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1.
Biotechnol J ; 18(9): e2300143, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37222181

RESUMO

A paper microfluidic device capable of conducting enzyme-linked assays is presented: a microfluidic enzyme-linked paper analytical device (µEL-PAD). The system exploits a wash-free sandwich coupling to form beads/analyte/enzyme complexes, which are subsequently added to the vertical flow device composed of wax-printed paper, waxed nitrocellulose membrane and absorbent/barrier layers. The nitrocellulose retains the bead complexes without disrupting the flow, enabling for an efficient washing step. The entrapped complexes then interact with the chromogenic substrate stored on the detection paper, generating a color change on it, quantified with an open-source smartphone software. This is a universal paper-based technology suitable for high-sensitivity quantification of many analytes, such as proteins or nucleic acids, with different enzyme-linked formats. Here, the potential of the µEL-PAD is demonstrated to detect DNA from Staphylococcus epidermidis. After generation of isothermally amplified genomic DNA from bacteria, Biotin/FITC-labeled products were analyzed with the µEL-PAD, exploiting streptavidin-coated beads and antiFITC-horseradish peroxidase. The µEL-PAD achieved a limit of detection (LOD) and quantification <10 genome copies/µL, these being at least 70- and 1000-fold lower, respectively, than a traditional lateral flow assay (LFA) exploiting immobilized streptavidin and antiFITC-gold nanoparticles. It is envisaged that the device will be a good option for low-cost, simple, quantitative, and sensitive paper-based point-of-care testing.


Assuntos
Técnicas de Química Analítica , Microfluídica , Papel , Microfluídica/instrumentação , Colódio/química , Compostos Cromogênicos/química , Aplicativos Móveis , Proteínas/análise , Ácidos Nucleicos/análise , Limite de Detecção , Técnicas de Química Analítica/instrumentação , Técnicas de Química Analítica/métodos
2.
Chemosphere ; 323: 138285, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36868424

RESUMO

Anion exchange resin is responsible for removing harmful anionic contaminants in drinking water treatment, but it may become a significant source of precursors for disinfection byproducts (DBPs) by shedding material during application without proper pretreatment. Batch contact experiments were performed to investigate the dissolution of magnetic anion exchange resins and their contribution to organics and DBPs. Dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) released from the resin were highly correlated with the dissolution conditions (contact time and pH), in which 0.7 mg/L DOC and 0.18 mg/L DON were distributed at exposure time of 2 h and pH 7. The formation potential of four DBPs in the shedding fraction was also revealed that trichloromethane (TCM), dichloroacetonitrile (DCAN), nitrosodimethylamine (NDMA), and dichloroacetamide (DCAcAm) concentrations could reach 21.4, 5.1, 12.1 µg/L, and 69.6 ng/L, respectively. Furthermore, the hydrophobic DOC that preferred to detach from the resin mainly originated from the residues of crosslinkers (divinylbenzene) and porogenic agents (straight-chain alkanes) detected by LC-OCD and GC-MS. Nevertheless, pre-cleaning inhibited the leaching of the resin, among which acid-base and ethanol treatments significantly lowered the concentration of leached organics, and formation potential of DBPs (TCM, DCAN, and DCAcAm) below 5 µg/L and NDMA dropped to 10 ng/L.


Assuntos
Resinas de Troca Aniônica , Técnicas de Química Analítica , Purificação da Água , Resinas de Troca Aniônica/química , Purificação da Água/instrumentação , Matéria Orgânica Dissolvida/análise , Matéria Orgânica Dissolvida/química , Clorofórmio/análise , Clorofórmio/química , Dimetilnitrosamina/análise , Dimetilnitrosamina/química , Concentração de Íons de Hidrogênio , Microscopia Eletrônica de Varredura , Técnicas de Química Analítica/instrumentação , Técnicas de Química Analítica/métodos
3.
Anal Biochem ; 639: 114520, 2022 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-34896376

RESUMO

Long non-coding RNAs (lncRNAs) are transcripts longer than 200 nt without evident protein coding function. They play important regulatory roles in many biological processes, e.g., gene regulation, chromatin remodeling, and cell fate determination during development. Dysregulation of lncRNAs has been observed in various diseases including cancer. Interacting with proteins is a crucial way for lncRNAs to play their biological roles. Therefore, the characterization of lncRNA binding proteins is important to understand their functions and to delineate the underlying molecular mechanism. Large-scale studies based on mass spectrometry have characterized over a thousand new RNA binding proteins without known RNA-binding domains, thus revealing the complexity and diversity of RNA-protein interactions. In addition, several methods have been developed to identify the binding proteins for particular RNAs of interest. Here we review the progress of the RNA-centric methods for the identification of RNA-protein interactions, focusing on the studies involving lncRNAs, and discuss their strengths and limitations.


Assuntos
RNA Longo não Codificante/metabolismo , Proteínas de Ligação a RNA/metabolismo , Animais , Técnicas de Química Analítica/instrumentação , Técnicas de Química Analítica/métodos , Humanos , Ligação Proteica , RNA Longo não Codificante/análise , Proteínas de Ligação a RNA/análise
4.
Mikrochim Acta ; 189(1): 34, 2021 12 23.
Artigo em Inglês | MEDLINE | ID: mdl-34940928

RESUMO

DNA is recognized as a powerful biomarker for clinical diagnostics because its specific sequences are closely related to the cause and development of diseases. However, achieving rapid, low-cost, and sensitive detection of short-length target DNA still remains a considerable challenge. Herein, we successfully combine the catalytic hairpin assembly (CHA) technique with capillary action to develop a new and cost-effective method, a target DNA- and pH-responsive DNA hydrogel-based capillary assay, for the naked eye detection of 24 nt short single-stranded target DNA. Upon contact of target DNA, three individual hairpin DNAs hybridize with each other to sufficiently amplify Y-shaped DNA nanostructures (Y-DNA) until they are completely consumed via CHA cycling reactions. Each arm of the resultant Y-DNA contains sticky ends with i-motif DNA structure-forming sequences that can be self-assembled in an acidic environment (pH 5.0) to form target DNA- and pH-responsive DNA hydrogels by means of i-motif DNA-driven crosslinking. When inserting a capillary tube in the resultant solution, the liquid level inside clearly reduces due to the decrease in capillary force induced by the gels. In this way, the developed assay demonstrates sensitive and quantitative detection, with a detection limit of approximately 10 pM of 24 nt short complementary DNA (cDNA) targeting SARS-CoV-2 RNA genes at room temperature within 1 h. The assay is further shown to successfully detect target cDNA in serum, and it is also applied to detect several types of target sequences. Requiring no analytic equipment, precise temperature control, or enzymatic reactions, the developed DNA hydrogel-based capillary assay has potential as a promising naked eye detection platform for target DNA in resource-limited clinical settings.


Assuntos
Técnicas de Química Analítica/métodos , DNA Catalítico/química , DNA Complementar/análise , Hidrogéis/química , RNA Viral/genética , SARS-CoV-2/química , Ação Capilar , Técnicas de Química Analítica/instrumentação , DNA Catalítico/genética , DNA Complementar/genética , Concentração de Íons de Hidrogênio , Sequências Repetidas Invertidas , Limite de Detecção , Técnicas de Amplificação de Ácido Nucleico , Hibridização de Ácido Nucleico
5.
PLoS Negl Trop Dis ; 15(9): e0009539, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34591842

RESUMO

Substandard and falsified (SF) antimalarials have devastating consequences including increased morbidity, mortality and economic losses. Portable medicine quality screening devices are increasingly available, but whether their use for the detection of SF antimalarials is cost-effective is not known. We evaluated the cost-effectiveness of introducing such devices in post-market surveillance in pharmacies in Laos, conservatively focusing on their outcome in detecting SF artemisinin-based combination therapies (ACTs). We simulated the deployment of six portable screening devices: two handheld near-infrared [MicroPHAZIR RX, NIR-S-G1], two handheld Raman [Progeny, TruScan RM]; one portable mid-infrared [4500a FTIR] spectrometers, and single-use disposable paper analytical devices [PADs]. We considered two scenarios with high and low levels of SF ACTs. Different sampling strategies in which medicine inspectors would test 1, 2, or 3 sample(s) of each brand of ACT were evaluated. Costs of inspection including device procurement, inspector time, reagents, reference testing, and replacement with genuine ACTs were estimated. Outcomes were measured as disability adjusted life years (DALYs) and incremental cost-effectiveness ratios were estimated for each device compared with a baseline of visual inspections alone. In the scenario with high levels of SF ACTs, all devices were cost-effective with a 1-sample strategy. In the scenario of low levels of SF ACTs, only four devices (MicroPHAZIR RX, 4500a FTIR, NIR-S-G1, and PADs) were cost-effective with a 1-sample strategy. In the multi-way comparative analysis, in both scenarios the NIR-S-G1 testing 2 samples was the most cost-effective option. Routine inspection of ACT quality using portable screening devices is likely to be cost-effective in the Laos context. This work should encourage policy-makers or regulators to further investigate investment in portable screening devices to detect SF medicines and reduce their associated undesired health and economic burdens.


Assuntos
Antimaláricos/química , Técnicas de Química Analítica/instrumentação , Técnicas de Química Analítica/métodos , Medicamentos Falsificados/análise , Medicamentos Fora do Padrão/análise , Antimaláricos/economia , Técnicas de Química Analítica/economia , Serviços Comunitários de Farmácia , Análise Custo-Benefício , Medicamentos Falsificados/economia , Humanos , Laos/epidemiologia , Malária/tratamento farmacológico , Malária/economia , Malária/epidemiologia , Vigilância de Produtos Comercializados , Medicamentos Fora do Padrão/economia
6.
PLoS Negl Trop Dis ; 15(9): e0009360, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34591844

RESUMO

BACKGROUND: Post-market surveillance is a key regulatory function to prevent substandard and falsified (SF) medicines from being consumed by patients. Field deployable technologies offer the potential for rapid objective screening for SF medicines. METHODS AND FINDINGS: We evaluated twelve devices: three near infrared spectrometers (MicroPHAZIR RX, NIR-S-G1, Neospectra 2.5), two Raman spectrometers (Progeny, TruScan RM), one mid-infrared spectrometer (4500a), one disposable colorimetric assay (Paper Analytical Devices, PAD), one disposable immunoassay (Rapid Diagnostic Test, RDT), one portable liquid chromatograph (C-Vue), one microfluidic system (PharmaChk), one mass spectrometer (QDa), and one thin layer chromatography kit (GPHF-Minilab). Each device was tested with a series of field collected medicines (FCM) along with simulated medicines (SIM) formulated in a laboratory. The FCM and SIM ranged from samples with good quality active pharmaceutical ingredient (API) concentrations, reduced concentrations of API (80% and 50% of the API), no API, and the wrong API. All the devices had high sensitivities (91.5 to 100.0%) detecting medicines with no API or the wrong API. However, the sensitivities of each device towards samples with 50% and 80% API varied greatly, from 0% to 100%. The infrared and Raman spectrometers had variable sensitivities for detecting samples with 50% and 80% API (from 5.6% to 50.0%). The devices with the ability to quantitate API (C-Vue, PharmaChk, QDa) had sensitivities ranging from 91.7% to 100% to detect all poor quality samples. The specificity was lower for the quantitative C-Vue, PharmaChk, & QDa (50.0% to 91.7%) than for all the other devices in this study (95.5% to 100%). CONCLUSIONS: The twelve devices evaluated could detect medicines with the wrong or none of the APIs, consistent with falsified medicines, with high accuracy. However, API quantitation to detect formulations similar to those commonly found in substandards proved more difficult, requiring further technological innovation.


Assuntos
Técnicas de Química Analítica/instrumentação , Técnicas de Química Analítica/métodos , Medicamentos Falsificados/análise , Avaliação Pré-Clínica de Medicamentos/instrumentação , Medicamentos Fora do Padrão/análise , Avaliação Pré-Clínica de Medicamentos/métodos , Dispositivos Lab-On-A-Chip , Controle de Qualidade , Sensibilidade e Especificidade
7.
PLoS Negl Trop Dis ; 15(9): e0009674, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34591852

RESUMO

BACKGROUND: Medicine quality screening devices hold great promise for post-market surveillance (PMS). However, there is little independent evidence on their field utility and usability to inform policy decisions. This pilot study in the Lao PDR tested six devices' utility and usability in detecting substandard and falsified (SF) medicines. METHODOLOGY/PRINCIPAL FINDINGS: Observational time and motion studies of the inspections by 16 Lao medicine inspectors of 1) the stock of an Evaluation Pharmacy (EP), constructed to resemble a Lao pharmacy, and 2) a sample set of medicines (SSM); were conducted without and with six devices: four handheld spectrometers (two near infrared: MicroPHAZIR RX, NIR-S-G1 & two Raman: Progeny, Truscan RM); one portable mid-infrared spectrometer (4500a), and single-use paper analytical devices (PAD). User experiences were documented by interviews and focus group discussions. Significantly more samples were wrongly categorised as pass/fail with the PAD compared to the other devices in EP inspections (p<0.05). The numbers of samples wrongly classified in EP inspections were significantly lower than in initial visual inspections without devices for 3/6 devices (NIR-S-G1, MicroPHAZIR RX, 4500a). The NIR-S-G1 had the fastest testing time per sample (median 93.5 sec, p<0.001). The time spent on EP visual inspection was significantly shorter when using a device than for inspections without devices, except with the 4500a, risking missing visual clues of samples being SF. The main user errors were the selection of wrong spectrometer reference libraries and wrong user interpretation of PAD results. Limitations included repeated inspections of the EP by the same inspectors with different devices and the small sample size of SF medicines. CONCLUSIONS/SIGNIFICANCE: This pilot study suggests policy makers wishing to implement portable screening devices in PMS should be aware that overconfidence in devices may cause harm by reducing inspectors' investment in visual inspection. It also provides insight into the advantages/limitations of diverse screening devices in the hands of end-users.


Assuntos
Anti-Infecciosos/química , Técnicas de Química Analítica/instrumentação , Medicamentos Falsificados/análise , Medicamentos Fora do Padrão/análise , Técnicas de Química Analítica/métodos , Humanos , Laos/epidemiologia , Projetos Piloto , Pilotos , Sensibilidade e Especificidade
9.
J Chromatogr A ; 1654: 462448, 2021 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-34392123

RESUMO

Although, in general, the application of coated capillaries is recommended for the separation of intact proteins, bare silica capillary is still the most often used capillary due to its simplicity and cheapness. In this work, the performance of bare fused silica capillary for intact protein analysis was compared to that of different (dynamically coated polybrene (PB) and permanently coated linear polyacrylamide (LPA)) coated capillaries using capillary zone electrophoresis - mass spectrometry (CZE-MS). In cases where low pH (pH=1.8) was used in bare silica capillaries, good precision (0.56-0.78 RSD% and 1.7-6.5 RSD% for migration times and peak areas, respectively), minimal adsorption and separation efficiency (N= 27 000/m - 322 000/m) similar to or even better than those obtained with the coated capillaries (created by an intricate multi-step process) was achieved. The PB and the LPA capillaries demonstrated their slightly better resolving power in terms of separating the different forms/variants of the same protein (e.g., hemoglobin subunits). Among the studied capillaries the one with LPA coating showed the most stable separations in the long term (n=25: 0.18-0.49 RSD% and 3.1-4.9 RSD% for migration times and peak areas, respectively). For the separation of a few proteins or even a larger number of proteins in biological samples (e.g., snake venom) the application of the simple and cheap bare fused silica capillary can be considered as an efficient choice.


Assuntos
Técnicas de Química Analítica , Eletroforese Capilar , Espectrometria de Massas , Proteínas , Resinas Acrílicas/química , Técnicas de Química Analítica/instrumentação , Técnicas de Química Analítica/métodos , Eletroforese Capilar/instrumentação , Brometo de Hexadimetrina/química , Proteínas/química , Proteínas/isolamento & purificação
10.
Mikrochim Acta ; 188(8): 258, 2021 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-34268648

RESUMO

Pathogenic bacteria have become a huge threat to social health and economy for their frighteningly infectious and lethal capacity. It is quite important to make a diagnosis in advance to prevent infection or allow a rapid treatment after infection. Noble metal nanoparticles, due to their unique physicochemical properties, especially optical properties, have drawn a great attention during the past decades and have been widely applied into all kinds of fields related to human health. By utilizing these noble metal nanoparticles, optical diagnosis platforms towards pathogenic bacteria have emerged continually, providing highly sensitive, selective, and particularly facile detection tools for clinic or point-of-care diagnosis. This review summarizes the recent development in this field. It begins with a brief introduction of pathogenic bacteria and noble metal nanoparticles. And then, optical detection methods are systematically discussed in three distinct aspects. In addition to these proof-of-concept methods, corresponding algorithms and point-of-care detection devices are also described. Finally, the review ends up with subjective views on present limitations and some appropriate advice for future research directions.


Assuntos
Bactérias/isolamento & purificação , Técnicas Bacteriológicas/métodos , Técnicas de Química Analítica/métodos , Nanopartículas Metálicas/química , Técnicas Bacteriológicas/instrumentação , Técnicas de Química Analítica/instrumentação , Metais Pesados/química , Testes Imediatos
11.
J Chromatogr A ; 1652: 462372, 2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-34246058

RESUMO

In the present work, three-dimensional (3D) and flower-like SnS2 materials were coated on the surface of Fe3O4@nSiO2 through an in-situ growth method. The 3D architecture could avoid the accumulation and reaggregation with better stability and was beneficial for the exposure of more active sites. The prepared magnetic SnS2 composites were used for the enrichment of sulfonamide antibiotics (SAs), and various experimental parameters affecting the extraction efficiency were investigated. The results showed the equilibrium of extraction and desorption towards target SAs could be reached within 3 min by using the Fe3O4@nSiO2-SnS2 composites. Under optimized conditions, the proposed approach possessed good linearity in the range of 0.1-200 ng·mL-1 with correlation coefficients r2 above 0.9964 and low limits of detection (LODs) from 0.025 to 0.250 ng·mL-1 for the five target SAs. Moreover, good repeatability was obtained with the intra-day and inter-day precision in terms of relative standard deviations (RSDs) within 1.1%-10.8% and 7.4%-13.1%, and the recoveries under three spiked concentrations were between 81.8% and 119.7% with adequate accuracy. Different samples including tap water, milk and honey were collected for magnetic solid-phase extraction and determination of target SAs by using the obtained Fe3O4@nSiO2-SnS2 composites to demonstrate the utility. All the results indicated that the proposed method had great potential for effective preconcentration and determination of SAs in complex samples.


Assuntos
Antibacterianos , Técnicas de Química Analítica , Dissulfetos , Extração em Fase Sólida , Antibacterianos/isolamento & purificação , Técnicas de Química Analítica/instrumentação , Técnicas de Química Analítica/métodos , Limite de Detecção , Fenômenos Magnéticos
12.
J Chromatogr A ; 1650: 462251, 2021 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-34062482

RESUMO

The carbon dots (CDs) from natural nanographite oxide mixture (NGO-MIX) and from its fraction NGO (3.5-10K) recovered after ultrafiltration and dialysis were analyzed by 3D-excitation/emission matrix and high-performance size exclusion chromatography (HPSEC) combined with online fluorescence and absorbance detections. HPSEC chromatograms obtained simultaneously with absorption within the wavelength range 200-500 nm and fluorescence detection at λexc/λem = 270/450 nm/nm showed that NGO-MIX sample is not homogeneous and consist of well resolved CDs fractions with different sizes, absorption spectra and distinct fluorescence and non-fluorescence properties. Despite the twice higher fluorescence intensity of fraction NGO (3.5-10K) compared to the NGO-MIX, some impurity of non-fluorescent components was detected by HPSEC. The absorbance spectra of chromatographic peaks, extracted from the data of multi-wavelength absorbance detector, demonstrated different combinations of absorbance maxima. It means that different chromatographic peaks correspond to sized and chemically different CDs fractions. This study demonstrated for the first time the possibility of separating oxidized nanographite into homogeneous free from non-fluorescent material CDs fractions with their simultaneous spectroscopic characterization.


Assuntos
Carbono , Técnicas de Química Analítica , Cromatografia em Gel , Pontos Quânticos , Carbono/análise , Carbono/isolamento & purificação , Técnicas de Química Analítica/instrumentação , Técnicas de Química Analítica/métodos , Fluorescência , Compostos Orgânicos/análise , Pontos Quânticos/análise , Análise Espectral
13.
J Chromatogr A ; 1650: 462256, 2021 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-34082187

RESUMO

An analytical method based on low-temperature partitioning extraction (LTPE) followed by high performance liquid chromatography coupled to triple quadrupole mass spectrometry analysis was developed and validated for the determination of eight multiclass antibiotics in wastewater. The analyzed target antibiotics included one ß-lactam, two sulfonamides, three fluoroquinolones, one macrolide and one diaminopyrimidine. LTPE parameters such as sample pH, volume ratio between sample and extractor solvent, ultra-sonic extraction time, extraction tube material, solvent and volume to reconstitute the sample extracts, were optimized. Additionally, the influence of solids on extraction efficiency was evaluated. Quantification of the target antibiotics was performed by double consecutive injection method, without the use of a labeled compound, in order to correct matrix effects. The whole samples were analyzed, including, liquid and solid fractions of wastewater. The results revealed that the filtration step can underestimate the total antibiotics concentration, particularly to the hydrophobic compounds that have higher affinity for solids, indicating that the suspended wastewater particulate should not be neglected. The method detection limit ranged from 18.54 ng L-1 (trimethoprim) to 78.49 ng L-1 (ciprofloxacin). Intra-day precision of less than 12.3% was achieved. The recoveries values ranged from 13.9% (sulfadiazine) to 48.9% (erythromycin) in influent samples and from 19.1% (sulfadiazine) to 57.2% (ciprofloxacin) in effluent samples. The method was applied to the measurement of antibiotic residues in influent and effluent from wastewater treatment plants. The majority target antibiotics were detected in wastewater samples. Their concentrations ranged from 237 to 9553 ng L-1 in influent and from 212 to 1660 ng L-1 in effluent. This work provides new insights on the applicability of LTPE for antibiotic residues extraction from wastewater. In addition, the performed analysis highlights the importance of measuring total concentrations of analytes in whole sample.


Assuntos
Antibacterianos , Técnicas de Química Analítica , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas em Tandem , Águas Residuárias , Poluentes Químicos da Água , Antibacterianos/análise , Técnicas de Química Analítica/instrumentação , Técnicas de Química Analítica/métodos , Temperatura Baixa , Interações Hidrofóbicas e Hidrofílicas , Extração em Fase Sólida , Águas Residuárias/química , Poluentes Químicos da Água/análise
14.
J Chromatogr A ; 1651: 462303, 2021 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-34147834

RESUMO

The advantages of using mixtures of organic solvents for the separation of labeled oligosaccharides on the amide stationary phase under hydrophilic interaction liquid chromatography conditions are presented. The effect of the type of buffer as well as solvent or their mixtures on retention of uracil, saccharide labeling reagents (2-aminobenzoic acid, 2-aminobenzamide, ethyl 4-aminobenzoate, procainamide), and corresponding labeled saccharides were evaluated. The successful isocratic separation of labeled isomeric trisaccharides (maltotriose, panose, and isomaltotriose) was achieved in the mobile phase consisting of a 90% (v/v) mixture of organic solvents (methanol/acetonitrile 60:40) and 10% (v/v) 30 mM ammonium formate, pH 3.3. Changing the volume ratio between methanol/acetonitrile from 60:40 to 50:50 (v/v) allowed to obtain the separation of di-, tri-, and tetrasaccharides labeled by ethyl 4-aminobenzoate in less than 10.5 min.


Assuntos
Amidas/química , Técnicas de Química Analítica/métodos , Cromatografia Líquida , Oligossacarídeos/isolamento & purificação , Acetonitrilas/química , Carboidratos/química , Técnicas de Química Analítica/instrumentação , Formiatos/química , Interações Hidrofóbicas e Hidrofílicas , Isomerismo , Solventes/química , ortoaminobenzoatos/química
15.
J Chromatogr A ; 1651: 462337, 2021 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-34157476

RESUMO

Here, a m-xylene bisphosphonate immobilized tentacle-type cellulose monolith (BP-PCM) is prepared by atom transfer radical polymerization for lysozyme purification. In the preparation, the m-xylene bisphosphonate was anchored glycidyl methacrylate and then polymerized to enhance the flexibility of the ligands to improve lysozyme adsorption capacity, and glycerol monomethacrylate serves as spacer to further optimize the layers structure and ligands density of the grafted tentacles for satisfactory adsorption capacity. The maximum static and dynamic adsorption capacity (10% breakthrough) of BP-PCM reach to 169.6 and 102.6 mg mL-1, respectively. Moreover, BP-PCM displays weak nonspecific adsorption and is able to successfully enrich lysozyme from diluted chicken egg white, indicating the excellent selectivity. The results demonstrated that BP-PCM is promising for use as high-capacity protein chromatography.


Assuntos
Celulose/química , Técnicas de Química Analítica/métodos , Cromatografia , Difosfonatos/química , Muramidase/isolamento & purificação , Adsorção , Técnicas de Química Analítica/instrumentação , Compostos de Epóxi/química , Ligantes , Metacrilatos/química , Muramidase/química , Polimerização , Porosidade
16.
J Chromatogr A ; 1651: 462339, 2021 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-34161838

RESUMO

A new sensitive and selective analytical methodology to quantify glyphosate (GLY), aminomethylphosphonic acid (AMPA), and glufosinate (GLU) in both soil and earthworms (Allolobophora chlorotica) was developed. The extraction and purification methods were optimized. The samples were extracted with various aqueous solutions (HNO3, H2O, KOH and borate buffer) and derivatized with 9-Fluorenylmethyl chloroformate (FMOCCl). To optimize the extraction step, a method to remove the excess FMOCCl was applied based on liquid-liquid extraction with diethyl ether. The purification of derivatized extracts was carried out using XLB solid phase extraction (SPE) cartridges before internal standard quantification by liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS). The elution step was optimized to obtain the best recoveries possible, which was with acidic methanol (1% formic acid) (67% for GLY, 70% for GLU and 65% for AMPA). The extraction and purification method followed by analysis of the two herbicides and AMPA in soils using LC/MS/MS determined limit of quantification (LOQ) values of 0.030 µg g - 1 for GLY, 0.025 µg g - 1 for AMPA and 0.020 µg g - 1 for GLU . For earthworms, LOQ were 0.23 µg g - 1 for GLY, 0.20 µg g - 1 for AMPA and 0.12 µg g - 1 for GLU. . The developed method was applied to determine these compounds in natural soils and earthworms.


Assuntos
Aminobutiratos/análise , Técnicas de Química Analítica/métodos , Glicina/análogos & derivados , Oligoquetos/química , Organofosfonatos/análise , Solo/química , Aminobutiratos/isolamento & purificação , Animais , Técnicas de Química Analítica/instrumentação , Cromatografia Líquida , Glicina/análise , Glicina/isolamento & purificação , Herbicidas/análise , Herbicidas/isolamento & purificação , Organofosfonatos/isolamento & purificação , Extração em Fase Sólida , Espectrometria de Massas em Tandem , Glifosato
17.
J Chromatogr A ; 1651: 462304, 2021 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-34118531

RESUMO

Hemp and cannabis industry is undergoing a renewed interest due to legalization of marijuana (a topic that all countries are discussing, especially in recent years) and the growing importance of therapeutic properties of cannabinoids. Together with an increment in the production of hemp and recreational cannabis, there has been an increasing demand for accurate potency testing of products (i.e. quantification of main cannabinoids present in the plant in terms of weight percentage) prior commercialization. This translates in an urgent need of reliable analytical methods to characterize cannabis and hemp samples. Cannabis and hemp preparations are commercialized under various forms (e.g., flowers, oils, candies or even baked goods) usually containing a large number of often very similar compounds making their separation very challenging. Strictly connected to this, another emerging topic concerns the need for the developing of large scale separation techniques for the purification of cannabinoids from complex matrices and for the preparation of analytical-grade standards (including the chiral ones). This paper reviews the most recent achievements in both these aspects. Cutting-edge applications and novel opportunities in potency testing by high performance liquid chromatography (HPLC) with UV detection (which is becoming the golden standard, according to several pharmacopeias, for this kind of measurements) are discussed. The focus has been given to the very important topic of enantio-discrimination of chiral cannabinoids, for which supercritical fluid chromatography (SFC) appears to be particularly suitable. The last part of the work covers the purification of cannabinoids through preparative chromatography. In this regard, particular attention has been given to the most innovative multi-column techniques allowing for the continuous purification of target molecules. The most recent advancements and future challenges in this field are discussed.


Assuntos
Canabinoides/análise , Técnicas de Química Analítica/métodos , Cromatografia Líquida de Alta Pressão , Cromatografia com Fluido Supercrítico , Cannabis/química , Técnicas de Química Analítica/instrumentação , Flores/química , Extratos Vegetais/química
18.
J Chromatogr A ; 1650: 462232, 2021 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-34051578

RESUMO

Comprehensive two-dimensional gas chromatography with parallel mass spectrometry and flame ionization detection (GC × GC-MS/FID) enables effective chromatographic fingerprinting of complex samples by comprehensively mapping untargeted and targeted components. Moreover, the complementary characteristics of MS and FID open the possibility of performing multi-target quantitative profiling with great accuracy. If this synergy is applied to the complex volatile fraction of food, sample preparation is crucial and requires appropriate methodologies capable of providing true quantitative results. In this study, untargeted/targeted (UT) fingerprinting of extra-virgin olive oil volatile fractions is combined with accurate quantitative profiling by multiple headspace solid phase microextraction (MHS-SPME). External calibration on fifteen pre-selected analytes and FID predicted relative response factors (RRFs) enable the accurate quantification of forty-two analytes in total, including key-aroma compounds, potent odorants, and olive oil geographical markers. Results confirm good performances of comprehensive UT fingerprinting in developing classification models for geographical origin discrimination, while quantification by MHS-SPME provides accurate results and guarantees data referability and results transferability over years. Moreover, by this approach the extent of internal standardization procedure inaccuracy, largely adopted in food volatiles profiling, is measured. Internal standardization yielded an average relative error of 208 % for the fifteen calibrated compounds, with an overestimation of + 538% for (E)-2-hexenal, the most abundant yet informative volatile of olive oil, and a -89% and -80% for (E)-2-octenal and (E)-2-nonenal respectively, analytes with a lower HS distribution constant. Compared to existing methods based on 1D-GC, the current procedure offers better separation power and chromatographic resolution that greatly improve method specificity and selectivity and results in lower LODs and LOQs, high calibration performances (i.e., R2 and residual distribution), and wider linear range of responses. As an artificial intelligence smelling machine, the MHS-SPME-GC × GC-MS/FID method is here adopted to delineate extra-virgin olive oil aroma blueprints; an objective tool with great flexibility and reliability that can improve the quality and information power of each analytical run.


Assuntos
Técnicas de Química Analítica , Análise de Alimentos , Cromatografia Gasosa-Espectrometria de Massas , Azeite de Oliva , Microextração em Fase Sólida , Aldeídos/análise , Inteligência Artificial , Técnicas de Química Analítica/instrumentação , Técnicas de Química Analítica/métodos , Ionização de Chama , Análise de Alimentos/instrumentação , Análise de Alimentos/métodos , Odorantes/análise , Azeite de Oliva/química , Padrões de Referência , Reprodutibilidade dos Testes , Compostos Orgânicos Voláteis/análise
19.
ACS Appl Mater Interfaces ; 13(16): 19342-19350, 2021 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-33848121

RESUMO

Discriminative and sensitive detection of environmentally important and health-related trichloroacetic acid (TCA) suffers from various problems such as bulky instruments and time-consuming operation as well as complex sample processing. Herein, we present a rapid, sensitive, and specific method for the detection of gaseous TCA using a fluorescent single-molecule array. An o-carborane-based benzothiazole derivative (CB-BT-OCH3) with specific fluorescence properties was specifically designed and utilized to fabricate a film-based single-molecule array. It was revealed that the fluorescent film is photochemically stable and extremely sensitive to TCA vapor, depicting an observable fluorescence color change from green to blue. The experimental detection limit is 0.2 ppm, which is lower than the safety limit (1 ppm) required by the threshold limit values and biological exposure indices. In addition, the film could show detectable intensity change within 0.2 s. On the basis of multiple signal responses, a conceptual two-channel-based fluorescent TCA sensor was developed. Importantly, the proposed conceptual sensor paves a new route to the development of specific fluorescent film-based sensor arrays with a single fluorophore as sensing units.


Assuntos
Compostos de Boro/química , Técnicas de Química Analítica/instrumentação , Corantes Fluorescentes/química , Ácido Tricloroacético/análise , Ácido Tricloroacético/química , Ligação de Hidrogênio , Limite de Detecção , Fatores de Tempo
20.
Food Chem ; 351: 129248, 2021 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-33640766

RESUMO

Iron-based metal-organic framework, NH2-MIL-101(Fe), was doped with different dosages of cobalt phthalocyanine nanoparticles (CoPc) to synthesize a series of NH2-MIL-101(Fe)@CoPc nanocomposites. The NH2-MIL-101(Fe)@CoPc nanocomposites were then employed to construct novel impedimetric aptasensors for the detection of ochratoxin A (OTA). Combining the intrinsic advantages of NH2-MIL-101(Fe) (highly porous structure and excellently electrochemical activity) and CoPc (good physiochemical stability and strong bioaffinity), the NH2-MIL-101(Fe)@CoPc nanocomposites show promising properties, which are beneficial for immobilizing OTA-targeted aptamer strands. Amongst, the developed impedimetric aptasensor based on NH2-MIL-101(Fe)@CoPc6:1, prepared using the mass ratio of NH2-MIL-101(Fe):CoPc of 6:1, exhibits the best amplified electrochemical signal and highest sensitivity for detecting OTA. The detection limitation is 0.063 fg·mL-1 within the OTA concentration of 0.0001-100 pg·mL-1, accompanying with high selectivity, good reproducibility and stability, acceptable regenerability, and wide applicability in diverse real samples. Consequently, the proposed sensing strategy can be applied for detecting OTA to cope with food safety.


Assuntos
Técnicas de Química Analítica/instrumentação , Indóis/química , Estruturas Metalorgânicas/química , Nanopartículas/química , Ocratoxinas/análise , Compostos Organometálicos/química , Limite de Detecção , Reprodutibilidade dos Testes
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