Instrumental Analysis and Forensic Chemistry

Hossein Ahmadzadeh mhahmadzadeh@csupomona.edu

1- Quantitative Chemical Analysis (6th Ed.) Daniel C. Harris 2- Fundamentals of Analytical Chemistry (7th Ed.) Skoog, West and Holler 3- Principles of Instrumental Analysis, (5th Ed.) Skoog, Holler, Nieman

The Chemical Analysis

Two general areas of analysis:
Qualitative analysis attempting to identify what materials are present in a sample. Quantitative analysis determining how much of a material is present in a sample.

Types of Analysis
Complete analysis - The goal is to determine the amount of each component in a sample. Partial analysis -The goal is to determine the amount of one or a limited number of components without regard to total composition.

Cocaine

in a sample. Alcohol level in blood Presence of lead in a sample

Steps in a Chemical Analysis

Skoog & West

Methods of Analysis

Gravimetry - Methods based on measured weight. Titrimetry - Methods based on a measured volume. Electrochemical - Approaches that rely on measurement of potential, current, resistance, charge, Spectral Methods - Interaction of an analyte with Electro-Magnetic (EM) radiation. Chromatography - Separation of a material due to its interaction with two different phases. Chemometrics - The statistical treatment of data

Spectroscopy
Light Source Wavelength Selector Sample

Absorption Emission Fluorescence

Wavelength Selector

Light Detector

Spectroscopy
IR spectrum IS CONCLUSIVE IR can specifically identify a substance IR is a definitive identification technique Measures a fingerprint of the particular dug Disadvantage The drug must be pure for a definitive match May have to perform purification steps

A person with phenobarbital overdose show a marked decrease in electrical activity, to the point of mimicking brain death.

Phenobarbitone Seizure control

Caffeine

InfraRed (IR) Microscopy

Hair analysis overcomes drug testing problems associated with urinalysis. Contamination of the exterior surface of the hair = false positive IR microscopy examines the interior of the hair and differentiate contamination from drugs absorbed into the hair from ingestion. Microtom the hair, cross-sectional or lateral, with spatial resolution of 10 m. 3D IR imaging is possible with Fourier transform (FT-IR) microscopy

Molecular Fluorescence

Optical emission from molecules that have been excited to higher energy levels by absorption of electromagnetic radiation.

Dkkdj

From a German stamp

Hair Elemental Analysis

Issac Newton Napoleon Beethoven King Charles II Hg As Pb Hg

1- R. Passwater, and E. Cranton, Trace Elements, Hair Analysis and Nutrition, Keats Pub, 1983, p 272 2- Atomic Absorption Spectrometry in Occupational And Environmental Health Practice, Vol. 1, D.L. Tsallev And Z.K. Zaprianov

Atomic Spectroscopy
Atomization is by far the most critical step in atomic spectroscopy. Atomic Absorption Atomic Emission Atomic Fluorescence

Gas flow rate = Burning velocity Flashback

Flame Atomization

Energy level diagrams

Atomic spectra: predominantly resonance lines The higher the atomic number, the more complicated is the spectra: Li (30 Lines), Cs (645), Mg (173), Ca (662), Ba (472), Cr (2277), Fe (4757),Ce (5755), .. Resolution of the mono-chromator is very important

Energy Level Diagram

Hollow Cathode Lamps

Tungsten anode Cylindrical cathode - made of the element of interest Sealed glass tube - Ar or Ne at 1-5 torr 300 V, 5-15 mAmps

Schematic of Instruments

Sample Preparation

For flame AAS, sample must be in solution

hot mineral acids oxidation (wet ashing) combustion fusion with with boric oxide sodium carbonate

For GFAAS, sample can be solid, but calibration can be difficult

How to Introduce the Sample?

Solutions

Pneumatic nebulization Ultrasound nebulization Electrothermal vaporization Hydride generation Electrothermal vaporization Direct insertion Laser ablation Spark or arc ablation Glow discharge sputtering

Solids

Properties of Flames

Electrothermal Atomizers
Graphite furnace atomic absorption spectrometry (GFAAS) is also known as electrothermal (ETAAS). An ideal graphite furnace:

A constant temperature in time and space during the interval in which free atoms are produced Quantitative atom formation regardless of the sample composition Separate control of the volatilization and atomization processes High sensitivity and good detection limits A minimum of spectral interferences

Temp. Programming for GFAA

Separation Techniques

Extraction Distillation Electrophoresis

Capillary Electrophoresis Thin Layer Chromatography (TLC) Gas Chromatography (GC) High Pressure Liquid Chromatography (HPLC)

Chromatography

Distillation

Thin Layer Chromatography (TLC)

What is the composition of Unknown From Case

The drug works directly on the brain and spinal cord by interfering with normal neurotransmission

Capillary Electrophoresis

Ephedrine *does* increase your heart rate and constrict blood vessels. If you are careless, you *can* have a heart attack and/or a stroke.

Zones of sampling for gunshot residues: (A) web and (B) palm

Faber-Casell fine Metal point

Micro Fasion Roller

Pilot Precise V5 Extra fine point

Tombo Roll pen

Sanford Uniball Vision micro

Sanford Uniball fine

Ion Mobility Spectrometer

Gas Chromatography (GC)

In GC, gaseous analyte is transported through a column by a gaseous mobile phase (carrier gas).

Carrier gas is usually He, N2 or H2.

Stationary phase is usually a nonvolatile liquid, but sometimes a solid. Analyte can be either a gas or volatile liquid.

Columns

15 100 meters long

Usage

Qualitative analysis

Mass spectrometer IR spectrometer Compare retention times The area under a chromatographic peak is proportional to the quantity of that component. For very narrow peaks, peak height can be substituted for area.

Quantitative analysis

Flame Ionization Detectors

Eluate is burned in H2 and air. CH radicals which react with O radicals to produce CHO+ and electrons. CH + O CHO+ + e Only about 1 in 105 carbon atoms produce an ion, but ion production is strictly proportional to the number of carbon atoms entering the flame. Electrons flow from the anode to the cathode, where they neutralize CHO+ in the flame. This current is the detector signal.

Diagram of a Mass Spectrometer

GC separation of Gasoline (0.1 uL) components (Fossil Fuels)

GC separation of 0.05 uL of steam distilled wood turpentine, a naturally occurring background found in many wood samples.

Arson: A wide range of low boiling point components present in standard accelerants

High-Performance Liquid Chromatography (HPLC)

Most compounds are not sufficiently volatile for GC analysis. HPLC uses high pressure to force solvent through closed columns containing very fine particles that give high-resolution separations. Smaller particles give more theoretical plates.

Typical equipment for HPLC

The Column
The

stationary phase is usually highly pure, spherical, microporous particles of silica that are permeable to solvent and have a high surface area

The Elution Process

Solvent molecules compete with solute molecules for sites on the stationary phase. The more polar the solvent, the greater its eluent strength, and solutes will be more rapidly eluted from the column.

Elution occurs when solvent displaces solute from the stationary phase

Detection of a drug and its glucuronide metabolite(s) is great important in interpretive forensic and clinical toxicology. Metabolite/drug ratios provide information about route, dose and time of exposure. In instances where the parent drug is biotransformed quickly, the detection of metabolites allows for the identification of parent drugs.