Analysis of Triphenylmethane Dye Residues and their Leuco-Forms in Frozen Fish by LC-MS/MS, Fish Microbial Quality, and Effect of Immersion in Whole Milk on Dye Removal.

A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was validated and used to quantify crystal violet (CV), leucocrystal violet (LCV), malachite green (MG), leucomalachite green (LMG), and brilliant green (BG) residues in frozen fish (121 samples) from various countries, in order to detect the use of prohibited antibiotic dyes in fish for human consumption. The microbial quality of the fish was also assessed along with the effectiveness of a simple treatment with whole fat milk to reduce the levels of CV and LCV contamination. CV and LCV were the only two residues detected. They were found in farmed Pangasius (0.362 to 41.34 µg/kg and 0.178 to 10.58 µg/kg, respectively) and Tilapia (1.24 to 9.48 µg/kg and 1.29 to 2.81 µg/kg). Based on aerobic plate count (APC), 74%, 59%, and 55% of the samples of Tilapia fillets (from China) and Pangasius fillets (United Arab Emirates and Vietnam), and 100% and 50% of the skin samples of Hake (Argentina and U.S.A.) were of unacceptable microbial quality (APC > 107 cfu/g). Human pathogens, namely Escherichia coli, Staphylococcus aureus, and Vibrio spp., were detected in most fish. A significant reduction in CV and LCV concentrations by more than a third was achieved after immersing Pangasius and Tilapia fillets in whole fat milk for 120 minutes. These findings support the necessity of regular inspections and monitoring of CV and other antibiotic dye residues in fish, along with routine assessments of fish microbial quality, in order to protect public health. PRACTICAL APPLICATION: The described LC-MS/MS method can be used to rapidly and simultaneously quantify antibiotic dye residues in frozen fish. CV and LCV were detected in farmed Pangasius and Tilapia fillets and their concentrations was reduced by more than one third after immersing the fillets in whole milk for 120 min, a treatment which is not intended to replace safe fish farming practices upstream to artificially lower the level of banned dyes in fish. The findings support the necessity of regular inspections and monitoring of CV and other antibiotic dye residues in fish, along with assessments of fish microbial quality, to protect public health.

Biofilm formation of filamentous fungi Coriolopsis sp. on simple muslin cloth to enhance removal of triphenylmethane dyes.

The isolate Coriolopsis sp. (1c3) was cultured on muslin cloth to induce formation of filamentous biofilm. The biofilm and the free-mycelium forms (control) were then used to treat two triphenylmethane dyes; Cotton Blue (CB) and Crystal Violet (CV). The biofilm comprised primarily of a compact mass of mycelium while sparse mycelium network was detected in free-mycelium forms. Results revealed significant decolourization activities by filamentous biofilm of 1c3 for CB (79.6%) and CV (85.1%), compared to free-mycelium forms (72.6 and 58.3%, for CB and CV, respectively). Biodegradation occurred in both biofilm and free-mycelium forms. FTIR spectra revealed that biofilm formation (stacking of mycelium), did not have severe implications to the number and types of functional groups available for dye biosorption. The findings here suggested that formation of biofilm in 1c3 was induced effectively on muslin cloth, leading to enhanced decolourization activities. This technology is simple, feasible and can be adopted and further improved to obtain biofilm to enhance their dye removal efficiency in aqueous solutions.

Chemical Compositions and Antioxidant Activities of Essential Oils Extracted from the Petals of Three Wild Tree Peony Species and Eleven Cultivars.

The aim of this study was to investigate the essential oil (EO) compositions and antioxidant activities from petals of three wild tree peony species (Paeonia delavayi, P. lutea, and P. rockii) and eleven P. suffruticosa cultivars from different cultivar groups. The EOs yields varied from 0.63% to 1.25% (v/v) among samples when using supercritical CO2 extraction. One hundred and sixty-three components were detected by GC/MS; and among them, linalool oxide, (Z)-5-dodecen-1-yl acetate, nonadecane, (Z)-5-nonadecene, heneicosane, phytol, and linoleic acid ethyl ester were dominant. According to hierarchical cluster analysis, principal component analysis and correspondence analysis, P. lutea, P. delavayi, and 'High Noon' were clustered in a group described as having a refreshing herbal-like note due to high rates of phytol and linalool oxide. Notably, P. lutea and P. delavayi also had strong DPPH and ABTS radical scavenging activities. These results suggest that P. lutea and P. delavayi are the most promising candidates as useful sources of fragrances and natural antioxidants.

Volatile organic compounds of Thai honeys produced from several floral sources by different honey bee species.

The volatile organic compounds (VOCs) of four monofloral and one multifloral of Thai honeys produced by Apis cerana, Apis dorsata and Apis mellifera were analyzed by headspace solid-phase microextraction (HS-SPME) followed by gas chromatography and mass spectrometry (GC-MS). The floral sources were longan, sunflower, coffee, wild flowers (wild) and lychee. Honey originating from longan had more VOCs than all other floral sources. Sunflower honey had the least numbers of VOCs. cis-Linalool oxide, trans-linalool oxide, ho-trienol, and furan-2,5-dicarbaldehyde were present in all the honeys studied, independent of their floral origin. Interestingly, 2-phenylacetaldehyde was detected in all honey sample except longan honey produced by A. cerana. Thirty-two VOCs were identified as possible floral markers. After validating differences in honey volatiles from different floral sources and honeybee species, the results suggest that differences in quality and quantity of honey volatiles are influenced by both floral source and honeybee species. The group of honey volatiles detected from A. cerana was completely different from those of A. mellifera and A. dorsata. VOCs could therefore be applied as chemical markers of honeys and may reflect preferences of shared floral sources amongst different honeybee species.

Pd nanoparticles supported on MIL-101/reduced graphene oxide photocatalyst: an efficient and recyclable photocatalyst for triphenylmethane dye degradation.

To improve the photocatalytic efficiency of chromium-based metal-organic framework (MIL-101) photocatalyst, Pd nanoparticles and reduced graphene oxide were used to modify the MIL-101 via a facile method. The resulting novel photocatalyst was characterized by UV-vis diffuse reflectance spectra (DRS), powder X-ray diffraction (XRD), transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). It was indicated that the photocatalyst afforded high photocatalytic efficiency for degradation of two triphenylmethane dyes, brilliant green and acid fuchsin, under exposure to visible light irradiation. Cyclic experiments demonstrated that the photocatalyst showed good reusability and stability for the dye degradation.

Identification of Aroma-active Compounds in Essential Oil from Uncaria Hook by Gas Chromatography- Mass Spectrometry and Gas Chromatography-Olfactometry.

The chemical composition of essential oil extracted from Uncaria Hook ("Chotoko" in Japanese), the branch with curved hook of the herbal medicine Uncaria rhynchophylla has been investigated by GC and GC-MS analyses. Eighty-four compounds, representing 90.8% of the total content was identified in oil obtained from Uncaria Hook. The main components i were (E)-cinnamaldehyde (13.4%), α-copaene (8.0%), methyl eugenol (6.8%), δ-cadinene (5.3%), and curcumene (3.6%). The important key aroma-active compounds in the oil were detected by gas chromatography-olfactometry (GC-O) and aroma extract dilution analysis (AEDA), using the flavor dilution (FD) factor to express the odor potency of each compounds. Furthermore, the odor activity value (OAV) has been used as a measure of the relative contribution of each compound to the aroma of the Uncaria Hook oil. The GC-O and AEDA results showed that α-copaene (FD = 4, OAV = 4376), (E)-linalool oxide (FD = 64, OAV = 9.1), and methyl eugenol (FD = 64, OAV = 29) contributed to the woody and spicy odor of Uncaria Hook oil, whereas furfural (FD = 8, OAV = 4808) contributed to its sweet odor. These results warrant further investigations of the application of essential oil from Uncaria Hook in the phytochemical and medicinal fields.

Degradation of triphenylmethane dyes using a temperature and pH stable spore laccase from a novel strain of Bacillus vallismortis.

The characterization of a spore laccase from Bacillus vallismortis fmb-103, isolated from textile industry disposal sites, is described. The activity was 6.5 U/g of dry spore with ABTS as the substrate. The enzyme was quite stable at high temperature. It retained more than 90% of its initial activity after 10h at 70 °C. The enzyme demonstrated broad pH stability in both acidic and alkaline conditions. There was almost no activity loss at pH 3 over an extended period of time, and the relative activity remained at 82% and 38% at pH 7 and pH 9 after 10 days. NaN(3), SDS, L-cysterine, Dithiothreitol, EDTA and NaCl inhibit the enzyme activity. Triphenylmethane dyes, including malachite green, brilliant green and aniline blue were efficiently degraded by the enzyme after 24h in combination with a mediator with efficiencies of 76.84%, 96.56% and 81.17%, respectively. The reusability of spore laccase for decolorization dyes was also examined.

Preparation and characterization of porous cross linked laccase aggregates for the decolorization of triphenyl methane and reactive dyes.

The production of porous cross-linked enzyme aggregates (p-CLEAs) is a simple and effective methodology for laccase immobilization. A three-phase partitioning technique was applied to co-precipitate laccase and starch, followed by cross-linking with glutaraldehyde and removal of starch by α-amylase to create pores in the CLEAs. Scanning electron microscopy revealed a very smooth spherical structure with numerous large pores. The half-life of free laccase at 55°C was calculated to be 1.3h, while p-CLEAs did not lose any activity even after 14 h. p-CLEAs also exhibited improved storage stability, catalytic efficiency and could be recycled 15 times with 60% loss of activity. The catalysts decolorized triphenylmethane and reactive dyes by 60-70% at initial dye concentrations of 2 and 0.5 g L(-1), respectively, without any mediators. These results suggest the potential of CLEA technology in waste-water treatment.

Phytodegradation of the triphenylmethane dye Malachite Green mediated by cell suspension cultures of Blumea malcolmii Hook.

Cell cultures of Blumea malcolmii Hook., developed in the laboratory, rapidly decolorized textile industry effluent along with a variety of dyes with diverse structural properties. Most rapid decolorization was observed in case of Malachite Green (93.41% decolorization within 24 h). The cells were capable of tolerating and degrading high concentrations of the dye, thus making them remarkable systems for phytoremediation studies. The enzymatic analysis during decolorization of Malachite Green showed the induction of enzymes such as laccase, veratryl alcohol oxidase and DCIP reductase indicating the involvement of these enzymes in the degradation of the dye. The cell cultures also mediated the remediation of textile industry effluent by bringing about a decrease in the BOD, COD and ADMI values of the effluent within 48 h. Phytotransformation was confirmed with the help of HPLC and the probable fate of metabolism of the dye was predicted with the help of GCMS analysis.

Structure determination of selaginellins G and H from Selaginella pulvinata by NMR spectroscopy.

Selaginellins G (1) and H (2), two new selaginellin derivatives, were isolated from the whole plant of Selaginella pulvinata. Their structures were elucidated, and complete assignments of the (1)H and (13)C NMR spectroscopic data were achieved by 1D and 2D NMR experiments (HSQC, HMBC, COSY and ROESY). Compound 1 displayed good antifungal activity against Candida albicans with an IC(50) value of 5.3 microg/ml.

Multiresidue method for the triphenylmethane dyes in fish: Malachite green, crystal (gentian) violet, and brilliant green.

Liquid chromatographic methods are presented for the quantitative and confirmatory determination of crystal violet (CV; also known as gentian violet), leucocrystal violet (LCV), brilliant green (BG), and leucobrilliant green (LBG) in catfish. LCV and LBG were oxidized to the chromic CV and BG by reaction with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, and residues were measured as the combined CV+/-LCV and BG+/-LBG. These methods are extensions of published methods for malachite green (MG) analysis to allow simultaneous determination of MG, CV, and BG. Residues were extracted from muscle with ammonium acetate buffer and acetonitrile, and extracts cleaned up using dichloromethane partitioning and solid-phase extraction. Extracts were analyzed by liquid chromatography with visible detection (LC-VIS). The method was validated for catfish fortified with LCV over the range 0.25-10 ngg(-1) and CV at 2 ngg(-1). Average recoveries were 90.6% (+/-8.1% R.S.D., n=45) for LCV and 84.4% (+/-4.2% R.S.D., n=6) for CV. The average recovery for samples fortified with BG or LBG over the range 0.5-10 ngg(-1) was 67.2% (+/-14.8% R.S.D., n=31). CV and BG were confirmed in fish extracts by ion trap LC-mass spectrometry (LC-MS(n)) with no discharge-atmospheric pressure chemical ionization. Average LC-MS(n) recoveries were 96.5, 96.6, and 70.2% for samples fortified with CV, LCV, and BG or LBG. The limits of detection for CV, BG, and MG were in the range of 0.07-0.24 ngg(-1) (ppb) for the two different instrumental methods. This methodology was applied to the analysis of catfish treated with CV and BG.

H2SO4-silica-promoted 'on-column' removal of benzylidene, isopropylidene, trityl and tert-butyldimethylsilyl groups.

H(2)SO(4)-silica-promoted removal of benzylidene, isopropylidene, trityl and tert-butyldimethylsilyl groups from sugar derivatives was accomplished by following an 'on-column' protocol in a virtually waste-free condition.

Efficient removal of triphenylmethane dyes from aqueous medium by in situ electrogenerated Fenton's reagent at carbon-felt cathode.

Fenton's reagent (Fe2+ +H2O2) has been electrogenerated in situ in an undivided electrolytic cell from the effective reduction of Fe3+ and O2 at carbon-felt cathode for the treatment of aqueous solutions of four triphenylmethane dyes (TPMs), namely malachite green (MG), crystal violet (CV), methyl green (MeG) and fast green FCF (FCF), at pH 3.0 and room temperature. MG has been used as a model among them to study the influence of some experimental parameters on the decay kinetics, COD removal and current efficiency. The results in such electro-Fenton system are explained in terms of the many parasitic reactions involving .OH. Higher efficiency values are obtained with rising organic content and decreasing applied current. The first stage of the mineralization process, involving aromatic by-products, leads to fast decoloration as well as quick initial COD removal that fit well to a pseudo-first-order kinetics. At prolonged electrolysis time, the mineralization rate and efficiency decrease due to the formation of hardly oxidizable compounds and the enhancement of wasting reactions. Solutions of all four TPMs are quickly degraded following a pseudo-first-order decay kinetics. The absolute rate constant (kTPM) for their reaction with .OH increases in the order MeG

The first occurrence in nature of two compounds from hops.

Two compounds, (p-methoxyphenyl) diphenylmethanol (1) and tribenzylamine (2), were isolated from Humulus lupulus. Their structures were established on the basis of spectral evidence (MS, IR, NMR, HMBC, HMQC, 1H-1H COSY experiments). Compounds 1 and 2 were found as natural products at the first time.

Isolation, identification and determination of a magenta subsidiary colour in food blue no. 1 (brilliant blue FCF).

A magenta subsidiary colour was isolated from commercial Food Blue No. 1 (B-1; Brilliant Blue FCF). The absorption maximum for this subsidiary colour at 580 nm is outside of the range of 614-628 nm found for other subsidiary colours and m,m-B-1. On the basis of MS and NMR analyses, the structure of the subsidiary colour was elucidated as the disodium salt of 2-[[4-[N-ethyl-N-(3-sulphophenylmethyl)amino]phenyl][4-oxo- 2,5-cyclohexadienylidene]methyl]benzenesulphonic acid. HPLC analyses revealed that 24 batches of commercial Food Blue No. 1 (three manufacturers) contain 0.1-0.8% (average: 0.5%) of the magenta subsidiary colour.

Bis(4-hydroxyphenyl)[2-(phenoxysulfonyl)phenyl]methane: isolation and structure elucidation of a novel estrogen from commercial preparations of phenol red (phenolsulfonphthalein)

Commercial preparations of phenolsulfonphthalein (Phenol Red), a pH indicator dye widely added to cell culture media, have weak estrogenic activity that can be accounted for by a minor lipophilic impurity (ca. 0.002%). We have isolated this impurity, determined its structure to be bis(4-hydroxyphenyl)[2-(phenoxysulfonyl)phenyl]methane, and synthesized it from phenolsulfonphthalein. This compound binds to the estrogen receptor with an affinity 50% that of estradiol; it stimulates the proliferation and increases the progesterone receptor content of estrogen-responsive breast cancer cells in vitro, and it stimulates uterine weight gain in rats in vivo, but shows a potency in these assays only 0.1-0.2% that of estradiol. We suggest how this novel estrogen may be generated during the preparation of phenolsulfonphthalein.

Polymer support synthesis. XII. Derivatization with the 4-decyloxytrityl group as an aid in the affinity chromatography of oligo- and polynucleotides.

In a continuation of previous studies on trityl groups substituted with long alkyl chains as affinity-protecting groups for oligonucleotides, the use of the (4-decyloxyphenyl)diphenylmethyl (DTr) group as an aid in the separation of solid-phase products has been investigated. This substituent, which can be introduced and removed from the 5'-position in a similar manner to the di-p-anisylphenylmethyl group, helps in the purification of deoxypolynucleotides with up to 140 bases. Their retention time was shown to depend not only on the length of the nucleotide chain and alkyl substituent, but also on the elution gradient rate. In oligoribonucleotide synthesis, the DTr group allows the purification of partially protected sequences, which are more stable to enzymatic degradation and, therefore, more suitable for handling and storage.