Synthesis and evaluation of ninhydrin analogues as reagents for the development of latent fingerprints on paper surfaces

Thirteen ninhydrin analogues have been synthesized and evaluated as amino acid-specific fingerprint reagents. All of the ninhydrin analogues tested developed latent fingerprints on paper surfaces in a manner similar to ninhydrin itself. The photoluminescent enhancement of prints developed with each reagent was assessed after treatment with zinc(II) or cadmium(II). The results obtained have been related to the effects of substitution on the ninhydrin skeleton. Two compounds in particular, 5-methoxyninhydrin and benzo[f]ninhydrin, were shown to offer operational advantages over ninhydrin on a number of difficult surfaces. Prints developed with either reagent show stronger room temperature luminescence after zinc(II) or cadmium(II) treatment in comparison with equivalent prints developed with ninhydrin. Both 5-methoxyninhydrin and benzo[f]ninhydrin have been shown to be effective on surfaces where background luminescence precludes the successful enhancement of ninhydrin-developed prints.     

Reagents for the chemical development of latent fingerprints: scope and limitations of benzo[f]ninhydrin in comparison to ninhydrin

Benzo[f]ninhydrin was compared to ninhydrin for fingerprint development on paper. Overall, the performance of ninhydrin on exhibits was slightly better than that of benzo[f]ninhydrin. The significant advantages of the benzo[f]ninhydrin over ninhydrin were the much stronger fluorescence it gave after treatment with zinc salts and a slightly quicker reaction under ambient conditions. This fluorescence is, however, similar to that obtained with other reagents, such as DFO or ninhydrin analogs. These advantages apparently are not sufficient to justify regular usage of benzo[f]ninhydrin, especially when one considers its low solubility and high cost.     

Dual fingerprint reagents with enhanced sensitivity: 5-methoxy- and 5-methylthioninhydrin

"Dual fingerprint reagents" are chemical formulations which produce with latent fingerprints in a single step, impressions that are both colored and fluorescent. Pre-mixed solutions of the two commercially available ninhydrin analogues, 5-methoxyninhydrin (MN) and 5-methylthioninhydrin (MTN) with zinc or cadmium salts, are true dual reagents. They are much more sensitive than the parent dual reagent, ninhydrin/ZnCl(2). The main advantage of the new formulations is that they can be used at room temperature, with no need to cool the sample to liquid nitrogen temperature. At 0.05% concentration, which is 10-fold lower than the common ninhydrin working solution, MTN/ZnCl(2) is as sensitive as DFO in the fluorescence mode and considerably more sensitive in the color mode. MTN is also slightly cheaper than DFO.     

Genipin, a novel fingerprint reagent with colorimetric and fluorogenic activity, part II: optimization, scope and limitations

Genipin, a hydrolytic product of geniposide extracted from gardenia fruit, was thoroughly studied as a potential fingerprint reagent, and optimal conditions for fingerprint development have been determined. Latent fingerprints on paper items that have been treated with a non-ink running formulation containing 0.17% of the reagent, showed up as both colored and fluorescent images. On brown wrapping paper and on papers with highly luminescent backgrounds, genipin developed more visible and clearer prints than did classical reagents such as ninhydrin or DFO. Another potential advantage of genipin is that it is totally harmless and an environmentally friendly reagent.     

防止茚三酮溶液扩散纸张字迹的研究

目的 解决茚三酮溶液在显现手印同时对纸张上字迹产生扩散作的问题;方法 分别采用茚三酮丙酮溶液,茚三酮氟利昂溶液,茚三酮石油谜溶液,茚三酮庚烷溶液作比较。结果 钢笔字迹用4种溶液均不易扩散;纸张上其它成份字迹用后两种方法显现手印时可有效保存纸张上文字原貌。     

Fingerprint reagents with dual action: color and fluorescence

We define "dual fingerprint reagents" as chemical formulations that produce with latent fingerprints in one stage impressions that are both colored and fluorescent. Solutions containing ninhydrin and group IIb metal salts appear to be true dual reagents. Application of these formulations to latent fingerprints on paper is as efficient as the two-step process beginning with ninhydrin and followed by treatment with metal salt. In the color mode, fingerprint detectability with the two ninhydrin-metal salt reagents (one with zinc chloride and the other with cadmium chloride) is comparable with that of ninhydrin itself, in spite of the difference in color. The sensitivity is significantly higher in the fluorescence mode. To view the latent impressions the exhibits are treated with ninhydrin-metal salt reagents and observed under white light illumination and under fluorescence conditions. Cooling to liquid nitrogen temperature enhances the fluorescence considerably. In the shorter wavelength domain, ninhydrin-metal salt reagents exhibit higher sensitivity than the recently reported dual reagent, genipin. The latter is advantageous, however, in the longer wavelength domain, on paper items with strong self-fluorescence, such as brown wrapping paper or paper printed with fluorescent ink. Upon reduction of the ninhydrin concentration 10-fold, ninhydrin-metal salt formulations become purely fluorogenic reagents; no color is noticed but the fluorescence is as intense as with concentrated solutions. Working at lower concentrations is an advantage from ecological and economical viewpoints.     

Latent Fingerprint Development on Thermal Paper Using Traditional Ninhydrin and 1,2‐indanedione

Thermal paper poses a significant challenge to latent print development as it tends to change color when traditional fingerprint development formulations are applied to it. In this study, the optimal components of ninhydrin, 1,2‐indanedione, 1,8‐diazafluoren‐9‐one (DFO), and 5‐methylthioninhydrin (5‐MTN) for yielding clear fingerprints on thermal paper were determined by systematically adjusting the relative amounts of the reagents, polar solvents, and the nonpolar diluent petroleum ether, followed by validation on text‐printed thermal paper. Specifically, 3.0% ethyl acetate as the polar solvent in petroleum ether was found to be the optimal combination; the optimal dilution ratios of ninhydrin, DFO, and 5‐MTN original solutions with petroleum ether were 1 to 2, 11, and 7, respectively. The optimal concentration of 1,2‐indanedione in petroleum ether was 0.125 g/L, with a string of 0.5% ethyl acetate in petroleum ether.     

Ninhydrin Thiohemiketals: Basic Research Towards Improved Fingermark Detection Techniques Employing Nano‐Technology*

Abstract: In the first part of a comprehensive research project towards more efficient application of nano‐technology to fingerprint visualization, we investigated the possibility of more selective binding of gold nanoparticles (NP) to fingerprint material. We synthesized derivatives of ninhydrin and 1,2‐indanedione containing loosely bound thiol groups. In particular: thiohemiketals (THK) of ninhydrin, and thioketals of 1,2‐indanedione were prepared and tested as potential fingerprint reagents. By reacting ninhydrin with various thiols we were able to produce a series of novel THK, bearing the SR group always at C2. Ninhydrin THK reacted with amino acids to produce the expected Ruhemann’s purple, and they also developed latent fingermarks on paper in a similar manner to ninhydrin. Ketals and thioketals derived from 1,2‐indanedione reacted neither with amino acids nor with latent fingermarks. In the second part of the research, the thiols which are formed on the ridges as byproducts of the reaction with amino acids will be tested for their potential as stabilizers for gold NP that will become covalently bound to the fingerprint ridges.     

Chemical enhancement of footwear impressions in blood on fabric — Part 3: Amino acid staining

Enhancement of footwear impressions, using ninhydrin or ninhydrin analogues is not considered common practice and such techniques are generally used to target amino acids present in fingermarks where the reaction gives rise to colour and possibly fluorescence. Ninhydrin and two of its analogues were used for the enhancement of footwear impressions in blood on various types, colours and porosities of fabric. Test footwear impressions on fabric were prepared using a specifically built rig to minimise the variability between each impression. Ninhydrin enhancement of footwear impressions in blood on light coloured fabric yielded good enhancement results, however the contrast was weak or non-existent on dark coloured fabrics. Other ninhydrin analogues which have the advantage of fluorescence failed to enhance the impressions in blood on all fabrics. The sequential treatment of impressions in blood on fabric with other blood enhancing reagents (e.g. protein stains and heme reagents) was also investigated.     

Determination of efficacy of fingermark enhancement reagents; the use of propyl chloroformate for the derivatization of fingerprint amino acids extracted from paper

The analysis of the constituents of fingerprints has been described numerous times, mainly with the purpose of determining the aging effect on fingerprints or showing the differences between donors or groups of donors. In this paper we describe the use of derivatized amino acids to determine the efficacy of the visualization reagents 1,8-diazafluoren-9-one (DFO) and ninhydrin. At present certain conditions are used for the application of these reagents, as determined by trial-and-error investigations, to the effect on fingerprints. The recovery of amino acids from a porous surface can be used as a measure for the efficacy of a visualization agent.In this paper we describe a method for the determination of the amount of amino acid left after reaction with well known fingerprint visualization reagents. This will allow a more scientific approach to method development for fingermark enhancement techniques. Furthermore, investigations on the influence of the concentration of fingermark amino acids, the order of application of and exposure time to reagents and the influence of age of the amino acids were carried out. These studies have resulted in a broader understanding of the mechanism involved in visualization of fingermarks using DFO and ninhydrin.     

手印相纸转印显现法的研发与应用

目的 在现有手印转印显现法研究基础上进行改进创新,找到一种操作简便快捷、适用手印种类和客体更广泛、更方便于在勘查现场实施的新型方法.方法 选择滤纸、胶带、相纸分别作为转印载体针对非渗透性客体上手印进行转印显现提取,对比选择出效果较好的转印载体,并检验其应用条件和效果.结果 研究出新型手印转印显现方法——相纸转印显现法....     

Amino acid alanine reactivity with the fingerprint reagent ninhydrin. A detailed ab initio computational study

Ninhydrin is one of the most widely used reagents for chemical development of fingerprints on porous surfaces. The detection is based on the reaction of ninhydrin with a monoacidic component of the fingerprint to form an intensively colored compound named Ruhemann's Purple. A computational study of the mechanisms and reaction energetics of the formation of Ruhemann's Purple from ninhydrin and alanine is presented. Such a study is significant from a forensic science point of view because of the strong interest in the forensic chemistry and law enforcement communities in developing alternatives to the current generation of ninhydrin like chemicals for the detection and development of latent fingerprints. Information about the mechanism of reaction between ninhydrin and amino acids can ultimately help to design compounds with stronger chromo-fluorogenic properties in aid of detecting fingerprints at crime scenes. The three most accepted mechanisms of formation have been considered using ab initio quantum mechanical calculations. At relatively high temperature ( approximately 100 degrees C) all three mechanisms are energetically feasible. However since it is recommended that forensic analyses be performed at room temperature, a revised mechanism is proposed for the formation of Ruhemann's Purple under this condition.     

纸张上潜在手印三种前处理方法对DNA检验的影响

目的通过比较常见纸张上潜在手印盲提法与显现后精准提取法的接触DNA检出率,探讨常见纸张上接触DNA前处理的优选方案。方法比较五种常见纸张上使用盲提法和显现潜在手印(茚二酮显现法、茚三酮熏显法)后精准提取法采集的接触DNA样本检出率。结果粗糙日历纸盲提的接触DNA检出率为17.8%,通过茚二酮法和茚三酮法显现的潜在手印所提取的DNA检出率分别为75.6%、77.8%;光滑日历纸三种方法所提取的接触DNA检出率为4.4%、11.1%、11.1%;A4复印纸三种方法接触DNA检出率为20%、37.8%、66.7%;牛皮纸三种方法接触DNA检出率为20%、68.9%、64.4%;快递纸袋的三种方法接触DNA检出率为2.2%、6.7%、46.7%。结论不同纸张上潜在手印经显现后接触DNA检出率不同,通过茚二酮或茚三酮显示潜在手印后精准提取DNA的前处理方法相较于盲提法的接触DNA检出率高。实战中可应用此类方法同时获得手印与DNA分型,以有效提高证据力。     

茚三酮显现汗潜指印的三种操作方法对DNA检测的影响

目的探讨并比较茚三酮显现汗潜指印的3种操作方法,即溶液浸泡法、涂抹法和喷雾法对后续DNA检测带来的影响。方法取16名志愿者的指印分4组,分为茚三酮浸泡法、涂抹法和喷雾法以及空白组,然后提取DlNA进行定量和STR分型检测。结果3种操作方法都会减少汗潜指印DNA的量,喷雾法的损失量最大,浸泡法和涂抹法结果比较接近。结论现场可疑指印检材,应根据检验需要决定指印显现和DNA提取的先后顺序。     

Thermal development of latent fingermarks on porous surfaces—Further observations and refinements

In a further study of the thermal development of fingermarks on paper and similar surfaces, it is demonstrated that direct contact heating of the substrate using coated or ceramic surfaces at temperatures in excess of 230 °C produces results superior to those obtained using hot air. Fingermarks can also be developed in this way on other cellulose-based substrates such as wood and cotton fabric, though ridge detail is difficult to obtain in the latter case. Fluorescence spectroscopy indicates that the phenomena observed during the thermal development of fingermarks can be reproduced simply by heating untreated white copy paper or filter paper, or these papers treated with solutions of sodium chloride or alanine. There is no evidence to suggest that the observed fluorescence of fingermarks heated on paper is due to a reaction of fingermark constituents on or with the paper. Instead, we maintain that the ridge contrast observed first as fluorescence, and later as brown charring, is simply an acceleration of the thermal degradation of the paper. Thermal degradation of cellulose, a major constituent of paper and wood, is known to give rise to a fluorescent product if sufficient oxygen is available [1], [2], [3], [4], [5]. However, the absence of atmospheric oxygen has only a slight effect on the thermal development of fingermarks, indicating that there is sufficient oxygen already present in paper to allow the formation of the fluorescent and charred products. In a depletion study comparing thermal development of fingermarks on paper with development using ninhydrin, the thermal technique was found to be as sensitive as ninhydrin for six out of seven donors. When thermal development was used in sequence with ninhydrin and DFO, it was found that only fingermarks that had been developed to the fluorescent stage (a few seconds of heating) could subsequently be developed with the other reagents. In the reverse sequence, no useful further development was noted for fingermarks that were treated thermally after having been developed with ninhydrin or DFO. Aged fingermarks, including marks from 1-year-old university examination papers were successfully developed using the thermal technique.     

Chemical development of latent fingerprints: 1,2-indanedione has come of age

The performance of 1,2-indanedione as a latent fingerprint reagent on some types of paper was found to exceed that of DFO, the leading fluorogenic fingerprint reagent. It even exceeds the performance of the sequence, DFO, followed by ninhydrin. No new prints could be observed when ninhydrin was applied after indanedione. On a large number of actual exhibits (used checks) indanedione developed 46% more identifiable prints than the sequence DFO-ninhydrin. A standard procedure for fingerprint development by indanedione is proposed. Best results are obtained with a 0.2% indanedione solution in HFE7100 solvent containing 7% ethyl acetate, but no acetic acid. It can be recommended to start using 1,2-indanedione, which is already commercially available, in actual fingerprint casework.     

The application of infrared chemical imaging to the detection and enhancement of latent fingerprints: method optimization and further findings

Fourier transform infrared (FTIR) chemical imaging allows the collection of fingerprint images from backgrounds that have traditionally posed problems for conventional fingerprint detection methods. In this work, the suitability of this technique for the imaging of fingerprints on a wider range of difficult surfaces (including polymer banknotes, various types of paper, and aluminum drink cans) has been tested. For each new surface, a systematic methodology was employed to optimize settings such as spectral resolution, number of scans, and pixel aggregation in order to reduce collection time and file-size without compromising spatial resolution and the quality of the final fingerprint image. The imaging of cyanoacrylate-fumed fingerprints on polymer banknotes has been improved, with shorter collection times for larger image areas. One-month-old fingerprints on polymer banknotes have been successfully fumed and imaged. It was also found that FTIR chemical imaging gives high quality images of cyanoacrylate-fumed fingerprints on aluminum drink cans, regardless of the printed background. Although visible and UV light sources do not yield fingerprint images of the same quality on difficult, nonporous backgrounds, in many cases they can be used to locate a fingerprint prior to higher quality imaging by the FTIR technique. Attempts to acquire FTIR images of fingerprints on paper-based porous surfaces that had been treated with established reagents such as ninhydrin were all unsuccessful due to the swamping effect of the cellulose constituents of the paper.     

ESDA processing and latent fingerprint development: the humidity effect

The influence of humidification in the ESDA process on subsequent development of fingerprints on paper items was studied. It was found that, while the DFO process is nearly insensitive to previous humidification, fingerprint development with ninhydrin or with indanedione can be significantly affected by previous humidification of the paper.     

Optimisation and evaluation of 1,2-indanedione for use as a fingermark reagent and its application to real samples

1,2-indanedione is an emerging fingermark reagent used on porous surfaces. The general consensus is that this reagent is at least as sensitive as DFO, with some research showing higher sensitivity for 1,2-indanedione as opposed to DFO. However, a number of discrepancies existed in the literature as to which formulation and which development procedure produces optimal results. This project set out to investigate the best formulation and development procedure under Australian conditions, encompassing all published recommendations as well as some novel approaches. 1,2-indanedione formulations were compared with respect to initial colour, fluorescence, concentration of reagent, acetic acid concentration, and the effect of different carrier solvents. Numerous development conditions were investigated, including a conventional oven, a heat press and humidity. Further enhancement using metal salt treatment and liquid nitrogen was also evaluated. The heat press set at 165 degrees C for 10s proved to give the best initial colour and most intense luminescence. Secondary metal salt treatment improved initial colour and luminescence. The Polilight, the VSC 2000, and the Condor Chemical Imaging macroscope have been used to detect the fingerprints developed with 1,2-indanedione on a variety of high- and low-quality porous and semi-porous surfaces, with impressive results overall. Laboratory and field tests were conducted to compare 1,2-indanedione with DFO and ninhydrin as well as to investigate the position of 1,2-indanedione in the sequence of reagents for fingermark detection on porous surfaces. Overall, 1,2-indanedione proved to be a viable alternative to traditional methods for the detection of fingermarks on porous surfaces, with more fingermarks being developed using this reagent on real samples than both DFO and ninhydrin and a combination of the two reagents.     

An investigation into the enhancement of fingermarks in blood on paper with genipin and lawsone

The abilities of two natural products, genipin and lawsone, to enhance blood contaminated fingermarks on papers of various porosities and colour were investigated and compared to the routinely used amino acid reagents, ninhydrin and 1,8-diazafluoren-9-one (DFO).Fingermarks in blood were deposited as a split depletion series on various paper types and colours for ageing periods of 6 weeks, 4 weeks, 2 weeks and 1 week before enhancement. The developed marks were observed under different lighting conditions, recorded and graded by way of attributing quantitative data to each series.Results indicated that while genipin showed some potential as a reagent for the enhancement of latent fingermarks, it displayed no suitability for the enhancement of fingermarks in blood on paper. Lawsone also failed to successfully enhance either type of fingermark. Upon comparison of the results with those of ninhydrin and DFO it was found that ninhydrin displayed the highest success rate of development of these marks.