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Salas A Prieto L Montesino M Albarrán C Arroyo E Paredes-Herrera MR Di Lonardo AM Doutremepuich C Fernández-Fernández I de la Vega AG Alves C López CM López-Soto M Lorente JA Picornell A Espinheira RM Hernández A Palacio AM Espinoza M Yunis JJ Pérez-Lezaun A Pestano JJ Carril JC Corach D Vide MC Alvarez-Iglesias V Pinheiro MF Whittle MR Brehm A Gómez J 《Forensic science international》2005,150(2-3):191-198
A qualitative and quantitative analytical method was developed and validated for the determination of 49 licit and illicit drugs in oral fluid. Small oral fluid samples, volume 1mL, were collected from volunteers using a modified Omni-Sal device and the analytes were extracted from an oral fluid/buffer mixture using a single Bond Elut Certify solid phase extraction cartridge. Liquid chromatography-tandem mass spectrometry (LC-MS-MS) and gas chromatography-repetitive full scan mass spectrometry (GC-MS) were used in parallel to analyze the extracts for the targeted drugs. Extracts were analyzed by GC-MS in their underivatized form and as their pentafluoropropionyl derivatives. Deuterated internal standards were used for quantification of drugs of abuse by LC-MS-MS to minimize matrix effects. Methadone-d(9) and tumoxetine were used as the internal standards for quantification of non-derivatized and derivatized analytes respectively by GC-MS. Linearity was demonstrated over the range 5-200 ng/mL and limits of detection were less than 4 ng/mL for each drug analyzed. The method demonstrated acceptable recoveries for most of the analytes and good intra- and inter-day precision. Acquisition of data by repetitive full scan GC-MS allows the addition of further analytes to the target menu. 相似文献
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Crespillo M Paredes MR Prieto L Montesino M Salas A Albarran C Alvarez-Iglesias V Amorin A Berniell-Lee G Brehm A Carril JC Corach D Cuevas N Di Lonardo AM Doutremepuich C Espinheira RM Espinoza M Gómez F González A Hernández A Hidalgo M Jimenez M Leite FP López AM López-Soto M Lorente JA Pagano S Palacio AM Pestano JJ Pinheiro MF Raimondi E Ramón MM Tovar F Vidal-Rioja L Vide MC Whittle MR Yunis JJ Garcia-Hirschfel J 《Forensic science international》2006,160(2-3):157-167
We report here a review of the seventh mitochondrial DNA (mtDNA) exercise undertaken by the Spanish and Portuguese working group (GEP) of the International Society for Forensic Genetics (ISFG) corresponding to the period 2003-2004. Five reference bloodstains from five donors (M1-M5), a mixed stain of saliva and semen (M6), and a hair sample (M7) were submitted to each participating laboratory for nuclear DNA (nDNA; autosomal STR and Y-STR) and mtDNA analysis. Laboratories were asked to investigate the contributors of samples M6 and M7 among the reference donors (M1-M5). A total of 34 laboratories reported total or partial mtDNA sequence data from both, the reference bloodstains (M1-M5) and the hair sample (M7) concluding a match between mtDNA profiles of M5 and M7. Autosomal STR and Y-STR profiling was the preferred strategy to investigate the contributors of the semen/saliva mixture (M6). Nuclear DNA profiles were consistent with a mixture of saliva from the donor (female) of M4 and semen from donor M5, being the semen (XY) profile the dominant component of the mixture. Strikingly, and in contradiction to the nuclear DNA analysis, mtDNA sequencing results yield a more simple result: only the saliva contribution (M4) was detected, either after preferential lysis or after complete DNA digestion. Some labs provided with several explanations for this finding and carried out additional experiments to explain this apparent contradictory result. The results pointed to the existence of different relative amounts of nuclear and mtDNAs in saliva and semen. We conclude that this circumstance could strongly influence the interpretation of the mtDNA evidence in unbalanced mixtures and in consequence lead to false exclusions. During the GEP-ISFG annual conference a validation study was planned to progress in the interpretation of mtDNA from different mixtures. 相似文献
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Regueiro M Carril JC Pontes ML Pinheiro MF Luis JR Caeiro B 《Forensic science international》2004,143(1):61-63
A genetic study of 15 autosomal STRs is carried out (D2S1338, D3S1358, D5S818, D7S820, D8S1 79, D13S317, D16S359, D18S51, D19S433, D21S11, CSF1PO, FGA, TPOX, THO1, VWA) in a sample of unrelated Tutsis. The molecular phenotypes were determined by means of multiplex strategies (AmpFlSTR Identifiler PCR Amplification Kit, Applied Biosystems) followed by capillary electrophoresis. 相似文献
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The aim of this study is to assess the utility of the STR D5S373 in human identification. PCR amplification and electrophoretic separation were optimized in order to achieve unambiguous phenotyping. We concluded that primer concentration and annealing temperature are the main factors affecting the specificity of PCR. In our population survey including three human major groups (Europe, Sub-Saharan Africa, and Asia), up to six alleles and six interalleles have been found ranging in size from 86 to 101 bp. The phenotypes were determined using horizontal polyacrylamide gel electrophoresis, a technique which has turned out to be suitable for separating fragments as close as 1 bp. In each population, the genotype frequencies conformed to the expectations of genetic equilibrium. Sequence studies were carried out to make the allele nomenclature fit to ISFH recommendations. Results from our population analysis of D5S373 show clear differences in allelic frequency patterns among the three major human groups examined. Human identification parameters estimated from our study are similar to those obtained for other STRs currently used in DNA testing. 相似文献
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