Color tests

Investigations
in a case of
poisoning

Toxicological Non toxicological

screening confirmatory
 Aims for
toxicological
analysis

To estimate
To identify
its level In blood
the poison
and or urine
Laboratories Must use at least
two methods
one screening and the other
confirmatory for the positive
results.
Screening tests :

1) Color tests.
2) Spectophotometry.
3) Immunologic methods:
(RIA) Radio-Immuno Assays
(EMIT) Enzyme Multiplied Immuno assay Technique
4) Chromatography:
(TLC) Thin layer chromatography.
Confirmatory tests:

1) GC in volatiles, gases and pesticides.

2) HPLC. Analgesics ,drug of abuse and
psychotropic drugs.
3) GC/MS . This is the most sensitive and
reliable method.
4) Atomic absorption spectrophotometry
used for heavy metals
SPECTROPHOTOMETER
EMIT
HPLC
Atomic absorption spectrometry
Samples used in toxicological analysis.

I. Urine samples.
II. Blood samples.
III. Serum samples.
IV. Hair samples.
V. Other biological sample used e.g.
gastric contents, saliva and sweat.

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 Urine samples :

Preferred more than blood

samples for detection of drugs of
abuse ( opiates , barbiturates )

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 Advantages of urine sampling:
 1- Concentration of the drugs and their
metabolites are relatively high
compared with blood.
 2- Drugs may be detected for longer
periods after intake.
 3- Urine sampling is non-invasive.
 4- No sample preparation and easy
analysis.
Disadvantages of urine sampling:

 Substitution
 Dilution
 Adulteration.
 Blood samples

Requires qualified personnel for taking

the sample.
Commonly used for estimation
 Blood alcohol level
 Co Hb %
 met Hb%.
COLOR TEST
Principle of color tests

 Many drugs and poisons give

characteristic colors with appropriate
reagents if present in sufficient
concentration.

 The color produced usually varies in

intensity proportional to the
concentration.
Samples used in color tests:

 Urine sample
 Blood sample
 Gastric contents.
 Advantages

 Simple
 easy to perform (bed-side tests)
 Carried directly on the available
sample.
Disadvantages

 Screening test only

 Rapid color changes
 Low sensitivity
  1-salisalyate
 2-phenothiazine
Color test
 3-barbiturate
 4-ethylalcohol
Comment
(ethanol)
 1-color
 5-methyl alcohol
 2-poison
(methanol)
 3-reagent
 Color test for Salicylate

1ml urine + 3 drops of 5% ferric

chloride

Violet color.
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Color test for Salicylate
Color test of Phenothiazine

 1ml urine + 1ml FPN reagent

FPN reagent
5ml 5% Ferric chloride
45ml 20% Perchloric acid
50ml 50% Nitric acid
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Color test of Phenothiazine

Pink color
Color test of Barbiturate
 1mlurine + 1ml 0.2% Cobalt
acetate in methanol warm then
add small amount of powdered
borax

Bluish color with precipitations.

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 Color test of Barbiturate

Bluish color with

precipitations
Color tests of Ethyl alcohol

(Potassium Permanganate Method)

 1ml urine + 1ml potassium permanganate
0.1% + 2 drops of acid + heat

 Pink color of K permanganate disappears.

(Color fade gradually with heating)

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 KMNO4 act as oxidizing agent

Ethyl Acetal- Acetic

dehyde acid
alcohol
heat

Acid to get rid off excess KMNO4

Water +
Co2
Color tests of Ethyl alcohol

Water like
(colorless)
Color test of Methyl alcohol
Chromotropic acid test (Ring test)
1ml urine
+ 5 drops Potassium Permanganate
+ 5 drops oxalic acid
+ 5 drops chromotropic acid
+ 2 cc sulphuric acid
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KMNO4 act as oxidizing agent

Methyl Formal- Formic

alcohol dehyde acid

Oxalic acid to get rid off excess KMNO4

FORMIC acid
+ compound
Chromotropic acid

We add Few drops of sulphuric acid on inner

wall of the tube
sulphuric acid responsible for
configuration of ring
Potassium Permanganate : oxidizing agent

oxalic acid : remove excess K perminganate

chromotropic acid :conjugate with formic acid

sulphuric acid :configuration of the ring

Color test of Methyl alcohol

Violet ring
in the interface
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Thank you for your attention!
Any Questions?