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It is a common practice in forensic casework to use presumptive tests for blood stains before DNA extraction and testing. Stains are usually swabbed and then the swabs are sent for analysis. The Kastle-Meyer (KM) and Leucomalachite green (LMG) presumptive tests for blood are widely used, and their sensitivities have been thoroughly tested in the literature in solution and directly on stains, but not on swabbed stains to mimic casework. In this study, the sensitivity of the KM and LMG tests was tested on eight blood dilutions on cotton fabric and ceramic tile that were stained and subsequently swabbed. Both tests showed sensitivity up to 1:5000, which is slightly lower than reported values in solution or directly on stain but still highly effective in most cases. Stains were also cleaned with common agents, then swabbed and re-tested. Stained ceramic tiles cleaned with soap/water or bleach gave mixed positive and negative results for the 1:10 dilution, presumably due to variance in how thoroughly each investigator cleaned the stain, and other dilutions were undetectable after cleaning. The LMG test gave false positives for bleach cleaned stains, due to reagent reactivity with bleach. Surprisingly, blood was detectible up to the 1:100 dilution with both tests on stained cotton fabric that was cleaned in a washing machine with detergent and dried. Ultimately the KM and LMG presumptive tests remain effective tools for swabbed blood stains, and their practicality for cleaned stains is dependent on material containing the stain, cleaning agent and processing. [ABSTRACT FROM AUTHOR]

Leucomalachite green (LMG), a derivative antimicrobial agent, can accumulate in animal tissues for long periods and is categorized as a carcinogen affecting consumer health. Therefore, determining the contamination of aquatic products by LMG is necessary to prevent residual contamination. In the work described herein, the detection of LMG based on a colorimetric method using gold nanoparticles (AuNPs) was developed. The specificity of isolated single-stranded DNA aptamers to LMG was proven by using LMG and other antibiotics. The binding affinity of the LMG-aptamer showed a Kd value of 1.94 pM. Furthermore, a colorimetric detection module was successfully developed, showing a limit of detection (LOD) value of LMG at 250 nM using standard LMG substance. Taken together, this research demonstrates a rapid and sensitive method to detect chemical residues, reducing the possibility of contamination in fishery products. Considering these findings, the aptamer-based colorimetric approach might be feasible to utilize as a biosensing module, providing a rapid, inexpensive analysis method for detecting deposited chemical residues. [ABSTRACT FROM AUTHOR]

Application of malachite green (MG) and leucomalachite green (LMG) in fish farm water causes an environmental problem. This study proposes for the first time a sensitive and convenient electrochemical impedance spectroscopy (EIS) method for determining MG and LMG by a bovine serum albumin-decorated gold nanocluster (BSA-AuNC)/antibody composite film-based immunosensor. In order to improve the analytical performance, the glassy carbon electrode (GCE) was modified with 1, 4-phenylenediamine to form a stable layer, and then, BSA-AuNCs were covalently bound to the GCE. An adequate quantity of the polyclonal antibody of LMG was immobilized onto the surface of the BSA-AuNCs by the chemical reaction of EDC/NHS. The sensors can respond to the specific target based on specific covalent bonding. The experimental parameters, such as the pH, incubating concentration, and time, have been investigated and optimized. The calibration curve for LMG was linear in the range of 0.1~10.0 ng/mL with the limit of detection (LOD) 0.03 ng/mL. Furthermore, the sum of MG and LMG was detected in fish farm water by MG reduction. The recovery was between 89.7 % and 99.2 % in spiked samples. The EC sensor method was also compared with the ELISA method and validated by the LC-MS/MS method, which proves its great promise as a field instrument for the rapid monitoring of MG and LMG pollution. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Background Bloodstains often constitute the major physical evidence in criminal investi-gations. In many cases, the bloodstains found by the crime scene examiner are minute, possibly because of dissimulation efforts by the perpetrator to eliminate evidence that reveals his identity. In such cases, short tandem repeat (STR) detection procedures must be performed using the same minute bloodstain evidence on which presumptive tests had been performed earlier. In the present study, two of the most often used presumptive test reagents, phenolphthalein and leucomalachite green, were tested to determine their effects on the ability to obtain STR profiles from minute bloodstains. Methods Dried minute bloodstains obtained from 10 donors were treated with phenolphthalein and leucomalachite green. After various times, genomic DNA was extracted from the treated samples using a QIAamp DNA Micro Kit. DNA was quantified with real-time PCR using a Quantifiler Kit. STR loci were amplified using an AmpFLSTR Identifiler Plus Kit, and the amplified products were separated via capillary electrophoresis in a 3130 Genetic Analyzer. Results Full DNA profiles were obtained from all minute bloodstain samples treated with phenolph-thalein when extracted after intervals ranging from 1 h to 1 week. In contrast, the DNA in minute bloodstain samples treated with leucomalachite green was severely degraded, especially after relatively long intervals, leading to poor partial DNA profiles. Conclusion Phenolphthalein is recommended as a safe presumptive test reagent for the detection of blood evidence recovered from crime scenes that might subsequently undergo DNA profiling analysis. [ABSTRACT FROM AUTHOR]

Farmed fish are exposed to risks from feed-borne chemical contamination, such as leucomalachite green (LMG) in fish meal. Consequently, the use of malachite green is prohibited in aquacultural practice. An improved understanding of the toxicity of dietary LMG provided to farmed fish is needed in order to manage risk. Oreochromis niloticus specimens were fed experimental diets containing 0, 100, 500, and 2500 μg/kg LMG for 28 days. On sampling days 7, 14, and 28, fish in the exposed groups had detectable levels of LMG. Accumulation levels were approximately 12.2 % (in the liver) and 6.2 % (in the muscle) of the LMG concentration in the feed (104, 510, and 2200 μg/kg). Hematocrit and mean corpuscular hemoglobin concentrations in the 500 and 2500 μg/kg groups were significantly elevated at 7 and 14 days. Hemoglobin in the group that received the highest dose was significantly higher than that in the control group. Significant increases in the activities of alanine aminotransferase and alkaline phosphatase were also detected in the group receiving the highest dose. Total cholesterol concentrations in all of the exposed groups were significantly higher than those in the control group. These observations of toxicity were dose dependent. Histological changes in gills and livers were observed in LMG-exposed fish. [ABSTRACT FROM AUTHOR]

The persistence of malachite green (MG), and its metabolite leucomalachite green (LMG), in fish tissues is still unclear, leading to many trade disputes. In this research, we established and evaluated an HPLC method that could detect MG and LMG simultaneously, and then investigated the persistence of these two toxins in the tissues of juvenile perch ( Lateolabrax japonicus) post sub-chronic MG exposure at 1.0 mg/L. Exposure lasted for 2 h everyday and was repeated six times. The perch were then placed in MG-free seawater for 100 d to eliminate the toxins. Results show that MG accumulated in the tissues, including the gills, liver, muscle, blood and viscera, and then was metabolized rapidly to LMG. The concentrations of these two toxins increased significantly with the accumulation process. In general, the highest concentrations of MG and LMG in all tissue exceeded 1 000 μg/kg, except for MG in the muscle. The order of accumulation levels (highest to lowest) of MG was gill>blood>liver>viscera>muscle, while that of LMG was liver>blood>gill>viscera>muscle. High levels of MG or LMG could persist for several hours but decreased rapidly during the elimination process. The concentration of LMG was much higher than that of MG during the experiment, especially in the gill, liver and blood. Therefore, the three tissues play important roles in toxin accumulation, biotransformation, and elimination. Although the MG and LMG concentrations in muscle were much lower than in other tissues, the content still exceeded the European minimum required performance limit (MRPL), even after 2 400 h (100 d) of elimination. This demonstrates that it is extremely difficult to eliminate MG and LMG from tissues of perch, and therefore use of these toxins is of concern to public health. [ABSTRACT FROM AUTHOR]

Malachite green oxalate (MG oxalate) and leucomalachite green (LMG) have been prepared and certified as pure reference materials. The purities of MG oxalate and LMG were assessed by high-performance liquid chromatography–diode array detection (HPLC–DAD), nuclear magnetic resonance (NMR) spectroscopy, differential scanning calorimetry (DSC), Karl Fischer titration, ashing and thermogravimetric analysis (TGA). MG oxalate was purified by supercritical fluid extraction (SFE). Prior to purification, commercial MG oxalate purity was estimated to be about 90%. The main impurities present in SFE-purified MG oxalate were identified and quantified using HPLC–DAD. The main impurities were found to be monode-MG (monodemethylated MG oxalate synthesis impurity), 4-(dimethylamino)benzophenone (4-DMABP), MG-carbinol and LMG. The homogeneity of both reference materials was also determined. Issues associated with the stability of LMG and MG oxalate in solution forced an extensive study investigating different parameters i.e. solvent, acid, analyte concentration and temperature. MG oxalate (100 μg/mL) was found to be stable in acetonitrile containing 1% v/v glacial acetic acid for at least 155 days and LMG (100 μg/mL) was stable in acetonitrile for at least 133 days. The final purity value for MG oxalate was 94.3 ± 1.4% m/m at the 95% confidence interval (or 67% m/m if MG cation is reported). For LMG, the certified purity was found to be 98.8 ± 0.8% m/m at the 95% confidence interval. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]