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Lecture: Forensic Evidence Physical Evidence Any material either in gross or trace quantities that can establish through scientific examination and analysis that a crime has been committed. Forensic laboratories Items of physical evidence identification evaluation individualization Classification of Physical Evidence • Trace evidence extremely small items • Direct evidence stands on its own to prove an alleged fact • Prima facie evidence evidence established by law • Circumstantial evidence incriminates a person • Exculpatory evidence helps to prove that an accused individual is not guilty Physical evidence utilization in other areas of forensic investigation • Provides investigative leads for a case • Ties one crime to a similar crime or connects one suspect with another • Corroborates statements from witnesses to or victims of a crime • The elements of a crime help to determine what will be useful as evidence. • Besides knowing what types of evidence to search for, it is necessary to know where evidence is most likely to be found. Characteristics of evidence Class characteristics features that place the item into a specific category Individual characteristics features that distinguish one item from another of the same type E x a m in a t io n a n d a n a l y s is o f p h y s ic a l e v id e n c e H ig h e s t d e g r e e o f s c ie n t ific c e r ta in t y p o s s ib le w ith c u rre n t te c h n o lo g y p h y s ic a l id e n tific a tio n c h e m ic a l id e n tific a tio n b io lo g ic a l id e n tific a tio n Evidence Analyte/Characteristic Techniques Blood Ethanol Drugs of abuse Composition Color Fabric Fibers Glass Shoes Powder Soil Saliva stain Hair Composition Physical properties Refractive index Magnesium Miscellaneous Drugs of abuse pH Iron Proteins DNA Appearance Headspace analysis GC GC/MS FT-Raman spectroscopy Visible, diffuse reflectance spectroscopy FT-IR microscopy Solubility, melting point Microscopy Atomic absorption spectrophotometry Solid-phase extraction; LC FT-IR Potentiometry UV-Visible Spectr. Immunological tests Short tandem repeat DNA analysis Microscopy Evidence Techniques Gunshot residue Atomic absorption spectrophotometry, scanning electron microscopy Visible reflectance, FT-IR microscopy, FTRaman UV–vis, LC FT-IR, UV–vis Thermal analysis, FT-IR Liquid- and solid-phase extraction, GC/MS Fluorescent visualization Atomic absorption spectrophotometry, titrations Clothing Pen inks Plastic fragments Tire fragments Food (poisoned) Fingerprints Metals Arson samples GC, GC/MS X-Ray Diffraction 13-8 Crystal Structures X-Ray Diffraction One morning in the summer of 1961, hundreds of crazed birds attacked the seaside town of Capitola, California. The birds "cried like babies" as they dove into streetlamps, crashed through glass windows, and attacked people on the ground. Most of the birds were sooty shearwaters, a normally nonaggressive species that feeds on small fish and comes ashore only to breed. The incident fascinated Alfred Hitchcock, who frequently vacationed in nearby Santa Cruz. He included newspaper clippings about the Capitola attack in his studio proposal for The Birds, which appeared in cinemas two years later. In the winter of 1987, the agent that is now believed to be responsible for the Capitola incident struck on the opposite shore of the continent. This time, it struck higher on the food chain. Over a hundred people became extremely ill within hours after dining on cultured blue mussels in restaurants around Prince Edward Island in Canada. It quickly became apparent that this was no ordinary outbreak of food poisoning. Vomiting, cramps, diarrhea, and incapacitating headaches were followed by confusion, loss of memory, disorientation, and (in severe cases) seizures and coma. A few exhibited emotional volatility, with uncontrolled crying or aggressiveness. Three elderly victims died. [Perl]. A tragic symptom of poisoning was the destruction of short term memory in about one quarter of the survivors. They could remember nothing that happened after the poisoning. Some were unable to recognize their surroundings or relatives. They could learn no new facts or skills. The most severely affected lost memories several years old. For twelve of the victims, the loss of short term memory was permanent. Figure 1. General strategy for isolation of the toxin responsible for amnesic shellfish poisoning. Based on a diagram by M. Quilliam and J. L. C. Wright (Analytical Chemistry, 61, 1054 (1989)). A band very close to the band for glutamic acid was observed in the electrophoresis of the toxic XAD-2 fraction, but not in the control fraction. It stained a distinctly different color from the glutamic acid. When the material in the band was collected and injected onto the HPLC column, it took exactly the same amount of time to move through the column as the toxic component found by the HPLC analysis. It also produced exactly the same amount of toxicity as the HPLC fraction had. Mass spectrometry was used to determine the compound's molecular weight (312 g/mol) and molecular formula (C15H22NO6). Spectroscopic analysis revealed the presence of conjugated double bonds and features characteristic of an amino acid. By matching the spectra with those from STN International's Registry system, the compound was unambiguously identified as domoic acid, an triprotic amino acid: Domoic acid in acidic solution. Glutamic acid in acidic solution. Glutamate (the ionized form of glutamic acid) is a neurotransmitter, so glutamate plays a fundamental role in thought, learning and memory. It is possible to have too much of a good thing, however. Glutamate at high concentrations acts as an excitotoxin -a compound that kills cells by literally exciting them to death. Excess glutamate keeps the gates that allow calcium ions across the cell membrane open too long. Calcium ions flood into the cell, causing it fire uncontrollably. The neuron swells and eventually bursts. The damage cascades to nearby neurons because the damaged and ruptured neurons release their glutamate and other excitatory amino acids, overstimulating nearby cells. The excess calcium inside the cell stimulates certain protein-cutting enzymes, which produce large quantities of free radicals as a by-product. The free radicals are extremely reactive, and damage any biochemical structure they come into contact with [Berman]. This excitotoxic cascade is thought to play an important role in brain injury and neurodegenerative diseases. Domoic acid is a molecular Trojan Horse. Nerve cells mistakenly recognize domoic acid as glutamic acid- a fatal error. Domoic acid's structure is obviously similar to glutamic acid. But its five-sided ring makes it less flexible than glutamate, which causes it to bind very tightly to glutamate receptors. As a result, the excitatory effect of domoate is 30 to 100 times more powerful than that of glutamate [Perl]. How did the domoic acid get into the shellfish (and the anchovies eaten by the birds at Capitola)? Remember that phytoplankton pigments were found in the aqueous layer after solvent extraction. This wasn't quite a smoking gun, but it was definitely a fingerprint of the killer. An extensive investigation traced the domic acid to an obscure species of needle-like diatom*, called Pseudonitzschia pungens (shown in the title banner at the top of this page). Pseudonitzschia has been found in oceans around the world, so further outbreaks are possible in many locations. Commercial shellfish and seafood is now monitored regularly for domoic acid, using HPLC to identify the toxin. The screening and testing procedures have so far been successful- not a single instance of domoic acid poisoning in humans has been reported since the 1987 outbreak.