Blocks of colour IV: the evidential value of blue and red cotton fibres.

Samples of blue and red cotton fibres were examined using light and fluorescence microscopy as well as UV/VIS microspectrophotometry and fluorescence microspectroscopy. The degree of fluorescence and spectral variation was recorded. Particular attention was paid to the recurrence of certain spectral patterns. The importance of spectral information in the UV range is emphasized again. Colour plays a critical role in the comparison of cotton fibres in forensic sciences. Normally, fibres producing spectral patterns that are frequently seen will tend to have a lower evidential value in criminal cases as the choice of putative sources is theoretically greater and vice versa. Besides black cotton, blue and red cotton fibres are very frequent in fibre casework. The very high discriminating power using a combination of light microscopy, fluorescence microscopy and UV/VIS microspectrophotometry shows that even blue and red cotton fibres can provide excellent evidence when involved in fibre transfer cases.     

The discrimination of (non-denim) blue cotton

This study was conducted to determine the degree of discrimination obtained between non-denim blue cotton fibres using visible–UV range microspectrophotometry alone. To this end, samples of fibres were taken from 100, non-denim, blue cotton, outer garments, including t-shirts, trousers and jumpers and subjected to analysis by both visible and UV range microspectrophotometry. The results obtained from the samples of each garment were compared to determine if they ‘matched’ or not. From an initial visual comparison of the garments it was possible to subdivide the samples into two populations consisting of 73 ‘dark blue’ garments and 27 ‘mid-blue’ garments. It was found that of the 73 ‘dark blue’ garments, 22 distinct sub-populations could be distinguished using visible range MSP, this figure being increased to 43 when the analysis was extended into the UV range. In the case of the 27 ‘mid-blue’ garments, 9 distinct sub-populations were discriminated using visible range MSP, this figure being increased to 17 when the analysis was extended into the UV range. The discriminating power (i.e. the number of discriminated pairs divided by the number of possible pairs) of visible range microspectrophotometry was calculated as 0.89 for ‘mid-blue’ garments and 0.87 for ‘dark blue’ garments. Extending microspectrophotometry into the UV range increased discrimination by 7%, giving a discriminating power of 0.96 for both mid and dark blue cotton fibres which was similar to that reported by a previous study where this method was combined with light and fluorescence microscopy.Intra-garment variation was found to be negligible. The implications of this study for casework are discussed and a revised analytical pathway for the comparison of this fibre type/colour combination using microspectrophotometry as a primary screening tool, is proposed.     

Enzymatic extraction of dyes for differentiation of red cotton fibres by TLC coupled with VSC

The purpose of this work was to assess the usefulness of thin layer chromatography (TLC) for discriminating single cotton fibres dyed with red reactive dyes. An effective enzymatic extraction procedure with the use of cellulase for the red reactively-dyed cotton fibres was developed and used for the discrimination of fibres derived from 21 garments purchased commercially. Discrimination of the fibres relied on the separation of the extracted dyes by thin layer chromatography (TLC). Four eluents were used to develop the plates with the extracted dyes, and the obtained results were analysed using, among others, video spectral comparator (VSC). Observation of TLC plates in visible, ultraviolet and infrared light allowed unambiguous discrimination of 5 and probable discrimination of 6 of the 21 fibres tested. The remaining fibres were divided into several groups. Comparison of the acquired results with those obtained for the same examination material by standard non-destructive methods used in forensic fibres examinations (transmitted light microscopy, fluorescence microscopy, UV–Vis microspectrophotometry and Raman spectroscopy) has shown that efficiency in fibres differentiation is similar for all methods. TLC coupled with VSC was even found to be more effective in differentiation of red cotton fibres. The chemometric analysis was helpful to discriminate dyed cotton fibres, characterized by very similar colour.     

A further study of dye batch variation in textile and carpet fibres.

Four sets of acrylic fibre samples were obtained from a company that dyes fabrics for the fashion industry. Between seven and ten different batches of fibres constituted each set. Comparison microscopy, visible and UV range microspectrophotometry and thin layer chromatography (TLC) were used to compare the dyes on each batch of fibres within the sets. Only one of the four sets exhibited variation within the batches. The differences were seen when both microscopical and analytical techniques were used. In addition, two further sets of samples had been obtained from a company that produces carpets for the car industry. The first set consisted of 26 batches of acid dyed orange nylon fibres. The second consisted of 21 batches of acid dyed mustard coloured nylon and direct dyed brown viscose fibres blended together. When the first set was viewed under UV light one batch had more pale orange fibres present and they fluoresced more brightly than the other fibres. This could be due to the blending with a different dye batch of fibre or due to poor dye uptake--the latter being more likely. When tested using visible and UV range microspectrophotometry and TLC, further dye batch variation was not detected. The second set was examined after separating the nylon and viscose fibres from each other. The nylon fibres were indistinguishable when a range of microscopical and analytical techniques were employed; however, the viscose fibres showed dye batch variation when TLC was used.     

High-performance liquid chromatography-ultraviolet-visible spectroscopy-electrospray ionization mass spectrometry method for acrylic and polyester forensic fiber dye analysis

A critical point of comparison between a fiber collected from a crime scene and a fiber from a known source is the color. Fiber dye analysis using thin-layer chromatography or ultraviolet (UV)-visible (Vis) microspectrophotometry provides useful, although limited, data for comparison. High-performance liquid chromatography-electrospray ionization mass spectrometry (LC/MS) overcomes these limitations by integrating chromatography, ultraviolet-visible spectroscopy, and mass spectrometry into a single instrument. In order to evaluate the applicability of the LC/MS to forensic fiber dye analysis, a multi-stage chromatographic method using acidified water and acidified acetonitrile was developed that separated and identified a mixture of 15 basic and 13 disperse dye standards. The LC/MS also detected and analyzed dyes extracted from individual 0.5 cm acrylic and polyester fibers, demonstrating its applicability to this type of analysis. With regard to the analysis of disperse dyes in polyester fibers, the replacement of pyridine with acetonitrile in the extraction system allowed direct injection of the extracts into the LC/MS. The advantage of the LC/MS over other instrumental methods of textile dye analysis is demonstrated by the analysis and differentiation of three black acrylic fibers: two fibers had similar UV-Vis spectra but were differentiated with chromatography and two had similar UV-Vis spectra and chromatograms but were differentiated using the mass spectrometer.     

The Discrimination of Colored Acrylic,Cotton, and Wool Textile Fibers Using Micro‐Raman Spectroscopy. Part 1: In situ Detection and Characterization of Dyes

Raman spectroscopy has been applied to characterize fiber dyes and determine the discriminating ability of the method. Black, blue, and red acrylic, cotton, and wool samples were analyzed. Four excitation sources were used to obtain complementary responses in the case of fluorescent samples. Fibers that did not provide informative spectra using a given laser were usually detected using another wavelength. For any colored acrylic, the 633‐nm laser did not provide Raman information. The 514‐nm laser provided the highest discrimination for blue and black cotton, but half of the blue cottons produced noninformative spectra. The 830‐nm laser exhibited the highest discrimination for red cotton. Both visible lasers provided the highest discrimination for black and blue wool, and NIR lasers produced remarkable separation for red and black wool. This study shows that the discriminating ability of Raman spectroscopy depends on the fiber type, color, and the laser wavelength.     

The Analysis of Metameric Blue Fibers and Their Forensic Significance

Abstract:  Metamerism is a phenomenon where two or more colored items with different colorant chemistries appear to the observer to be the same color. Those differences should result in different UV-visible spectra. Additionally, the literature on color science states that metameric samples will have spectra that intersect at three or more loci. Metameric samples of blue textile fibers, which were created using different coloring agents or different relative concentrations of the coloring agents, were studied to demonstrate that they could be differentiated by obtaining their spectra between 350 and 800 nm using UV-visible microspectrophotometry. However, while some of the metameric samples tested did intersect at three or more loci, others did not intersect at all. In the spectra that did intersect, no correlation was found between either the dye chemistries or the relative component concentrations.     

The Analysis of Colored Acrylic,Cotton, and Wool Textile Fibers Using Micro‐Raman Spectroscopy. Part 2: Comparison with the Traditional Methods of Fiber Examination

In the second part of this survey, the ability of micro‐Raman spectroscopy to discriminate 180 fiber samples of blue, black, and red cottons, wools, and acrylics was compared to that gathered with the traditional methods for the examination of textile fibers in a forensic context (including light microscopy methods, UV‐vis microspectrophotometry and thin‐layer chromatography). This study shows that the Raman technique plays a complementary and useful role to obtain further discriminations after the application of light microscopy methods and UV‐vis microspectrophotometry and assure the nondestructive nature of the analytical sequence. These additional discriminations were observed despite the lower discriminating powers of Raman data considered individually, compared to those of light microscopy and UV‐vis MSP. This study also confirms that an instrument equipped with several laser lines is necessary for an efficient use as applied to the examination of textile fibers in a forensic setting.     

A study to investigate the evidential value of blue and black ballpoint pen inks in Australia

The aim of this project was to investigate the evidential value of blue and black ballpoint pen inks in Australia. For this purpose, 49 blue and 42 black ballpoint pen inks, of different brands, models and batches, representative of those ballpoint pens available on the Australian market at the time of the study, were analysed by three techniques: filtered light examination (FLE); reflectance visible microspectrophotometry (MSP); and thin-layer chromatography (TLC). The results showed that the power of the individual techniques to discriminate inks between and within brands, models and batches varied, the most informative techniques being TLC>FLE>MSP. The greatest degree of differentiation was achieved when using a combined sequence of techniques, the discriminating power being 0.99 and close to 1 for blue and black inks, respectively (different brands and models). In respect to the discrimination between different batches of a same brand and model, it was shown that 14 out of the 33 pairs compared could be discriminated. Overall, this study confirmed the high value of the examination of inks when applied to ballpoint pens available in Australia, especially when a combined sequence of techniques is applied.     

The analysis and comparison of blue wool fibre populations found at random on clothing.

Fifty-eight garments were taped and searched for mid to dark blue wool fibres. These were then removed from the tapings, mounted on slides and examined using a high-power microscope (400x). A total of 2,740 blue wool fibres were identified and visible range microspectrophotometry (MSP) was performed on them. Three hundred independent blue wool populations were identified on 56 of the 58 garments searched. The lack of control fibres meant the spectral range of each population was unknown. The number of populations may have been underestimated by grouping together the fibres that had broad single peaks and a lack of distinguishing features in the spectra. Although blue wool is considered to be a common fibre type, 300 unique spectral shapes were identified by the use of microspectrophotometry alone. This demonstrates that the dyes used in the dyeing of blue wool are variable. Showing that many different populations of blue wool occur on a range of garments should ensure that the forensic scientist does not underestimate or understate the strength of evidence in cases where blue wool is found. Hopefully this work will enlighten scientists and enable them to also assess the true value of their findings when other commonly occurring fibres are encountered.     

Forensic Discrimination of Dyed Hair Color: I. UV‐Visible Microspectrophotometry*†

Abstract: Current protocols for examining hair do not attempt to differentiate hair color using instrumental analysis. In this study, hair samples treated with 55 different red hair dyes were analyzed using UV‐visible microspectrophotometry between 200 and 700 nm. Using air as a background reference gave the best results, although mounting media such as glycerin could also be used. The contribution of the hair substrate is predominantly observed in the range of 300–400 nm while the dye peak is evident in the range of 425–550 nm. It was found that the presence of hair dye reduces the overall intrasample variability of the hair color. In addition, visual inspection and spectral interpretation showed that dyed hair exhibits distinct and discernable shades. The color of all samples was stable during storage and while all hair dyes fade with washing, significant fading of the color was only evident after daily washing for 3 weeks.     

Forensic Discrimination of Dyed Hair Color: II. Multivariate Statistical Analysis*,†

Abstract: This research is intended to assess the ability of UV–visible microspectrophotometry to successfully discriminate the color of dyed hair. Fifty‐five red hair dyes were analyzed and evaluated using multivariate statistical techniques including agglomerative hierarchical clustering (AHC), principal component analysis (PCA), and discriminant analysis (DA). The spectra were grouped into three classes, which were visually consistent with different shades of red. A two‐dimensional PCA observations plot was constructed, describing 78.6% of the overall variance. The wavelength regions associated with the absorbance of hair and dye were highly correlated. Principal components were selected to represent 95% of the overall variance for analysis with DA. A classification accuracy of 89% was observed for the comprehensive dye set, while external validation using 20 of the dyes resulted in a prediction accuracy of 75%. Significant color loss from successive washing of hair samples was estimated to occur within 3 weeks of dye application.     

The kinetics of colour change in textiles and fibres treated with detergent solutions: Part I—Colour perception and fluorescence microscopy analysis

The aim of this study was to assess colour changes that occur in several types of commonly available textiles as a result of the long-term effects of various popularly used laundry detergents. A 14 day experiment was conducted using blue, red and grey/black cotton, wool, acrylic and polyester textiles. Colour changes were evaluated through the visual comparison of the colour of the textile samples against that of the untreated (control) material.The kinetics of the changes in the colour of the fibres were monitored using fluorescence microscopy (UV excitation filter). The conclusions include an assessment of the observed changes from a fibre analysis expert's point of view, as well as that of an average user/consumer of the products involved.     

Raman spectroscopy and microspectrophotometry of reactive dyes on cotton fibres: Analysis and detection limits

A collaborative study on Raman spectroscopy and microspectrophotometry (MSP) was carried out by members of the ENFSI (European Network of Forensic Science Institutes) European Fibres Group (EFG) on different dyed cotton fabrics. The detection limits of the two methods were tested on two cotton sets with a dye concentration ranging from 0.5 to 0.005% (w/w). This survey shows that it is possible to detect the presence of dye in fibres with concentrations below that detectable by the traditional methods of light microscopy and microspectrophotometry (MSP). The MSP detection limit for the dyes used in this study was found to be a concentration of 0.5% (w/w). At this concentration, the fibres appear colourless with light microscopy. Raman spectroscopy clearly shows a higher potential to detect concentrations of dyes as low as 0.05% for the yellow dye RY145 and 0.005% for the blue dye RB221. This detection limit was found to depend both on the chemical composition of the dye itself and on the analytical conditions, particularly the laser wavelength. Furthermore, analysis of binary mixtures of dyes showed that while the minor dye was detected at 1.5% (w/w) (30% of the total dye concentration) using microspectrophotometry, it was detected at a level as low as 0.05% (w/w) (10% of the total dye concentration) using Raman spectroscopy. This work also highlights the importance of a flexible Raman instrument equipped with several lasers at different wavelengths for the analysis of dyed fibres. The operator and the set up of the analytical conditions are also of prime importance in order to obtain high quality spectra. Changing the laser wavelength is important to detect different dyes in a mixture.     

运用光谱成像技术区分同色棉纤维

目的研究同色棉纤维的检验方法。方法运用成像光谱法对红、蓝、黑3个颜色组的棉纤维进行区分。结果红色棉纤维的鉴别率为62．2％，蓝色棉纤维的鉴别率为77．8％，黑色棉纤维的鉴别率为69．6％。结论实验结果表明，光谱成像技术可以区分大部分同色棉纤维。拓宽了纤维检验的方法，解决了同色棉纤维的区分问题，该方法能够用于对微量同色棉纤维的快速鉴别。     

Chemical identification and optimization of the 4-aminophenol colorimetric test for the differentiation between hemp-type and marijuana-type cannabis plant samples

The 4- Aminophenol (4-AP) colorimetric test is a fast, easy-to-use, and cost-effective presumptive assay of cannabis plant material producing different chromophores with THC-rich cannabis (blue color) and with CBD-rich cannabis (pink color). The main drawback of the 4-AP test is a brief observation window where the color rapidly changes to black, limiting the utility of the test. We now report for the first time, the identification of the product chromophores between 4-AP and CBD/THC as well as propose an explanation and a solution for the color degradation of the chromophores. The identification of the chromophores is provided by spectroscopic (UV–Vis), chromatography, and mass spectrometry (TLC and LC-QToF-MS). Oxidation of excess 4-AP (Reagent A) in the presence of NaOH (Reagent B) produces the black color observed for the previously reported 4-AP tests and reported in the literature. The adjustment of reactants concentrations and volumes of 4-AP:THC/CBD to a 1:1 ratio significantly reduces the black oxidation by-product and increases the observation window up to 2 h instead of the previously reported 5–10 min. For the first time, mass spectrometry and chromatography confirmed that the reaction of THC and CBD with 4-AP produced chromophores with m/z (M + H) = 420, consistent with proposed indophenol structures. The TLC method developed confirmed the separation between CBD and THC chromophores. The specificity of the test is also reported, showing false positive results for the presence of THC (blue color) for samples of thyme and oregano. LDA and SIMCA models showed that the optimized 4-AP procedure performs better than the previously reported 4-AP color test.     

Validation of LAB color mode as a nondestructive method to differentiate black ballpoint pen inks

Nondestructive digital processing methods such as lab color mode (available in Adobe Photoshop) are emerging as alternative methods for forensic document examiners to use when attempting to differentiate writing instrument inks. Although these techniques appear to be viable, little data currently exists regarding the known or potential error rates associated with these techniques. Without adequate data, the validity and reliability of these techniques, including lab color, can not be established. In an attempt to begin to address these issues, 44 black ballpoint ink pens were obtained and used to create 990 pen-pair samples for analysis using established lab color mode techniques. No erroneous findings of "different" were reported following the examination of the known pen-pair combinations in which the same pen was used to create the samples (n = 44). Of the remaining 946 samples, 737 pen-pair samples were differentiated using the lab color mode method, while 209 samples were unable to be differentiated and were recorded as either being "similar" (n = 153) or "unsure" (n = 56). Comparison of the lab color mode results with the results obtained through additional testing using traditional infrared reflectance and infrared luminescence test methods showed that lab color differentiated 102 pen-pair samples (11%; 102/946) that were not differentiated using a VSC-4C.     

The evidential value of black cotton fibres

Forensic scientists are faced with the problem of estimating the frequency of cotton fibres recovered in casework, in relation to those in the general population. One way of doing this is to consider the degree of spectral variation that occurs within a "block of colour". When a spectral pattern occurs very frequently, the evidential value of the fibres may be so low, that it is not worth considering them as target fibres. Using UV-visible range microspectro-photometry (MSP) spectra were recorded from 88 known black cotton dyes and 225 samples of black cotton taken from various textiles. UV-visible spectra originating from sulphur dyes and from the great majority of reactive and direct dyes can be easily recognised. Vat dyes present a little more difficulty. The degree of spectral variation and consequent discriminating power of MSP varied according to the dye class, from 0.13 for sulphur dyes to 0.93 for reactive dyes. From 99 textiles dyed with reactive dyes, the spectra could be divided into at least 40 varieties showing that these fibres have a high degree of individuality. Within the few direct dyes (11.5%) that were encountered, one basic spectral form predominated, but a number of minor variations were observed. Spectral information below 400 nm (UV-range) is important for making distinctions and is critical in the case of direct dyes.     

The discrimination potential of ultraviolet-visible spectrophotometry, thin layer chromatography, and Fourier transform infrared spectroscopy for the forensic analysis of black and blue ballpoint inks

The knowledge of the discriminating power of analytical techniques used for the differentiation of writing inks can be useful when interpreting results. Ultraviolet-visible (UV-VIS) spectrophotometry, thin layer chromatography (TLC), and diffuse reflectance Fourier transform infrared spectroscopy (FT-IR) were used to examine a population of 21 black and 12 blue ballpoint writing inks. Based on corroborative results of these methods, the discrimination power for UV-VIS, TLC, and FT-IR was determined to be 100% and 98% for the black and blue inks, respectively. Generally, TLC and UV-VIS can be used to differentiate the colorant components (i.e., dyes and some pigments) found in inks. As FT-IR can be utilized to identify some of the noncolorant components, it was determined to be an excellent complementary technique that can be implemented into an analytical scheme for ink analysis.     

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.