Search

Laccases are of particular interest in addressing environmental challenges, such as the degradation of triphenylmethane (TPM) dyes, including crystal violet (CV) and Coomassie Brilliant Blue (CBB), which are commonly used in SDS-PAGE for protein visualization. However, these dyes present significant environmental concerns due to their resistance to degradation, which makes their removal from industrial wastewater a major challenge. To address this, the current study investigates the potential of a novel CotA laccase derived from Bacillus sp. PAMC28748, an Antarctic bacterial isolate, for decolorizing these stubborn dyes. The CotA gene was successfully cloned and expressed, and the enzyme demonstrated optimal activity at pH 3 and 50 °C, which favors its maximum catalytic performance. The recombinant Bacillus sp. PAMC28748 rBCLac effectively decolorized CBB without additional mediators, whereas the degradation of CV required the use of the redox mediator ABTS. With ABTS, over 90 % decolorization was achieved at a 0.35 % concentration of CV after 240 min of incubation. Further investigation through molecular docking studies revealed that hydrogen bonding and hydrophobic interactions between the enzyme and the dye molecules are critical for effective degradation, highlighting the enzyme's specific interaction mechanisms. In addition to its catalytic effectiveness, the study also demonstrated the practical potential of the rBCLac system by recovering and reusing both ABTS and rBCLac through ultracentrifugation and acetone precipitation. The process maintained over 75 % efficiency across three cycles, despite a slight decline in enzyme activity, thus showcasing the system's sustainability and reusability. These findings collectively suggest that rBCLac, isolated from an extreme Antarctic environment, holds considerable promise as a candidate for the removal of industrial wastewater containing persistent dyes, with the added potential for cost-effective and sustainable water treatment through the reuse of both the enzyme and its mediator., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Ltd. All rights reserved.)

Cystic fibrosis results from mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) anion channel, ultimately leading to diminished transepithelial anion secretion and mucociliary clearance. CFTR correctors are therapeutics that restore the folding/trafficking of mutated CFTR to the plasma membrane. The large-conductance calcium-activated potassium channel (BKCa, KCa1.1) is also critical for maintaining lung airway surface liquid (ASL) volume. Here, we show that the class 2 (C2) CFTR corrector VX-445 (elexacaftor) induces K+ secretion across WT and F508del CFTR primary human bronchial epithelial cells (HBEs), which was entirely inhibited by the BKCa antagonist paxilline. Similar results were observed with VX-121, a corrector under clinical evaluation. Whole-cell patch-clamp recordings verified that CFTR correctors potentiated BKCa activity from both primary HBEs and HEK cells stably expressing the α subunit (HEK-BK cells). Furthermore, excised patch-clamp recordings from HEK-BK cells verified direct action on the channel and demonstrated a significant increase in open probability. In mouse mesenteric artery, VX-445 induced a paxilline-sensitive vasorelaxation of preconstricted arteries. VX-445 also reduced firing frequency in primary rat hippocampal and cortical neurons. We raise the possibilities that C2 CFTR correctors gain additional clinical benefit by activation of BKCa in the lung yet may lead to adverse events through BKCa activation elsewhere.

Malachite Green (MG) is a widely used industrial dye that is hazardous to health. Herein, the decolourisation and detoxification of MG were achieved using the engineered Saccharomyces cerevisiae expressing novel thermostable laccase lcc1 from Trametes trogii. The engineered strain RCL produced a high laccase activity of 121.83 U L -1 . Lcc1 was stable at temperatures ranging from 20 ℃ to 60 ℃ and showed a high tolerance to organic solvents. Moreover, Lcc1 could decolorize different kinds of dyes (azo, anthraquinone and triphenylmethane), among which, the decolorization ability of MG is the highest, reaching 95.10 %, and the decolorization rate of other triphenylmethane dyes also over 50 %. The RCL decolorized about 95 % of 50 mg L -1 of MG dye in 10 h at 30 ℃. The MG degradation products were analyzed. The industrial application potential of the RCL was evaluated by treating industrial wastewater and the decolourisation rates were over 90 %., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study aimed to produce linalool loaded zinc oxide nanocomposite (LZNPs) and assess its in vitro and in vivo antileishmanial effects against Leishmania major. LZNPs was produced through the synthesis of an ethanolic solution containing polyvinyl alcohol. The average size of LZNPs was determined to be 105 nm. The findings indicated that LZNPs displayed significant (p < 0.01) antileishmanial effects on promastigotes and amastigotes. Following exposure of promastigotes to LZNPs, there was a notable rise in the percentage of early and late apoptotic cells from 9.0 to 57.2 %. The gene expression levels of iNOS, IFN-γ, and TNF-α in macrophages were upregulated in a dose-dependent approach following exposure to LZNPs. LZNPs alone and in conjunction with glucantime (Glu) resulted in a reduction in the diameter and parasite load of CL lesions in infected mice. Treatment of the CL-infected mice with LZNPs at 25 and 50 mg/kg mainly in combination with Glu-reduced the tissue level of malondialdehyde (MDA), increased both gene and protein expression of the antioxidant enzymes as well as raised the expression level of IFN-γ and IL-12 cytokines, whereas caused a significant reduction in the expression level of IL-4. The present study shows that LZNPs has potent antileishmanial effects and controls CL in a mice model through its antioxidant and immunomodulatory properties. Further investigation, especially in clinical trials, could explore the potential use of this nanocomposite in managing and treating CL., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Background: Aroma is one of the most important quality criterion of different honeys and even defines their merchant value. The composition of volatile compounds, especially the characteristic odor-active compounds, contributes significantly to the aroma of honey. Evodia rutaecarpa (Juss) Benth honey (ERBH) is a special honey in China with unique flavor characteristics. However, no work in the literature has investigated the volatile compounds and characteristic odor-active compounds of ERBHs. Therefore, it is imperative to conduct systematic investigation into the volatile profile, odor-active compounds and odor properties of ERBHs., Results: The characteristic fingerprint of ERBHs was successfully constructed with 12 characteristic peaks and a similarity range of 0.785-0.975. In total, 297 volatile compounds were identified and relatively quantified by headspace solid-phase microextraction coupled with gas chromatography quadrupole time-of-flight mass spectrometry, of which 61 and 31 were identified as odor-active compounds by relative odor activity values and GC-olfactometry analysis, respectively, especially the common 22 odor-active compounds (E)-β-damascenone, phenethyl acetate, linalool, cis-linalool oxide (furanoid), octanal, hotrienol, trans-linalool oxide (furanoid), 4-oxoisophorone and eugenol, etc., contributed significantly to the aroma of ERBHs. The primary odor properties of ERBHs were floral, followed by fruity, herbaceous and woody aromas. The partial least-squares regression results showed that the odor-active compounds had good correlations with the odor properties., Conclusion: Identifying the aroma differences of different honeys is of great importance. The present study provides a reliable theoretical basis for the quality and authenticity of ERBHs. © 2023 Society of Chemical Industry., (© 2023 Society of Chemical Industry.)

Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. We wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome. RIFM staff are employees of the Research Institute for Fragrance Materials, Inc. (RIFM). The Expert Panel receives a small honorarium for time spent reviewing the subject work.

In the last few decades, Brilliant green (BG) dye is widely employed to colour the fabric materials in various industries (e.g. textile, pulp and paper, etc.). The wastewater containing BG dye emerges as a major challenge among the researchers due to its toxic, mutagenic, and carcinogenic effects on human beings and marine life. In this context, the present study is mainly focused on the biodegradation of BG dye present in wastewater. The biodegradation of BG dye was performed in an indigenously designed recirculating packed bed bioreactor (RPBBR). Modified Polypropylene-Polyurethane foam (PP-PUF), a support packing material, was immobilised with a newly isolated bacterial consortium of Enterobacter asburia e strain SG43 (BGT1) and Alcaligenes sp. SY1 (BGT2). The bioreactor was operated under various organic loading rates (OLRs) of 2.7, 1.27, 0.93, 0.71, and 0.53 kg COD/m 3 .d -1 with a hydraulic retention time (HRT) of 4 days. The bioreactor exhibited the maximum BG dye removal efficiency of 91%. Proton Nuclear Magnetic Resonance ( 1 H NMR), UV-Vis spectroscopy, Gas chromatography-mass spectrometry (GC-MS), and Fourier Transform Infrared Spectroscopy (FTIR) depicted the biodegradation of BG dye. Phaseolus mungo seeds germinated in BG dye biodegraded wastewater was significantly high (83.56%) than the untreated wastewater (32.4%), which was reasonably subjected to the detoxification of treated wastewater.

To fully harness the potential of laccase in the efficient decolorization and detoxification of single and mixed dyes with diverse chemical structures, we carried out a systematic study on the decolorization and detoxification of single and mixed dyes using a crude laccase preparation obtained from a white-rot fungus strain, Pleurotus eryngii . The crude laccase preparation showed efficient decolorization of azo, anthraquinone, triphenylmethane, and indigo dyes, and the reaction rate constants followed the order Remazol Brilliant Blue R > Bromophenol blue > Indigo carmine > New Coccine > Reactive Blue 4 > Reactive Black 5 > Acid Orange 7 > Methyl green. This laccase preparation exhibited notable tolerance to SO 4 2- salts such as MnSO 4 , MgSO 4 , ZnSO 4 , Na 2 SO 4 , K 2 SO 4 , and CdSO 4 during the decolorization of various types of dyes, but was significantly inhibited by Cl - salts. Additionally, this laccase preparation demonstrated strong tolerance to some organic solvents such as glycerol, ethylene glycol, propanediol, and butanediol. The crude laccase preparation demonstrated the efficient decolorization of dye mixtures, including azo + azo, azo + anthraquinone, azo + triphenylmethane, anthraquinone + indigo, anthraquinone + triphenylmethane, and indigo + triphenylmethane dyes. The decolorization kinetics of mixed dyes provided preliminary insight into the interactions between dyes in the decolorization process of mixed dyes, and the underlying reasons and mechanisms were discussed. Importantly, the crude laccase from Pleurotus eryngii showed efficient repeated-batch decolorization of single-, two-, and four-dye mixtures. This crude laccase demonstrated high stability and reusability in repeated-batch decolorization. Furthermore, this crude laccase was efficient in the detoxification of different types of single dyes and mixed dyes containing different types of dyes, and the phytotoxicity of decolorized dyes (single and mixed dyes) was significantly reduced. The crude laccase efficiently eliminated phytotoxicity associated with single and mixed dyes. Consequently, the crude laccase from Pleurotus eryngii offers significant potential for practical applications in the efficient decolorization and management of single and mixed dye pollutants with different chemical structures.

We describe the synthesis, characterization, and application of an isotopologue of the trityl radical OX071, labeled with 13 C at the central carbon ( 13 C 1 ). This spin probe features large anisotropy of the hyperfine coupling with the 13 C 1 ( I = 1/2), leading to an EPR spectrum highly sensitive to molecular tumbling. The high biocompatibility and lack of interaction with blood albumin allow for systemic delivery and in vivo measurement of tissue microviscosity by EPR.

In this paper, we present the synthesis of C@Fe 3 O 4 -MoO 3 binary composite were prepared through the facile hydrothermal process. The ultrasonic aided adsorption efficacy was evaluated by studying triphenylmethane dye's adsorption potential. The ultrasonic aided adsorption capacity towards crystal violet was 993.6 mg/g, which is remarkably higher and best fitted with the Langmuir isotherm model and followed pseudo-second-order kinetics. The electrochemical studies working electrode have been prepared with 80 wt% active material, 10 wt% carbon black, and 10% polyvinylidene difluoride to evaluate energy storage characteristics. The C@Fe 3 O 4 -MoO 3 demonstrated an excellent specific capacitance of 40.94 F/g with better retention and stability, making it a potential cathode material for next-generation electrochemical energy storage devices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Linalool, which is one of the most representative aroma substances in tea, is transformed into other aroma-related compounds, including linalool 3,6-oxides and linalool 3,7-oxides. The objective of this study was to elucidate the linalool oxide synthesis pathway and its response to stress in tea. By feeding experiment, chemical synthesis, and compound analysis, it was found that linalool can be transformed to linalool oxides via 6,7-epoxylinalool. The conversion rate from 6,7-epoxylinalool to linalool oxides was relatively high under acidic conditions. Four linalool oxide glucosides obtained from tea were structurally characterized. Additionally, tea green leafhopper infestation was observed to activate the whole metabolic flow from linalool into linalool oxides and their glucosides (p < 0.01). Moreover, light treatments further increased the accumulation of linalool oxides and their glucosides (p < 0.05). These results will be useful for elucidating the mechanism mediating linalool oxides content changes in response to stress in tea., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Herein, we found that anthraquinone (AQ) acted as a catalyst for the rapid and effective removal of triphenylmethane dye containing tertiary amino group (TDAG). Results showed that AQ had an enhanced catalytic reactivity towards the removal of TDAG compared to hydro-quinone, which was further proved and explained using density functional theory (DFT) calculations. AQs could achieve a TDAG removal efficiency and rate of approximately 100% and 0.3583 min -1 , respectively, within 20 min. Quenching experiments and electron paramagnetic resonance (EPR) tests indicated that the superoxide radical (O 2 •- ) generated through the catalytic reduction of an oxygen molecule (O 2 ) by AQ contributed to the effective removal of the TDAG. In addition, it was found that the electrophilic attack of the O 2 •- radical on the TDAG was the driving force for the dye degradation process. Decreasing the pH led to protonation of the substituted group of AG, which resulted in formation of an electron deficient center in the TDAG molecule (TDAG-EDC + ) through delocalization of the π electron. Therefore, the possibility of the electrophilic attack for the dye by the negative O 2 •- radical was significantly enhanced. This study revealed that the H + and the O 2 •- generated by the catalytic reduction of O 2 have synergistic effects that led to a significant increase in the dye removal rate and efficiency, which were higher than those obtained through persulfate oxidation., (Copyright © 2022. Published by Elsevier B.V.)

Fenghuang Dancong, Tieguanyin, and Dahongpao teas are belonged to semi-fermented oolong teas and are famous for their unique aroma. However, reports regarding the systematic comparison, differentiation, and classification of the volatile components of these three types of oolong teas are lacking. In this study, we aimed to establish a method for distinguishing these three types of oolong teas. The volatile components in a total of 21 tea samples of these three types of oolong teas were extracted, determined, and identified by headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS). In addition, chemometric methods such as hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were used for distinguishing and classifying the three types of oolong teas on the basis of the similarities and differences in the volatile components. The results showed that 125 volatile components were extracted and identified from the three types of oolong teas, among which 53 volatile components overlapped among the samples. The results of HCA indicated that the samples of each of the three types of oolong teas could be placed in one category when the t value was 220. The results of PCA and OPLS-DA showed that the volatile components such as dehydrolinalool, linalool oxide II, linalool, α-farnesene, linalool oxide I, β-ocimene, nerolidol, cis-3-butyric acid folate, myrcene, and (Z)-hexanoic acid-3-hexenyl ester are the characteristic components, which can be used to distinguish the three types of oolong teas. We developed a simple, fast, and efficient method for distinguishing three types of oolong teas and provided a feasible technique for the identification of oolong tea types., Competing Interests: The authors have declared that no competing interests exist.

This study revealed Streptomyces bacillaris as an efficient biological agent for the removal of triphenylmethane (TPM) dyes. The isolate decolorized Malachite Green (MG), Methyl Violet (MV), Crystal Violet (CV), and Cotton Blue (CB) effectively. S. bacillaris in the treated dye solutions were analyzed for enzyme production, and the cell biomass was observed for functional groups and cell morphology. The treated dye solutions were also analyzed for degraded compounds and their toxicity. Results revealed high decolorization activities for MG (94.7%), MV (91.8%), CV (86.6%), CB (68.4%), attributed to both biosorption and biodegradation. In biosorption, dye molecules interacted with the hydroxyl, amino, phosphoryl, and sulfonyl groups present on the cell surface. Biodegradation was associated with induced activities of MnP and NADH-DCIP reductase, giving rise to various simpler compounds. The degraded compounds in the treated dyes were less toxic, as revealed by the significant growth of Vigna radiata in the phytotoxicity test. There were no significant changes in cell morphology before and after use in dye solutions, suggesting S. bacillaris is less susceptible to dye toxicity. This study concluded that S. bacillaris demonstrated effective removal of TPM dyes via biosorption and biodegradation, rendering the treated dyes less toxic than untreated dyes. Findings in this study enabled further explorations into the potential application of lesser-known actinobacteria (i.e. Streptomyces sp.) for dye removal., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The toxic wastewater effluents from textile dyes have been a significant environmental threat worldwide in recent decades. Against this backdrop, this study investigates the performance of C@Fe 3 O 4 -MoO 3 -rGO as a sonoadsorbent to ameliorate crystal violet (CV) dye from the aqua matrix and further explores its potential as an electrode in supercapacitor applications. The phase purity, crystal structure, surface morphology, thermal stability and magnetic behaviour characteristics of the composite were studied using various characterisation techniques such as powder X-ray diffraction (XRD), Raman Spectroscopy, Fourier transform infrared spectroscopy (FTIR), Field emission scanning electron microscopy (FESEM), High-resolution transmission electron microscopy (HRTEM), Thermogravimetric analysis (TGA) and Vibrating-sample magnetometry (VSM). From the Langmuir isotherm model, the synthesised sonoadsorbent exhibited a maximum adsorption capacity of 1664.26 mg/g for crystal violet, which is remarkably high. Further, to its inherited magnetic characteristics, the composite can be easily separated from the solution by using an external magnet. Furthermore, the working electrode was synthesised with 80% active material, 10% carbon black, and 10% polyvinylidene difluoride to investigate its suitability in supercapacitor applications. The C@Fe 3 O 4 -MoO 3 -rGO composite exhibited an excellent capacitance value of 180.36 F/g with commendable cycling stability, making it suitable as a potential cathode material for the next generation supercapacitors., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: The aim of the current study was to determine if treatment with senicapoc, improves the PaO 2 /FiO 2 ratio in patients with COVID-19 and severe respiratory insufficiency., Methods: Investigator-initiated, randomized, open-label, phase II trial in four intensive care units (ICU) in Denmark. We included patients aged ≥18 years and admitted to an ICU with severe respiratory insufficiency due to COVID-19. The intervention consisted of 50 mg enteral senicapoc administered as soon as possible after randomization and again after 24 h. Patients in the control group received standard care only. The primary outcome was the PaO 2 /FiO 2 ratio at 72 h., Results: Twenty patients were randomized to senicapoc and 26 patients to standard care. Important differences existed in patient characteristics at baseline, including more patients being on non-invasive/invasive ventilation in the control group (54% vs. 35%). The median senicapoc concentration at 72 h was 62.1 ng/ml (IQR 46.7-71.2). The primary outcome, PaO 2 /FiO 2 ratio at 72 h, was significantly lower in the senicapoc group (mean 19.5 kPa, SD 6.6) than in the control group (mean 24.4 kPa, SD 9.2) (mean difference -5.1 kPa [95% CI -10.2, -0.04] p = .05). The 28-day mortality in the senicapoc group was 2/20 (10%) compared with 6/26 (23%) in the control group (OR 0.36 95% CI 0.06-2.07, p = .26)., Conclusions: Treatment with senicapoc resulted in a significantly lower PaO 2 /FiO 2 ratio at 72 h with no differences for other outcomes., (© 2022 The Authors. Acta Anaesthesiologica Scandinavica published by John Wiley & Sons Ltd on behalf of Acta Anaesthesiologica Scandinavica Foundation.)

Widespread application of synthetic dyes could generate colored wastewaters causing a range of serious environmental problems. Due to the complex nature of effluents from textile industries, it is difficult to obtain satisfactory treatment of dyes-contaminated wastewater using one single method. Biohybrids coupling of photocatalysts and biocatalysts have great potential in environmental purification. However, how to select suitable organisms and enhance the hybrid's catalytic activities remain challenging. Here, a novel biohybrid system (Clostridium thermocellum-CdS), created for light-driven biodecolorization under thermophilic treatment by using non-photosynthetic microorganism C. thermocellum self-photosensitized with CdS nanoparticles was established. The biohybrids exhibited remarkable decolorization effects on triphenylmethane dyes. The highest decolorization rate was 0.206 min -1 . More importantly, enhanced catalytic activities of cadmium sulfide (CdS)-based biohybrids by controlling the particle sizes of semiconductors were demonstrated. Biohybrids systems (Clostridium thermocellum-CdS) through the self-precipitation of CdS with different particle sizes not only showed dramatic changes in the optical properties but also exhibited a very different decolorization rate. This work can not only further broaden targeted applications of CdS-based biohybrids but also demonstrate a promising route for improving biohybrids corresponding photocatalytic capabilities through in situ precipitation CdS with different particle sizes., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Triphenylmethane (tpm) derivatives (e.g. tpm CV ) have threatened the safety of the aquatic environment due to the potential toxicity and carcinogenicity. In this study, the novel ultrasonic/persulfate/chlorite (US/S 2 O 8 2- /ClO 2 - ) oxidation process was developed for the effective removal of tpm CV in wastewater. The apparent non-integer kinetics (n around 1.20) of tpm CV degradation under different factors (R 2 Adj > 0.990) were investigated, respectively. Inhibiting effects of anions were greater than those of cations (except Fe(II/III)). The adding of micromolecule organic acids could regulate degradation towards positive direction. The double response surface methodology (RSM) was designed to optimize tpm CV removal process, and the acoustic-piezoelectric interaction was simulated to determine the propagation process of acoustic wave in the reactor. The possible degradation pathway was explored to mainly include carbonylation, carboxylation, and demethylation. The estimated effective-mean temperature at the bubble-water interface was calculated from 721 to 566 K after introducing the ClO 2 - , however, the adsorption or partitioning capacity of tpm CV in the reactive zone was widened from 0.0218 to 0.0982. The proposed co-catalysis of US/S 2 O 8 2- /ClO 2 - was based on the determined active species mainly including ClO 2 , SO 4 ⋅ - , and ⋅OH. Compared with other US-based processes, the operating cost (3.97 $/m 3 ) of US/S 2 O 8 2- /ClO 2 - with the EE/O value (16.8 kWh/m 3 ) was relatively reduced., (Copyright © 2021. Published by Elsevier B.V.)

Tetrathiatriarylmethyl (trityl) radicals have been recently shown to react with biological oxidoreductants including glutathione (GSH), ascorbic acid (Asc), and superoxide anion radical (O 2 •- ). However, how the substituents affect the reactivity of trityl radicals is still unknown. In this work, five asymmetric trityl radicals were synthesized and their reactivities with GSH, Asc, and O 2 •- investigated. Under aerobic conditions, GSH induces fast decays for the thioether- (TSA) and N -methyleneglycine-substituted (TGA) derivatives and slow decay for the 4-carboxyphenyl-containing one (TPA). Under anaerobic conditions, the direct reduction of these radicals by GSH also occurs with rate constants ( k GSH ) from 1.8 × 10 -4 M -1 s -1 for TPA to 1.0 × 10 -2 M -1 s -1 for TGA. Moreover, these radicals can also react with O 2 •- with rate constants ( k SO ) from 1.2 × 10 3 M -1 s -1 for ET-01 to 1.6 × 10 4 M -1 s -1 for TGA. Surprisingly, these radicals are completely inert to Asc in both aerobic and anaerobic conditions. Additionally, the substituents exert an important effect on redox potentials of these trityl radicals. This work demonstrates that the redox properties of the trityl radicals strongly depend on their substituents, and TPA with high stability toward GSH shows great potential for intracellular applications.

As an important natural product, the sufficient separation of plant essential oil (EO) is helpful to improve its utilization value. In this work, deep eutectic solvent-homogenate based microwave-assisted hydrodistillation (DES-HMAHD) was developed and applied to isolate EO from the fruits of Litsea cubeba (Lour.) Pers. Different types of DES were investigated in terms of the EO kinetics and composition, among which oxalic acid/choline chloride (OA/ChCl) had obvious advantages. Following, molar ratio of OA and ChCl (1:1), water content (50%), liquid-solid ratio (12.5:1 mL/g), homogenate time (2 min), and microwave power (700 W) were found to be the optimum conditions. Gas chromatography-mass spectrometer (GC-MS) analysis showed that the EO isolated from DES-HMAHD contained a large proportion of m-cymene and trans-linalool oxide, which were quite different from the conventionally reported L. cubeba EO. In addition, the proposed DES-HMAHD resulted in higher separation efficiency and economic value, as well as lower environmental impact, as compared with other techniques. Afterwards, the EO isolated by different methods was evaluated from the perspective of biological activity. The EO obtained by DES-HMAHD showed higher antioxidant activity (DPPH and ABTS) but lower antifungal activity, which was related to its chemical composition. In general, DES-HMAHD produced a kind of L. cubeba EO with different components, which provided a scientific foundation for the sufficient isolation of plant EO and its application in the natural products., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier B.V.)

In this work, the Ni-doped anatase TiO 2 single crystals loaded on activated carbon (Ni-T/AC) were synthesized by a sol-gel method. The chemical compositions and physical properties of as-prepared materials were analyzed by XRD, TEM, BET, FTIR, XPS, and PL characterizations. The obtained results implied that all of samples presented anatase phase with a clear mesoporous structure. The photocatalytic properties of nanocomposites were evaluated through photodegradation of crystal violet (CV), basic fuchsine (BF), and malachite green (MG). The results revealed that the catalyst Ni-T/AC-loaded AC exhibited an excellent photocatalytic activity compared with the original TiO 2 , and the photodegradation efficiency for CV, BF, and MG is 99.00%, 94.85%, and 98.89% after 120 min of irradiation, respectively. This enhancement may be ascribed to the small crystallite size, large specific surface area, and pore volume of the photocatalysts. In addition, the possible degradation mechanism and pathway for triphenylmethane dyes (TPMs) were also well investigated. This work provides a new, low-cost, and effective route to improve the performance of TiO 2 for effective removing TPMs.

In this work, we employed the in situ synthesis method to implant Fe 3 O 4 into activated carbon (AC), in which the synthesis of the magnetic AC (MAC) was realized. Thence, Ni-doped anatase TiO 2 (NATiO 2 ) were anchored on different addition amount of MAC to synthesize the series of Ni-TiO 2 /MAC photocatalysts. The chemical compositions and physical properties of these nanocomposites were analyzed by various characterization technologies. The photocatalytic capabilities of as-produced materials were then investigated via adsorption and photodegradation of triphenylmethane dyes (TPMs) as crystal violet (CV), basic fuchsine (BF), and malachite green (MG) solution. The results revealed that the removal of Ni-TiO 2 /AC, Ni-TiO 2 /2MAC, Ni-TiO 2 /4MAC, and Ni-TiO 2 /8MAC on TPMs is a very fast process and the removal efficiency can almost reach to about 90% in 10 min, and the catalyst has good cycle stability and is easy to be reused. This work provides a novel, low-cost, and effective way to rationally design and synthesize TiO 2 -based photocatalysts for effective removal of TPMs.

The long-term stability of aerobic granules is critical for decolourization of different dyes in textile wastewater. Here, we investigated dye decolourization and the stability of acetate-cultivated granules after exposure to dyes. Results show that granules can maintain excellent structure stability with the presence of azo and triphenylmethane dyes during a 200-day operation period, achieving biomass concentrations as high as 8-12 g L -1 and 90% and 100% decolourization efficiency, respectively. Aerobic granules, however, partially disintegrated after exposure to anthraquinone, resulting in dye decolourization efficiency ranging from 50 to 80% and a biomass concentration as low as around 0.5 g L -1 due to biomass wash-out. The study indicates that long-term granule stability is much dependent on the dye classes. The enrichment of specific species in granules for dye decolourization has not been affected by the granule structure. The specific dye decolourization rate and dye to microorganism ratio for anthraquinone were 5-6.5 and 13.5-16.4 times, respectively, higher than those for azo and triphenylmethane dyes, but the total reactor performance for anthraquinone decolourization is much poorer than azo and triphenylmethane dyes due to low biomass retention in the reactor. The results suggest the importance of stability of aerobic granules for biomass retention to achieve better treatment performance of dye-containing wastewater. For the first time, the long-term stability and decolourization performance of aerobic granules for treating anthraquinone and triphenylmethane dyes are reported here and compared with azo dye, which can be used to guide the treatment of real textile wastewater containing azo, anthraquinone and triphenylmethane dyes by aerobic granules., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Deep eutectic solvents (DESs) have received significant attention as potential extracting agents in recent years due to their favorable characteristics including low cost, easy preparation and environmentally safe starting materials. Experimentally screening for highly efficient DESs meeting various requirements for natural gas sweetening remains a challenging task. Thus, an extensive database of estimated Henry's law constants (Hi) and solubilities (xi) of CO2 in 170 different DESs at 25°C has been constructed using the COSMO-RS method to select potential DESs. Based on the COSMO-RS study, three DESs, namely tetrabutylammonium bromide (TBAB)+polyethylene glycol (PEG-8) (on a molar basis 1:4), TBAB+octanoic acid (OCT) (1:4), and methyltriphenylphosphonium bromide (MTPB)+PEG-8 (1:10), were chosen for further experimentation up to 2 bar at 25°C using a vapor-liquid equilibria (VLE) apparatus. Reliable thermophysical properties were determined experimentally, and a detailed equilibrium-based model was developed for one of the glycol-based DESs (i.e., TBAB+PEG-8 (1:4)). This information is an essential prerequisite for carrying out process simulations of natural gas sweetening plants using ASPEN PLUS. The simulation results for the proposed DES were compared to those of monoethylene glycol (MEG). Here, we find that the aqueous TBAB+PEG-8 (1:4) solvent shows ~60% lower total energy consumption and higher CO2 removal when compared to those using the MEG solvent., Competing Interests: The authors have declared that no competing interests exist.

Stable tetrathiatriarylmethyl radicals have significantly contributed to the recent progress in biomedical electron paramagnetic resonance (EPR) due to their unmatched stability in biological media and long relaxation times. However, the lipophilic core of the most commonly used structure (Finland trityl) is responsible for its interaction with plasma biomacromolecules, such as albumin, and self-aggregation at high concentrations and/or low pH. While Finland trityl is generally considered inert toward many reactive radical species, we report that sulfite anion radical efficiently substitutes the three carboxyl moieties of Finland trityl with a high rate constant of 3.53 × 10 8 M -1 s -1 , leading to a trisulfonated Finland trityl radical. This newly synthesized highly hydrophilic trityl radical shows an ultranarrow linewidth (Δ B pp = 24 mG), a lower affinity for albumin than Finland trityl, and a high aqueous solubility even at acidic pH. Therefore, this new tetrathiatriarylmethyl radical can be considered as a superior spin probe in comparison to the widely used Finland trityl. One of its potential applications was demonstrated by in vivo mapping oxygen in a mouse model of breast cancer. Moreover, we showed that one of the three sulfo groups can be easily substituted with S-, N-, and P-nucleophiles, opening access to various monofunctionalized sulfonated trityl radicals.

Tetrathiatriarylmethyl (TAM, trityl) radicals have found wide applications in electron paramagnetic resonance (EPR) oximetry. However, the biomedical applications of TAM radicals were exclusively limited to an extracellular region owing to their negatively charged nature. The intracellular delivery of TAM radicals still remains a challenge. In the present work, we report a liposome-based method to encapsulate the water-soluble Finland trityl radical CT-03 for its intracellular delivery. Using the thin lipid film hydration method, CT-03-loaded liposomes were prepared from DSPC/cholesterol/DOTAP with a mean size of 167.5 ± 2.4 nm and a zeta potential of 27.8 ± 0.8 mV. EPR results showed that CT-03 was entrapped into the liposomes and still exhibited good oxygen (O 2 ) sensitivity. Moreover, CT-03 was successfully delivered into HepG2 cells and HUVECs using the CT-03-loaded liposomes. Importantly, the combination of the liposome-encapsulated radical CT-03 and the other TAM radical CT02-H enabled simultaneous measurements of the intracellular and extracellular O 2 concentrations and O 2 consumption rates in HepG2 cells. Our present study provides a new approach for intracellular delivery of TAM radicals and could significantly expand their biomedical applications.

As a demonstration of the application of rapid-scan EPR to imaging at low frequency and magnetic field, a multi-compartment phantom containing six different samples was imaged. The samples were nitroxide radicals, trityl (substituted triarylmethyl) radicals, and the oxygen-sensitive solid lithium phthalocyanine (LiPc), all of which are useful for in vivo imaging. The 2D spectral-spatial image demonstration was performed at 250 MHz, with samples in sealed tubes of various sizes arranged in a 3D-printed plastic holder. Maximum gradients of 10 G/cm gave a spatial resolution of about 0.1 mm for the narrow trityl and LiPc signals and about 1 mm for the nitroxide. The importance of proper selection of resonator bandwidth and scan rate for obtaining accurate linewidth information is demonstrated for a case in which the phantom is composed of species with signal linewidths and relaxation times that differ by more than a factor of 10., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Biosorption potential of novel lignocellulosic biosorbents Musa sp. peel (MSP) and Aegle marmelos shell (AMS) was investigated for the removal of toxic triphenylmethane dye malachite green (MG), from aqueous solution. Batch experiments were performed to study the biosorption characteristics of malachite green onto lignocellulosic biosorbents as a function of initial solution pH, initial malachite green concentration, biosorbents dosage, and temperature. Biosorption equilibrium data were fitted to two and three parameters isotherm models. Three-parameter isotherm models better described the equilibrium data. The maximum monolayer biosorption capacities obtained using the Langmuir model for MG removal using MSP and AMS was 47.61 and 18.86 mg/g, respectively. The biosorption kinetic data were analyzed using pseudo-first-order, pseudo-second-order, Elovich and intraparticle diffusion models. The pseudo-second-order kinetic model best fitted the experimental data, indicated the MG biosorption using MSP and AMS as chemisorption process. The removal of MG using AMS was found as highly dependent on the process temperature. The removal efficiency of MG showed declined effect at the higher concentrations of NaCl and CaCl 2 . The regeneration test of the biosorbents toward MG removal was successful up to three cycles.

Background: Recent malaria vector control measures have considerably reduced indoor biting mosquito populations. However, reducing the outdoor biting populations remains a challenge because of the unavailability of appropriate lures to achieve this. This study sought to test the efficacy of plant-based synthetic odor baits in trapping outdoor populations of malaria vectors., Methodology and Principal Finding: Three plant-based lures ((E)-linalool oxide [LO], (E)-linalool oxide and (E)-β-ocimene [LO + OC], and a six-component blend comprising (E)-linalool oxide, (E)-β-ocimene, hexanal, β-pinene, limonene, and (E)-β-farnesene [Blend C]), were tested alongside an animal/human-based synthetic lure (comprising heptanal, octanal, nonanal, and decanal [Blend F]) and worn socks in a malaria endemic zone in the western part of Kenya. Mosquito Magnet-X (MM-X) and lightless Centre for Disease Control (CDC) light traps were used. Odor-baited traps were compared with traps baited with either solvent alone or solvent + carbon dioxide (controls) for 18 days in a series of randomized incomplete-block designs of days × sites × treatments. The interactive effect of plant and animal/human odor was also tested by combining LO with either Blend F or worn socks. Our results show that irrespective of trap type, traps baited with synthetic plant odors compared favorably to the same traps baited with synthetic animal odors and worn socks in trapping malaria vectors, relative to the controls. Combining LO and worn socks enhanced trap captures of Anopheles species while LO + Blend F recorded reduced trap capture. Carbon dioxide enhanced total trap capture of both plant- and animal/human-derived odors. However, significantly higher proportions of male and engorged female Anopheles gambiae s.l. were caught when the odor treatments did not include carbon dioxide., Conclusion and Significance: The results highlight the potential of plant-based odors and specifically linalool oxide, with or without carbon dioxide, for surveillance and mass trapping of malaria vectors.

Triphenylmethanes - Malachite Green (MG), Crystal Violet (CV) and Brilliant Green (BG) are dyes with known genotoxic and carcinogenic properties. Apart from being illegally used in aquaculture for treatment of fish diseases they are also applied in industry such as paper production to colour paper towels widely used in hospitals, factories and other locations for hand drying after washing. The present study provides evidence that the triphenylmethane dye (BG) present in green paper towels can migrate through the skin even when the exposure time is short (30-300 s). The transfer of the dye from the towel to food (fish) was also studied and a high amount of colour was found to migrate during overnight exposure. The risk to humans associated with these two dye transfer studies was assessed using a 'margin of exposure approach' on the basis of the toxicological data available for the closely related dye MG and its metabolite Leucomalachite Green. The data indicated that the risk associated with the use of triphenylmethane containing paper towels is of a similar proportion to the risk associated with consumption of fish contaminated with these dyes due to the illegal application in aquaculture., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

An integrated Overhauser-enhanced MRI-Prepolarized MRI system was developed to obtain radiobiological information that could be accurately coregistered with diagnostic quality anatomic images. EPR and NMR images were acquired through the double resonance technique and field cycling of the main magnetic field from 5 mT to 0.5 T. Dedicated EPR and NMR coils were devised to minimize radiofrequency power deposition with high signal-to-noise ratio. Trityl and nitroxide radicals were used to characterize oxygen and redox sensitivities of multispin echo Overhauser-enhanced MRI. Oxygen resolution of 3 mmHg was obtained from 2 mM deoxygenated trityl phantoms. Trityl radicals were stable in reducing environments and did not alter the redox-sensitive decaying rate of the nitroxide signals. Nitroxide radicals had a compounding effect for the trityl oximetry. Tumor oxygenation and redox status were acquired with anatomical images by injecting trityl and nitroxide probes subsequently in murine tumors. The Overhauser-enhanced MRI-Prepolarized MRI system is ready for quantitative longitudinal imaging studies of tumor hypoxia and redox status as radiotherapy prognostic factors., (Copyright © 2010 Wiley-Liss, Inc.)

In rapid scan EPR the magnetic field is scanned through the signal in a time that is short relative to electron spin relaxation times. Previously it was shown that the slow-scan lineshape could be recovered from triangular rapid scans by Fourier deconvolution. In this paper a general Fourier deconvolution method is described and demonstrated to recover the slow-scan lineshape from sinusoidal rapid scans. Since an analytical expression for the Fourier transform of the driving function for a sinusoidal scan was not readily apparent, a numerical method was developed to do the deconvolution. The slow scan EPR lineshapes recovered from rapid triangular and sinusoidal scans are in excellent agreement for lithium phthalocyanine, a trityl radical, and the nitroxyl radical, tempone. The availability of a method to deconvolute sinusoidal rapid scans makes it possible to scan faster than is feasible for triangular scans because of hardware limitations on triangular scans., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Oligonucleotide microarray has become an important and powerful tool for various genomic analyses, where, unlike conventional methods, one can identify simultaneously a large number of targets in a given sample. Postsynthesis immobilization, the most widely used method, involves the noncovalent and covalent fixing of suitably modified oligonucleotides on the solid supports. Among the various functional groups aminoalkyl, hydroxyalkyl, mercaptoalkyl, aldehyde, epoxy, and carboxyl the most frequently used functional groups on the polymeric surfaces. Because glass and polypropylene, the most widely used materials, are nonporous in nature, the functional groups density on the surface remains very low. In order to know the exact concentration of a ligand to be immobilized, it is essential to estimate the accessible functional groups on these surfaces. For this purpose, sensitive methods are required to estimate exact density of available functional groups on the surfaces. Apart from physical methods, a number of sensitive chemical methods, by making use of high extinction coefficient of 4,4'-dimethoxytrityl cation (epsilon(498) = 70,000 L mol-1 cm-1), have been reported to estimate accessible functional groups on the glass based polymer supports. In this chapter, use of these reagents for spectrophotometric determination of functional group density on glass microslides and polypropylene film has been discussed.

Jones oxidation of Andrographolide (1), gave mixture of three products (3-dehydroandrographolide (5), 3,19-bis dehydroandrographolide (6) and 19-dehydroandrographolide (7). Tritylation of andrographolide at C19-OH resulted to products 8 and diene 9, which can be converted to its acetate 10 and oxidation product 11.

We report a staining method for cadaveric tissue using sodium rhodizonate as a skin marker for gunshot residues and a counterstain for the surrounding connective tissue. We studied six well preserved subjects who had died of close range gunshot injury. Skin fragments were removed from the bullet entrance hole including both the disrupted area and adjacent macroscopically intact tissue. Because microscopic examination of postmortem material is difficult after histomorphologic alterations already have occurred as a consequence of postmortem tissue changes, it is necessary to use a staining method that, while detecting gunshot residues, can also make skin cell constituents recognizable from both qualitative and quantitative perspectives. Triphenylmethane dyes (acid fuchsin, aniline blue WS, light green SF yellowish, brilliant green and ethyl green) have proven appropriate for the purpose.

Cyclotetrapeptides are constrained cyclic peptides whose synthesis is considered a difficult task. A methodology based on on-resin head-to-tail cyclization by anchoring the side chain of a trifunctional amino acid was investigated. A series of model cyclotetrapeptides containing the RGD sequence cyclo(Xaa-Arg-Gly-Asp) (Xaa = Ala, Phe, Phg, D-Ala, D-Phe, D-Phg) was synthesized with no cyclodimerization by-products. An evaluation and optimization study of all of the parameters directly involved in the ring closure was performed.

3-O-Mesyl-1,6-di-O-trityl-beta-D-fructofuranosyl-(2-->1)-6-O-trityl-alpha-D-glucopyranoside (3) was synthesized via stannylation of 6,1',6'-tri-O-tritylsucrose with dibutyltin oxide in benzene, followed by treatment of the crude product with methanesulfonyl chloride in the presence of triethylamine in dichloromethane at 0 degrees C. A similar treatment of the tri-tritylsucrose in toluene, instead of benzene, yielded 4-O-mesyl-1,6-di-O-trityl-beta-D-fructofuranosyl-(2-->1)-6-O-trityl-alpha-D-glucopyranoside (4) as the major product. The X-ray crystal structure of the corresponding acetyl derivative, 3-O-acetyl-4-O-mesyl-1,6-di-O-trityl-beta-D-fructofuranosyl-(2-->1)-2,3,4-tri-O-acetyl-6-O-trityl-alpha-D-glucopyranoside (5), confirms the position and stereochemistry of the methanesulfonyl group at C-4 of the fructofuranosyl ring.

One of the most convenient methods for generating oligonucleotides possessing intra- or interstrand cross-links is through incorporation of oligoethylene glycol bridges by solid-phase synthesis. The reagents are commercially available or can be synthesized in a few easy synthetic steps. Unlike many other DNA and RNA cross-links, aspects of the structural and thermodynamic impact of modifying nucleic acids with oligoethylene glycols have been studied. This unit covers protection, phosphitylation, and preparation of the glycol linker for oligonucleotide synthesis.

HPLC is a powerful and popular method for analyzing and purifying biomolecules. Reversed-phase HPLC allows a high-capacity method for purification, and uses volatile buffer systems that simplify product recovery. Anion-exchange HPLC provides better resolution and a more predictable elution pattern. This unit presents protocols that are optimized for HPLC of oligonucleotides. Because of the resolution limits of both reversed-phase and anion-exchange HPLC, it can be used for oligonucleotides of up to approximately 50 nt in length.

Unlabelled: In vitro studies have demonstrated the membrane potential-dependent enhanced uptake of phosphonium salts, including [3H]triphenylmethylphosphonium (TPMP), into mitochondria of carcinoma and glioma-derived tumor cells, suggesting the potential use of phosphonium salts as tracers for tumor imaging. This study characterizes the in vivo uptake of [11C]TPMP in canine brain glioma using PET., Methods: Dynamic paired PET studies of [11C]TPMP followed by [68Ga]ethylenediaminetetraacetic acid (EDTA) were performed 4 d before and 9 d after tumor cell inoculation. Graphical analysis was used to evaluate [11C]TPMP retention in tumor tissue. Distribution of tracer uptake was compared with tumor histological sections., Results: [11C]TPMP exhibited enhanced uptake and prolonged retention in tumor cells. Patlak plot was linear over the 20- to 95-min postinjection period (r = 0.97 +/- 0.1). [68Ga]EDTA exhibited a gradual washout from the tumor tissue. The tumor-to-normal brain uptake ratio at 55 to 95 min postinjection was 47.5 for [11C]TPMP and 8.1 for [68Ga]EDTA. Qualitative comparison with histological sections indicated that [11C]TPMP enhanced uptake was restricted to the tumor area., Conclusion: The enhanced uptake and prolonged retention in tumor suggest [11C]TPMP as a promising means for imaging of gliomas in dogs. The need for studies in humans is indicated.

Combinatorial libraries of oligonucleotides on beads were synthesised by a split-and-mix strategy using 5'-DMTr- or 5'-Fmoc- nucleoside phosphoramidites. Trityl moieties with different masses were used to encode for the bases coupled at each step in the synthesis of oligonucleotides selected by hybridisation from the libraries. Tags orthogonal to the nucleotides added were produced by coupling amines of different MW to an activated carboxyl group(s) on the trityl moiety. Tags can be released from the support by laser irradiation and measured directly by TOF without matrix. Alternatively, they may be released by an acidic treatment and then analysed by (MA)LDI-TOF.

The present study describes extraction fraction and uptake measurements of the [11C]methyl triphenyl phosphonium (11C-MTP), a promising positron emission tomography (PET) agent for cardiac imaging. PET imaging was performed in mongrel dogs. Under physiological flow conditions 11C-MTP uptake reached a maximum within the first 10 minutes after injection and remained constant during the entire observation period of 80 minutes. Over the same time period, the heart/blood ratio was 46-106:1, and the heart/lung ratio 14:1. Following permanent occlusion of the left anterior descending coronary artery, 11C-MTP uptake in the normally perfused myocardium also reached a maximum at 10 minutes after injection, whereas in the infarcted area there was no significant accumulation of 11C-MTP. For a time period of 80 minutes the noninfarcted/infarcted myocardium ratio was 12:1. Extraction was measured in anesthetized dogs with a double isotope method using 99mTc-HSA as the reference tracer. The extraction fraction was 91% at a flow of 69 mL/min/100g. As flow increased to five-fold (342 mL/min/100g) following administration of adenosine, extraction fell to 61%. Following coronary artery occlusion, the 11C-MTP content in the myocardium was highly correlated (r = 0.93, p < 0.01; y = 10.46 + 0.92x) with the microsphere determined regional myocardial blood flow.

We investigated the effects of bases, resins, and S-protecting groups on the extent of racemization at the C-terminal cysteine during Fmoc-based(Fmoc=fluoren-9-yl-methoxy-carbonyl) solid phase peptide synthesis. The use of 2-chlorotrityl resin was most effective in suppressing the racemization caused by the base treatment for Fmoc-cleavage. Somatostatin was successfully synthesized with practically no racemization using 2-chlorotrityl resin by Fmoc-chemistry.

The esterification of 2-chlorotrityl chloride resin with Fmoc-amino acids in the presence of DIEA is studied under various conditions. High esterification yields are obtained using 0.6 equiv. Fmoc-amino acid/mmol resin in DCM or DCE, in 25 min, at room temperature. The reaction proceeds without by product formation even in the case of Fmoc-Asn and Fmoc-Gln. The quantitative and easy cleavage of amino acids and peptides from 2-chlorotrityl resin, by using AcOH/TFE/DCM mixtures, is accomplished within 15-60 min at room temperature, while t-butyl type protecting groups remain unaffected. Under these exceptionally mild conditions 2-chlorotrityl cations generated during the cleavage of amino acids and peptides from resin do not attack the nucleophilic side chains of Trp, Met, and Tyr.

We have automated the trityl-on purification of oligonucleotides by use of an XYZ axis robotic solid-phase extraction system. This greatly decreased the preparation time required for oligonucleotide purification. After about 15 min for set up of the samples and instrument, the oligonucleotides are automatically purified with a 15-min run time per sample. Thus, for example, the purification of 15 oligonucleotides requires only about 15 min of preparation time and 4 h of machine time. Yields and purity are equivalent to manual methods.

A procedure is described for the large-scale purification of synthetic oligonucleotides using a polystyrene (PRP-1, Hamilton Co.) high-performance liquid chromatography (HPLC) column with a phosphate/methanol/acetonitrile solvent system. Pure oligonucleotides are obtained with a three-step procedure that involves only one column purification step. The dimethoxytrityl group is left on the oligomer for the HPLC purification. The use of the PRP-1 polystyrene column with a phosphate/methanol/acetonitrile solvent system provides excellent separation of the desired dimethoxytrityl-bearing oligonucleotide from failure sequences. The dimethoxytrityl group is removed by treatment with acetic acid and the oligonucleotide is desalted on a C-18 Sep-Pak cartridge. The oligodeoxynucleotides obtained are shown to be essentially pure by HPLC, polyacrylamide gel electrophoresis, and 500-MHzNMR spectroscopy. This procedure is especially useful for the large-scale purification of oligonucleotides required for NMR studies. The PRP-1 column and the phosphate/methanol/acetonitrile solvent system is useful for purifying modified oligonucleotides containing lipophilic groups such as the carcinogen 2-(acetylamino)fluorene.