Photodegradation and laser desorption mass spectrometry for the characterization of dyes used in red pen inks

Photodegradation and laser desorption mass spectrometry (LDMS) is a powerful combination of methods capable of characterizing dyes found in pen inks. Rhodamine dyes in pens that contain red ink were analyzed directly from paper (no extraction step is necessary). Inks exposed to incandescent light form photodegradation products (compounds with lower molecular weights than that of the intact dye) and in some instances, photoproducts (compounds with higher molecular weights than that of the intact dye). The degradation products and photoproducts can be detected with LDMS, and the results can be used for dye identification. Advantages include: (1) the instrumental analysis takes less than a minute; (2) sample preparation is minimal; (3) LDMS is a minimally destructive technique; (4) incandescent light sources are inexpensive, safe to use, and readily available; and (5) isomeric dyes can be distinguished.     

Evaluation of desorption/ionization mass spectrometric methods in the forensic applications of the analysis of inks on paper.

Fast atom bombardment and laser desorption mass spectrometry (LDMS) provide molecular level information concerning an ink's composition. Two ink-jet printer inks, Ink A containing the cationic dye Methyl Violet 2B, and Ink B containing the anionic dye, Solvent Black, were studied. Both positive and negative ion detection modes of the mass spectrometer were used. LD may be used for the analysis of inks on paper. Once on paper, the ink's solvent system has evaporated, leaving mainly the dyes behind, which are detected using LDMS. An ink fades with time, indicating that the dyes are degrading. Preliminary results from an accelerated aging study of ballpoint pen ink using UV irradiation confirm that dye degradation products are formed. The degradation chemistry follows an oxidative demethylation process for which all products formed are detected using LDMS. Results suggest that LDMS may be developed to determine the relative age of inks.     

The direct detection and identification of staining dyes from security inks in the presence of other colorants, on currency and fabrics, by laser desorption mass spectrometry

Laser desorption mass spectrometry (LDMS) is useful for the direct desorption and MS analysis of dyes off materials such as paper. Here it is shown that staining dyes, produced by currency degradation devices, such as those used by financial institutions or armored transport companies, can be detected by LDMS. These staining dye packs are commonly used in bank security programs to deter theft or unauthorized removal of currency from teller stations or from ATM cash cassettes. Working automatically, these technologies release a security dye to degrade the surface of the notes and possibly mark the assailant involved in the attack. These dyes can be characterized and identified directly from paper currency, and from fabrics, and uniquely identified in the presence of other dyes that are used in the printing and dying of such materials, by LDMS. In these experiments, no extraction step is required. A pulsed UV laser directly irradiates a paper or fabric sample--colorants are desorbed, ionized and detected using time-of-flight MS. Results shown here suggest that dyes such as Basic Red 1:1 and Basic Violet 11:1 are used in such devices.     

Analysis of Pigmented Inkjet Printer Inks and Printed Documents by Laser Desorption/Mass Spectrometry*,†

Abstract: Anyone with a computer, scanner, and color printer has the capability for creating documents such as identification cards, passports, and counterfeit currency. Laser desorption mass spectrometry (LDMS) has been demonstrated as a powerful tool for colorant analysis. Inkjet printers are now moving largely toward the use of pigments as colorants; their insolubility makes analysis by simpler methods such as thin‐layer chromatography no longer an option. Recent developments in pigmented inkjet printer inks, such as gloss optimizers that coat pigment particles, may prohibit colorant analysis by LDMS. We demonstrate here that pigments used in inks from two Epson printers can be detected and analyzed by LDMS. Also, LDMS spectra of various colors created using a 4‐cartridge (cyan/magenta/yellow/black, CMYK) inkset are evaluated, to begin to develop an approach for unraveling LDMS data from real samples, to determine the number of inks used by a printer, and the chemical composition of the colorants.     

Evaluation of laser desorption mass spectrometry and UV accelerated aging of dyes on paper as tools for the evaluation of a questioned document

Laser desorption mass spectrometry (LDMS) may be used for the detection and identification of dyes found in inks. Naturally-aged and artificially-aged blue and black ballpoint pen inks containing the cationic dye methyl violet were analyzed on paper. The average molecular weight of the dye sample was calculated from LD mass spectral data and plotted versus time. The resulting aging curves demonstrate that, as dye degradation increases, the average molecular weight of the dye decreases. Typical variables involved in ink aging, such as the type of paper and ink formulation, were investigated. Results show that these variables influence the rate of dye degradation. Furthermore, UV accelerated aging has been developed and tested as an alternative to thermal approaches.     

Identification of organic pigments in automotive coatings using laser desorption mass spectrometry

When one looks at an automotive coating, one sees color due to pigments. Modern organic pigments, with high molar absorptivities, may be only minor components of the mixture. Laser desorption mass spectrometry (LDMS) has been shown to be a useful tool for the analysis of colorants such as pen ink dyes. Here, LDMS is used to determine its utility for the identification of pigments, in simple media and in more complex paints. Small paint chips can be introduced into the LDMS instrument, and when an ultraviolet laser is focused on a portion of a chip, ions representative of the pigment(s) are selectively formed. Some pigments such as quinacridones and copper phthalocyanine are very stable and are desorbed and ionized intact. In contrast, benzimidazolones, which contain some single-skeletal bonds, form fragment ions. This method proves to be sensitive and convenient, as no sample preparation is required. The presence of inorganic pigments in addition to modern organic pigments can be determined, and pigments can be directly identified in actual automotive paint chip samples.     

Differentiation of blue ballpoint pen inks by laser desorption ionization mass spectrometry and high-performance thin-layer chromatography

The differentiation of inks on a questioned document can highlight a fraudulent insertion and is usually carried out by optical comparison and thin-layer chromatography (TLC). Laser desorption ionization mass spectrometry (LDI-MS) may also be used for the analysis of dyes from ink. This analytical technique was compared with a standard method of high-performance TLC (HPTLC) according to their capacity to differentiate blue ballpoint inks. Ink entries on paper from 31 blue ballpoint pens have been analyzed and their dye ink formulations compared. The pens were classified into 26 classes by LDI-MS against 18 for HPTLC. LDI-MS proved to be a more powerful method for differentiating ink formulations because it provides information about dye structures (molecular weights) and relative quantification of dye classes (peak areas). Sample preparation was minimal and analysis time was short in contrast to the more complex extraction, application, and development steps of the HPTLC method. However, only basic dyes and pigments were identified using positive mode LDI-MS, while HPTLC did yield additional information about acid dyes.     

Raman spectroscopy and laser desorption mass spectrometry for minimal destructive forensic analysis of black and color inkjet printed documents

Inkjet ink analysis is the best way to discriminate between printed documents, or even though more difficult, to connect an inkjet printed document with a brand or model of printers. Raman spectroscopy and laser desorption mass spectrometry (LDMS) have been demonstrated as powerful tools for dyes and pigments analysis, which are ink components. The aim of this work is to evaluate the aforementioned techniques for inkjet inks analysis in terms of discriminating power, information quality, and nondestructive capability. So, we investigated 10 different inkjet ink cartridges (primary colors and black), 7 from the HP manufacturer and one each from Epson, Canon and Lexmark. This paper demonstrates the capabilities of three methods: Raman spectroscopy, LDMS and MALDI-MS. Raman spectroscopy, as it is preferable to try the nondestructive approach first, is successfully adapted to the analysis of color printed documents in most cases. For analysis of color inkjet inks by LDMS, we show that a MALDI matrix (9-aminoacridine, 9AA) is needed to desorb and to ionize dyes from most inkjet inks (except Epson inks). Therefore, a method was developed to apply the 9AA MALDI matrix directly onto the piece of paper while avoiding analyte spreading. The obtained mass spectra are very discriminating and lead to information about ink additives and paper compositions. Discrimination of black inkjet printed documents is more difficult because of the common use of carbon black as the principal pigment. We show for the first time the possibility to discriminate between two black-printed documents coming from different, as well as from the same, manufacturers. Mass spectra recorded from black inks in positive ion mode LDMS detect polyethylene glycol polymers which have characteristic mass distributions and end groups. Moreover, software has been developed for rapid and objective comparison of the low mass range of these positive mode LDMS spectra which have characteristic unknown peaks.     

Identification of colorants in pigmented pen inks by laser desorption mass spectrometry

Pigments are rapidly replacing dyes as colorants in pen and printer inks, due to their superior colors and stability. Unfortunately, tools commonly used in questioned document examination for analyzing pen inks, such as TLC, cannot be used for the analysis of insoluble pigments on paper. Laser desorption mass spectrometry is demonstrated here as a tool for analyzing pigment-based pen inks. A pulsed nitrogen laser can be focused onto a pen stroke from a pigmented ink pen on paper, and positive and negative ions representative of the pigment can be generated for subsequent mass spectrometric analysis. Targeted pens for this work were a set of Uni-ball 207 pigmented ink pens containing blue, light blue, orange, green, violet, red, pink, and black inks. Copper phthalocyanine was identified as the pigment used to make both blue inks. A mixture of halogenated copper phthalocyanines were identified in the green ink. Unexpectedly, the pink ink was found to contain a red pigment, Pigment Red 12, treated with a mixture of water-soluble dyes. Each sample yielded ions representative of the pigments present.     

Direct identification of various copper phthalocyanine pigments in automotive paints and paint smears by laser desorption ionization mass spectrometry

Direct identification of copper phthalocyanine (CuPc) and chlorinated CuPcs in paints for discrimination between blue automobile paints by means of laser desorption mass spectrometry (LDMS) in the absence of a matrix is reported. The models consisted of eight commercially available CuPc pigments applied to a piece of plain white coating paper. The relationship between the peak intensity at m/z 575 of the CuPc, the number of pulsed laser shots, and laser power was compared to optimize laser abrasion. LDMS analysis of the model paints demonstrated that all characteristic components of the CuPc pigments in the paint films were in good agreement with those in the powder pigments. Further, the chlorinated CuPcs in the paint films could be distinguished. A quantity of 42 blue paint films, representing the paints used for painting Japanese domestic trucks, was examined by LDMS analysis. Results indicate that the paints can be classified into four categories based on the chlorinated CuPc components of the paints. Therefore, LDMS spectra of CuPc pigments would be useful for the identification of paints in forensic investigations. Herein, we report the successful identification of the CuPcs in a paint smear on the frame of a bicycle damaged in a hit-and-run accident, using the LDMS spectra.     

Carbon‐Based Fingerprint Powder as a One‐Step Development and Matrix Application for High‐Resolution Mass Spectrometry Imaging of Latent Fingerprints

Carbon‐based materials are often used as matrices for matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS) and its imaging (MALDI‐MSI). However, researchers have refrained from using carbon‐based fingerprint powder (CFP) as a matrix due to high background and contamination. In this work, the compatibility of CFP is reevaluated with MALDI‐MSI using a high‐resolution mass spectrometer (HRMS) and compared to traditional organic matrices. Relevant fingerprint compounds were easily distinguished from carbon cluster peaks when using HRMS. For fair comparison, half of a fingerprint was dusted with CFP while the other half was dusted with traditional organic matrices. All compounds studied had comparable, or higher, signal‐to‐noise (S/N) ratios when CFP was used as the matrix. Additionally, chemical image qualities closely followed the trend of S/N ratios. CFP proved to be an effective one‐step development and matrix application technique for MALDI‐MSI of latent fingerprints, when carbon cluster peaks are well separated by a HRMS.     

Ballpoint pen inks: characterization by positive and negative ion-electrospray ionization mass spectrometry for the forensic examination of writing inks

A method based on profiling of dye components by electrospray ionization mass spectrometry (ESI/MS) is described for the characterization of ballpoint pen inks. The method involves benzyl alcohol (30 microL) extraction of ink from paper. The extracts of ink lines 1 and 5 mm in length are used for direct ESI/MS analysis in positive and negative modes, respectively. The instrumental analysis takes 3 min. Basic and acid dyes in the inks are detected in the positive and negative modes, respectively, with each dye yielding one or two characteristic ion peaks. The mass spectrum, which is mainly a compositional signature of the dyes in the ink, was not affected by the type of paper from which the ink was extracted, or by natural ageing of the ink on document in the absence of light. However, exposure to fluorescent illumination caused dealkylation of polyalkylated basic dyes and resulted in changes in the homologous distribution of the dyes. In this study, a total of 44 blue inks, 23 black inks, and 10 red inks have been analyzed, and the mass spectra were used to establish a searchable library. ESI/MS analysis provides a simple and fast way to compare ink specimens and in combination with on-line library search permits rapid screening of inks for forensic document investigations.     

Application of MALDI‐TOF MS for Estimating the Postmortem Interval in Rat Muscle Samples

Estimating the postmortem interval (PMI) is very important in the forensic sciences. Although many approaches have been used for estimating the PMI, accurate PMI calculations are still difficult. In this study, four Sprague Dawley (SD) rats were sacrificed by suffocation, and muscle samples were collected by dissection at various time intervals (0, 48, 96, and 144 h) after death. All samples were probed using matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS) to obtain molecular images and data for principal component analysis (PCA). The results showed that the peaks at m/z 1511, 1543, 1564, 1586 clearly decreased in intensity from 0 to 144 h postmortem and that the time groups were separated from each other on the PCA score plot. The prediction model showed high recognition capability (95.93%) and cross‐validation (83.72%). Our work suggests that MALDI‐TOF MS can be used to determine the PMI.     

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.     

Evaluation of the Photodegradation of Crystal Violet upon Light Exposure by Mass Spectrometric and Spectroscopic Methods

Abstract: Crystal violet is a very common dye in ballpoint ink. Recent research suggests that the degradation of triarylmethane dyes gives an indication of the age of a ballpoint pen entry on a document. The main problem for the quantitative evaluation of the degradation is that it is highly dependent on the exposure to light. Moreover additional factors, such as additives and substrate play an important role in this process. The aim of this work is to compare the degradation pathways of the pure dye in water and ethanol upon exposure to xenon light by UV/VIS spectrophotometry and laser desorption ionization. Significant differences have been observed in the products and the kinetics of the degradation. N-demethylation, an expected decomposition process, was found to take place only in aqueous solution and kinetics calculations showed that the degradation occurred 2.5 times faster in ethanol compared to water. The degradation of crystal violet in inks from four ballpoint pens on paper was also studied for entries made over 2–3 years. It was observed that degradation reactions were quenched by the presence of another dye due to competitive absorption. It was also observed that the thickness of a stroke (concentration of ink) influenced the degradation process. In the absence of light only one ballpoint pen showed slight degradation. A better understanding of the influence of the paper, ink composition, and storage conditions is necessary to interpret correctly the age of an ink based on the degradation of dyes.     

涤纶纤维上分散染料的液-质分析

目的建立液-质联用技术对涤纶纤维上分散染料进行分析的方法。方法以N,N-二甲基甲酰胺为提取剂,在负离子模式下对12个黄色涤纶纤维上分散染料进行分析。结果12种分散染料在负离子模式质谱检测条件下均得到了有效的分析。根据一级负离子模式下总离子流图中准分子离子峰质荷比的不同,可将其中8种样品一一鉴别开。在此基础上,从染料离子化产物中选取响应信号较强的准分子离子为母离子,再对母离子进行二级质谱全扫描,依据二级质谱图中特征碎片峰的数目、质荷比的异同可对纤维上染料进一步进行鉴别。结论液-质联用法是检验有色纤维上染料的一种科学、有效的方法。     

Comparison of natural and artificial aging of ballpoint inks

Abstract:  Solvent evaporation caused by aging from ballpoint inks was measured by gas chromatography/mass spectroscopy (GC/MS). The sample preparation was carried out with two different thermal desorption systems. The results are compared. Thirteen inks were classified with regard to their solvents, polymers, and additives. The variation of the aforementioned compounds caused by aging was monitored for naturally and artificially aged samples. In this paper, the results are compared and discussed with respect to forensic casework.     

Age determination of ballpoint pen ink by thermal desorption and gas chromatography-mass spectrometry

Two main approaches can be used for determining the age of an ink: indirect dating and direct dating. Indirect dating is based on the chemical analysis of an ink followed by comparison with known samples in a reference collection. The collection should contain information about the inks including the market introduction dates. This approach may allow for an anachronism to be detected. The second concept is based on measuring ink components that change with age. The analysis of solvents in ballpoint inks may be a useful parameter for determining the age of ink on paper. In a previous study, the authors demonstrated that thermal desorption of ink directly from paper, followed by chemical analysis using gas chromatography-mass spectrometry (GC-MS), is a promising procedure for characterizing ink-binder resins and solvents. Preliminary tests showed that monitoring the evaporation of ink solvent from ink on paper is not a suitable method for ink dating. Thermal analysis of ink on paper in two steps revealed that fresh ink releases a relative amount of solvent at a certain low temperature in a defined period of time, which decreases as the ink ages. As a consequence, this relative amount of solvent released at a certain low temperature, and its decrease with time, can be used to estimate ink age. This age-dependent parameter was studied in 85 different inks ranging in age from 1 week to 1.5 years. It was found that some inks showed a significant decrease of this parameter up to an age of several months, and that the aging process can be monitored within this period. For other inks, however, the age-dependent parameter decreases relatively fast, e.g., within a few days, to a constant level, which can be too fast for casework. Based on these results, a general procedure for assessing the age of ballpoint pen inks on paper was developed.     

Some Examples of Applications of a Microthermal Desorption Device in the Forensic Laboratory

Abstract:  Several applications of a microthermal desorption device for analysis of small forensic samples are presented. The method uses a solid phase microextraction holder with the fiber removed. In addition to samples of inks on paper, this device was successfully used for analysis of toners, various stains on bank notes, and lipstick stains on paper. Other small items encountered in a forensic science laboratory were also analyzed: particles of smokeless powder, particles of coffee, and automotive clear topcoat layer. The desorbed compounds were analyzed by gas chromatography with a flame ionization detector or by gas chromatography–mass spectrometry. This device can be used in connection with any kind of gas chromatograph. By selection of different injector temperatures, fractionated thermal desorption of samples is achieved. The procedure was demonstrated on samples of ballpoint pen ink of various age.     

How Much Can a Forensic Laboratory Do to Discriminate Questioned Ink Entries?

Questioned document examiners are frequently required to determine whether the questioned ink entries on a document are written with the same ink, either for forged document identification or relative ink dating. How many methods are sufficient to discriminate potentially different inks? In this paper, 18 blue ballpoint inks were first nondestructively examined with a video spectral comparator. The ink entries were subsequently extracted with organic solvents for analysis of the volatile solvents and dyes by GC/MS, TLC, and LC‐MS/MS. The 18 ink samples were divided into 10, 9, 12, and 14 categories by these four methods, respectively. With the combined results, there were only two inks that remained indistinguishable, but they were further differentiated by microscopy. Therefore, to achieve effective discrimination of ballpoint ink entries, the authors suggest that a complete examination should include an analysis of their optical features, volatile solvents, and dyes as well as the ink quality.