Forensic Chemsitry CHEM 4204
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This 6 page Class Notes was uploaded by Saige Wolf on Monday October 5, 2015. The Class Notes belongs to CHEM 4204 at Clayton State University taught by Susan Hornbuckle in Fall. Since its upload, it has received 26 views. For similar materials see /class/219569/chem-4204-clayton-state-university in Chemistry at Clayton State University.
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Date Created: 10/05/15
CHEM 4204 Chapter Objectives Clayton State University Dr Susan F Hornbuckle Chapter 4 1quot 9 4 V39 0 gt1 9 0 0 Be able to de ne the following terms sample preparation partitioning internal standard surrogate spike matrix spike headspace method hydrophilic hydrophobic lipophilic lipophobic ionization centers dry extraction liquidliquid extraction LLE dynamic headspace DHS purge and trap PT solid phase extraction SPE normalphase SPE reversephase SPE Solid phase microextraction SPME solid phase stirbar extraction SPSE thinlayer chromatography TLC immunoassay antigen antibody immunogen hapten polyclonal antibody monoclonal antibody Hybridoma Cells r quotquot 39 r iti ve 39 Homogeneous Assay Heterogeneous Assay Radioimmunoassay RIA Fluorescent Polarization 39 FPIA quot391 quot J 39 Technique EMIT and EnzymeLinked Immunosorbent Assay ELISA Be able to explain the difference between extraction and digestion Be able to explain when and why an internal standard or spike might be added to a sample Be able to list the two fundamental requirements for a successful separation of an analyte from a matrix by any method Be able to explain which equilibrium is being exploited and how is that equilibrium is being manipulated in the following partitioning techniques headspace method SPE LLE immunoassays dry extraction Be able to write an equilibrium expression for the following terms Ka Kb Ksp Kow Log P and KD Be able to explain how the pH of the solution can change the KOW for many drugs and other larger analytes Be able to list six properties or interactions that are the basis for a compound s af nity for one partition phase layer etc over another Be able to list recognize and classify the basic and acidic functional groups that common occur in the structures of drugs 13 xv Lu J Know the relationship between pH and pKa HendersonHasselbalch Equation Given the pKa values for an analyte determine the pH necessary for the compound to be 100 ionized Also determine the pH necessary for the compound to be 100 unionized How does this differ for an acidic a basic or an amphoteric compound Be able to describe with words and diagrams an AcidBaseNeutral Preparation LLE and be able to predict which solutions will contain the neutral water soluble compounds neutral water insoluble compounds strong acids weak acids and the basic compounds Be able to list the ve factors to consider when choosing a solvent for LLE Be able to explain when and why solid phase extraction may sometimes be successful at separation of an analytes from the matrix when liquidliquid extraction would not Be able to describe the process and theory behind solid phase extraction Be able to list at least ve common interactions between an analyte and the solid phase in a SPE Know the most commonly used solid phases for SPE Be able to list three polar and one nonpolar solid phase CHEM 4204 Chapter Objectives Clayton State University Dr Susan F Hornbuckle 17 Give a developed TLC plate be able to calculate Rf values for each of the analytes 18 Be able to list common developing techniques for TLC plate Specify the analytes they are used to detect if they are exhibit specificity for one or a group of analytes Be able to describe at least two issues that might cause an analyte to strea on the TLC plate What can be done to prevent streaking Be able to describe the technique used to produce a polyclonal antibody Be able to describe the technique used to produce a monoclonal antibody Be able to describe the difference in a noncompetitive and a competitive immunoassay Describe the similarities and differences in the following techniques Radioimmunoassay RIA Homogeneous Assay Fluorescent Polarization Immunoassay FPIA EnzymeMultiplied Immunoassay Technique EMIT and EnzymeLinked Immunosorbent Assay ELISA NNN D I Nt IO O N L De nitions sample preparation 7 The process that isolates the analytes from a matrix partitioning The preference or affinity of a compound for one physical phase or state over another the basis of solvent extraction and chromatography internal standard pure organic compounds added to the sample that are similar in analytical behavior to the analyte and not affected by the matrix surrogate spike pure organic compounds that are similar to the analytes of interest in chemical r quot39 quot and 39 but which are not normally found in the sample matrix spike A representative matrix that is spiked with target analytes of interest prior to being taken through the entire analytical procedure in order to evaluate overall precision for an actual matrix headspace method 7 The partitioning of an analytes into the gas phase in the headspace CHEM 4204 Chapter Objectives Clayton State University Dr Susan F Hornbuckle of a closed container from a solid or liquid matrix Based on Henry s Law the concentration of the analyte in the headspace gases is proportional to the concentration of the analyte in the liquid or solid matrix Hydrophilic 7 water soluble polar Hydrophobic water insoluble nonpolar Lipophilic 7 fat soluble nonpolar Lipophobic 7 fat insoluble polar ionization centers 7 functional groups that maybe protonated or deprotonated to form an 1on dry extraction 7 the partitioning of an analytes from a solid matrix into a liquid phase solvent liquidliquid extraction LLE 7 the partitioning of analytes between a nonpolar solvent and an aqueous solution dynamic headspace DHS 7 a partitioning between a solid or liquid matrix and the gas phase where the gas phase is isolated concentrated and analyzed by GC GCMS or HPLC purge and trap PT 7 another name for dynamic headspace solid phase extraction SPE 7 partitioning an analytes between a stationary phase solid and a mobile phase liquid such as with column chromatography TLC and HPLC normalphase SPE 7 Solid Phase Extraction using a polar stationary phase and a nonpolar mobile phase reversephase SPE Solid Phase Extraction using a nonpolar stationary phase and a polar mobile phase Solid phase microextraction SPME Extraction into a solid phase coated on a micro ber The preconcentrated analytes are typically introduced directly in the injection port ofa GC solid phase stirbar extraction SPSE 7 solid phase extraction where the solid phase is coated on the surface of a stir bar thinlayer chromatography TLC 7 solid phase extraction where the solid phase is coated in a glass plate The sample is spotted on the solid phase and the mobile phase solvent is drawn up the plate due to capillary action Immunoassay An analytical technique based on antigenantibody reactions Antigen A substance that when introduced into an organism stimulates an immunological response and production of antibody Antibody A substance produced in an organism in response to the introduction of an antigen Immunogen The drughapten complex that stimulates the production of antibodies Hapten a large protein polyclonal antibody A mix of related antibodies produced when an antigen is introduced into an organism monoclonal antibody A nearly pure antibody produced by a several step procedure that includes the use of hybridoma cells Hybridoma Cells Cells created by a fusion of antibodyproducing cells and cancer cells used to produce monoclonal antibodies competitive immunoassay 7 an immunoassay in which the antigens compete for a limited CHEM 4204 Chapter Objectives Clayton State University Dr Susan F Hornbuckle number of antibody binding sites noncompetitive immunoassay also called immunometric methods 7 an immunoassay in which the antibody is usually present in excess Homogeneous Assay 7 the antibody and the antigen are both in a liquid solution Heterogeneous Assay 7 the antibody is bound to a surface and the antigen is in a liquid solution Radioimmunoassay RIA 7 a competitive heterogeneous immunoassay in which radioactive labeled drug is initially bound to the immobilized antibody If the drug is present in the sample it will replace the radioactive labeled drug there by reducing the radioactivity measurement measured by a Geiger counter Fluorescent Polarization Immunoassay FPIA 7 a competitive homogeneous immunoassay in which a uorescent labeled drug and the drug in the sample compete for binding sites on the antibody The higher the drug concentration in the sample the lower the concentration of labeled drugantibody complex therefore the lower the uorescence polarization measured by a uorimeter EnzymeMultiplied Immunoassay Technique EMIT a competitive homogeneous immunoassay in which a enzymelinked drug and the drug in the sample compete for binding sites on the antibody The free enzymelinked drug catalyzes a reaction that produces a colored product The antibody bound enzymelinked drug is inactive Therefore the higher the concentration of the drug in the sample the more free enzymelinked drug and the higher the concentration of the colored product measured by UVVIS spectroscopy EnzymeLinked Immunosorbent Assay ELISA a competitive heterogeneous immunoassay in which a enzymelinked drug is bound to the bound antibody Drugs in the sample displace the enzymelinked drug thereby increasing the enzyme activity and a color appears ELISA is typically not quantitative The observation of a color change supports the hypothesis that the drug of interest is present CHEM 4204 Chapter Objectives Clayton State University Dr Susan F Hornbuckle Chapter 5 99gt w 0 9 gt1 t O O D I D I D I U3 N D I oo1 uJ ND I 00 Be able to de ne the following terms refractive index numerical aperture empty magni cation spectroscopyand give four examples spectrometryand give one example atomic spectroscopyand give three examples molecular spectroscopyand give three examples Beer s Law radiationless transition emission uorescence phosphorescence singlet state triplet state metastable band A 39 39 39 quot J39 r 39 J quot 39 dispersion interferometry Fourier transform resolution Atomic Absorption AA microspectrophotometry GC GC MS thermal detector Know the approximate magnifying power for each of the following magnifying glass 39 39 39 39 r Be able to explain how each of the following microscopes function and what types of forensic evidence might be analyzed by each compound microscope comparison microscope stereoscopic microscope and polarizing microscope Be able to explain how the numerical aperture relates to the resolution of image Be able to discuss how a change in magnifying power affects the following field of View and depth of focus Be able to list the different regions of the electromagnetic spectrum in order of increasing energy and indicate the event triggered by absorption of energy in each region Be able to use Beer s Law given data ie I supply A and k and you calculate I supply a graph for you to interpret or you graph a few points and extrapolate Be able to describe the three methods in which an excited state species dissipate energy Be able to explain how the number of possible transitions effects band broadening r r r a C Know what types of spectroscopy typically have the least amount of band broadening Be able to draw the simple diagram for an Absorbancetransmittance spectrophotometer Be able to draw the simple diagram for an emission spectrometer and label the components Be able to discuss the similarities and differences of an Absorbancetransmittance spectrophotometer and an Emission spectrometer Be able to draw the simple diagram for an FTIR and label the components Be able to explain how simultaneous dispersion and sequential dispersion differ 16 Be able to explain the relationship between bandwidth and intensity for instruments equipped with a monochromator not FT Be able to describe how UV and VIS spectroscopy are used in today s crime labs Know what types of compounds functional groups will absorb energy in the UV and VIS regions Is UV and VIS spectroscopy considered a destructive or nondestructive method Be able to describe how IR spectroscopy is used in today s crime labs Are these analyses qualitative and or quantitative Is IR considered a destructive or nondestructive method CHEM 4204 Chapter Objectives Clayton State University Dr Susan F Hornbuckle Be able to explain how the data from a IR is interpreted Know the regions where the following bonds absorb CO CC sp2 CH sp3 CH OH Be able to describe the three subregions within the infrared region Which one is most commonly exploited in the forensics lab Be able to describe a hollow cathode lamp and the process of sputtering Know the difference between AA and Flameless AA Know whether the instrumental methods that we have covered in this chapter are qualitative and or quantitative Know how many elements can be measured at a time using AA Explain Is AA considered a destructive or nondestructive method Know the types of spectroscopy that are commonly used with microspectrophotometry Be able to list three examples of physical evidence that may be analyzed using microspectrophotometry Be able to explain how the data from a GC is interpreted Is analysis by GC qualitative andor quantitative Be able to list three types of forensic samples that might be tested by GC Be able to list the limitations of the GC method Be able to draw the simple diagram for a GC and label the components Be able to list and describe three types of detectors for a gas chromatograph Is GC considered a destructive or nondestructive method Do not focus on any of the problems at the end of Chapter 5
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