Common Reagents Research Articles

Resource utilization of hazardous Cr/Fe-rich sludge: synthesis of erdite flocculant to treat real electroplating wastewater.

Cr/Fe-bearing sludge is a hazardous solid waste, produced at mass production in smelting, plating and surface finishing industries. Such waste is commonly treated by chemical detoxification and safety landfill, whereas only a few Cr-rich sludge is recycled as a tanning reagent. In this study, a novel route was developed to recycle Cr/Fe-bearing sludge as erdite-bearing flocculant for wastewater treatment. Results showed that two sludges were irregular aggregates, one of which contained 1.6wt.% Cr (short for LS) and the other contained 4.2wt.% Cr (HS). After hydrothermal treatment, stable Cr(III)/S-bearing product was formed from the Cr(VI) reduction in the sludges. Conversely, erdite was generated in nanorod form with diameter and length of 200nm and 0.5-1μm from LS, respectively, whereas grew radially to 1.5-2.5μm for HS. The two erdite-bearing products were spontaneously hydrolysed to Fe/S-bearing flocs and showed similar performance in the treatment of real electroplating effluent with 91.55, 1.94 and 0.25mg/L of Zn, Ni and Cr, respectively. For instance, by adding 1g/L product of LS, the release of Cr from the products did not occur, and the residual Zn, Ni and Cr in the effluent was 0.25, 0.65 and 0.17mg/L, respectively, which met the discharge standard of the electroplating industry. With the two converted products, the residual Zn/Ni/Cr concentrations were apparently lower than those of the raw sludges and other common reagents (e.g. polymeric ferric sulphate, activated carbon and diatomite). Thus, such erdite-bearing products could serve as a flocculant and then be applied in electroplating wastewater treatment.

Development of a UHPLC-MS/MS method to enhance the detection of Glyphosate, AMPA and Glufosinate at sub-microgram / L levels in water samples

Glyphosate is the best-selling herbicide worldwide. The toxicity on ecosystems and the possible effects on human health have long been at the centre of a complex controversy concerning the authorisation for its use. The peculiar chemical-physical properties of glyphosate, AMPA and glufosinate make their determination at trace levels in the water a real analytical challenge. All three compounds can be derivatised to less polar ones and FMOC-Cl (9-fluorenylmethyl chloroformate) is the most common pre-column derivatisation reagent used for this analysis. It can be successfully combined with Ultra-High-Performance Liquid Chromatography with Mass Spectrometry (UHPLC-MS/MS) to determine all three analytes in one method as part of water monitoring programs. The developed method aims to determine glyphosate with AMPA and glufosinate at sub-micrograms/L levels in groundwater, surface and water by UHPLC-MS/MS after derivatisation with FMOC-Cl. The novelty of this method is its high simplicity, robustness and sensitivity allowing the identification and quantification of the compounds at the detection limits required by the European regulations (0.1 µg /L). No pre-concentration or purification steps (by using Solid Phase Extraction cartridges) are necessary for our method saving time and consumables costs. The method demonstrated an excellent linear relationship (R2 ≥ 0.999) in the concentration range from 0.025 to 10 µg /L for glyphosate and AMPA and 0.025 to 5 µg /L for glufosinate. The method Limit Of Quantification (LOQ) is 0.025 µg/L, the lowest among all previously published studies, and it was demonstrated according to the European SANTE guidelines.

Inactivation efficacy of H5N1 avian influenza virus by commonly used sample preparation reagents for safe laboratory practices

The objective of this study was to determine the inactivation efficiency of common sample preparation reagents against highly pathogenic avian influenza A (HPAI) H5N1 virus. HPAI H5N1 virus has caused infections in humans with a mortality rate of over 50%. Due to the high mortality and the risk of aerosol transmission of that virus to humans and birds, infectious HPAI H5N1 viruses are contained in a biosafety level 3 laboratory. However, many procedures for further molecular analyses would be easier in lower biosafety conditions. To ensure the laboratory safety the successful inactivation procedures should be demonstrated before the samples are transferred to a lower containment facility. We tested the inactivation capacity of commonly used cell lysis buffer radio-immuno precipitation assay (RIPA) buffer for protein samples, cell fixatives methanol (MeOH) and paraformaldehyde (PFA) and guanidine isothiocyanate-containing lysis buffer for RNA isolation (RLT, Qiagen) in H5N1-infected cells. Based on our results RLT buffer, 90% MeOH (20 min, −20 °C) and 4% PFA (30 min, RT) all completely inactivated the HPAI H5N1 virus. However, RIPA buffer alone was not sufficient to inactivate the HPAI H5N1 virus in infected cell samples but, instead, combining RIPA lysis buffer and boiling for 10 min the samples in Laemmli buffer led to complete inactivation of the virus.

Affordable IgY-based antiviral prophylaxis for resource-limited settings to address epidemic and pandemic risks.

BackgroundThe COVID-19 pandemic caused by SARS-CoV-2 exposed a global problem, as highly effective vaccines are challenging to produce and distribute, particularly in regions with limited resources and funding. As an alternative, immunoglobulins produced in eggs of immunized hens (IgY) can be a simple and inexpensive source for a topical and temporary prophylaxis. Here, we developed a method to extract and purify IgY antibodies from egg yolks of hens immunized against viral pathogen-derived proteins using low-cost, readily available materials, for use in resource-limited settings.MethodsExisting protocols for IgY purification and equipment were modified, including extraction from yolks and separation of water-soluble IgY using common household reagents and tools. A replacement for a commercial centrifuge was developed, using a home food processor equipped with a 3D printed adapter to enable IgY precipitation. IgY purification was verified using standard gel electrophoresis and Western blot analyses.ResultsWe developed a step-by-step protocol for IgY purification for two settings in low- and middle-income countries (LMIC): a local laboratory, where commercial centrifuges are available, or a more rural setting, where an alternative for expensive centrifuges can be used. Gel electrophoresis and Western blot analyses confirmed that the method produced highly enriched IgY preparation; each commercial egg produced ~ 90 mg of IgY. We also designed a kit for IgY production in these two settings and provided a cost estimate of the kit.ConclusionIgY purified from eggs of immunized local hens can offer a fast and affordable prophylaxis, provided that purification can be performed in a resource-limited setting. Here, we created a low-cost method that can be used anywhere where electricity is available using inexpensive, readily available materials in place of costly, specialized laboratory equipment and chemicals. This procedure can readily be used now to make an anti-SARS-CoV-2 prophylaxis in areas where vaccines are unavailable, and can be modified to combat future threats from viral epidemics and pandemics.

Trigonal Bipyramidal Rhodium(I) Methyl and Phenyl Complexes: Precursors of Oxidative Methyl and Phenyl Radical Generation

The new complexes [Rh(Me)(trop3P)] (2) and [Rh(Ph)(trop3P)] (3) (trop = 5H-dibenzo[a,d]cyclohepten-5-yl) were synthesised by addition of organolithium reagents (MeLi and PhLi) to the parent pentacoordinated chloride complex [RhCl(trop3P)]. The compounds have a trigonal bipyramidal structure with olefin-only ligands in the equatorial position and the methyl or phenyl substituent in the axial position. Oxidation of complexes 2 and 3 leads to the liberation of methyl and phenyl radicals, which were indirectly detected by reaction with common spin trapping reagents.

Recent Developments for the Deuterium and Tritium Labeling of Organic Molecules

Organic compounds labeled with hydrogen isotopes play a crucial role in numerous areas, from materials science to medicinal chemistry. Indeed, while the replacement of hydrogen by deuterium gives rise to improved absorption, distribution, metabolism, and excretion (ADME) properties in drugs and enables the preparation of internal standards for analytical mass spectrometry, the use of tritium-labeled compounds is a key technique all along drug discovery and development in the pharmaceutical industry. For these reasons, the interest in new methodologies for the isotopic enrichment of organic molecules and the extent of their applications are equally rising. In this regard, this Review intends to comprehensively discuss the new developments in this area over the last years (2017-2021). Notably, besides the fundamental hydrogen isotope exchange (HIE) reactions and the use of isotopically labeled analogues of common organic reagents, a plethora of reductive and dehalogenative deuteration techniques and other transformations with isotope incorporation are emerging and are now part of the labeling toolkit.

The potential of NO+ and O2 +• in switchable reagent ion proton transfer reaction time-of-flight mass spectrometry.

Selected ion flow tube mass spectrometry (SIFT-MS) and proton transfer reaction mass spectrometry with switchable reagent ion capability (PTR+SRI-MS) are analytical techniques for real-time qualification and quantification of compounds in gas samples with trace level concentrations. In the detection process, neutral compounds-mainly volatile organic compounds-are ionized via chemical ionization with ionic reagents or primary ions. The most common reagent ions are H3 O+ , NO+ and O2 +• . While ionization with H3 O+ occurs by means of proton transfer, the ionization via NO+ and O2 +• offers a larger variety on ionization pathways, as charge transfer, hydride abstraction and so on are possible. The distribution of the reactant into various reaction channels depends not only on the usage of either NO+ or O2 +• , but also on the class of analyte compounds. Furthermore, the choice of the reaction conditions as well as the choice of either SIFT-MS or PTR+SRI-MS might have a large impact on the resulting products. Therefore, an overview of both NO+ and O2 +• as reagent ions is given, showing differences between SIFT-MS and PTR+SRI-MS as used analytical methods revealing the potential how the knowledge obtained with H3 O+ for different classes of compounds can be extended with the usage of NO+ and O2 +• .

Modified Histone Peptides Linked to Magnetic Beads Reduce Binding Specificity.

Histone post-translational modifications are small chemical changes to the histone protein structure that have cascading effects on diverse cellular functions. Detecting histone modifications and characterizing their binding partners are critical steps in understanding chromatin biochemistry and have been accessed using common reagents such as antibodies, recombinant assays, and FRET-based systems. High-throughput platforms could accelerate work in this field, and also could be used to engineer de novo histone affinity reagents; yet, published studies on their use with histones have been noticeably sparse. Here, we describe specific experimental conditions that affect binding specificities of post-translationally modified histones in classic protein engineering platforms and likely explain the relative difficulty with histone targets in these platforms. We also show that manipulating avidity of binding interactions may improve specificity of binding.

An international, interlaboratory ring trial confirms the feasibility of an extraction-less "direct" RT-qPCR method for reliable detection of SARS-CoV-2 RNA in clinical samples.

Reverse transcription–quantitative polymerase chain reaction (RT-qPCR) is used worldwide to test and trace the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). “Extraction-less” or “direct” real time–reverse transcription polymerase chain reaction (RT-PCR) is a transparent and accessible qualitative method for SARS-CoV-2 detection from nasopharyngeal or oral pharyngeal samples with the potential to generate actionable data more quickly, at a lower cost, and with fewer experimental resources than full RT-qPCR. This study engaged 10 global testing sites, including laboratories currently experiencing testing limitations due to reagent or equipment shortages, in an international interlaboratory ring trial. Participating laboratories were provided a common protocol, common reagents, aliquots of identical pooled clinical samples, and purified nucleic acids and used their existing in-house equipment. We observed 100% concordance across laboratories in the correct identification of all positive and negative samples, with highly similar cycle threshold values. The test also performed well when applied to locally collected patient nasopharyngeal samples, provided the viral transport media did not contain charcoal or guanidine, both of which appeared to potently inhibit the RT-PCR reaction. Our results suggest that direct RT-PCR assay methods can be clearly translated across sites utilizing readily available equipment and expertise and are thus a feasible option for more efficient COVID-19 coronavirus disease testing as demanded by the continuing pandemic.

Tandem amide coupling and hydroamination: unexpected benzotriazole oxide addition to the propiolic acid triple bond

An unexpected tandem reaction consisting of amide coupling and hydroamination occurring with common triazole coupling reagents.

Comparison of in-house SARS-CoV-2 genome extraction procedures. A need for COVID-19 pandemic

Among the methods used to diagnose COVID-19, those based on genomic detection by q(RT)-PCR are the most sensitive. To perform these assays, a previous genome extraction of the sample is required. The dramatic increase in the number of SARS-CoV-2 detection assays has increased the demand for extraction reagents hindering the supply of commercial reagents. Homemade reagents and procedures could be an alternative.Nasopharyngeal samples were extracted by seven different methods as well as the automatic method MagNaPure96, to detect SARS-CoV-2. All protocols show sensitivity higher than 87 %, in comparison with reference method, for detecting SARS-CoV-2 as well as human β- globin.Our results support that these procedures, using common and cheap reagents, are effective to extract RNA (from SARS-CoV-2) or DNA (from human β-globin) genome from nasopharyngeal swabs. Furthermore, these procedures could be easily adopted by routine diagnostic laboratories to implement detection methods to help to fight against COVID-19 pandemic.

Validation of antibodies: Lessons learned from the Common Fund Protein Capture Reagents Program.

Large-scale generation of protein capture reagents remains a technical challenge, but their generation is just the beginning. Validation is a critical, iterative process that yields different results for different uses. Independent, community-based validation offers the possibility of transparent data sharing, with use case–specific results made broadly available. This type of resource, which can grow as new validation data are obtained for an expanding group of reagents, provides a community resource that should accompany future reagent-generating efforts. To address a pressing need for antibodies or other reagents that recognize human proteins, the National Institutes of Health Common Fund launched the Protein Capture Reagents Program in 2010 as a pilot to target human transcription factors. Here, we describe lessons learned from this program concerning generation and validation of research reagents, which we believe are generally applicable for future research endeavors working in a similar space.

A Carbohydrate-based Synthetic Approach to Diverse Structurally and Stereochemically Complex Chiral Polyheterocycles.

Chiral polyheterocycles are one of the most frequently encountered scaffolds in natural products and in current drugs repertoire. A carbohydrate-based diversity oriented synthetic (DOS) approach has been employed for gaining access to many structurally diverse and stereochemically complex rigid polyheterocyclic molecules with multiple chiral hydroxyl groups to enhance aqueous solubility. Inexpensive chiral pool of D-Glucose has been judiciously exploited to get access of complex chiral polyheterocyclic structures using inexpensive, common achiral reagents and domino-Knoevenagel hetero-Diels-Alder (DKHDA) reaction as one of the key synthetic tools. Stereochemistry of newly generated stereocenters of polycyclic structures are unambiguously determined through NMR and X-ray crystallographic study. A chemoinformatic comparison (PCA and PMI) with 40 branded blockbuster drugs showed that newly generated polyheterocycles have good three-dimensional scaffold diversity and most of these pass the Lipinski filter of drug-likeness.

A Two-step Electrodeposition of Pd-Cu/Cu2O Nanocomposite on FTO Substrate for Non-enzymatic Hydrogen Peroxide Sensor

Background: Hydrogen peroxide (H2O2) is a common reagent in the production and living, but excessive H2O2 may enhance the danger to the human body. Consequently, it is very important to develop economical, fast and accurate techniques for detecting H2O2. Methods: A simple two-step electrodeposition process was applied to synthesize Pd-Cu/Cu2O nanocomposite for non-enzymatic H2O2 sensor. Cu/Cu2O nanomaterial was firstly electrodeposited on FTO by potential oscillation technique, and then Pd nanoparticles were electrodeposited on Cu/Cu2O nanomaterial by cyclic voltammetry. The chemical structure, component, and morphology of the synthesized Pd-Cu/Cu2O nanocomposite were characterized by X-ray diffraction, scanning electron microscopy and X-ray photoelectron spectroscopy. The electrochemical properties of Pd-Cu/Cu2O nanocomposite were studied by cyclic voltammetry and amperometry. Results: Under optimal conditions, the as-fabricated sensor displayed a broad linear range (5-4000 μM) and low detection limit (1.8 μM) for the determination of H2O2. The proposed sensor showed good selectivity and reproducibility. Meanwhile, the proposed sensor has been successfully applied to detect H2O2 in milk. Conclusion: The Pd-Cu/Cu2O/FTO biosensor exhibits excellent electrochemical activity for H2O2 reduction, which has great potential application in the field of food safety.

Method transfer from ion pair chromatography to charged aerosol detector-compatible mixed-mode chromatography: A case study using carbocisteine

To take advantage of the charged aerosol detector (CAD) for sensitive LC analysis of analytes with weak chromophore, the volatility of the entire mobile phase is a prerequisite. Therefore, it is not feasible to use common non-volatile ion-pairing reagents such as tetrabutylammonium hydroxide for the analysis of polar, acidic amino acids.This case study presents a possible procedure for transferring an ion pair chromatography to an HPLCCAD method using the example of carbocisteine and its two main degradation products, carbocisteine sulfoxide and carbocisteine lactam. In screening experiments, substitution by various volatile alkylamines resulted in inferior chromatographic separation. As an alternative, mixed-mode chromatography using a column with strong anion exchange functionalities and a mobile phase containing 12 mM formic acid and 2 mM trifluoroacetic acid, led to a selectivity equivalent to ion pair chromatography with tetrabutylammonium hydroxide. A change in the level of background noise generated by non-volatile impurities eluting from the column during measurements – often called column bleeding – invalidated the settings predicted from response surface methodology experiments in terms of modelling chromatographic selectivity and CAD sensitivity, and a reduction of the acetonitrile ratio to 22% (v/v) was required. At a flow rate of 1.2 mL/min and an evaporation temperature of 65 °C, the experimentally determined LOQs of both degradation products were below 50 ng on column. The combination of mixed-mode chromatography with CAD offers a viable alternative to ion pair chromatography with UV detection in the analysis of acidic analytes with a weak chromophore such as amino acids and impurities thereof.

Metagenomic Identification of Viral Sequences in Laboratory Reagents.

Metagenomic next-generation sequencing has transformed the discovery and diagnosis of infectious disease, with the power to characterise the complete ‘infectome’ (bacteria, viruses, fungi, parasites) of an individual host organism. However, the identification of novel pathogens has been complicated by widespread microbial contamination in commonly used laboratory reagents. Using total RNA sequencing (“metatranscriptomics”) we documented the presence of contaminant viral sequences in multiple ‘blank’ negative control sequencing libraries that comprise a sterile water and reagent mix. Accordingly, we identified 14 viral sequences in 7 negative control sequencing libraries. As in previous studies, several circular replication-associated protein encoding (CRESS) DNA virus-like sequences were recovered in the blank control libraries, as well as contaminating sequences from the Totiviridae, Tombusviridae and Lentiviridae families of RNA virus. These data suggest that viral contamination of common laboratory reagents is likely commonplace and can comprise a wide variety of viruses.

A generic liquid chromatography-mass spectrometry method for monitoring bis(pinacolato)diboron mutagenic impurity in pharmaceutical compounds

Bis(pinacolato)diboron (B2Pin2) is a common reagent used to prepare pinacolboronate esters for Suzuki-Miyaura coupling reactions in pharmaceutical syntheses. As a known mutagenic impurity, it is challenging to develop analytical methods for the determination of B2Pin2 due to its low control limit, high reactivity, and lack of chromophore. Here, we report a generic liquid chromatography (LC)- mass spectrometry (MS) method to monitor B2Pin2 at low parts per million (ppm) levels in pharmaceutical compounds by single ion monitoring. Ammonium acetate (0.5 mM) was added in the mobile phases to form B2Pin2 gas-phase ammonium adduct, which eliminated MS instrumentation variances and achieved a quantification limit (QL) better than 10 ng/mL (corresponding to 1 ppm in substrate at 10 mg/mL) with optimized LC conditions. To accommodate diverse solubility profiles of pharmaceutical compounds, three diluents including acetonitrile, stabilized tetrahydrofuran, and dimethyl sulfoxide were recommended as the primary diluents based on B2Pin2 solution stability. Three alternative diluents with additives were proposed as backups. Three Bristol Myers Squibb compounds that required B2Pin2 control were spiked with 1–20 ppm B2Pin2 as the case studies. The QLs for all three materials were better than 1 ppm and the recovery values were within 100 ± 20% at 1, 10, and 20 ppm spiked levels.

Diethylaminosulfur Trifluoride: A Novel, Low-Cost, Stable Double Thiolation Reagent for Imidazo[1,2-α]pyridines.

We report a novel, inexpensive double thiolation reagent that sulfurizes a broad range of imidazo[1,2-α]pyridines under mild conditions. Importantly, diethylaminosulfur trifluoride, as a common nucleophilic fluorinating reagent, was utilized as a novel thiolation reagent.

Controllable Release of Povidone-Iodine from Networked Pectin@Carboxymethyl Pullulan Hydrogel.

Povidone-iodine (PI) is a common antiseptic reagent which is used for skin infections and wound healing. The control release of PI is quite important to heal the deep and intense wounds. Herein, the preparation of biodegradable pectin@carboxymethyl pullulan (Pe@CMP) hydrogel was carried out and applied for controllable release of PI. CMP was synthesized by interaction of monochloroacetic acid with pullulan at different ratios. The Pe@CMP hydrogel was then prepared by crosslinking of pectin with CMP in presence of glutaraldehyde as cross linker. After carboxymethylation, COOH contents were enlarged to be 24.2–51.2 mmol/kg and degree of substitution was 0.44–0.93. The rheological properties of Pe@CMP hydrogel were enlarged by increment of pectin ratio. Swelling ratio in water (16.0–18.0%) was higher than that of artificial sweat (11.7–13.2%). Pe@CMP hydrogel containing 20% pectin, exhibited the lowest release and 57.7% from PI was released within 360 min. The biological activity of the released PI was monitored to be highly efficient. The kinetic of release was fitted well to the first ordered reaction and Higuchi models. The mechanism of release was explained by the swelling of hydrogel. The networked structure of hydrogel was opened by swelling and PI was released from the outer pores followed by inner pores, achieving the controllable release.

Synthesis of closo-CB11H12- Salts Using Common Laboratory Reagents.

Lithium and sodium salts of the closo-carbadodecaborate anion [CB11H12]- have been shown to form stable solid-state electrolytes with excellent ionic conductivity for all-solid-state batteries (ASSB). However, potential commercial application is currently hindered by the difficult, low-yielding, and expensive synthetic pathways. We report a novel and cost-effective method to synthesize the [CB11H12]- anion in a 40% yield from [B11H14]-, which can be synthesized using common laboratory reagents. The method avoids the use of expensive and dangerous reagents such as NaH, decaborane, and CF3SiMe3 and shows excellent reproducibility in product yield and purity.