Chem 113 Spectroscopy and Spectrometry 1 Chapters 5 and 6 Forensic Science Copyright © James T. Spencer 2003-2010 All Rights Reserved Chem 113 Spectroscopy and Spectrometry 2 Spectroscopy: study of the interaction of radiation with matter Spectrometry: measurement of such interactions Used for analysis of the matter! Organic/inorganic Chemical Analysis What is this stuff? Chem 113 Organic/Inorganic Analysis 3 • Organic: a substance composed of carbon, hydrogen and often smaller amounts of oxygen, nitrogen, halogens, phosphorus, and sulfur • Inorganic: a chemical compound not based on carbon Chem 113 Qualitative vs. Quantitative 4 Qualitative: relates just to the identity of the material of question Example - identification of a powder may reveal the presence of heroin and quinine Quantitative: requires the determination of the percentage combination of the components of the substance Example - indicates 10% heroin and 90% quinine Chem 113 Combustion Reactions 5 Reaction of Hydrogen with Oxygen [COMBUSTION] (note precautions) » 2 H2(g) + O2(g) 2 H2O(g) H = 232 kJ/mol H2 » Ignition temperature = 580° - 590°C » Explosive [“when stuff gets really big really fast” Beakman’ World] » The rapid release of energy [-232 kJ/mol H2] into the surrounding air causes the air to very quickly expand. The explosion of pure H2 sounds quieter because the air expansion is slower. [Video No. 20-21; 4:42 +1:29 m] [Video No. 22; 2:50 m] Chem 113 Combustion Reactions Combustion of Alcohol (ethanol): C2H5OH(g) + 3 O2(g) 6 2CO2(g) + 3H2O(g) H = -1366.2 kJ mol-1 Tesla coil produces a high voltage electric spark. The spark is required to initiate this reaction. Conversion of chemical energy (PE stored in bonds) to mechanical energy. Questions for After Demonstration Are other types of energy are produced besides mechanical energy? Why can the reaction not be repeated without Chem 113 Combustion Analysis 7 • Empirical Formula from reaction with oxygen • Organic Compounds - C to CO2 and H to H2O • Use CO2 and H2O to determine the amount of C and H in original sample H2O furnace absorbant (Mg(ClO4)2) O2 flow sample contaminant catalyst (CuO); oxidizes traces of CO and C to CO2 CO2 absorbant (NaOH) Chem 113 Combustion Analysis 8 • Tells us how much of each element is present in unknown sample • Allows us to determine % of each element in the sample: % = [amount of element/amount of sample]100% • Compare with reference samples Chem 113 Mass Spectrometry • • • • Instrumental Principles and Design Spectral Features Spectral Interpretation and Comparison GC-MS and LC-MS 9 Chem 113 Mass Spectrometry Basic Ideas 10 Creates charged particles (ions) from gas phase molecules. Electron Ionization (EI)- Uses electron impact to ionize a molecule. Chemical Ionization (CI)- First ionizes a molecular gas which in turn ionizes the molecule of interest. A “gentler” method of ionization. Fast Atom Bombardment (FABS)- Mainly for non-volatile compounds - very harsh. The MS analyzes ions to provide information about the molecular weight of the compound and its chemical structure. Chem 113 Mass Spectrometry 11 Basic Ideas M ionization -e- M+ M+ M+ Chem 113 Mass Spectrometry 12 Basic Ideas M -e- M+ M+ acceleration M+ Chem 113 Mass Spectrometry 13 Basic Ideas M -e- M+ M+ acceleration M+ Chem 113 Mass Spectrometer Magnetic field deflection (quadrupole MS) 14 • Direct methods of measuring (separating) mass. • Sample molecules are ionized by e-beam to cations (+1 by “knocking off” one electron) which are then deflected by magnetic field - for ions of the same charge the angle of deflection in proportional to the ion’s mass vacuum chamber beam of pos. ions accelerating grid (-) sample Mass Spectrum Hg N 200 focusing slits ionizing e- beam S magnetic field Int. mass number (amu) detector Chem 113 Mass Spectrometer Atomic Spectra - Isotopic Abundance 15 Mass Spectrum Cl Int. 35 Mass Spectrum Int. C 12 37 mass number (amu) 35Cl: 75% abundant 37Cl: 24% abundant Mass Spectrum P Int. 31 13 mass number (amu) 12Cl: mass number (amu) 98.9% abundant 31P: 100% abundant 13Cl: 1.11% abundant Chem 113 Mass Spectrometry Molecules 16 Chem 113 Mass Spectrometry 18 Ionization produces singly charged ions. The intact charged molecule is the molecular ion. Energy from the electron impact and instability in a molecular ion can cause that ion to break into smaller pieces (fragments). The methanol ion may fragment in various ways, with one fragment carrying the charge and one fragment remaining uncharged. For example: CH3OH+. (molecular ion) (or) CH3OH+.(molecular ion) CH2OH+(fragment ion) + . H CH3+(fragment ion) + .OH Chem 113 Mass Spectrometry 19 Chem 113 Mass Spectrometry 20 169 Chem 113 Mass Spectrometer 21 Unknown white powdery substance ingested by unconscious patient. What do you do? Is it Heroin, Cocaine, Caffeine? Intensity Mass Spectrum of Unknown Compound Mass 25 50 75 100 125 150 175 200 225 250 275 300 Chem 113 Mass Spectrometer Intensity MS Library 43 Intensity Mass 25 50 75 67 55 42 50 other peaks at 327 and 369 268 204 215 94 Mass 25 Heroin Heroin 22 146 100 125 150 175 200 225 250 275 194 Caffeine 300 109 82 75 MS of Unknown 100 125 150 175 200 225 250 275 300 Chem 113 Intensity Mass Spectrometer MS Library 82 182 Cocaine Cocaine 303 42 122 25 Intensity Mass 23 Mass 25 50 75 67 55 42 50 150 272 100 125 150 175 200 225 250 275 194 Caffeine 300 109 82 75 MS of Unknown 100 125 150 175 200 225 250 275 300 Chem 113 Mass Spectrometer Intensity MS Library Intensity Mass 25 Mass 25 67 55 109 75 67 55 42 50 Caffeine Caffeine 82 42 50 194 24 100 125 150 175 200 225 250 275 194 Caffeine 300 109 82 75 MS of Unknown 100 125 150 175 200 225 250 275 300 Chem 113 Mass Spectrometer O 25 CH3 H 3C N N Mass Spectrum CH3 Caffeine Intensity Mass 25 N N O Mol. Wgt = 194 50 75 100 125 150 175 200 225 250 275 300 Chem 113 Sensitivity 10p g_be nzo phen one01 _020 2261 84800 26 02/2 6/2002 06:48 :0 0 PM RT: 5.99 - 7.99 NL: 2.94E4 m/z= 182.50183.50 MS 10pg_benzop henone01_0 2022618480 0 7.08 100 m/z 183 S/N 53:1 95 90 85 80 10 pg Benzophenone – S/N >10:1 75 Relative Abundance 70 65 1 pg (picogram) = 10-12 grams! 60 55 50 45 40 35 30 25 20 15 6.24 6.30 6.09 6.06 6.03 6.16 6.43 6.40 6.52 6.32 10 6.72 6.54 7.18 6.83 6.65 7.32 6.59 6.81 6.91 7.39 7.42 7.61 7.26 7.03 7.46 6.93 7.56 7.88 7.69 7.72 7.83 7.77 6.44 5 0 6.0 6.1 Base Peak Retention Time : 7.08 Scan : 243 Intensity : 29374 6.2 6.3 6.4 6.5 6.6 6.7 6.8 6.9 7.0 Ti me (mi n) 7.1 7.2 Calculated values Noise Range : 6.84 - 7.03 minutes (22 scans) Baseline : 1851 Sig nal To Noise : 53 Standard Deviation : 519 7.3 7.4 7.5 7.6 7.7 7.8 7.9 7.92 Chem 113 Chromatography 27 • Used to separate mixtures of compounds (most things in nature come as mixtures) • Based upon the differing interactions between the components of a mixture and some other substance. Chem 113 Chromatography See: http://antoine.frostburg.edu/chem/senese/101/matter/chromatography.shtml 28 Chem 113 Chromatography 29 Chem 113 Chromatography • • • • • 30 Chromatographic systems have a stationary phase (which can be solid or liquid) and a mobile phase (usually liquid or gas). The mixture to be separated is placed at the beginning of the chromatographic system (stationary phase). The mobile phase then “pushes” the components of the mixture through the system. Each component adsorbs on the stationary phase with a different strength (stronger means moves more slowly through the system). Each component comes out the end of the system at a different time (retention time). Chem 113 TLC Thin Layer Chromatography Spotting sample 31 Developing plate Chem 113 GC-Mass Spectrometry 32 A mixture is injected into the GC where the mixture is vaporized. The gas mixture travels through a GC column, where the compounds become separated. Those separated compounds then immediately enter the mass spectrometer. MS Chem 113 GC-Mass Spectrometry 33 The peak at 4.97 minutes is from dodecane, the peak at 6.36 minutes is from biphenyl, the peak at 7.64 minutes is from chlorobiphenyl, and the peak at 9.41 minutes is from hexadecanoic acid methyl ester. Chem 113 GC-Mass Spectrometry GC of a charcoal lighter fluid standard 34 Chem 113 GC-Mass Spectrometry MS-GC? 35 Chem 113 Street Drugs in Real Time- Know what the funny white powder is in less than 10 minutes.36 RT: 2.80 - 7.43 100 SM: 9G NL: 7.91E6 m/z= 43.50-44.50 F: MS level4 80 60 Amphetamine 40 20 0 100 NL: 1.18E7 m/z= 57.50-58.50 F: MS level4 Methamphetamine and MDMA 80 60 40 20 0 100 NL: 5.92E5 m/z= 298.50299.50 F: MS level4 80 60 40 Hydrocodone 20 0 100 NL: 2.17E6 m/z= 81.50-82.50 F: MS level4 80 60 Cocaine 40 20 0 100 NL: 2.40E5 m/z= 314.50315.50 F: MS level4 80 60 40 20 0 3.0 3.5 4.0 4.5 5.0 5.5 Time (min) 6.0 6.5 7.0 Oxycodone Chem 113 Cases for the GC-MS • Toxicology: – – – – – – Components in blood, etc. Toxin ID Quality control (Tylenol murders) Arson investigations (VOC) Alcohol Intox. Gunshot residues 37 Chem 113 Atomic and Molecular Spectroscopy 38 • Science: Atomic Theory – “The strength of a science is that its conclusions are derived by logical arguments from facts that result from well-designed experiments. Science has produced a picture of the microscopic structure of the atom so detailed and subtle of something so far removed from our immediate experience that it is difficult to see how its many features were constructed. This is because so many experiments have contributed to our ideas about the atom.” B. Mahan from University Chemistry Chem 113 Atomic and Molecular Spectroscopy 39 • Interaction of electromagnetic radiation (“light”) with atoms and molecules • Absorption, transmission and emission spectra Chem 113 Spectroscopy Background - Electromagnetic Radiation 40 = c where = wavelength, = frequency, c = light speed amplitude 1 cycle per sec = 1 hertz wavelength () Chem 113 Electromagnetic Radiation 41 Chem 113 Spectroscopy 42 • When electromagnetic radiation passes through a substance, it can either be absorbed or transmitted, depending upon the structure of the substance. Chem 113 Spectroscopy 43 When a molecule absorbs radiation it gains energy as it undergoes a quantum transition from one energy state (Einitial) to another (Efinal). The frequency of the absorbed radiation is related to the energy of the transition Efinal - Einitial = E = h = hc/ . emission Chem 113 Quantization 44 • Light energy may behave as waves or as small particles (photons). • Particles may also behave as waves or as small particles. • Both matter and energy (light) occur only in discrete units (quantized). Quantized (can stand only on steps) Non-Quantized (can stand at any position on the ramp) Chem 113 What is Quantization 45 • Examples of quantization (when only discrete and defined quantities or states are possible): Quantized Non-Quantized Piano Stair Steps Typewriter Dollar Bills Football Game Score Light Switch (On/Off) Energy Matter Violin or Guitar Ramp Pencil and Paper Exchange rates Long Jump Distance Dimmer Switch Chem 113 Atomic Spectroscopy 46 Chem 113 Flame Tests 47 Atomic Emission Chem 113 Atomic Emission 48 Chem 113 Red Blue 364.6 nm 410.2 nm 434.0 nm 486.1 nm 656.3 nm Hydrogen Emission Ultraviolet 49 Chem 113 50 364.6 nm 410.2 nm 434.0 nm 486.1 nm 656.3 nm Hydrogen Emission Ultraviolet Red Blue A Swiss schoolteacher in 1885 (J. Balmer) derived a simple formula to calculate the wavelengths of the emission lines (purely a mathematical feat with no understanding of why this formula worked). frequency = C ( 1 - 1 ) where n = 1, 2, 3, etc... 22 n2 C = constant Chem 113 Bohr’s Model “Microscopic Solar System” • Electrons in around nucleus with quantized (allowed) energy states • When in a state, no energy is radiated but when it changes states, energy is emitted or gained equal to the energy difference between the states • Emission from higher to lower, absorption from lower to higher 51 n=∞ n=4 n=3 n=2 electronic transitions n=1 Chem 113 Atomic Emission AES 52 • Atomic Emission (AE) - uses quantitative measurement of the optical emission from excited atoms to determine analyte concentration. Analyte atoms in solution are aspirated into the excitation region where they are atomized by a flame, discharge, or plasma. These high-temperature atomization sources provide sufficient energy to promote the atoms into high energy levels. The atoms decay back to lower levels by emitting light. Since the transitions are between distinct atomic energy levels, the emission lines in the spectra are narrow. Ch. 6: p 150. Chem 113 Atomic Emission AES • Russian Icon of St. Nicholas The pigments present on this mid-19th Century painting were characterized by AES spectroscopy (laser-induced breakdown spectroscopy, LIBS) and Raman microscopy. The identification of pigments on the original work along with those applied in restoration of cracks in the varnish and painting surface were analyzed. Art Restoration History and Art Forgery 53 Chem 113 Atomic Emission AES 54 • LIBS depth profile measurements leave a minute crater in the surface of the art object being studied. This allows stratagraphic information to be collected. A typical cross section of the icon is shown. Chem 113 Atomic Emission AES 55 • Several areas of the icon, where white paint was used, were analyzed. The LIBS spectrum showed strong peaks characteristic of lead. This was confirmed by the Raman spectrum, which verified the presence of lead carbonate, [2PbCO3·Pb(OH)2]. LIBS Raman Chem 113 Atomic Emission AES The brown pigment was characterized as an iron-based pigment mixed with lead white. The LIBS spectrum showed the presence of Fe and Al, corresponding to an iron oxide and an earth such as clay. Also present are emissions characteristic of magnesium, lead and calcium. The peak corresponding to iron at ~275 nm is characteristic of iron that has been observed in studies on pure iron oxide pigments (for example, Mars black, Fe3O4). 56 Chem 113 Atomic Absorption AAS • Atomic Absorption - Atomic-absorption (AA) 57 spectroscopy uses the absorption of light to measure the concentration of gas-phase atoms. Since samples are usually liquids or solids, the analyte atoms or ions must be vaporized in a flame or graphite furnace. The atoms absorb ultraviolet or visible light and make transitions to higher electronic energy levels. The analyte concentration is determined from the amount of absorption. Chem 113 Atomic Absorption AAS 58 Typical Problem - A child becomes quite ill and is taken to the hospital. It is found that the child is suffering from lead poisoning. A forensic laboratory is contacted and asked if it can determine the source of the lead which the child has ingested. No crime has been committed, per se, but the source must be eliminated to prevent future danger to the child. Paint samples from a number of objects with which the child has had repeated contact are collected. Paint on the child's crib, paint from his toys, and paint from the child's swing, to name a few, are sent to the laboratory. AA is the best method for these analyses. Chem 113 Neutron Activation Analysis NAA 59 Neutrons interact with a target nucleus to form a compound nucleus in an excited state. The compound nucleus will decay into a more stable configuration through emission of one or more gamma rays. This new configuration may yield a radioactive nucleus which also decays by emission of delayed gamma rays, but at a much slower rate according to the unique half-life of the radioactive nucleus. Chapt 6.p 163 Chem 113 Neutron Activation Analysis NAA 60 •About 70% of the elements have properties suitable for measurement by NAA. •Parts per billion or better. Gamma-ray spectrum in a sample of pottery irradiated for 24 hours, decayed for 9 days, and counted for 30 minutes. Chem 113 Neutron Activation Analysis NAA 61 An example of the gamma-ray spectrum from the activation of a human nail used as a biological monitor of trace-element status. Chem 113 Neutron Activation Analysis Arsenic in Hair Napoleon Bonaparte One of the most brilliant individuals in history, Napoleon Bonaparte was a masterful soldier, grand tactician, sublime statesman and exceedingly capable administrator. After an extraordinary career, he was finally defeated and exiled to Elba. He returned from Elba to be ultimately defeated at Waterloo. He was finally exiled to the remote tiny volcanic island of St. Helena, south of the Equator. The nearest land is Ascension Island, 700 miles to the north. 62 Chem 113 Neutron Activation Analysis Arsenic in Hair Murdered or Not? For years a controversy has raged about Napoleon being killed on St. Helena - either by French Royalists, persons in his exiled entourage or the British - and all have pointed to the high levels of arsenic in the emperor's body as being evidence of such behavior. The emperor's body contained some 15 parts per million of the poison, where the maximum safe limit is only three parts per million. The determination was by neutron activation analysis of his hair. 63 Chem 113 Neutron Activation Analysis Arsenic in Hair “So Who Done It?” (if it was done at all) British Authorities - The Allied heads of state had no greater wish than to ensure that Napoleon was permanently “out of the way”. Strong hatred by British local commander. Royalists - Revenge and insurance against Napoleon for declaring himself Emperor and dismantling the aristocracy. Exiled Entourage - Jealousy (romantic triangles), intrigue, revenge. 64 Chem 113 Neutron Activation Analysis Arsenic in Hair The wallpaper in his room was dyed with Scheele's Green (Paris Green), a coloring pigment that had been used in fabrics and wallpapers from around 1770. Named after the Swedish chemist who invented it, the dye contained copper arsenite. It was discovered that if wallpaper containing Scheele’s Green became damp, the mould converted the copper arsenite to a poisonous vapor form of arsenic. Breathing the arsenic on its own might not have been enough to kill Napoleon, but he was ill already with a stomach ulcer/cancer. On May 5, 1821, the arsenic tipped the scale against "the little corporal." 65 Chem 113 Molecular Spectroscopy 66 • Electronic Spectroscopy • Vibrational Spectroscopy • Nuclear Magnetic Resonance Spectroscopy (NMR or MRI) (All of these are absorption methods, but this list is far from comprehensive!) Chem 113 Electronic Spectroscopy UV-visible 67 When white light passes through or is reflected by a colored substance, a characteristic portion of the total wavelengths is absorbed. The remaining light will assume the complementary color to the wavelength(s) absorbed. Chem 113 Electronic Spectroscopy UV-visible 68 Visible region - photon energies to excite a electron to a higher energy level (orbital). Of the six transitions outlined, only the two lowest energy ones (leftmost, colored blue) are in the UV-visible region (200-800 nm). Chem 113 Electronic Spectroscopy UV-visible 69 Effect of Conjugation Chem 113 Vibrational Spectroscopy Infrared and Raman Spectroscopy 70 • Radiation from 500 to 4000 cm-1 (vibrational transitions in the molecules). • Vibrational “mode” must have a change in dipole moment in the transition. Energy of the transition is dependent upon the strengths of the bonds and geometric structure. Chem 113 Vibrational Spectroscopy Infrared and Raman Spectroscopy 71 • For the water molecule, there are three vibrational modes that may occur (are allowed). •The spacing between energy levels depends upon the vibration being considered. •Each spacing requires a photon of different energy to cause the transition •expect photons of three different energies to be absorbed by H2O. Chem 113 Vibrational Spectroscopy Infrared and Raman Spectroscopy 72 • For a molecule to directly absorb infrared electromagnetic radiation, the vibrational motion must produce a change in the dipole moment of the molecule. • There are many molecules which, although possessing no permanent dipole moment, still undergo vibrations which cause changes in the value of the dipole moment from 0 to some non-zero value. Consider the CO2 molecule: Chem 113 IR Spectrum of CO2 73 Calculated spectrum of co2 B3LYP/6-31G(d) 595 495 absorbance 395 O=C=O 295 O=C=O 195 95 -5 0 500 1000 1500 2000 wavenumber/cm-1 2500 3000 3500 4000 Chem 113 Vibrational Spectroscopy Infrared and Raman Spectroscopy 74 • Different types of bonds have characteristic regions of the spectrum where they absorb Chem 113 Vibrational Spectroscopy Infrared and Raman Spectroscopy 75 • Forensic Applications of Infrared Spectroscopy • Use of computer databases of IR’s of known compounds » Analyzing Alcohol - The breath is tested with a mechanism similar to a breathalyzer (chemical oxidation) but uses the infrared absorptions of alcohol. » Chem 113 Vibrational Spectroscopy Infrared and Raman Spectroscopy 76 • Forensic Applications of Infrared Spectroscopy • Use of computer databases of IR’s of known compounds » Analyzing Drugs - The drug's various chemical components absorb infrared light. The absorptions are compared to known samples using a database. Chem 113 Vibrational Spectroscopy Infrared and Raman Spectroscopy 77 • Forensic Applications of Infrared Spectroscopy • Use of computer databases of IR’s of known compounds » Analyzing Fibers - The expected identity of the fiber has been established by observing it under a microscope. Its IR spectrum can confirm the suspected identity. • Chem 113 Vibrational Spectroscopy Infrared and Raman Spectroscopy 78 • Forensic Applications of Infrared Spectroscopy • Use of computer databases of IR’s of known compounds » Analyzing Paint - Paint has been recovered from a crime scene. Since there is a limited amount of paint, the first tests to be done should be nondestructive. Colors, layers, texture, and other physical properties are recorded. The individual layers of paint are analyzed by infrared spectroscopy. The results can be compared to IR results of known paint samples. Chem 113 Infrared Spectroscopy 79 • “With infrared radiation, forensic scientists can determine the exact ink type and pen that a death threat was written in, or the very model and year of a suspect's automobile in a hit-and-run accident. Using infrared spectromicroscopy, forensic investigators have been able to identify samples from inks and paint chips to fibers and drugs. Researchers at Lawrence Berkeley National Laboratory have now expanded the boundaries of infrared forensics with the use of synchrotron radiation from the Lab's Advanced Light Source (ALS) facility…..” (Daily Californian, Wednesday, September 18, 2002) Chem 113 Magnetic Resonance Spectroscopy NMR (MRI) 80 • Visualize soft tissue by measuring proton (nuclear) magnetic alignments relative to an external magnetic field. Review Electron Spin Properties First. Chem 113 Nuclear Spin 81 • Like electrons, nuclei spin and because of this spinning of a charged particle (positively charged), it generates a magnetic field. Two states are possible for the proton (1H). N S + + S N Chem 113 Magnetic Fields 82 Chem 113 Nuclear Spin Similar to a canoe paddling either upstream or downstream 83 S Antiparallel Degenerate E N N N S Parallel S N External Magnetic Field S Chem 113 84 Magnetic Resonance Imaging MRI • Hydrogen atom has two nuclear spin quantum numbers possible (+1/2 and -1/2). • When placed in an external magnetic field, 1H can either align with the field (“parallel” - lower energy) or against the field (“antiparallel” - higher energy). • Energy added (E) can raise the energy level of an electron from parallel to antiparallel orientation (by absorbing radio frequency irradiation). • Electrons (also “magnets”) in “neighborhood” affect the value of E (i.e., rocks in stream). • By detecting the E values as a function of position within a body, an image of a body’s hydrogen atoms may be obtained. Chem 113 MRI 85 • Advantages (first three are not really important for forensics) – non-invasive. – no ionizing or other “dangerous” radiation (such as Xrays or positrons). – Can be done frequently to monitor progress of treatment. – images soft tissues (only those with hydrogen atoms (almost all “soft” tissues). – images function through the use of contrast media. • Disadvantages – Relatively expensive equipment. Chem 113 MRI; Hardware 86 Chem 113 MRI 87 Chem 113 MRI 88 Chem 113 MRI 89 Chem 113 MRI 90 Chem 113 MRI 91 Chem 113 Forensic MRI/CT 92 • Used to reconstruct facial images from skulls. Use for ancient mummies to modern skulls. • Allows a very fine discrimination between materials with different densities providing an enormous amount of information about the mummy and its skeleton. • The level of automation reached in building models from CT data, reconstruction, texture application and visualization allow to the user to complete whole process in 2-3 hours on a PC or graphic workstation. Chem 113 Forensic MRI and CT 93 • The “Virtopsy” focuses on four goals: • radiological digital imaging methods as main diagnostic tools in forensic pathology, ultimately leading to "minimally invasive autopsy" analogous to "keyhole surgery" in clinical medicine. • three-dimensional optical measuring techniques - a reliable, accurate geometric presentation of all forensic findings (the body surface as well as the interior). • 3D surface scanning in forensic reconstruction. • Producing and validating of a post-mortem biochemical profile to estimate the time of death. • The implementation of an imaging database as a technical basis of a "center for competence in virtual autopsy”. Chem 113 Forensic MRI 94 Virtopsy, a new imaging horizon in forensic pathology: virtual autopsy by postmortem multislice computed tomography (MSCT) and magnetic resonance imaging (MRI) - 40 forensic cases were examined and findings were verified by subsequent autopsy. Results were classified as follows: (I) cause of death, (II) relevant traumatological and pathological findings, (III) vital reactions, (IV) reconstruction of injuries, (V) visualization. In these 40 forensic cases, 47 partly combined causes of death were diagnosed at autopsy, 26 (55%) causes of death were found independently using only radiological image data. Radiology was superior to autopsy in revealing certain cases of cranial, skeletal, or tissue trauma. Some forensic vital reactions were diagnosed equally well or better using MSCT/MRI. Radiological imaging techniques are particularly beneficial for reconstruction and visualization of forensic cases. (J Forensic Sci. 2003, 48, 386-403) Chem 113 Forensic MRI Validating of a post-mortem analysis 95 Complex scull fracture system following motor vehicle accident (victim was overrun by automobile). 3D reconstructed MSCT image. Chem 113 Forensic MRI Validating of a post-mortem analysis 96 Injury due to vehicle impact in a motor vehicle accident (pedestrian). (right) finding at autopsy; right lower leg showing fracture of the fibula. (left) 3 D reconstructed MSCT; Chem 113 Forensic MRI and CT Facial Reconstructions Egyptian Mummy Head 97 The method uses the tables combined with the warping of a 3D model of a reference scanned head, until the relevant surface to bone distances are correct. Texture mapping is used to provide colors and aesthetic features. Chem 113 Forensic MRI and CT Mummy Facial Reconstruction 98 Model skin (blue) and mummy skull (white) Face shape generated Chem 113 Forensic MRI and CT Mummy Facial Reconstruction 99 Texturized model of reconstructed soft tissues of the mummy Chem 113 X-ray Methods • X-ray Diffraction (XRD and CT) • Energy Dispersive X-ray Fluorescence 100 Chem 113 Bragg’s Law and X-ray Diffraction 101 incoming light E D B d lattice in a crystal C Since BCD = 2d sin is the limiting condition for observing a reflection then because of wave addition and cancellation; Bragg’s Law: n = 2d sin where n = 1, 2, 3, etc... Chem 113 Energy-Dispersive X-ray Fluorescence (EDXRF) •Did your luxury purchase originate in a mine deep in the heart of Central America, or the bottom of a silty river tributary in Africa, or perhaps even a flask in a laboratory in Chicago or Minsk? •Metal ions such as V3+, Cr3+, Mn2+, Mn3+, Fe2+, Fe3+, Ni2+, Cu2+, and UO22+ are responsible for the colors of most common gemstones and minerals. •U.S. Federal Trade Commission says consumers must be informed of alterations in gemstones. 102 Chem 113 Energy-Dispersive X-ray Fluorescence (EDXRF) 103 •Among the most sensitive and popular of the nondestructive spectroscopic techniques used for trace-metal determination is EDXRF. In this technique, X-rays excite the gemstone to fluoresce and the fluorescent line spectrum indicates which chemical elements are present. EDXRF can also be used to differentiate freshwater from saltwater pearls on the basis of the greater concentration of magnesium present in the former. Chem 113 Energy-Dispersive X-ray Fluorescence (EDXRF) 104 •EDXRF has been called 'the curator's dream instrument' because measurements are non-destructive and usually the whole object can be analyzed, rather than a sample removed from one. The technique involves aiming an Xray beam at the surface of an object; this beam is about 2 mm in diameter. •The interaction of X-rays with an object causes secondary (fluorescent) X-rays to be generated. Each element present in the object produces X-rays with different energies. These X-rays can be detected and displayed as a spectrum of intensity against energy: the positions of the peaks identify which elements are present and the peak heights identify how much of each element is present. Chem 113 Energy-Dispersive X-ray Fluorescence (EDXRF) 105 An incoming X-ray ejects a K-shell electron from an atom of the target. An electron in the M or L-shell loses energy as it transitions to the vacant K-shell. It given off energy in the form of fluorescence. Chem 113 Energy-Dispersive X-ray Fluorescence (EDXRF) http://www.thebritishmuseum.ac.uk/science/techniques/sr-tech-xrf.html 106 Chem 113 Analytical Methods 107 • Questions to consider in choosing an analytical (chemical) method: – Quantitative or qualitative required – Sample size and sample preparation requirements – What level of analysis is required (e.g., ± 1.0% or ± 0.001%) – Detection levels and useful analytical concentration ranges – Destructive or non-destructive – Availability of instrumentation – Admissibility (e.g., are all lead pipes compositionally the same or are there sufficient variations among “known” Pb pipes of the world to link two samples) Chem 113 108