Determination of Time since Deposition of Fingerprints via Colorimetric Assays.

Past investigations involving fingerprints have revolved heavily around the image of the fingerprint-including the minutiae, scarring, and other distinguishing features-to visually find a match to its originator. Recently, it has been proven that the biochemical composition can be used to determine originator attributes, such as sex, via chemical and enzymatic cascades. While this provides pertinent information about the originator's identity, it is not the only piece of information that can be provided. This research was designed with three goals in mind: (1) identify how long it would take before an aged female fingerprint could no longer be differentiated from a male fingerprint, (2) identify a correlation between the data collected and a specific time since deposition (TSD) time point, and (3) identify if a specific amino acid could be contributing to the decreasing response seen for the aging fingerprints. Using ultraviolet-visible (UV-vis) spectroscopy, aged fingerprints were evaluated over the course of 12 weeks via three chemical assays previously used for fingerprint analysis-the ninhydrin assay, the Bradford assay, and the Sakaguchi assay. As fingerprints age, the conditions they are exposed to cause the biochemical composition to decompose. As this occurs, there is less available to be detected by analytical means. This results in a less intense color production and, thus, a lower measured absorbance. The results displayed here afforded the ability to conclude that all three goals set forth for this research were accomplished-a female fingerprint can be differentiated from a male fingerprint for at least 12 weeks, UV-vis data collected from aged fingerprints can be correlated to a TSD range but not necessarily a specific time point, and the decomposition of at least a single amino acid can afford the ability to estimate the TSD of the fingerprint.

Step toward Roadside Sensing: Noninvasive Detection of a THC Metabolite from the Sweat Content of Fingerprints.

The sudden increase in states legalizing marijuana has forced law enforcement into a situation where the use and consumption are legal, but there are no limitations for what is acceptable for driving or operating machinery. Using ultraviolet-visible (UV-vis) spectroscopy, fingerprints from volunteers who had used marijuana were analyzed via a competitive immunoassay for the detection of Δ9-tetrahydrocannabinol (Δ9-THC), the main psychoactive component of marijuana, and 11-nor-9-carboxy-THC (THC-COOH), one of the main metabolites produced in the body following the use/consumption of THC-related products. In this research, the THC-COOH metabolite and the enzyme-labeled conjugate compete against each other as the antigens for the system. The antibody used in this assay has a greater affinity for the metabolite; so, as its concentration increases, the absorbance of the system decreases due to less binding of the enzyme-labeled conjugate.

Metabolite Biometrics for the Differentiation of Individuals.

Sweat is a biological fluid present on the skin surface of every individual and is known to contain amino acids as well as other low molecular weight compounds. (1) Each individual is inherently different from one another based on certain factors including, but not limited to, his/her genetic makeup, environment, and lifestyle. As such, the biochemical composition of each person greatly differs. The concentrations of the biochemical content within an individual's sweat are largely controlled by metabolic processes within the body that fluctuate regularly based on attributes such as age, sex, and activity level. Therefore, the concentrations of these sweat components are person-specific and can be exploited, as presented here, to differentiate individuals based on trace amounts of sweat. For this concept, we analyzed three model compounds-lactate, urea, and glutamate. The average absorbance change from each compound in sweat was determined using three separate bioaffinity-based systems: lactate oxidase coupled with horseradish peroxidase (LOx-HRP), urease coupled with glutamate dehydrogenase (UR-GlDH), and glutamate dehydrogenase alone (GlDH). After optimization of a linear dependence for each assay to its respective analyte, analysis was performed on 50 mimicked sweat samples. Additionally, a collection and extraction method was developed and optimized by our group to evaluate authentic sweat samples from the skin surface of 25 individuals. A multivariate analysis of variance (MANOVA) test was performed to demonstrate that these three single-analyte enzymatic assays were effectively used to identify each person in both sample sets. This novel sweat analysis approach is capable of differentiating individuals, without the use of DNA, based on the collective responses from the chosen metabolic compounds in sweat. Applications for this newly developed, noninvasive analysis can include the field of forensic science in order to differentiate between individuals as well as the fields of homeland security and cybersecurity for personal authentication via unlocking mechanisms in smart devices that monitor metabolites. Through further development and analysis, this concept also has the potential to be clinically applicable in monitoring the health of individuals based on particular biomarker combinations.

Fingerprint Analysis: Moving Toward Multiattribute Determination via Individual Markers.

Forensic science will be forever revolutionized if law enforcement can identify personal attributes of a person of interest solely from a fingerprint. For the past 2 years, the goal of our group has been to establish a way to identify originator attributes, specifically biological sex, from a single analyte. To date, an enzymatic assay and two chemical assays have been developed for the analysis of multiple analytes. In this manuscript, two additional assays have been developed. This time, however, the assays utilize only one amino acid each. The enzymatic assay targets alanine and employs alanine transaminase (ALT), pyruvate oxidase (POx), and horseradish peroxidase (HRP). The other, a chemical assay, is known as the Sakaguchi test and targets arginine. It is important to note that alanine has a significantly higher concentration than arginine in the fingerprint content of both males and females. Both assays proved to be capable of accurately differentiating between male and female fingerprints, regardless of their respective average concentration. The ability to target a single analyte will transform forensic science as each originator attribute can be correlated to a different analyte. This would then lead to the possibility of identifying multiple attributes from a single fingerprint sample. Ultimately, this would allow for a profile of a person of interest to be established without the need for time-consuming lab processes.

Bioaffinity-based assay for the sensitive detection and discrimination of sweat aimed at forensic applications.

Sweat is a well-known piece of biological evidence that is actually used much less than expected. Biological samples are important because their components can often provide some type of information about a person-of-interest. Sweat, in particular, is important because of its DNA content which can be extracted and analyzed to provide information that can be imperative to a criminal investigation. While it is a very important source of forensic information, the methods for detection and discrimination of sweat are limited, causing it to be overlooked during evidence collection. This manuscript presents a biocatalytic method for sweat detection that utilizes an enzyme cascade system that has the capability to detect trace amounts of sweat and distinguish it from saliva, even after the sample has dried. The results show the initial calibration studies performed to insure that the cascade performs well using both mimicked and authentic sweat samples which have components that could negatively affect the enzymes needed for the analysis. The method presented here also has the potential to be adapted for on-site analysis. The initial results of the development of a sweat-sensitive strip are shown here.

Coomassie Brilliant Blue G-250 Dye: An Application for Forensic Fingerprint Analysis.

The Bradford reagent, comprised of the Coomassie Brilliant Blue G-250 dye, methanol, and phosphoric acid, has been traditionally used for quantifying proteins. Use of this reagent in the Bradford assay relies on the binding of the Coomassie Blue G-250 dye to proteins. However, the ability of the dye to react with a small group of amino acids (arginine, histidine, lysine, phenylalanine, tyrosine, and tryptophan) makes it a viable chemical assay for fingerprint analysis in order to identify the biological sex of the fingerprint originator. It is recognized that the identification of biological sex has been readily accomplished using two other methods; however, both of those systems are reliant upon a large group of amino acids, 23 to be precise. The Bradford assay, described here, was developed specifically to aid in the transition from targeting large groups of amino acids, as demonstrated in the previous studies, to targeting only a single amino acid without compromising the intensity of the response and/or the ability to differentiate between two attributes. In this work, we aim to differentiate between female fingerprints and male fingerprints.

Ages at a Crime Scene: Simultaneous Estimation of the Time since Deposition and Age of Its Originator.

Blood is a major contributor of evidence in investigations involving violent crimes because of the unique composition of proteins and low molecular weight compounds present in the circulatory system, which often serve as biomarkers in clinical diagnostics. It was recently shown that biomarkers present in blood can also identify characteristics of the originator, such as ethnicity and biological sex. A biocatalytic assay for on-site forensic investigations was developed to simultaneously identify the age range of the blood sample originator and the time since deposition (TSD) of the blood spot. For these two characteristics to be identified, the levels of alkaline phosphatase (ALP), a marker commonly used in clinical diagnostics corresponding to old and young originators, were monitored after deposition for up to 48 h to mimic a crime scene setting. ALP was chosen as the biomarker due to its age-dependent nature. The biocatalytic assay was used to determine the age range of the originator using human serum samples. By means of statistical tools for evaluation and the physiological levels of ALP in healthy people, the applicability of this assay in forensic science was shown for the simultaneous determination of the age of the originator and the TSD of the blood spot. The stability of ALP in serum allows for the differentiation between old and young originators up to 2 days after the sample was left under mimicked crime scene conditions.

New Horizons for Ninhydrin: Colorimetric Determination of Gender from Fingerprints.

In the past century, forensic investigators have universally accepted fingerprinting as a reliable identification method via pictorial comparison. One of the most traditional detection methods uses ninhydrin, a chemical that reacts with amino acids in the fingerprint content to produce the blue-purple color known as Ruhemann's purple. It has recently been demonstrated that the amino acid content in fingerprints can be used to differentiate between male and female fingerprints. Here, we present a modified approach to the traditional ninhydrin method. This new approach for using ninhydrin is combined with an optimized extraction protocol and the concept of determining gender from fingerprints. In doing so, we are able to focus on the biochemical material rather than exclusively the physical image.

Forensic Identification of Gender from Fingerprints.

In the past century, forensic investigators have universally accepted fingerprinting as a reliable identification method, which relies mainly on pictorial comparisons. Despite developments to software systems in order to increase the probability and speed of identification, there has been limited success in the efforts that have been made to move away from the discipline's absolute dependence on the existence of a prerecorded matching fingerprint. Here, we have revealed that an information-rich latent fingerprint has not been used to its full potential. In our approach, the content present in the sweat left behind-namely the amino acids-can be used to determine physical such as gender of the originator. As a result, we were able to focus on the biochemical content in the fingerprint using a biocatalytic assay, coupled with a specially designed extraction protocol, for determining gender rather than focusing solely on the physical image.