Reactions of latent prints exposed to blood

We explored whether an undeveloped latent print (fingermark) exposed to blood and later developed by enhancement with blood reagents such as amido black (AB) or leucocrystal violet (LCV) could appear as a genuine blood mark. We examined three different experimental conditions. In Experiment I, fingermark residue only was tested, as a control to confirm that fingermark residue alone does not react with the blood reagents AB and LCV. Experiment II investigated whether latent fingermarks exposed to blood dilutions could be treated with AB or LCV and subsequently appear as a genuine blood mark enhanced with AB or LCV. Experiment III tested whether latent fingermarks exposed to whole blood could be processed with AB or LCV and subsequently appear as a genuine blood mark enhanced with AB or LCV.The present study found that indeed, fingermark residue alone does not react with the blood reagents AB and LCV. In Experiment II, an interaction occurred between the fingermark residue and the diluted blood that caused the ridges to appear a red color. In the present study, this interaction is called a faux blood mark. While the faux blood mark phenomenon occurred most often following exposure to diluted blood, it did not occur consistently, and a predictable pattern could not be established. However, the reaction occurred more frequently following extended fingermark residue drying times. Faux blood marks are distinguishable from genuine blood marks prior to enhancement with blood reagents. Following treatment with blood reagents, it became increasingly difficult to determine whether the enhanced mark was a genuine blood print or a latent fingermark exposed to diluted blood. Latent fingermarks exposed to whole blood often resulted in a void prior to enhancement, but following treatment with blood reagents, were difficult to distinguish from a genuine blood mark enhanced with blood reagents.     

A method for impregnating nylon transfer membranes with leucocrystal violet for enhancing and lifting bloody impressions

The objective of this research project was to demonstrate a quick and easy method for impregnating nylon transfer membranes with leucocrystal violet (LCV) for the purpose of lifting and enhancing impressions made in blood. A stamp that would simulate fine detail found in fingerprints or footwear was used to create impressions on a variety of substrates. Four different LCV formulations were tested to determine the effectiveness of the prepared membranes in lifting and enhancing the impressions. Further investigation involved the feasibility of using the LCV membranes in the field by studying the shelf life and storage of the impregnated membranes and the longevity of the lifted impressions. One of the formations studied demonstrated superior lifting and enhancing capabilities, as well as a prolonged shelf life and a resilience of the lifted impressions, thus proving LCV to be an extremely valuable technique.     

Fluorescence spectra and images of latent fingerprints excited with a tunable laser in the ultraviolet region

Fluorescence spectra of sebum-rich latent fingerprints were studied with a tunable laser for non-destructive fingerprint detection without chemical treatment. The tunable laser consists of a nanosecond pulsed Nd-YAG laser and an optical parametric oscillator (OPO) crystal. The fluorescence spectra and images were measured at various excitation wavelengths in the ultraviolet region by the time-resolved fluorescence method. We have previously reported that a typical fluorescence spectrum of fingerprints consists of two peaks located at c. 330 and 440 nm. In order to determine the wavelength of optimal excitation, excitation spectra were measured at wavelengths ranging from 220 to 310 nm. The fluorescence intensity of the 330 nm peak became maximal with excitation at 280 nm. The images of latent fingerprints on white papers were also measured and the clearest image was obtained with excitation at 280 nm. The influence of continuous irradiation on the fluorescence of fingerprints was measured at the optimal excitation wavelengths. The 330 nm peak was strong at first and decreased with continuous irradiation, whereas the 440 nm peak, which was weak at first, increased gradually.     

A rapid method to detect dried saliva stains swabbed from human skin using fluorescence spectroscopy

Saliva on skin is important in forensic trace evidence. If areas where saliva is present can be outlined, this may lead to DNA analysis and identification. This study describes a rapid and non-destructive method to detect dried saliva on the surface of the skin by fluorescence spectroscopy. Eighty-two volunteers deposited samples of their own saliva on the skin of their ventral forearm. A control sample of water was deposited at three different sites on the contralateral arm. Saliva and water control were then allowed to air-dry. Swab samples were taken from dried saliva and control sites and were dissolved in 0.1M KCl solution. Emission spectra were obtained from the solution and were characterized by a principal maximum at 345-355nm with excitation at 282nm. The fluorescence emission intensity was greater than background readings obtained from the control swab site in 80 of 82 volunteers (approximately 97.6%). The fluorescence profile of saliva samples were similar to those obtained from aqueous samples of pure amylase and tryptophan, an endogenous fluorophore in alpha-amylase. The presence of an emission peak at 345-355nm with excitation at 282nm could provide a strong presumptive indication of saliva deposition.     

Photoluminescence of blood by acidic hydrogen peroxide—A preliminary test

In this study, the authors found that treating blood with 1 M HCl and 2% (w/v) 5-sulfosalicylic acid (SSA) in 1% (v/v) hydrogen peroxide mixture can produce photoluminescence of blood. SSA was added as a blood fixer. The photoluminescence was induced by irradiation of a forensic light source at 505 nm, which was detected using a 550 nm barrier filter. In this experiment, various level of acid and hydrogen peroxide were tested to find the optimal formulation of reagents, spot tests were conducted with diluted blood to test the sensitivity of this reagent, and impressions in blood left on porous/nonporous surfaces were enhanced. The sensitivity of this solution was slightly lower than Bluestar and was similar to leucocrystal violet or leucomalachite green on both porous/non-porous surfaces. The photoluminescence of blood treated with this reagent has been observed over 2 months. Using this reagent, it was possible to observe fingermarks or footwear impressions in blood on a black porous/non-porous surface. Through this, it was found that using this reagent could enhance bloodstains regardless of the porosity or color of the surface.     

Chemical enhancement of footwear impressions in blood on fabric — Part 2: Peroxidase reagents

This study investigates the optimisation of peroxidase based enhancement techniques for footwear impressions made in blood on various fabric surfaces. Four different haem reagents: leuco crystal violet (LCV), leuco malachite green (LMG), fluorescein and luminol were used to enhance the blood contaminated impressions.The enhancement techniques in this study were used successfully to enhance the impressions in blood on light coloured surfaces, however, only fluorescent and/or chemiluminescent techniques allowed visualisation on dark coloured fabrics, denim and leather. Luminol was the only technique to enhance footwear impressions made in blood on all the fabrics investigated in this study.     

Exploring the recovery and detection of messenger RNA and DNA from enhanced fingermarks in blood

Often in the examination of bloodstained fingermarks discussion occurs around whether to prioritise the fingerprint evidence or focus on the biological evidence. Collecting a sample for genetic profiling may result in the loss of ridge detail that could have been used for fingerprint comparison. Fingermark enhancement and recovery methods along with sample collection methods could also compromise downstream genetic analysis. Previous forensic casework has highlighted circumstances where, after enhancement had been performed, it would have been extremely valuable to both identify the body fluid and generate a DNA profile from the same sample.We enhanced depletion series of fingermarks made in blood, using single treatments consisting of aqueous amido black, methanol-based amido black, acid yellow and leucocrystal violet, and exposure to long wave UV light. We then extracted the DNA and RNA for profiling, to assess the recovery and detection of genetic material from the enhanced fingermarks.We have shown that genetic profiling of bloodstained fingermarks can be successful after chemical enhancement; however it may still be necessary to prioritise evidence types in certain circumstances. From our results it appears that even with visible bloodstained fingermarks, leucocrystal violet can reduce the effectiveness of subsequent messenger RNA profiling. Aqueous amido black and acid yellow also have adverse effects on messenger RNA profiling of depleted fingermarks with low levels of cellular material. These results help with forensic decision-making by expanding knowledge of the extent of the detrimental effects of blood-enhancement reagents on both DNA profiling and body fluid identification using messenger RNA profiling.     

Fluorometric determination of pseudocholinesterase activity in postmortem blood samples

A fluorometric assay using 3-(p-hydroxyphenyl) propionic acid (HPPA) was conducted to determine the activity of pseudocholinesterase (ChE) [Enzyme Commission (EC) No. 3.1.1.8] in postmortem blood samples so as to test for organophosphate poisoning. By the enzymatic reaction of ChE, its substrate, benzoylcholine, produces choline, which is oxidized by choline oxidase to generate hydrogen peroxide. HPPA is oxidized by hydrogen peroxide and peroxidase to become the fluorogenic dimer whose concentration is measured fluorometrically at an excitation emission wavelength of 320 nm and an elimination emission wavelength of 404 nm. The selectivity and sensitivity of the present method were found to be superior to those of conventional pH and spectrophotometric methods.     

Improved method for DFO development of latent fingerprints

A new formulation has been developed for DFO stock solution. The working DFO solution, based on the new stock solution, appears to be more stable, has a longer shelf life, and has little effect on inks (and therefore does not cause inks to run). Photoluminescence spectra of latent fingerprints developed with DFO reagent have been measured and the choice of filters for excitation and emission (barrier) have been derived from these measurements. Comparisons of latent fingerprints developed with ninhydrin and DFO have been made using various papers and at various intensities. These comparisons show the much greater sensitivity of DFO developed latent fingerprints. Although enhanced ninhydrin and DFO develop latent fingerprints with similar sensitivity, the DFO process is much simpler.     

Evaluation of Raman spectroscopy for the analysis of colored fibers: a collaborative study

A collaborative study on Raman spectroscopy was carried out by members of the ENFSI (European Network of Forensic Science Institutes) European Fibres Group (EFG) on three dyed fibers: two red acrylics and one red wool. Raman instruments from six different manufacturers were tested as well as nine different laser wavelengths ranging from blue (lambda = 458 nm) to near infrared-NIR (lambda = 1064 nm). This represents the largest comparison study of Raman analytical parameters carried out on identical fiber samples. For the chosen fiber and dye samples, red lasers (lambda = 633 and 685 nm) gave the poorest spectral quality whereas blue (458 nm), green (514 nm) and near infrared lasers (785, 830 and 1064 nm) provided average results. Blue (488 nm) and green lasers (532 nm) globally gave the best quality spectra. Fluorescence problems were often encountered with some of the excitation wavelengths and therefore a flexible Raman instrument equipped with different lasers can be recommended to measure forensic fiber samples. The instrument should also be equipped with a Raman microscope in order to be able to focus on a single fiber. This study shows that Raman spectroscopy usually enables the identification of the main dye present in a colored fiber; however, minor dye components are much more difficult to detect. SERRS (Surface Enhanced Resonance Raman Scattering) techniques give an improvement of the dye's spectral intensity but no spectral improvement was observed for the two red acrylic and red wool fibers tested.     

Determination of MDMA, MDA, MDEA and MBDB in oral fluid using high performance liquid chromatography with native fluorescence detection

This paper describes the analytical methodology for the determination of MDMA, MDA, MDEA and MBDB in oral fluid. After a liquid–liquid extraction, the analysis was carried out by high performance liquid chromatography (HPLC), with fluorescence detection. The detector wavelength was fixed at 285 nm for excitation and 320 nm for emission. The mobile phase, a mixture of phosphate buffer (pH = 5) and acetonitrile (75:25), and the column, Kromasil 100 C8 5 μm 250 mm × 4.6 mm, allowed good separation of the compounds in an isocratic mode in only 10 min. The method was validated and showed good limits of detection (2 ng/mL) and quantitation (10 ng/mL) for all the amphetamine derivatives. No interfering substances were detected. A stability study of these compounds in oral fluid stored at three different temperatures (−18, 4 and 20 °C) over 10 weeks was conducted, showing a time-dependent degradation of the four compounds.     

Application of mist to fingermark detection: Misting with high-boiling-point liquid containing p-dimethylaminocinnamaldehyde and cyanoacrylate

In order to detect latent fingerprints that could be damaged by liquid or powder reagents, non-destructive processes such as gaseous reagents have been developed. In this report, we propose the use of fine mist generated when hot vapor of high-boiling-point liquids is rapidly cooled by surrounding air for fingermark detection. Octyl acetate (OA), 2-phenoxyethanol (2PE), and methyl decanoate (MD) were found to efficiently produce mist when heated to 230°C. By combining these liquids with p-dimethylaminocinnamaldehyde (DMAC) and cyanoacrylate (CN), our team demonstrated effective fluorescence staining of cyano-treated fingermarks using DMAC/OA misting or DMAC/2PE misting, and one-step fluorescence detection of latent fingermarks without cyanoacrylate treatment using DMAC/OA/CN misting or DMAC/MD/CN misting. Fingermark fluorescence was efficiently observed by excitation with a blue LED light (max. wavelength 470 nm) equipped with an interference filter and passing through a 520 nm long-pass filter. We successfully obtained fluorescent images from fingermarks on several substrate materials using the developed misting method.     

A rapid screening procedure for drugs and poisons in gastric contents by direct injection-HPLC analysis

A high performance liquid chromatographic method for toxicological drug screening of gastric content has been developed. The samples were diluted (1:3-1:30) in 0.01 N hydrochloric acid and injected into a reverse phase column for separation by gradient elution. Mobile phase consisted of solvent A (acetonitrile/water 90:10, 0.01 M sodium dodecylsulphate, 0.5% v/v glacial acetic acid) and solvent B (water/acetonitrile 90:10, 0.01 M sodium dodecylsulphate, 0.5% v/v glacial acetic acid); the gradient was programmed from 20 to 80% A in 30 min. The flow was kept constant at 1.5 ml/min. Two home-made internal standards, butyrylsalicylic acid and diacetyltubocurarine with retention times of 5.6 and 21.4 min, respectively, were used. Drugs are identified by matching their relative retention times and UV spectra (200-400 nm) with those contained in a home made library of more than 340 reference compounds (9 analgesics, 22 antidepressants, 30 antihistamines, 14 antihypertensives, 21 antirheumatics, 15 beta-blockers, 9 bronchodilators, 10 Ca antagonists, 14 diuretics, 26 neuroleptics, 25 tranquilizers, and other significant xenobiotic compounds). The fluorometric (FLD) emission spectrum (280-700 nm; excitation wavelength, 230 nm) was used as a further identification. At 50mg/l analyte concentrations, the injection of gastric content after dilution (1:3) produced S/N ratios in the range 8-140. The method is simple, rapid, rather inexpensive and proved to be a useful means of investigation if used in combination with GC-MS screening in blood. On the other hand, the system suffers from a relatively limited sensitivity for compounds with a low UV absorption and from interferences due to the presence in the matrix of some highly UV- and FL-responsive compounds (e.g. tryptophan).     

Visual detection of trace nitroaromatic explosive residue using photoluminescent metallole-containing polymers

The detection of trace explosives is important for forensic, military, and homeland security applications. Detection of widely used nitroaromatic explosives (trinitrotoluene [TNT], 2,4-dinitrotoluene [DNT], picric acid [PA]) was carried out using photoluminescent metallole-containing polymers. The method of detection is through the quenching of fluorescence of thin films of the polymer, prepared by spray coating organic solutions of the polymer, by the explosive analyte. Visual quenching of luminescence (lambda(em) approximately 400-510 nm) in the presence of the explosive is seen immediately upon illumination with near-UV light (lambda(ex)=360 nm). Detection limits were observed to be as low as 5 ng for TNT, 20 ng for DNT, and 5 ng for PA. In addition, experiments with normal production line explosives and their components show that this technology is also able to detect composition B, Pyrodex, and nitromethane. This method offers a convenient and sensitive method of detection of trace nitroaromatic explosive residue.     

Spectrofluorimetric and Spectrophotometric Analysis of Two Analgesic Drugs in Pharmaceutical Formulations and Biological Fluids

Simple, reliable, sensitive, and accurate spectrofluorimetric and spectrophotometric methods were proposed for the determination of two selected analgesic drugs, namely tramadol and morphine, in pharmaceuticals and biological fluids. The proposed methods were based on the oxidation of the studied drugs by Cerium (Ce)IV in an acidic medium. The spectrofluorimetric method is based on measuring the relative fluorescence intensity of Ce(III) arising from Ce(IV) at 350 nm with an excitation wavelength at 250 nm. The spectrophotometric method involves on addition of a known excess of Ce(IV) to the studied drugs in an acid medium, followed by the determination of residual Ce(IV) by reacting with a fixed amount of methyl orange and measuring the absorbance at 510 nm. Different variables affecting the reaction conditions, such as the concentrations of Ce(IV), type and concentration of the acid medium, reaction time, temperature, and the diluting solvents, were carefully studied and optimized.     

二阶导数光谱双波长法测定血中碳氧血红蛋白饱和度的研究

本文将双波长法用于二阶导数光谱中,提出一种新的COHb饱和度测定方法。用本法测定了已知COHb饱和度的标准血样和未知COHb饱和度的检血,都得到了比较满意的结果。     

全血中苯骈二氮杂类药物的胶束电动色谱法分析

目的建立用固相萃取胶束电动毛细管色谱法测定人体全血中苯骈二氮杂艹卓类药物的方法。方法全血以Oasis小柱提取,以克仑特罗为内标,采用未涂层毛细管(75μmID×50.2cm,有效分离长度为40cm),缓冲液为30mmol/LSDS→15mmol/L硼砂→15mmol/L磷酸盐(pH8.2)→18%甲醇。进样条件:压力进样0.5psi×10s,分离电压为25kV,柱温25℃,检测波长为230nm。结果本法分离效率高,9种苯骈二氮杂艹卓类药物的最低检测浓度为5～50ng/ml;血药浓度的线性范围为0.02～1.6μg/ml,日内、日间精密度<12%。结论本法简便、高效、可靠。     

A practical method for the accurate determination of methemoglobin in blood containing carboxyhemoglobin

Kinetics of the oxidation of carboxyhemoglobin (HbCO) by potassium ferricyanide was studied photometrically in a weakly acid solution. An increase in the absorbance at 630 nm reached a maximum within 10 min when over a 100-fold excess of ferricyanide to hemoglobin iron was used. A slight decrease in the absorbance was observed after completion of the reaction when over a 500-fold excess of the reagent was used. In the presence of 0.4% Sterox SE, the absorbance began to decrease without complete oxidation. From these findings, a simple, rapid and accurate method for the determination of methemoglobin (Met-Hb) in blood was devised. The method was compared with two other methods, using 11 blood samples containing various amounts of HbCO, and proved to be suitable for blood containing elevated HbCO as well as for ordinary blood.     

The effect of mark enhancement techniques on the presumptive and confirmatory tests for blood

An investigation into the effects of physical and chemical enhancement on subsequent presumptive and confirmatory tests for human blood is presented. Human blood was deposited onto porous (white 80 gsm paper and brown envelope) and non-porous (tile and linoleum) substrates in a depletion series (30 depletions on non-porous and 20 on porous) and subjected to three ageing periods; 1, 7 and 28?days. A number of enhancement techniques were tested [fluorescence, black magnetic powder (BMP), iron-oxide black powder suspension (PS), cyanoacrylate (CA) fuming, acid violet 17 (AV17), acid yellow 7 (AY7), ninhydrin, DFO and Bluestar Forensic Magnum (BFM) luminol] to evaluate their potential effects on subsequent presumptive and confirmatory tests. AV17 and Bluestar provided the best enhancement and fully enhanced all depletions in the series. The sensitivity of the Kastle-Meyer (KM) (presumptive), Takayama and RSID-Blood tests (confirmatory) was initially investigated to determine the range of detectable depletions. The KM test detected all depletions, whereas the Takayama test detected up to depletion 6 and RSID-Blood detected up to depletion 20 (paper), 10 (envelope), 15 (tile) and 9 (lino). The abilities of these tests to detect blood after enhancement were then observed.A number of techniques resulted in little to no effect on any of the blood tests, whereas adverse effects were observed for others. Ninhydrin and CA fuming caused weak but instantaneous positive KM results whereas methanol-based AV17 and AY7 delayed the reaction by as much as 1?min. The Takayama test was not very sensitive, therefore, its performance was easily affected by enhancement and negative results were often observed. RSID-Blood tests were largely unaffected by chemical enhancement although a drop in positive results was observed for some of the techniques when compared to positive controls.Using a standard procedure for DNA extraction, all the tested blood samples (before and after enhancement) gave a detectable quantity of DNA and were successfully profiled. Out of the 45 samples processed for DNA profiling, 41 gave full profiles, while the remaining showed allele drop out in one or two loci.     

An investigation into the enhancement of fingermarks in blood on fruit and vegetables

A number of studies have reported the successful enhancement of latent fingermarks on fruit and vegetables. A study was set up to identify the most effective technique for the enhancement of fingermarks in blood on various fruit and vegetables. The enhancement techniques targeted different components in blood and consisted of protein stains (e.g. acid black 1), peroxidase reagents (e.g. leuco crystal violet) and amino acid stains (e.g. ninhydrin). Different variables such as the ageing periods of the marks and a diminishing series were employed to assess the suitability and sensitivity of the enhancement techniques.Overall, the use of different protein stains appeared to be the most effective techniques for the enhancement of fingermarks in blood on fruit and vegetables. In addition, the aubergine and cucumber skins appeared to be the most responsive surface to the different chemical techniques during enhancement. On the contrary, little or no enhancement was achieved for fingermarks in blood on the nectarine fruit.