用引物Y_3,Y_4和DNA扩增技术作血液(痕)和毛根性别鉴定的研究

作者用引物Y_3、Y_4和DNA聚合酶链式反应(PCR)作微量人类血液(痕)和毛根的性别鉴定。扩增的靶序列位于Y染色体DNA特异3.4kb重复序列中,扩增产物为460bp。检材用量为:新鲜血液0.5μl、血痕纱纤维1mm、毛根单个。20例保存4个月的血痕与2例保存6年半的血痕性别判定结果均正确,无性别记载的保存9～11年的3例血痕显现了清晰的460bpY特异DNA扩增带。15例保存20天的自然脱落毛根性别判定结果均正确。本法省略了检材处理中的酚-氯仿抽提DNA等纯化步骤,既简化了实验操作,又减少了检验过程中外源DNA的污染机会和样品DNA的损耗,使这一性别鉴定方法更符合法医学实践的需要。     

Hair analysis: a worthless tool for therapeutic compliance monitoring

Hair analysis has been presented by some authors as a possible tool of investigation for estimating patients' compliance to long-term therapies. This paper summarises the different publications that have been devoted to this topic and highlights the available statistical data presented to support this proposition. Qualitative results of such determinations may be of some interest but due to the enormous interindividual variations of quantitative data, the idea of using hair analysis to ascertain whether a patient has taken his treatment exactly as prescribed, clearly appears to be inapplicable.     

Arsenic speciation of two specimens of Napoleon's hair

Since 1960, it has been demonstrated by various analytical procedures that high concentrations of arsenic were present in Napoleon's hair. Various authors, indicating that the detected arsenic levels are a consequence of external contamination, have challenged the results of these examinations. We have tested two strands of hair, referenced as Noverraz and Grand Maréchal Bertrand. Samples were incubated 6 h in water at 90 °C, and arsenic speciation was carried out by HPLC–ICP/MS. The residue was injected on a cation-exchange PRP-X200 column that allows the detection of arsenobetaine and on an anion-exchange PRP-X100 column to test for the mineral forms. In these conditions, the inorganic species As(III), As(V) and their metabolites (DMA and MMA) were separated. Analysis of hair samples highlighted massive amounts of total arsenic (42.1 and 37.4 ng/mg). Arsenical species found in the two samples of analyzed hair are distributed in the following: As(III) 31.1 and 44.7%; As(V) 66.3 and 53.2%; DMA 0.42 and 0.15%. Traces of MMA were detected, and 2% of the species could not be identified. These results prove that more than 97% of the arsenic found in the hair of Napoleon is in inorganic form, which is consistent with a chronic intoxication to arsenic.     

Simultaneous analysis of six amphetamines and analogues in hair, blood and urine by LC-ESI-MS/MS: Application to the determination of MDMA after low Ecstasy intake

A rapid and sensitive method using LC-MS/MS triple stage quadrupole for the determination of traces of amphetamine (AP), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA, “ecstasy”), 3,4-methylenedioxyethamphetamine (MDEA), and N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MBDB) in hair, blood and urine has been developed and validated. Chromatography was carried out on an Uptisphere ODB C₁₈ 5 μm, 2.1 mm × 150 mm column (Interchim, France) with a gradient of acetonitrile and formate 2 mM pH 3.0 buffer. Urine and blood were extracted with Toxitube A^® (Varian, France). Segmented scalp hair was treated by incubation 15 min at 80 °C in NaOH 1 M before liquid–liquid extraction with hexane/ethyl acetate (2/1, v/v). The limits of quantification (LOQ) in blood and urine were at 0.1 ng/mL for all analytes. In hair, LOQ was <5 pg/mg for MA, MDMA, MDEA and MBDB, at 14.7 pg/mg for AP and 15.7 pg/mg for MDA. Calibration curves were linear in the range 0.1–50 ng/mL in blood and urine; in the range 5–500 pg/mg for MA, MDMA, MDEA and MBDB, and 20–500 pg/mg for AP and MDA. Inter-day precisions were <13% for all analytes in all matrices. Accuracy was <20% in blood and urine at 1 and 50 ng/mL and <10% in hair at 20 and 250 pg/mg. This method was applied to the determination of MDMA in a forensic case of single administration of ecstasy to a 16-year-old female without her knowledge during a party. She suffered from hyperactivity, sweating and agitation. A first sample of urine was collected a few hours after (T + 12 h) and tested positive to amphetamines by immunoassay by a clinical laboratory. Blood and urine were sampled for forensic purposes at day 8 (D + 8) and scalp hair at day 60 (D + 60). No MDMA was detected in blood, but urine and hair were tested positive, respectively at 0.42 ng/mL and at 22 pg/mg in hair only in the segment corresponding to the period of the offence, while no MDA was detectable. This method allows the detection of MDMA up to 8 days in urine after single intake.     

Hair analysis for cocaine: Factors in laboratory contamination studies and their relevance to proficiency sample preparation and hair testing practices

Hair samples were contaminated by rubbing with cocaine (COC) followed by sweat application, multiple shampoo treatments and storage. The samples were then washed with isopropanol for 15 min, followed by sequential aqueous washes totaling 3.5 h. The amount of drug in the last wash was used to calculate a wash criterion to determine whether samples were positive due to use or contamination. Analyses of cocaine and metabolites were done by LC/MS/MS. These procedures were applied to samples produced by a U.S. government-sponsored cooperative study, in which this laboratory participated, and to samples in a parallel in-house study. All contaminated samples in both studies were correctly identified as contaminated by cutoff, benzoylecgonine (BE) presence, BE ratio, and/or the wash criterion. A method for determining hair porosity was applied to samples in both studies, and porosity characteristics of hair are discussed as they relate to experimental and real-world contamination of hair, preparation of proficiency survey samples, and analysis of unknown hair samples.     

液氮冷冻单根毛发根直接PCR进行DNA分型

首次用液氮冷冻单根毛发根，再经三次94℃高温处理，直接PCR扩增，从68根毛发根中，成功地检测了62根毛发根的人类vWF基因40内含子VNTR，检出率达91．2％。本方法简便、快速、实用，在法医学检验及群体遗传学研究中有重要应用价值。     

Blood and sperm traces on human hair. A study on preservation and detection after 3-month outdoor exposure

Hair can retain important biological traces for forensic investigations. Forensic scientists are used to looking for such traces on clothing and skin of victims, however, when decomposition kicks in and all that remains of the victims is the skeleton, hair may be the only tissue representing the surface of the body at the time of a crime on which biological traces of an aggressor may have been left and still be detectable. Given the lack of research on this topic, this pilot study aims to assess the capacity of hair to retain semen and blood in hair, and the possibility to detect these fluids with well-known techniques and to obtain a useful genetic profile even when exposed to environmental conditions (Open Natural Environment (woods), Open Man Made Environment (urban)) for three months.Results showed that both traces were always visible and detectable with almost all techniques in the Control Environment, while in the two open environments some difficulties arose. However, biomolecular analysis was effective up to three months on both fluids in the Natural Environment and up to two months and one week respectively on blood and semen in the Man Made Environment. The Combur Test, OBTI, and Luminol were effective on blood up to three months in both environments while Sperm-HY-Liter and observation of cellular components were effective on semen up to at least 1 month and PSA testing was positive up to 1 week in both environments.The present work can be considered an encouraging starting point for the analysis of biological traces on hair in forensic contexts, regardless of the PMI, since blood and semen related to a crime may survive.     

Capillary electrophoresis: a new tool in forensic toxicology. Applications and prospects in hair analysis for illicit drugs

Capillary electrophoresis, the modern approach to instrumental electrophoresis, is probably the most rapidly expanding analytical technique that has appeared in recent years. In the hands of forensic toxicologists, capillary electrophoresis (CE) represents a powerful new analytical tool, which has proved suitable for the investigation of illicit drugs in seized preparations and also in complex biological matrices, among which is hair. CE can be applied according to different separation mechanisms, and among those that are toxicologically relevant are capillary zone electrophoresis and micellar electrokinetic capillary chromatography, which display different selectivities. For the investigation of hair for drugs of abuse, capillary electrophoresis proved effective, providing simultaneous determinations of different drugs without derivatization, with acceptable sensitivity (typically better than 1 ng of drug per mg of hair). The possibility of carrying out determinations of the same analytes, based on different separation mechanisms (capillary zone electrophoresis and micellar electrokinetic chromatography) with the same instrumentation, simply changing the buffer composition, provides an interesting possibility of ‘internal’ confirmation of the results.     

The determination of cocaine in hair: a review

The explosion of literature related to the analysis of hair for cocaine and its products is reviewed. In the commonly accepted applications of hair testing for cocaine, those related to criminal or civil investigations and pharmacotoxicologic studies occupy most of the relevant published work. This review uses detailed, ‘binary’ (yes/no) tables to demonstrate trends in the literature, and allows researchers and caseworkers quick access to the literature most important for answering a variety of questions.     

Segmental hair analysis for cocaine and heroin abuse determination

Segmental hair analysis was performed to obtain information about the history of drug abuse of subjects in a rehabilitation programme. The analytical data from hair samples were correlated, when possible, with urine analysis and to toxicological anamnesis. Toxicological analysis of hair seems to be a valid tool in this specific field.