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" Development of a combined DNA and drug extraction methodology for forensic toxicology application "
Kashkary, Loay M. T.
Haswell, S. J.; Greenway, Gillian; Pamme, Nicole
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Latin Dissertation
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| Record Number
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831681
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TLets668206
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| Main Entry
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Kashkary, Loay M. T.
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| Title & Author
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Development of a combined DNA and drug extraction methodology for forensic toxicology application\ Kashkary, Loay M. T.Haswell, S. J.; Greenway, Gillian; Pamme, Nicole
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| College
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University of Hull
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| Date
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2014
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2014
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Thesis (Ph.D.)
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| Abstract
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Biological samples recovered at crime scenes may contain unsuspected and valuable evidence, such as illicit drugs, in addition to nucleic acids. Deoxyribonucleic acid (DNA) analysis provides valuable information to identify a suspect or victim, as well as to exclude an innocent individual as the perpetrator of a crime. Identification of drugs can also be very informative for forensic investigation to determine whether a perpetrator committed a crime under the influence of illicit substances. In the field of forensic analysis, sample preparation for identifying both DNA and drugs of abuse represents a challenge due to limited sample quantity and only trace levels of target analytes present in the matrices. As a result, an analytical approach has been developed to enable the combined extraction of DNA and four amphetamines (amphetamine [AM], methamphetamine [MA], 3,4-methylenedioxyamphetamine [MDA], and 3,4-methylenedioxymethamphetamine hydrochloride [MDMA]) from a small amount of sample (50 μl) using a single extraction procedure. This study has focused on solid-phase extraction (SPE) using inorganic silica-based matrices as sorbents to facilitate such sample processing. The advantages of using inorganic silica-based monoliths are due to the simple fixation of the material in a column or within a microfluidic device, their mechanical stability with organic solvents, the availability of simple surface modifications to enable the desired chemical interaction with the target molecules, and a unique bimodal structure that allows a large surface area with minimum back pressure. A dual-phase SPE method was developed consisting of silica beads modified with octadecyl groups packed inside a luer lock adapter for amphetamine extraction coupled in series with a silica-based monolith for DNA extraction within a microfluidic system for a fully combined genetic and drug extraction system. The proposed method was effective for the extraction of the target drugs from a spiked buffer and artificial urine giving an average recovery greater than 70% and 50%, respectively, with high reproducibility (˂ 15% RSD). The limits of detection were 0.6 μg ml⁻¹ for AM and MA, 0.7 μg ml⁻¹ for MDA, and 0.8 μg ml⁻¹ for MDMA with linear calibration curves between 0.625 and 20 μg ml⁻¹. The method was also able to extract DNA from the spiked TE buffer and urine sample with average extraction efficiencies of 36% and 30%, respectively, which were successfully amplified via the polymerase chain reaction (PCR). The proposed method is not only suitable for the combined extraction of DNA and amphetamines from a limited sample size, but also reduces sample handling and potential contamination. This method could, in future, be applied to anti-doping analysis for the detection of doping agents and conducting DNA profiling as evidence to ascertain whether samples belong to the right athletes.
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Chemistry
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Haswell, S. J.; Greenway, Gillian; Pamme, Nicole
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University of Hull
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