Toxicology Analysis
of
Salicylic Acid and its Derivate in
Urine
Year 4, semester 1/2023
INTRODUCTION
• Salicylic acid is well‐known for its pharmacological effects:
• peripheral analgesic,
• antipyretic,
• anti‐ inflammatory,
• antiplatelet agents.
• Salicylic acid and Acetylsalicylic acid are eliminated primarily as salicyluric acid (combination with
glycocolle).
• This eliminating form maintains the main properties of the original acid, in particular ferric
complex formation.
• Toxic dose: 10 g adult; 100 mg/Kg child.
SALICYLIC ACID AND
SALICYLIC ACID
2‐Hydroxybenzoic acid;
DERIVATE ACETYLSALICYLIC ACID 4‐AMINOSALICYLIC ACID
p‐Aminosalicylic acid; PAS;
Aspirin;
4‐ amino‐2‐hydroxybenzoic
C 7H 60 3
C 9H 80 4 acid;
C7H7 N03
METHYL SALICYLATE SALICYLAMIDE
Methyl 2‐ 2‐Hydroxybenzamide;
hydroxybenzoate; salicylic C7H7N02
acid methyl ester;
C 8 H 8 03
THE PRATICE IN TOXICOLOGICAL
LABORATORY
Qualitative
Assay
OBJECTIVE and
• Objective: PRINCIPLE
•Identify the presence of Salicylic acid or its derivate via qualitative test in human sample such
as urine collected from patient suspected of taking this kind of substance.
•Case: Urine sample was taken from a coma person in his house. The doctor of
emergency unit in a hospital is asking if the sample contains salicylates.
• you need to establish the assay protocol.
• you must ensure the quality of your results.
Principle: The salicylic ions contain trivalent iron salts (ferric nitrate solution) that yield the
formation of a grey complex chelated.
Applicable to urine, stomach contents and scene residues.
PRINCIPLE
• The salicylic ions contain trivalent iron salts (ferric nitrate solution) that yield the formation of a
grey complex chelated.
Applicable to urine, stomach contents and scene residues.
MATERIALS and
REAGENTS
Materials: Reagents:
Beaker
• Preparation of ferric nitrate
Volumetric flask
solution (TRINDER’s reagent)
Test tubes
Tube rack • HCl 37%.........................................10
ml
Pipette
• ED…………………………………..………..110
Pro‐pipette ml
Dropping pipette • Ferric nitrate Fe(NO3)3.9H2O……...40
Lab spatula g
• Mercury(II) chloride
HgCl2………….40g
• ED……………………………………. for 1000
ml
IDENTIFICATION METHOD
• Add 0.1 ml of Trinder's reagent to 2 ml of sample and mix for 5 seconds.
• To test for acetylsalicylic acid or methyl salicylate in stomach contents
or scene residues, and to test for salicylamide in urine, stomach contents or
scene residues, first boil 1ml of sample with 1ml of aqueous hydrochloric
acid (0.1 mol/l) for 10 minutes, cool, filter if necessary, and then neutralize
with 1ml of aqueous sodium hydroxide (0.1 mol/l).
RESULT
A strong grey colour indicates the presence of salicylates. Azide preservatives
read strongly in this test, and weak false positives can be given by urine
specimens containing high concentrations of ketones (ketone bodies).
This test is sensitive and will detect therapeutic dosage with salicylic acid,
acetylsalicylic acid, 4‐aminosalicylic acid, methyl salicylate and salicylamide.
Sensitivity: Salicylate, 10 mg/I.
Quantitative
Assay
REAGENTS and
1. Reagents STANDARD
• Trinder's reagent (see above).
2. Standards
•Aqueous solutions containing salicylic acid at concentrations of 0, 200, 400 and 800
mg/I. Store at 4 °C when not in use.
Applicable to plasma or serum (1 ml).
METHOD and
1. Method: RESULT
I. Add 5 ml of Trinder's reagent to I ml of sample or standard.
2. Vortex‐mix for 30 seconds and centrifuge for 5 minutes.
3. Measure the absorbance of the supernatant at 540 nm against a plasma blank.
2. Result:
• Calculate the plasma salicylate concentration from the graph obtained on analysis of the
salicylate standards. Some salicylate metabolites interfere, but plasma concentrations of these
compounds are usually low.
• Oxalates, for example, from fluoride/oxalate blood tubes, also interfere in this test.
Sensitivity: Salicylate, 50 mg/I.
CLINICAL INTERPRETATION
The topical use of salicylic acid and methyl salicylate and ingestion of salicylates may give
rise to features of salicylism. Respiratory alkalosis followed by metabolic acidosis is characteristic,
although in practice a mixed acid‐base disturbance is usually seen. The results of blood gas analyses
are an important guide to the severity of poisoning (see section i. below). If acute poisoning
is suspected, the plasma salicylate concentration should be measured using the method
described above.
Active measures to correct acid‐base status and urinary alkalinization to
enhance elimination of the poison may be considered, depending on the patient's condition and
the plasma salicylate concentration. Repeated oral activated charcoal may also be employed.
CLINICAL INTERPRETATION
Serial plasma salicylate and urine pH measurements are valuable in monitoring active
treatment. A guide to the interpretation of plasma salicylate results is given in Fig.
13. Concentrations of up to 300 mg/I may be encountered during therapy in adults.
CLINICAL INTERPRETATION
•Results obtained within 6 hours of ingestion are unreliable and should be used with caution,
since absorption may not be complete.
•If acute poisoning is suspected, the plasma salicylate concentration should be measured
using the method described above.
•Active measures to correct acid‐base status and urinary alkalinization to enhance elimination
of the poison may be considered, depending on the patient's condition and the plasma
salicylate concentration. Repeated oral activated charcoal may also be employed.
•Serial plasma salicylate and urine pH measurements are valuable in monitoring
active treatment. A guide to the interpretation of plasma salicylate results is given in Fig. 13.
Concentrations of up to 300 mg/I may be encountered during therapy in adults.
The
Ends.
