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.     

Fingermarks development on gloves: Relative efficiency of 1,2 Indanedione/ZnCl2, ninhydrin and wet powder

Powder suspensions such as wet powder are used to develop fingermarks on adhesive surfaces. They can also be used on gloves but these ready-to-use commercial pre-mixed solutions are not entirely satisfactory. Among other things, they generate some background noise that can sometimes be substantial and make the exploitation of the fingermarks difficult. Recent studies have proven the effectiveness of a HFE ninhydrin solution on some types of gloves. Ninhydrin and 1,2-indanedione are reagents that are commonly used for the visualisation of fingermarks on porous and semi-porous substrates. Applying them on gloves might help to overcome the issue of background noise being produced by powder suspensions. In this study, several formulations of ninhydrin and 1,2-indanedione are compared to wet powder on a sample of nitrile, latex and vinyl gloves. The subsequent results show a greater effectiveness with the use of wet powder.     

Evaluation of 1,2-indanedione and 5,6-dimethoxy-1,2-indanedione for the detection of latent fingerprints on porous surfaces

The ability of 1,2-indanedione and 5,6-dimethoxy-1,2-indanedione to detect latent prints on porous surfaces, as compared to DFO and ninhydrin, has been evaluated. Comparisons of prints developed under various conditions determined the optimum development conditions for the new reagents. The indanediones tested were found to have lower detection limits for glycine. The carrier solvent used was found to affect the quality of the prints developed. In Arklone, the new reagents developed prints that displayed superior luminescence to those developed with DFO. In HFE 7100, 1,2-indanedione and 5,6-dimethoxy-1,2-indanedione gave superior luminescence to DFO after zinc salt treatment and cooling with liquid nitrogen, both of which improve the luminescence of prints developed with 1,2-indanediones. 1,2-Indanediones could offer less expensive but effective alternatives to DFO. With further optimization, the new reagents may supersede DFO as the method of choice for the detection of latent fingerprints on porous surfaces.     

Printed artificial sweat as replacement for natural fingermarks: Qualitative and quantitative approach considering an amino acid reagent

The study presented in this paper aims at assessing how printed fingermarks can be used to generate realistic latent marks bearing varying quantities of materials to be detected. Considering dilution series of artificial sweat (eccrine secretion) and 1,2-indanedione/zinc as amino acid reagent, we assessed how printed marks behave in comparison to natural fingermarks provided by a set of 30 donors. The results were assessed in terms of relative intensity (contrast, luminescence) and expert grading (ridge details, overall quality). With regards to the set of 30 donors, this study brought a quantitative look to the influence of intra- and inter-variability on the relative intensity values observed when processing natural fingermarks. This provided new data to further understand the concept of “donorship”. With regards to the use of printed marks, it has been illustrated how dilution series of a concentrated solution allows covering a range of cases: unnatural marks (intensity values well above those obtained with donors), rich marks (corresponding to fingermarks left by good donors), and faint marks (associated with the kind of results observed with poor donors). Such a range of detection performance offers the possibility to generate fine-tuned detection exercises of varying difficulty levels. Printed items made of artificial sweat could hence constitute a valuable alternative to natural secretions in the context of education and proficiency testing.     

Gamma Irradiation as a Biological Decontaminant and Its Effect on Common Fingermark Detection Techniques and DNA Profiling

Abstract:  The use of disease-causing organisms and their toxins against the civilian population has defined bioterrorism and opened forensic science up to the challenges of processing contaminated evidence. This study sought to determine the use of gamma irradiation as an effective biological decontaminant and its effect on the recovery of latent fingermarks from both porous and nonporous items. Test items were contaminated with viable spores marked with latent prints and then decontaminated using a cobalt 60 gamma irradiator. Fingermark detection was the focus with standard methods including 1,2-indanedione, ninhydrin, diazafluoren-9-one, and physical developer used during this study. DNA recovery using 20% Chelex extraction and quantitative real-time polymerase chain reaction was also explored. Gamma irradiation proved effective as a bacterial decontaminant with D -values ranging from 458 to 500 Gy for nonporous items and 797–808 Gy for porous ones. The results demonstrated the successful recovery of latent marks and DNA establishing gamma irradiation as a viable decontamination option.     

An evaluation of the effect of incorporating metal salts into 1,8 diazafluoren-9-one (DFO) formulations for fingermark enhancement

A study into the modification of 1,8-diazafluoren-9-one (DFO) formulations by the additions of metal salts into the working solution is reported. Similar additions have been found to increase the fluorescence of marks developed using other amino acid reagents including 1,2-indandione and the ninhydrin analogue 5-methylthioninhydrin. It was found that adding zinc chloride to give a 1:1 ratio of zinc ions:DFO molecules gave optimum fluorescence, and improvements in performance over the standard DFO formulation were achieved. Attempts to produce equivalent formulations with iron, nickel and palladium chlorides were unsuccessful. In a comparative trial with a 1,2-indandione-zinc formulation on brown paper and cardboard substrates, 1,2-indandione-zinc gave superior results and it was decided to focus further research on this reagent instead of DFO-zinc.     

Enhanced Visualization of Latent Fingermarks on Rough Aluminum Surfaces Using Sequential Au and Zn/ZnS/ZnO Depositions

Detection and visualization of fingermarks on rough and diffuse surfaces is a relatively challenging task. We succeeded in developing latent fingermarks on scratched and rough aluminum surfaces by sequential deposition of a thin layer of gold followed by one of zinc or zinc‐based compounds on the fingermarks. The best image enhancement was achieved with sequential Au and ZnS depositions. Using this combination, we could enhance the visualization of latent fingermarks aged over 65 days in normal conditions. The optical reflectance from the fingermarks with the deposited layers of metal/dielectric is analyzed as a stratified medium. Significant contrast in the reflectance from the regions of the ridges and the valleys of the fingermark would enhance the visualization. Our results show that the Au and ZnS bi‐layer combination can have a large reflection contrast and improved fingermark visualization at wavelengths corresponding to the green light for specific thickness of ZnS.     

Mass spectra and time-resolved fluorescence spectroscopy of the reaction product of glycine with 1,2-indanedione in methanol

The reaction products of 1,2-indanedione (a new fluorescent fingerprint reagent) with glycine in methanol, at room temperature have been studied using excitation and emission and time-resolved fluorescence spectroscopy. Gas chromatography-mass spectroscopy (GC/MS) has also been used to determine which compounds are formed. Reaction products were identified using GC/MS as 2-carboxymethyliminoindanone (MW=203 g) and 1,2-di(carboxymethylimino)indane (MW=260 g). Identified compounds show room temperature fluorescence lifetimes of tau(1)=7.69 ns and tau(2)=1.27 ns. It is not clear yet which compound is having fluorescence lifetime of 7.69 ns and which one is showing 1.27 ns.     

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 effect of 1,2-indanedione,a latent fingerprint reagent on subsequent DNA profiling

The compound 1,2-indanedione was recently introduced in our laboratory as an operational reagent for developing latent fingerprints on porous surfaces. As part of the reagent implementation, a study was carried out in order to determine whether either of the two operational 1,2-indanediones formulations interferes with further DNA profiling. Both formulations are based on HFE7100 solvent. One is acidic and the other neutral. In a controlled experiment, known donors attached stamps to envelopes by licking them. The stamped envelopes were initially treated with either one indanedione formulation or the other, and DNA was then extracted for STR typing. No differences were observed between the STR profiles obtained from treated and untreated stamps and envelopes, indicating that 1,2-indanedione does not adversely affect the extraction and subsequent amplification of the STRs examined. However, preliminary results indicate that potential DNA analysis depends on the time interval between the indanedione treatment and DNA extraction as no DNA can be recovered six days following treatment. For this reason, it is strongly recommended to extract DNA from treated items of evidence as soon as possible after indanedione treatment.     

Latent fingermark development on a range of porous substrates using ninhydrin analogs--a comparison with ninhydrin and 1,8-diazofluoren

Three relatively new reagents for developing latent fingermarks on porous substrates, 1,2-indandione (IND), 5-methylthioninhydrin (5-MTN), and lawsone, are compared with the more widely used ninhydrin and 1,8-diazofluoren (DFO). Developed latent fingermark visualization on 10 different substrates comprising colored papers, cardboard, and cellophane rather than conventional printer and writing/notepad paper is assessed using latent fingermark deposits from 48 donors. Results show improved fluorescent fingermark visualization using IND compared with DFO on a range of colored cardboards and thick white paper, thus extending the range of substrates known to yield improved visualization with IND. Adding zinc chloride to IND failed to yield any further improvement in fluorescent fingermark visualization. 5-MTN (with and without zinc chloride posttreatment) showed no improvement in visualization compared with ninhydrin and DFO although visible fingermarks were developed. Lawsone produced fluorescent visible fingermarks only with white substrates, which were inferior to those produced with DFO.     

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.     

Visualization of Fingermarks Deposits on Untreated Thermal Paper Exploiting the Near Infrared Luminescence

Thermal paper is widely used as a print medium for different applications but it constitutes a tricky substrate for fingermark visualization. An earlier work (J Forensic Sci 2015; 60 :1034) reported how to visualize fingermarks on untreated thermal paper by illuminating the item with a UV-A light source. In the present paper, the potential of the near infrared (NIR) luminescence has been tested on thermal paper compared to the mentioned method. A controlled study was carried out utilizing eccrine enriched fingermarks. The promising outcomes obtained were further confirmed by performing a pseudo-operational trial. Data clearly showed that the use of the NIR filter gave better results. Finally, preliminary tests suggested a different mechanism of reaction induced by fingermarks with respect to the one behind the thermal printing. Thus, NIR luminescence represents a refinement to the suite of optical examination processes, including the potential to increase the number of marks recovered in a noncontact, nondestructive way.     

Performance evaluation of automated fingerprint identification systems for specific conditions observed in casework using simulated fingermarks

Few studies have been reported on the performance evaluation of automated fingerprint identification systems (AFIS) for fingermark-to-fingerprint comparisons. This paper aims to illustrate to fingerprint examiners the relevance of evaluating the AFIS performance under specific conditions by carrying out five types of performance tests. The conditions addressed are the number of minutiae assigned to a fingermark, manual and automatic assignment of the minutiae, the finger region from which the fingermark originates, the degree of distortion in the fingermark, and the difference in orientation between fingermarks and fingerprints. In these tests, the magnitude of the influence for each condition was quantified. The comparisons were performed using a research AFIS technology with simulated fingermarks. Simulated fingermarks provide a practical way to create fingermarks for specific conditions in large quantities. The results showed that each condition influences the performance significantly, emphasizing the relevance of developing, and applying performance tests for specific conditions.     

Refining Fingermark Development using Diacetylene Copolymers on Difficult Surfaces,

In further work investigating the intriguing application of diacetylene copolymers in fingermark detection, methods were developed to control (inhibit or enhance) the diacetylene polymerization reaction in fingermarks treated with a mixture of the monomers 2,4‐hexadiyne‐1,6‐bis‐(phenylurethane) (HDDPU) and 2,4‐hexadiyne‐1,6‐bis(p‐chlorophenylurethane) (HDDCPU) in acetone solution. These methods included the use of a humidity chamber to reduce the amount of background development while promoting development on the fingermark, subjecting developed fingermarks to freezing temperatures and using a solvent to remove unreacted monomer in order to inhibit the polymerization reaction. Developed fingermarks were enhanced by conventional lighting (white light, filtered light) and fast Raman mapping, which was shown to be advantageous over FTIR imaging. This study also demonstrated the applicability of diacetylene copolymer solutions in the covert detection of fingermarks on difficult surfaces. Furthermore, fingermarks were successfully developed with good ridge detail on pig skin (used as a model for human skin, a notoriously difficult surface on which to develop fingermarks).     

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.     

Preparation of Artificial Blood from the Extract of Legume Root Nodules,and the Creation of Artificial Latent Fingermarks in Blood Using Artificial Blood,

Distribution of homogeneous fingermarks in blood is essential for conducting proficiency tests in forensic science. Hence, the artificial blood was prepared using the root nodule extract of Glycine max plants. The reactivity of the artificial blood with widely used human blood detection reagents was tested. Artificial latent fingermarks in blood were printed using an inkjet cartridge case filled with artificial blood solution. The artificial latent fingermarks in blood were developed with amino acid‐sensitive reagents and could obtain development as prominent as the image of the master fingermark saved on the computer. Therefore, it has been confirmed that the extract of legume root nodules can be used as artificial blood, and the artificial blood can be used for the preparation of artificial latent fingermarks or footmarks in blood.     

Development of latent fingermarks on thermal paper: Preliminary investigation into use of iodine fuming

Thermal paper finds its extensive use in the modern day life and could act as a vital piece of physical evidence carrying latent fingermarks. A large number of citations are available in literature suggesting various techniques to develop these marks but all are suffering with one or the other drawbacks such as complex and cumbersome procedure, pre- or post-treatment, background coloration and efficiency to develop aged fingermarks. In present study, a very simple and novel method involving iodine fuming has been suggested to develop fingermarks which were not only permanent but also without any background coloration. The suggested method does not involve any pre- or post-treatment of the substrate and was able to develop very old fingermarks (upto >1 year). In this study an attempt has been made to explain the reaction mechanism of the process. In case of different types of thermal papers, presence of different substituents on leuco dye (lactone ring) structure resulted in development of different colored fingermarks upon reaction with iodine. Sebaceous material laden marks have been found to be more intensely developed as compared to eccrine marks, and the difference was more pronounced in case of aged fingermarks.     

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.