Analytical Toxicology Methods Guide
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This document outlines various analytical toxicology methods used to identify toxins, including qualitative and quantitative tests. Qualitative tests described include color tests that use specific reagents to produce distinct colors with different toxins. Thin layer chromatography is also discussed as a method to separate mixtures into components using a mobile and stationary phase. Quantitative methods presented are ultraviolet spectrophotometry, gas chromatography, high pressure liquid chromatography, mass spectrometry, and immunoassays. Each method is briefly described in one or two sentences.
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Analytical Toxicology Methods Guide
- 1. ANALYTICAL TOXICOLOGY DR. SAIKAT SAHA TUTOR 1
- 2. INDEX SLIDE 3- METHODS USED IN TOXICOLOGY SLIDE4 TO 6 – COLOUR TESTS SLIDE 7 TO 10- THIN LAYER CHROMATOGRAPHY SLIDE 11- UV SPECTROMETRY SLIDE 12- GAS CHROMATOGRAPHY SLIDE 13- HPLC SLIDE 14- MASS SPECTROMETRY SLIDE 15 - IMMUNOASSAYAS 2
- 3. METHODS USED IN TOXICOLOGY Qualitativ e Test Bedside Test Color Test Other Test Thin Layer Chromato graphy QUALITATIVE TEST QUANTITATIVE TEST Quantitativetest Ultraviolet visible spectromet ry Gas chromatog raphyHigh power Liquid chromategr aphyMass Spectromet ry 3
- 4. QUALITATIVE TESTS • Trinder’s test- Trinder’s reagent + 2ml urine violet purple colour (salicylates) • Ferric chloride test- 5% ferric chloride (1ml)+ 2ml urine purple colour Phenol • FPN test- FPN reagent + 1ml urine Pink, red, blue Phenothiazines • O-Cresol test-.5 ml conc. HCl + .5 ml urine 1ml aq O-Cresol+2 ml ammonium hydroxide Blue Black colour Paracetamol • Marquis test- 3 ml conc. H2SO4 + 3ml COLOUR TEST SPECIFIC REAGENT REACTS WITH THE TOXIN PRESENT IN THE SAMPLE AND PRODUCES SPECIFIC COLOUR. 4
- 5. QUALITATIVE TESTS • Lee-Jones Test- ferrous sulfate + 2% NaHCO3 +gastric f • Reinsch test- stomach content or urine+ HCL + Cu silvery deposit over copper Black deposit • Meixner Test- diluted sample 1-2 drops at a piece of paper, add conc HCl blue colour indicates amatoxin COLOUR TEST SPECIFIC REAGENT REACTS WITH THE TOXIN PRESENT IN THE SAMPLE AND PRODUCES SPECIFIC COLOUR. Mercury Boiling for an hour in conical flask 5
- 6. QUALITATIVE TESTS • Forrest test- 1 ml Forrest reagent + .5 ml sample yellow green colour • Fujiwara test- COLOUR TEST SPECIFIC REAGENT REACTS WITH THE TOXIN PRESENT IN THE SAMPLE AND PRODUCES SPECIFIC COLOUR. 5 secs 1st tube • sample • 1ml sodium hydroxide and pyridine 2nd tube • Purified water • 1ml sodium hydroxide and pyridine 3rd tube • Aq trichloroacetic acid • 1ml sodium hydroxide and pyridine 6
- 7. QUALITATIVE TESTS THIN LAYER CHROMATOGRAPHY CHROMATOGRAPHY IS USED TO SEPARATE MIXTURES OF SUBSTANCES INTO THEIR COMPONENTS. 7
- 8. QUALITATIVE TESTS THIN LAYER CHROMATOGRAPHY • A MOBILE PHASE (DEVELOPING SOLVENT) • A STATIONARY PHASE (A PLATE OR STRIP COATED WITH A FORM OF SILICA GEL) • ANALYSIS IS PERFORMED ON A FLAT SURFACE UNDER ATMOSPHERIC PRESSURE AND ROOM TEMPERATURE 8
- 9. QUALITATIVE TESTS THIN LAYER CHROMATOGRAPHY DISTANCE TRAVELLED BY SOLUTE FRONT DIATANCE TRAVELLED BY SOLVENT FRONT 9
- 10. QUALITATIVE TESTS • PURITY OF SAMPLE • EXAMINATION OF REACTION • IDENTIFICATION OF COMPOUNDS • BIOCHEMICAL ANALYSIS • IN PHARMACEUTICAL INDUSTRY • SEPARATION OF MULTICOMPONENT PHARMACEUTICAL FORMULATIONS • IN FOOD AND COSMETIC INDUSTRY THIN LAYER CHROMATOGRAPHY APPLICATIONS 10
- 11. QUANTITATIVE ASSAYS ULTRA VIOLET VISIBLE SPECTROPHOTOMETRY • SPECTROPHOTOMETRY IS A METHOD TO MEASURE HOW MUCH A CHEMICAL SUBSTANCE ABSORBS LIGHT BY MEASURING THE INTENSITY OF LIGHT AS A BEAM OF LIGHT PASSES THROUGH SAMPLE SOLUTION. • THE BASIC PRINCIPLE IS THAT EACH COMPOUND ABSORBS OR TRANSMITS LIGHT OVER A CERTAIN RANGE OF WAVELENGTH. • THIS MEASUREMENT CAN ALSO BE USED TO MEASURE THE AMOUNT OF A KNOWN CHEMICAL SUBSTANCE. 11
- 12. QUANTITATIVE ASSAYS GAS CHROMATOGRAPHY • FIRST, A VAPORIZED SAMPLE IS INJECTED ONTO THE CHROMATOGRAPHIC COLUMN. SECOND, THE SAMPLE MOVES THROUGH THE COLUMN THROUGH THE FLOW OF INERT GAS. • THIRD, THE COMPONENTS ARE RECORDED AS A SEQUENCE OF PEAKS AS THEY LEAVE THE COLUMN. 12
Editor's Notes
- Purple colour gradually turns blue indicates opioid derivetives
- Boiling for an hour in conical flask
- No change
- Solute front