11 g·mol −1
• Appearance transparent crystalline
solid odor sweet and tarry
• Density 1.07 g/cm 3
• Melting point 40.5 °c (104.9 °f;
313.6 k)
• Boiling point 181.7 °c (359.1 °f;
454.8 k)
• lethal dose: 2g
• use: drug precursor, disinfectant
(protoplasmic poison)
• effect: burning sensation
• tx: gastric lavage and activated
charcoal
• mot: cellular uptake of phenol is
due to its lipophilic character. It
denatures proteins. Phenol is
known to disrupt disulphide
bridges inp keratin in the skin
Tests
 bromine water test sx-> +
excess strong br water (+)
yellowish white precipitate
 ferric cl test sx-> + very dilute
fecl3 (drop by drop) -> + diluted
hcl or h2so4 (+) blue violet
 hypochlorite test sx-> + few
drops nh4oh -> + 2-4 drops
freshly prep. Na/ca hypochlorite
(+) blue
 melzer’s benzaldehyde test sx->
+2 ml conc. H2so4 -> heat for
2mins. -> +10 ml water in
cooled solution-> + koh (+)
violet-blue
 millon’s test (plugge’s rxn) sx->
+ few drops millon’s reagent->
heat (+) red
Kimberly Ann A. Vipinosa Clinical Toxicology Laboratory

Expt 6: chloroform/ Tests

trichloromethane/ methyl trichloride  isocyanide test sx-> + 1-2 drops
Chloroform properties aniline & few drops alcoholic/aq
• Molecular formula chcl3 koh-> warm for 2 mins. (+) very
• Molar mass 119.38 g·mol −1 penetrating repulsive odor
• Appearance colorless liquid odor  shwartz’s resorcinol test dissolve
heavy, ethereal odor 0.1g resorcinol in 2ml water -> +
• Density 1.564 g/cm 3 (-20 °c) few drops naoh-> + sx -> heat to
1.489 g/cm 3 (25 °c) 1.394 g/cm 3 boiling (+) yellowish
(60 °c) redprecipitate
• Melting point −63.5 °c (−82.3 °f;  lustgarten’s naphthol test sx-> +
209.7 k) few drops a-naphthol solution (+)
• Boiling point 61.15 °c (142.07 °f; blue to green to brown
334.30 k) decomposes at 450 °c  fujiwara’s pyridone test mix 2 ml
• fatal dose: 10 ml of pyridine with 3 ml 10% naoh
• use: freon solution-> heat to boiling -> + sx
• effect: hepatic and renal damage, (+) red/purple
cns depression, arrhythmia reduction test
• tx: n-acetylcysteine a. Fehling’s test sx-> + few drops
• mot: chloroform causes fehling’s reagent (+) red
progressive depression of the precipitate
central nervous system, ultimately b. Tollen’s test excess nh4oh
producing deep coma and solution to agno3-> sx -> heat
respiratory centre depression. It is to boiling (+) silver mirror
also hepatotoxic and nephrotoxic,
although liver and kidney damage
can be influenced by various
treatments which affect hepatic
drug- metabolizing enzymes;
induction of hepatic enzymes with
barbiturates, ddt or ethanol
potentiates hepatic cell necrosis
and kidney damage (niosh,
1979b). The reactive intermediates
of chloroform metabolism
(phosgene, carbene and cl) which
bind covalently and irreversibly to
cellular macromolecules are
believed to account for cellular
damage within the liver and
kidney.
Kimberly Ann A. Vipinosa
Expt 7: carbon tetrachloride/
etrachloromethane/ pyrine/ carbona
Carbon tetrachloride properties
• Molecular formula ccl4
• Molar mass 153.82 g·mol −1
• Appearance colourless liquid odor
ether-like odor
• Density 1.5867 g cm −3 (liquid)
1.831 g cm −3 at −186 °c (solid)
1.809 g cm −3 at −80 °c (solid)
• Melting point −22.92 °c (−9.26 °f;
250.23 k)
• Boiling point 76.72 °c (170.10 °f;
349.87 k)
• lethal dose: 3-5 ml
• use: propellant, antifreeze
• effect: lung, liver, heart and kidney
toxicity
• tx: n-acetylcysteine
• mot: ccl4 is activated by
cytochrome (cyp)2e1, cyp2b1 or
cyp2b2, and possibly cyp3a, to
form the trichloromethyl radical,
ccl3*. This radical can bind to
cellular molecules (nucleic acid,
protein, lipid), impairing crucial
cellular processes such as lipid
metabolism, with the potential
outcome of fatty degeneration
(steatosis)
Tests:
 fujiwara’s pyridine test mix 2 ml
pyridine with 3ml 10% naoh
solution-> heat to boiling-> + sx
(+) red
 isonitrile rxn sx-> + 2 drops
aniline & few drops of
alcoholic/aq koh-> warm for 2
mins (+) foul odor
 schwartz’s resorcinol test
dissolve 0.1g resorcinol in 2 ml
water-> + few drops naoh
solution-> +sx (+) red
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 lustgarten’s naphthol test sx-> +
few drops a-naphthol solution
(+) brown
Expt 8: carbon disulfide
Carbon disulfide properties
• Molecular formula cs2 molar
mass 76.14 g·mol −1
• Appearance colorless liquid
impure: light-yellow odor
chloroform (pure) foul
(commercial)
• Density 1.539 g/cm 3 (-186°c)
1.2927 g/cm 3 (0 °c) 1.266 g/cm
3 (25 °c)
• Melting point −111.61 °c
(−168.90 °f; 161.54 k)
• Boiling point 46.24 °c (115.23 °f;
319.39 k)
• lethal dose: 0.1mg/ml
• use: metal rust remover,
fumigant, pesticide
• effect: cns depression, peripheral
neuropathy, hemolysis and
methemoglobinemia
• tx: methylene blue
• mot: the mechanisms of carbon
disulfide toxicity can be divided
into two categories;
nonmicrosomal and microsomal.
The nonmicrosomal pathway
involves nonenzymatic
spontaneous reaction of carbon
disulfide with amino or thiol
groups that leads to formation of
dithiocarbamates or gsh
conjugates as well as inhibition of
certain enzymes such as
dopamine beta-hydroxylase.
These reactions primarily lead to
neurotoxic effects. The second
mechanism of carbon disulfide
toxicity involves its metabolism by
hepatic microsomal enzymes to
Kimberly Ann A. Vipinosa
two reactive sulfur atoms that
bind covalently to cell
macromolecules causing
hepatotoxicity. This oxidative
metabolism of carbon disulfide
has been suggested to be
responsible for much of the liver
pathology and impairment of liver
metabolism of other endogenous
substrates as well as exogenous
compounds entering the body.
Tests
 lead acetate test sx-> few drops
lead acetate solution -> observe-
> + koh & heat to boiling (+) black
ppt (with hcl: white precipitate)
 sulfocyanate test sx-> 5 drops
nh4oh -> +5 drops ethanol ->
heat to concentrate its volume to
1 ml -> acidify with dil. Hcl -> +
few drops fecl3 (+) deep red color
 xanthogenate test sx-> shake
with 3 ml sat. Koh in absolute
alcohol -> acidify mixture with2
ml diluted acetic acid and 2 drops
copper sulfate solution (+) yellow
precipitate
Clinical Toxicology Laboratory
Expt 9: aniline/
phenylamine/aminobenzene/blue oil
Aniline properties
• Molecular formula c6h5nh2
• Molar mass 93.13 g/mol
• Appearance colorless to yellow
liquid density 1.0217 g/ml, liquid
• Melting point −6.3 °c (20.7 °f;
266.8 k)
• Boiling point 184.13 °c (363.43 °f;
457.28 k)
• lethal dose: 15-30g
• use: drug precursor, explosive,
herbicide, synthetic dyes
• effect: methemoglobinemia,
convulsions
• tx. Methylene blue
• mot: it is proposed that
compound-derived toxicity to
erythrocytes results in
scavenging of damaged red
blood cells by the spleen,
initiating a series of events which
may contribute to the
development of spleen tumours
Tests
 hypochlorite test sx-> + drop by
drop aq na/ca hypochlorite-> + aq
phenol solution containing few
drops nh3 (+) blue
 phenyl isocyanide test sx-> heat
with few drops of chloroform and
koh (+) repulsive odor
 bromine water test sx-> + few drops
of strong bromine solution-> shake
(+) flesh precipitate
 chromic acid test rub in porcelain
dish a trace of pure, absolute
aniline-> + 4-5 drops of conc. H2so4
& few drops of aq k dichromate (+)
deep blue
Kimberly Ann A. Vipinosa
Expt 10: picric acid/ 4,4,6-
trinitrophenol/ phenoltrinitrate
Pocric acid properties
• Molecular formula c6h3n3o7
• Molar mass 229.10 g·mol −1
• Appearance colorless to yellow
solid
• Density 1.763 g·cm −3 , solid
• Melting point 122.5 °c (252.5 °f;
395.6 k)
• Boiling point > 300 °c (572 °f; 573
k) explodes
• fatal dose: 1-3g po
• use: explosive, dyes, fixatives
(forensic)
• effect: sensitization dermatitis,
yellow skin
• tx: methylene blue, activated
charcoal
Tests
a. Acidify sx with hcl -> extract 5
ml ether-> place drop by drop of
ether in a piece of linen-> wash
linen & moistened w/ little
ammonium sulfide (+) yellow
stain
b. Sx-> + lead acetate & h2so4 ->
extract with 5 ml chloroform->
mix with 1 ml chloroform &
enough nh4oh to form 2nd
layer-> + 0.5 ml of solution of 2g
ferrous sulfate & 10g tartaric
acid in 100ml dw (+) yellow
upper layer & red lower layer
c. Sx-> + few drops methylene
blue (+) green solution with
black residue
Clinical Toxicology Laboratory
Expt 11: naphthalene/ moth ball/
albocarbon/ tar camphor
Naphthalene properties
• Molecular formula c10h8
• Molar mass 128.17 g·mol −1
• Appearance white solid crystals/
flakes
• Odor strong odor of coal tar
• Density 1.145 g/cm 3 (15.5 °c) [1]
1.0253 g/cm 3 (20 °c) [2] 0.9625
g/cm 3 (100 °c)
• Melting point 78.2 °c (172.8 °f;
351.3 k) 80.26 °c (176.47 °f;
353.41 k) at 760 mmhg
• Boiling point 217.97 °c (424.35 °f;
491.12 k) at 760 mmhg
• lethal dose: 1-2g (4-8 moth balls)
• use: moth repellant, toilet
deodorizer
• effect: cns stimulation, hemolysis
& methemoglobinemia
• tx: methylene blue, activated
charcoal
Tests
a. Sx-> +h2so4 (+) clear/colorless
b. Sx-> + few drops chlorinated
lime & few drops conc. Hcl->
observe-> + few drops 1%
solution of resorcinol & few
drops nh3-> observe-> + few
drops nitric acid (+) blood red
bottom
Kimberly Ann A. Vipinosa
Expt 12: aspirin/acetylsalicylic acid
(asa)/ 2-acetoxybenzoic acid
Aspirin chemical data
• Formula c9h8o4
• Molecular mass 180.157 g/mol
• Physical data
• Density 1.40 g/cm 3
• Melting point 136 °c (277 °f)
• Boiling point 140 °c (284 °f)
(decomposes)
• Solubility in water 3 mg/ml (20
°c)
• acute ingestion
• mild intoxication: 150-200
mg/kg
• severe intoxication: 300-500
mg/kg
• chronic intoxication:
100mg/kg/day
• use: antiplatelet, analgesic,
antipyretic, anti-inflammatory
• effect: salicylism (tinnitus,
decrease hearing, vertigo,
vomiting)
• tx: iv na bicarbonate, iv
crystalloid solution
• mot
o interferes with aerobic
metabolism by means of
uncoupling of mitochondrial
oxidative phosphorylation.
o interruption of a series of
enzyme‐mediated
mitochondrial functions and
increased anaerobic
metabolism with cellular
conversion of pyruvate to
lactate and rapid
development of lactic
acidosis
o the inefficiency of anaerobic
metabolism results in less
energy being used to create
atp and release of the energy
Clinical Toxicology Laboratory
created during the
metabolism of glucose in the
electron transport chain as
heat, so salicylate poisoned
patients may become febrile
Tests
 Heat 0.5g aspirin -> +2 ml naoh
-> cool in ice water-> + 5 drops
h2so4-> divide into 2 portions
crystallized portion -> + fecl3 (+)
violet (salycylic acid) aqueous
portion -> + few drops alcohol &
5 drops conc. H2so4 (+) white
(acetic cid)
Expt 13: salycylic acid
Salycylic acid properties
• Molecular formula c7h6o3
• Molar mass 138.12 g·mol −1
• Appearance colorless to white
crystals
• Odor odorless
• Density 1.443 g/cm 3 (20 °c)
• Melting point 158.6 °c (317.5 °f;
431.8 k)
• Boiling point 200 °c (392 °f; 473
k)
• Decomposes 211 °c (412 °f; 484
k) at 20 mmhg
• use: keratolytic
• effect: burning of skin, salicylism
Tests
a. Sx-> +0.6 ml 30% crystalline na
carbonate solution-> + 1ml 1n
iodine solution-> heat (30
mins.)-> remove excess iodine
with little thiosulfate solution (+)
violet
b. Sx-> +0.5 ml h2so4 -> + 3 ml
ether -> ether layer: shake with
dw containing 1 dropof neutral
fecl3 solution (+) light violet at
lower layer
Kimberly Ann A. Vipinosa Clinical Toxicology Laboratory

Expt 14: acetanilide

Acetanilide properties
• Molecular formula c8h9no
• Molar mass 135.16 g·mol −1
• Odor odorless
• Density 1.219 g/cm 3
• Melting point 114.3 °c (237.7 °f;
387.4 k)
• Boiling point 304 °c (579 °f; 577
k)
• use: h2o2 decomposition
inhibitor, for synthesis of camphor
• effects: methemoglobinemia
• tx: methylene blue
Tests
 indosalycylic rxn. Sx-> heat with
25% of its vol of hcl ->
neutralize with chalk-> extract
with ether and evaporate-> take
residue with 25% hcl-> +5 drops
3% aq salycylic acid-> +5 drops
hydrogen peroxide-> observe->
small amount-> + small amount
of nh3 (+) bluish purple