Metal-organic frameworks as solid-phase microextraction adsorbents for the determination of triacetone triperoxide by gas chromatography-mass spectrometry.

Triacetone triperoxide (TATP) is a high-power explosive which is often used by criminals. The detection of TATP is of great significance for solving the explosion cases. However, the preconcentration and analysis of trace levels of TATP still pose challenges for analytical researchers. In this study, metal-organic frameworks (MOFs), including IRMOF-8, MOF-5, UIO-66, ZIF-8, and MIL-101(Cr), were immobilized on a stainless steel wire using a physical adhesive method as a solid-phase microextraction (SPME) fiber coating. The prepared fibers with a controllable thickness were used for the extraction of TATP followed by gas chromatography-mass spectrometry (GC-MS) analysis. Under the identical experimental conditions, the IRMOF-8-coated fiber exhibited higher extraction efficiency for TATP than the other fibers. The IRMOF-8-coated fiber was then characterized using scanning electron microscopy and thermogravimetric analysis. The results indicated that the IRMOF-8-coated fiber not only had good thermal and chemical stabilities but also afforded a high TATP extraction efficiency. Under the same extraction conditions, the extraction efficiency of the IRMOF-8-coated fiber was 2-8 times higher than those of commercial fibers. The limit of detection was 13 ng/mL, and linearity was observed in the range of 50-5000 ng/mL with a correlation coefficient greater than 0.998. The intraday repeatability (n = 6), interday repeatability (n = 3), and fiber-to-fiber reproducibility (n = 3), were 4.1 %, 4.8 %, and 8.0 %, respectively. The recoveries of TATP from the simulated tap water and soil samples were 87.32-90.57 % and 88.76-100.93 %, respectively, with relative standard deviations lower than 11.11 % (n = 3). The above method was successfully applied for the detection of TATP transferred from a finger to a paper surface, demonstrating its good application prospects in the analysis of trace TATP.

Application of lead isotope signature and likelihood ratio evaluation in a shooting incident investigation.

Bullet projectiles are often found in shooting crime scenes and can help identify the cartridges used in shooting events. The most conclusive way to link a fired projectile to a firearm is by identifying the unique markings of the barrel on the projectile or case. But in circumstances where the investigation of striation marks is impossible, lead isotope ratio signature analysis of projectile fragments can facilitate building the linkage of the suspect, the cartridge used and the victim. This work presents a shooting incident happened in a hunting field, where a hunting guide was shot to death. A tiny lead projectile fragment was collected from the victim's head. Pb isotope ratios measured by laser ablation multicollector inductively coupled plasma mass spectrometry (LA-MC-ICPMS) were used to make comparisons between the fragment and the shots seized from two suspects. Additionally, a reference dataset comprising 50 lead bullet projectiles from various manufacturers was characterized using LA-MC-ICPMS. The evaluation of the comparison was carried out by multivariate likelihood ratio (LR) computations based on the reference dataset. The lead isotope ratio and likelihood ratio evaluation methods helped infer the source of the fragment and were highly supportive in the shooting incident investigation.

[Applications of functional materials-based solid phase microextraction technique in forensic science].

Sample extraction is a crucial step in forensic analysis, especially when dealing with trace and ultra-trace levels of target analytes present in various complex matrices (e. g., soil, biological samples, and fire debris). Conventional sample preparation techniques include Soxhlet extraction and liquid-liquid extraction. However, these techniques are tedious, time-consuming, labor-intensive and require large amounts of solvents, which poses a threat to the environment and health of researchers. Moreover, sample loss and secondary pollution can easily occur during the preparation procedure. Conversely, the solid phase microextraction (SPME) technique either requires a small amount of solvent or no solvent at all. Its small and portable size, simple and fast operation, easy-to-realize automation, and other characteristics thus make it a widely used sample pretreatment technique. More attention was given to the preparation of SPME coatings by using various functional materials, as commercialized SPME devices used in early studies were expensive, fragile, and lacked selectivity. Examples of those functional materials include metal-organic frameworks, covalent organic frameworks, carbon-based materials, molecularly imprinted polymers, ionic liquids, and conducting polymers, all widely used in environmental monitoring, food analysis, and drug detection. However, these SPME coating materials have relatively few applications in forensics. Given the high potential of SPME technology for the in situ and efficient extraction of samples from crime scenes, this study briefly introduces functional coating materials and summarizes the applications of SPME coating materials for the analysis of explosives, ignitable liquids, illicit drugs, poisons, paints, and human odors. Compared to commercial coatings, functional material-based SPME coatings exhibit higher selectivity, sensitivity, and stability. These advantages are mainly achieved through the following approaches: First, the selectivity can be improved by increasing the π-π, hydrogen bonds, and hydrophilic/hydrophobic interactions between the materials and analytes. Second, the sensitivity can be improved by using porous materials or by increasing their porosity. Third, thermal, chemical, and mechanical stability can be improved by using robust materials or fixing the chemical bonding between the coating and substrate. In addition, composite materials with multiple advantages are gradually replacing the single materials. In terms of the substrate, the silica support was gradually replaced by the metal support. This study also outlines the existing shortcomings in forensic science analysis of functional material-based SPME techniques. First, the application of functional material-based SPME techniques in forensic science remains limited. On one hand, the analytes are narrow in scope. As far as explosive analysis is concerned, functional material-based SPME coatings are mainly applied to nitrobenzene explosives, while other categories, such as nitroamine and peroxides, are rarely or never involved. Research and development of coatings is insufficient and the application of COFs in forensic science has not yet been reported. Second, functional material-based SPME coatings have not been commercialized as they don't yet have inter-laboratory validation tests or established official standard analytical methods. Therefore, some suggestions are proposed for the future development of forensic science analyses of functional material-based SPME coatings. First, research and development of functional material-based SPME coatings, especially fiber coatings with broad-spectrum applicability and high sensitivity, or outstanding selectivity for some compounds, is still an important direction for SPME future research. Second, a theoretical calculation of the binding energy between the analyte and coating was introduced to guide the design of functional coatings and improve the screening efficiency of new coatings. Third, we expand its application in forensic science by expanding the number of analytes. Fourth, we focused on the promotion of functional material-based SPME coatings in conventional laboratories and established performance evaluation protocols for the commercialization of functional material-based SPME coatings. This study is expected to serve as a reference for peers engaged in related research.

Lead isotope measurement of primer gunshot residues and likelihood ratio predictions for forensic cartridge discrimination and individualization in China.

Gunshot residues (GSR), cartridge projectiles, and casings are frequently encountered evidence in gun-related forensic investigations. However, in circumstances where the investigation of striation marks is impossible, such as unrecovered or deformed projectiles and cartridge casings, GSR deposited on the hands or clothes of the shooter and victim-related items can provide information to establish a link between the suspect, the firearms used, and the victim. Since the formula of primers used by all cartridge manufacturers in China is identical, links based on the conventional morphological and compositional analysis of GSR are difficult to establish. However, the abundance of lead isotopes in primer components of lead styphnate varies significantly, and a fundamental understanding of these differences may facilitate the validation of primer (p)GSR evidence in forensic investigations. Here, 44 pGSR samples were characterized by Pb isotope ratios of 206 Pb/204 Pb, 207 Pb/204 Pb, and 208 Pb/204 Pb using laser ablation multicollector inductively coupled plasma mass spectrometry. There was no obvious mass fractionation of the lead isotope ratios of the primers from individual cartridges analyzed before and after the shooting process, thereby establishing a basis for the comparison of pGSR and unfired cartridges. Evaluation of the results using univariate likelihood ratio (LR) computations revealed low rates of misleading evidence (<0.53%) The results demonstrated that lead isotope ratio analysis of pGSR and LR predictions can provide a practicable method for forensic cartridge discrimination and individualization.

Determination of disperse dyes on polyester fibers by UHPLC-Orbitrap MS.

The determination of fiber dyes is important in forensic investigations. Although a variety of fiber dyes detection methods have been established, the sensitive and accurate determination of trace fiber dyes remains a challenge due to the possible interferences caused by complex environmental matrix and various fiber additives. Orbitrap mass spectrometry (Orbitrap MS) is a type of high-resolution mass spectrometry with high qualitative accuracy and detection sensitivity which highly meet the identification requirements of fiber dyes in real cases. However, the application of Orbitrap MS in fiber dye analysis is limited. In this regard, this study used polyester fiber, which is the most commonly-found fiber in forensic cases, as a model and established a UHPLC-Orbitrap MS method to analyze disperse dyes on polyester fibers. Using the optimized UHPLC-Orbitrap MS method, nine disperse dyes were accurately identified and well separated, and the limits of detection ranged between 0.1 ng/mL and 5.0 ng/mL. The developed method was applied to analyze actual fiber samples, and dyes from single fibers of 1 mm in length could be accurately detected. The established method is sensitive, accurate, and demonstrates good application prospects.

Desulfurization of black powder for isotopic profiling using isotope ratio mass spectrometry.

The stable isotope analysis of black powder is of utmost significance with regard to its further comparison and traceability in explosion-related cases. To date, only a limited number of studies have reported the stable isotope analysis of black powder using isotope ratio mass spectrometry (IRMS). However, because the sulfur content in black powder is high, sulfur deposition is extremely likely during the analysis process, thereby damaging the instrument and affecting the reproducibility of the results. Therefore, desulfurization of black powder samples prior to IRMS analysis is extremely desirable. Herein, based on the difference in the solubility of black powder components, chloroform was selected to desulfurize black powder. The results demonstrated that chloroform extraction could effectively eliminate sulfur from the black powder. The isotope fractionation effect caused by desulfurization was also studied. The changes in δ13C, δ15N, and δ18O after desulfurization were close to the standard deviation values and did not affect the analysis of black powder. Desulfurization and stable isotope analysis were performed on 12 black powder samples obtained from cases that happened in eight different areas in China, and the carbon, nitrogen, and oxygen isotope ratios in all the samples were effectively distinguished. Therefore, the established method can successfully distinguish the stable isotope ratios in black powder samples using IRMS after desulfurization while avoiding damage to the instrument and enhancing the stability of the results.

Mineralogical and elemental data for soil discriminating and geolocation tracing.

One of the key tasks of soil analysis in forensic sciences is to provide information about its diversities and geolocation. In fact, soil analysis is relevant for forensic geologists. In this study, a total of 80 soil samples were collected from eight Chinese cities (10 samples per city). Different minerals and their relative percentages were analyzed by the X-ray diffraction (XRD) method. In addition, the relative amounts of montmorillonite, kaolinite, amphibole, feldspar, calcite, and dolomite provided information about the origin of a soil, either if it came from a northern or southern city of China. The oxide weight percentages of 10 elements of Al2O3, SiO2, Fe2O3, K2O, Na2O, MgO, CaO, P2O5, MnO, and TiO2 were also obtained by using X-ray fluorescence (XRF) from the 80 soil samples. Moreover, principal component analysis (PCA) and hierarchical clustering analysis (HCA) methods were performed for dimensionality reduction, elemental marker identification and soils classification to the city they came from purposes. The eighty soils analyzed in this study could be tracked correctly to their city of origin. The K-Nearest Neighbors (KNN) model was done to evaluate the prediction ability based on the soil elemental composition, and it was confirmed by cross validation methods. The results demonstrated that mineralogical and elemental composition can provide powerful information for soil discrimination and source tracing.

Determination of stable nitrogen isotopic ratios of nitrate ions in ammonium nitrate.

Ammonium nitrate (AN) is one of the most commonly used explosives in criminal cases. The comparison and source-tracing of AN is important for investigation of attribution and fingerprinting of an explosive used at different events. The stable isotope signature of AN is an important index for comparison and tracing. However, the characteristics of the stable nitrogen isotopic ratios of AN (δ15 NNH4NO3 ) alone are not sufficient to achieve a fine comparison between different AN samples. To increase the comparison index and further improve the discriminability between stable nitrogen isotopic ratios of different ANs, a method of isolation and analysis of nitrate ions in AN was established using stable-isotope-ratio mass spectrometry (IRMS). The method was based on the principle that strong alkali react with AN to produce ammonia and nitrate. After the isolation, stable nitrogen isotopes of nitrate ions (δ15 NNO3 ) were obtained using IRMS, and then the stable nitrogen isotopes of ammonium ions from AN (δ15 NNH4 ) was calculated according to the principle of mass balance. The results show that the method is effective for the isolation of nitrate ions without notable isotope fractionation. The developed method was applied to analyze and discriminate AN samples from eight different cities in China. Three samples out of the initial eight AN samples with similar δ15 NNH4NO3  values were further distinguished by their δ15 NNH4 and δ15 NNO3  values. The isolation and stable-nitrogen isotopic analysis method developed for nitrate ions in AN is simple and effective, thereby increasing the discriminability of the stable isotope ratios in AN.

A case study in forensic soil comparison.

Soil examination can provide useful forensic information about the spatial location and suspect's activities. Many techniques have been applied for soil comparison and provenance determination in criminal investigations. Pollen and diatom identification, which has the potential to provide an independent ecological assessment of soil evidence, is currently underused in forensic soil analysis. This work presents a case study of application of these methods to help criminal investigation in a murder case, which happened in an irrigation ditch in Hunan Province, southern China. Soils from the suspect's clothes, the exact crime scene spot in the irrigation ditch, along the ditch and the reference ditches were collected and analyzed. In addition to the element and mineral analysis, pollen and diatom assemblages were analyzed for further comparison. The statistical methods of hierarchical cluster and cosine similarity analysis were carried out to assist in soil comparison and provenance determination. The results showed that soil on the suspect's clothes had a high probability to share the same source with the soil from the crime scene in the irrigation ditch. The suspect confessed to murder based largely on the soil examination result even without other evidences.

Purification of ammonium nitrate via recrystallization for isotopic profiling using isotope ratio mass spectrometry.

Stable-isotope analysis of ammonium nitrate (AN) plays an important role for comparing and tracing the sources of AN samples. Numerous studies have reported the application of isotope ratio mass spectrometry (IRMS) for the stable-isotope analysis of AN. However, in real cases, AN is often mixed with liquid hydrocarbons, organic explosives, wood flour, and sodium chloride. Therefore, the purification of AN samples prior to IRMS analysis is essential. Based on the different solubilities of AN at different temperatures, AN samples were purified by recrystallization. The results show that recrystallization can effectively purify AN, and the isotope fractionation effect caused by recrystallization increases the stable-isotope ratio (δ15N increased by 0.07-0.14‰, δ18O increased by 0.15-0.33‰, and δ2H increased by 4.61-18.16‰). The change of δ15N and δ18O were close to the standard deviation values, and did not affect the differentiation of AN. δ2H was deemed unsuitable for AN differentiation, owing to isotopic exchange during recrystallization. Eight AN samples from different cities and four AN samples produced in different years and batches from the same manufacturer in China were analyzed after recrystallization. Both groups of AN samples could be distinguished by combining the nitrogen and oxygen isotope ratios. Therefore, recrystallization can be used as a means of AN purification for the accurate determination of AN in ammonium nitrate fuel oil and explosive residues.

[Recent advances in stable isotope ratio analysis of common explosives].

The ratio of stable isotopes of the elements in explosives differs depending on the raw materials obtained from different geographical sources or the production processes adopted. Hence, this ratio can be used as an important index for the comparison and trace of explosives. Isotope ratio mass spectrometry (IRMS), a high-precision method for the analysis of stable isotope ratios, has evolved into a mature tool in this regard. In combination with elemental analysis, gas chromatography, liquid chromatography, etc., IRMS is widely used in food safety, environmental protection, forensic science, and other fields. IRMS also plays an important role in the comparison and trace of explosives. Since its application to distinguish trinitrotoluene (TNT) produced in different countries in 1975, IRMS has been successfully used in the analysis of various explosives. However, there is no systematic summary on the research progress on the stable isotope ratio analysis of common explosives. This paper provides a brief description of the related principle, instrumental composition, and characteristics of stable isotope ratio analysis. Methods for the stable isotope ratio analysis of common explosives such as ammonium nitrate, black powder, TNT, pentaerythritol tetranitrate (PETN), and cyclotrimethylene trinitroamine (RDX) are reviewed. The bulk stable isotopic ratio analysis method was used in most of the studies to determine the total isotope ratio of the sample. A compound-specific isotope analysis method was also employed to determine the isotope ratio of organic explosives in a complex matrix. The reported stable isotope ratios of explosives such as ammonium nitrate, black powder, and TNT produced in different countries are summarized. The discrimination ability of the stable isotope ratio for explosives is discussed. Based on the stable isotope ratio, explosives from different sources can be distinguished effectively. By combining the results of elemental analysis with the ICP-MS results, the discrimination efficiency of different samples could be further improved. The influence of relevant factors on the isotope ratio during the production and storage of explosives are collated. There is a strong correlation between the stable isotope ratios of explosives and raw materials. The stable isotope ratios of TNT, PETN, and other explosives are related to that of nitric acid used in the production. The stable isotope ratios of nitrogen and oxygen in the explosive are relatively stable and almost unchanged within one year of production. The complexity of the environmental matrix at the explosion site and the low concentration of explosive residues make the stable isotope analysis of explosive residues challenging. However, the changes in the stable isotope ratio before and after the explosion are discussed. Since there is no information on the application of stable isotope analysis to the traceability of explosives, the paper mentions that the standardized explosive sample pretreatment, stable isotope analysis method, collection and analysis of large amounts of explosive samples, and explosive stable isotope database are the basis of explosive traceability. This paper also outlines the existing challenges in the analysis of the stable isotope ratios of explosives, including the small number of explosive samples, lack of a stable isotope explosive analysis database, and difficulty in the stable isotope analysis of explosive residues. Possible solutions to these problems are proposed, followed by suggestions for the future development of the stable isotope ratio analysis of common explosives. The suggestions include establishing an effective extraction and enrichment method for explosive residues, combining IRMS with GC or LC for analyzing explosives, establishing a comprehensive process for the analysis of the stable isotope ratios of inorganic and organic explosives, and comparison and analysis of the stable isotope analysis data using statistical methods.

Forensic comparison analysis of plastic vehicle headlamp lens by gel permeation chromatography and statistical methods.

Comparison analysis of vehicle headlamp lens fragments can help establish links between the relevant vehicle with crime scenes or provide useful information to search the related vehicles. Headlamp lenses are mainly made of polycarbonate with very few signature additives, which makes them difficult to be discriminated by commonplace examination methods of infrared spectroscopy and scanning electron microscope coupled with energy dispersive X-ray spectroscopy. In this study, molecular weights (Mw) and the polydispersity index (PDI) value of 50 vehicle headlamp lens fragments from different vehicles were measured by gel permeation chromatography, and Hotelling's T2 statistical method was applied to facilitate interpretation of the data. Among the total C 50 2 = 1225 pairs of comparisons between 50 samples, 62 pairs cannot be distinguished, resulting in a discrimination rate of 94.94%. It suggested that the method of gel permeation chromatography and Hotelling's T2 statistical analysis were powerful to make further discrimination between plastic vehicle headlamp lens fragments.

Prediction of iron content in soil based on microspectrophotometry analysis.

Soil is a very important type of trace evidence. The iron content of soil is of great significance in distinguishing soil types, discriminating among different soils, and tracing soils. However, conventional methods for analyzing the iron content of soil are expensive, laborious, and time-consuming. Previous studies have shown that the color of soil correlates well with its hematite content. This article thus deals with the indirect determination of iron content using soil color as a proxy. Soil color measurements were conducted using microspectrophotometry (MSP), and resulting data were transformed into chromaticity value (L*, a*, and b*). Predictions using the redness index in conjunction with a linear regression model were compared with those using the chromaticity value and a back propagation neural network (BPNN) model. The influences of different modeling conditions on the modeling accuracy were compared, and more accurate predictions were achieved when the iron content was higher than 2.13%. The BPNN model produced predictions with R2 and RMSE values of 0.955 and 0.336%, which were better than the predictions of the linear regression model (R2: 0.859, RMSE, 1.07%). We thus demonstrated that MSP can be used for fast, accurate, and non-destructive measurements of soil color and prediction of its iron content. Although the results may not be as precise as conventional laboratory analysis, they still provide more information with acceptable accuracy, which should have promising applications in forensic applications.

The analysis of soil evidence to associate criminal tool and location.

The analysis of soil evidence is crucial for the association of crime scenes, tools, and people involved. This work presents a case of excavating ancient tombs to steal antiques. The soil from the criminal tools, tombs, and an antique were collected and analyzed. The color, element, and mineral composition of soil evidence were detected through microspectrophotometry, X-ray fluorescence, and X-ray diffraction. Chemometrics, including hierarchical cluster analysis and K-means clustering, was also utilized to assist in the differentiation and classification. The source of the antique was successfully inferred. The criminal tool and the location of the crime scene were also associated correctly. The soil analysis was highly supportive in reconstructing the crime scene.

Characterization of automotive paint by optical coherence tomography.

Automotive paint is common trace evidence that plays a significant role in many vehicle-related criminal cases. However, the conventional methods of obtaining tomographic images tend to damage the samples. Optical coherence tomography (OCT) is a novel method to obtain high-resolution and cross-sectional images of the automotive paints in a non-destructive, and high-speed manner. In this study, OCT was applied to image and analyze the automotive paint, using scanning electron microscope (SEM) as reference. Eight automotive paint samples of different brands were examined. The images of multi-layer structures provided by OCT system with 5µm depth resolution were consistent with those by SEM. To distinguish different paints with similar visual appearance, we extracted internal structural features from the images using peak analysis and optical attenuation fit. Six characterized parameters were found to distinguish the samples including the optical path length (OPL) of base coat, the optical attenuation coefficient (OAC) of base coat, the OPL of clear coat, the back-scattering ratio (BSR) of clear coat and base coat, the OPL of primer surfacer, and the BSR of base coat and primer. Statistical differences were evaluated by an independent t-test with p<0.05. OCT was applied to analyze repainted paint as well. Three-dimensional OCT reconstruction of the paints was also implemented to create en face (transverse section) images for morphology examination and comparison. These results suggest that OCT imaging can provide additional new features for analyzing the automotive paints and thereby may be a promising supplement to traditional methods. Meanwhile, the OCT system is favorable for achieving in-situ and real-time examination at the scene of crime.

G-quadruplex DNA aptamers for zeatin recognizing.

Zeatins, a major type of cytokinin, are ubiquitous in higher plants, and involve in regulating a wide range of developmental processes. The development of highly specific ligands to zeatins would be very useful in plant biological research. Here we describe a group of oligonucleotide ligands (aptamers) generated against trans-zeatin. The optimized aptamers possess strong affinity to trans-zeatin and trans-zeatin riboside (Kd=3-5 µM), and relatively weak affinity (Kd=27-30 µM) to cis-zeatin and dihydrozeatin. These aptamers adopt a hairpin-G-quadruplex structure for binding to zeatin. A fluorescence turn-on aptasensor based on graphene oxide (GO) was developed for the recognition of zeatins. The specificity assay of this aptasensor shows good response to zeatins, and no response to the adenine derivatives (analog of zeatins) abundantly existing in biological samples. These results show the great potential of these aptamers in chemical analysis and biological investigation of zeatins.

Rational design of Hg2+ controlled streptavidin-binding aptamer.

Through substituting two canonical base pairs of a streptavidin-binding aptamer with T-T mismatched base pairs, a new aptamer was constructed. Its binding ability could be controlled by Hg(2+) through the formation of T-Hg-T metal-base pairs.

Functional-group specific aptamers indirectly recognizing compounds with alkyl amino group.

Aptamers are usually generated against a specific molecule. Their high selectivity makes them only suitable for studying specific targets. Since it is nearly impossible to generate aptamers for every molecule, it can be of great interest to select aptamers recognizing a common feature of a group of molecules in many applications. In this paper, we describe the selection of aptamers for indirect recognition of alkyl amino groups. Because amino groups are small and positive charged, we introduced a protection group, p-nitrobenzene sulfonyl (p-nosyl) to convert them into a form suitable for aptamer selection. Taking N(ε)-p-nosyl-L-lysine (PSL) as a target, we obtained a group of aptamers using the SELEX technique. Two optimized aptamers, M6b-M14 and M13a exhibit strong affinity to PSL with the K(d) values in the range of 2-5 µM. They also show strong affinity to other compounds containing p-nosyl-protected amino groups except those also possessing an α-carboxyl group. Both aptamers adopt an antiparallel G-quadruplex structure when binding to targets. An aptamer beacon based on M6b-M14 showed good selectivity toward the reaction mixture of p-nosyl-Cl and alkyl amino compounds, and could recognize lysine from amino acid mixtures indirectly, suggesting that aptamers against a common moiety of a certain type of molecules can potentially lead to many new applications. Through this study, we have demonstrated the ability to select aptamers for a specific part of an organic compound, and the chemical conversion approach may prove to be valuable for aptamer selection against molecules that are generally difficult for SELEX.

Selective isolation of G-quadruplexes by affinity chromatography.

G-quadruplex (G4) is a characteristic secondary structure of nucleic acids containing repetitive tandem guanines. G4-forming sequences are found prevalent in the human genome by bioinformatics analysis. Accumulating evidence has suggested that G4s are involved in many biological processes. Selective isolation of G4s would be an effective tool in the study of G4s. In this paper, we prepared four affinity matrixes using hemin or a perylene derivative (N,N'-Bis-(2-(amino)ethyl)-3,4,9,10-perylenetetracarboxylic acid diimide, Pery01) as ligand, and investigated the retention behaviors of different G4s on these matrixes. Our experimental results suggest that the π-π stacking interaction between ligand and G-tetrad plays a key role in the selective isolation of G4s, whereas the electrostatic interaction between DNA and matrix causes the nonspecific binding. One matrix prepared by immobilizing Pery01 on polyglycidylmethacrylate (PGMA) beads through an aminocaproic acid spacer exhibits good selectivity for parallel structure G4s and has been successfully used to directly isolate a spiked parallel G4 from plasma.

Characterization of G-quadruplex/hemin peroxidase: substrate specificity and inactivation kinetics.

Recently, G-quadruplex/hemin (G4/hemin) complexes have been found to exhibit peroxidase activity, and this feature has been extensively exploited for colorimetric detection of various targets. To further understand and characterize this important DNAzyme, its substrate specificity, inactivation mechanism, and kinetics have been examined by comparison with horseradish peroxidase (HRP). G4/hemin DNAzyme exhibits broader substrate specificity and much higher inactivation rate than HRP because of the exposure of the catalytic hemin center. The inactivation of G4/hemin DNAzyme is mainly attributed to the degradation of hemin by H(2)O(2) rather than the destruction of G4. Both the inactivation rate and catalytic oxidation rate of G4/hemin DNAzyme depend on the concentration of H(2)O(2), which suggests that active intermediates formed by G4/hemin and H(2)O(2) are the branch point of catalysis and inactivation. Reducing substrates greatly inhibit the inactivation of G4/hemin DNAzyme by rapidly reacting with the active intermediates. A possible catalytic and inactivation process of G4/hemin has been proposed. These results imply a potential cause for the hemin-mediated cellular injury and provide insightful information for the future application of G4/hemin DNAzyme.