GC-MS/MS测定血液中巴比妥类安眠药物

建立GC-MS/MS测定血液中巴比妥类安眠药物的分析方法。方法通过固相萃取提取并富集血液样品中常见巴比妥类安眠药物,采用离子阱二级质谱定性并定量检测其含量,并优化萃取溶液pH值与气相色谱/二级质谱联用分析条件,对巴比妥类安眠药物进行定量分析。结果巴比妥类安眠药物检出限为0.04μg/mL～0.10μg/mL,回收率为80.3%～92.6%。结论该方法高效、简单,灵敏度高,可用于血液中巴比妥类安眠药物同时定性定量检测。     

双溶剂提取法判定印泥印文形成时间的初探

目的通过双溶剂提取法判定印泥印文的形成时间,为判定文件的真伪提供重要依据。方法选用6种不同品牌的印泥和4种纸张分别制作印泥印文的历时性样本,通过实验确定适用于双溶剂提取法判定印泥印文形成时间的弱提取剂和强提取剂;确定薄层扫描时狭缝的大小和扫描波长;绘制盖印时间与提取率的关系曲线。结果弱提取剂为50％的甲酸溶液,强提取剂为N,N-二甲基甲酰胺,狭缝大小为12．00mm×0．60mm,扫描波长为485nm,得到了盖印印文形成后半年内印泥提取率随印文形成时间变化的关系曲线。结论双溶剂提取法是判定印泥印文形成时间的一种有效方法。     

DNA Typing for the Identification of Old Skeletal Remains from Korean War Victims*

Abstract: The identification of missing casualties of the Korean War (1950–1953) has been performed using mitochondrial DNA (mtDNA) profiles, but recent advances in DNA extraction techniques and approaches using smaller amplicons have significantly increased the possibility of obtaining DNA profiles from highly degraded skeletal remains. Therefore, 21 skeletal remains of Korean War victims and 24 samples from biological relatives of the supposed victims were selected based on circumstantial evidence and/or mtDNA‐matching results and were analyzed to confirm the alleged relationship. Cumulative likelihood ratios were obtained from autosomal short tandem repeat, Y‐chromosomal STR, and mtDNA‐genotyping results, and mainly confirmed the alleged relationship with values over 10⁵. The present analysis emphasizes the value of mini‐ and Y‐STR systems as well as an efficient DNA extraction method in DNA testing for the identification of old skeletal remains.     

Simple and highly effective DNA extraction methods from old skeletal remains using silica columns

The recovery of DNA data from old skeletal remains is often difficult due to degraded and very low yield of extracted DNA and the presence of PCR inhibitors. Herein, we compared several silica-based DNA extraction methods from artificially degraded DNA, DNA with PCR inhibitors and DNA from old skeletal remains using quantitative real-time PCR. We present a modified large-scale silica-based extraction combined with complete demineralization, that enables maximum DNA recovery and efficient elimination of PCR inhibitors. This is performed with high concentration of EDTA solution for demineralization of bone powder followed by QIAamp^® spin columns and buffers from the QIAquick^® PCR purification kit. We have successfully used this modified technique to perform STR analysis for 55-year-old skeletal remains. The results of this study will contribute to solve the forensic cases dealing with skeletal remains.     

The Use of Hemastix® and the Subsequent Lack of DNA Recovery Using the Promega DNA IQTM System

Abstract:  Following implementation of our automated process incorporating the Promega DNA IQ^TM system as a DNA extraction method, a large number of blood-containing exhibits failed to produce DNA. These exhibits had been tested with the Hemastix^® reagent strip, commonly used by police investigators and forensic laboratories as a screening test for blood. Some exhibits were even tainted green following transfer of the presumptive test reagents onto the samples. A series of experiments were carried out to examine the effect of the Hemastix^® chemistries on the DNA IQ^TM system. Our results indicate that one or more chemicals imbedded in the Hemastix^® reagent strip severely reduce the ability to recover DNA from any suspected stain using the DNA IQ^TM magnetic bead technology. The 3,3',5,5'-tetramethylbenzidine (TMB) used as the reporting dye appears to interact with the magnetic beads to prevent DNA recovery. Hydrogen peroxide does not seem to be involved. The Hemastix^® chemistries do not interfere in any way with DNA extraction performed using phenol-chloroform. The incompatibility of the Hemastix^® chemistries on the DNA IQ^TM system forced us to adopt an indirect approach using filter paper to carry out the presumptive test.     

枪击案件中射击残留物提取方法的探讨

目的结合现场勘查提取和实验研究,探讨了几种提取射击残留物的不同方法。方法采用富集浓缩、颗粒收集等方法。结果运用上述方法提取了附着在玻璃、汽车、塑料和织物上弹孔周围的射击残留物。结论收到良好的效果,大大提高了射击残留物提取率和检出率。     

Combined DNA Typing and Protein Identification from Unfired Brass Cartridges,,,

Biological evidence analysis from contact traces is adversely affected by low quantity and quality of DNA. Proteins in these samples contain potentially individualizing information and may be particularly important for difficult surfaces such as brass, where DNA may yield incomplete profiles. In this study, touched unfired brass cartridges were sampled using dry tape or wet swabs and analyzed by separating DNA and protein from the same collected material, thus producing both genomic and proteomic information. DNA recovery was similar for both collection methods, with tape yielding an average of 1.36 ± 1.87 ng and swabs, 1.34 ± 3.04 ng. Analysis by mass spectrometry identified 95 proteins, with the two collection methods showing no significant difference (p = 0.76) in the average number of collected proteins: 44.5 ± 10.9, (tape) versus 47.9 ± 20.4 (swabs). Proteins can be collected from fingerprints at levels necessary to provide identifying information, thus expanding information obtained from challenging evidence.     

Implementation of Prep-n-Go™ Buffer for DNA extraction from buccal swabs

The efficacy of two extraction methods; room temperature and heat protocols was assessed for buccal swabs using the Prep-n-Go™ Buffer. DNA was extracted from buccal swabs using both extraction methods and their effectiveness to produce good quality DNA profiles was evaluated. Heat protocol was found to yield more DNA, however room temperature protocol produced better quality DNA profiles with fewer artefacts when the samples from both extraction methods were amplified directly without any normalisation with the VeriFiler™ Express PCR Amplification Kit.     

室外温度与室内恒温下EC随死后经历时间的变化

目的在室外环境和室内恒温条件下测定昼夜不同时间猪后腿肌肉电导率(electrical conductivity,EC)值,分析并比较两种环境条件下,EC值随死后经历时间(time since death, TSD)变化的规律。方法取5块即刻屠宰的猪后腿肌肉,均分为两份,随机分成两组,分别置于秋季室外环境和室内18℃恒温环境中。在死后10d内分别于早晚8时(每隔12h)取样,测定其浸渍液EC值。结果两种环境条件下,EC随TSD变化的趋势在整体上是一致的,二者相关性均较好(R2室外=0.971,P室外=0.004,R2室内=0.98,P室内=0.002)。室外环境温度下,肌肉的EC值在白天增长明显,夜间增长不明显,尤其环境温度低于13℃时,出现明显的平台现象,与室内恒温条件下肌肉EC值持续上升的变化趋势有显著的差异。结论无论室外环境温度还是室内恒温下,肌肉EC与TSD的相关性均较好,但在具体分析时,应考虑到夜间低温平台期造成的时间延搁。     

组织切片DNA提取的探讨

在医患关系纠纷中,病理组织切片经常被作为至关重要的证据,但对它的检验是目前法医DNA检验的难点。因样本制作固定的时间、保存环境等不同因素的影响,病理组织切片DNA检出率并不高。本文从实际检案角度出发,详细介绍了病理组织切片DNA提取的过程,并分析了其可能影响因素。