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1.
Biomed Chromatogr ; 34(1): e4714, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31633806

RESUMO

Eucommia ulmoides Oliv. (E. ulmoides) is a valuable and nourishing medicinal herb in China that has been used in the treatment of hypertension. Given the fact that most traditional Chinese medicine is mainly used to treat disease, investigating the pharmacokinetics of traditional Chinese medicines in the pathological state is more useful than that in the normal state. However, the differences in the absorption kinetics of active ingredients of E. ulmoides extract between pathological and physiological conditions have not been reported. Therefore, in this study, the rat intestinal in situ circulatory perfusion model was used to investigate the differences in absorption kinetics of seven active ingredients of E. ulmoides extract in normal and spontaneously hypertensive rats, namely, genipinic acid, protocatechuic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, (+)-pinoresinol di-O-ß-D-glucopyranoside and (+)-pinoresinol 4'-O-ß-D-glucopyranoside. Our results indicate that the pathological state of spontaneous hypertension may change the absorption of active components of E. ulmoides extracts, and these findings may provide a reference for improving the rational use of E. ulmoides in the clinic.


Assuntos
Eucommiaceae , Absorção Intestinal , Extratos Vegetais , Animais , Anti-Hipertensivos/análise , Anti-Hipertensivos/farmacocinética , Líquidos Corporais/química , Ácido Clorogênico/análogos & derivados , Ácido Clorogênico/análise , Ácido Clorogênico/farmacocinética , Furanos/análise , Furanos/farmacocinética , Hidroxibenzoatos/análise , Hidroxibenzoatos/farmacocinética , Lignanas/análise , Lignanas/farmacocinética , Extratos Vegetais/análise , Extratos Vegetais/farmacocinética , Ratos , Ratos Endogâmicos SHR , Ratos Wistar
2.
Biochem Med (Zagreb) ; 30(1): 010502, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31839720

RESUMO

Extravascular body fluids (EBF) analysis can provide useful information in the differential diagnosis of conditions that caused their accumulation. Their unique nature and particular requirements accompanying EBF analysis need to be recognized in order to minimize possible negative implications on patient safety. This recommendation was prepared by the members of the Working group for extravascular body fluid samples (WG EBFS). It is designed to address the total testing process and clinical significance of tests used in EBF analysis. The recommendation begins with a chapter addressing validation of methods used in EBF analysis, and continues with specific recommendations for serous fluids analysis. It is organized in sections referring to the preanalytical, analytical and postanalytical phase with specific recommendations presented in boxes. Its main goal is to assist in the attainment of national harmonization of serous fluid analysis and ultimately improve patient safety and healthcare outcomes. This recommendation is intended to all laboratory professionals performing EBF analysis and healthcare professionals involved in EBF collection and processing. Cytological and microbiological evaluations of EBF are beyond the scope of this document.


Assuntos
Líquidos Corporais/química , Técnicas de Laboratório Clínico/normas , Líquidos Corporais/metabolismo , Exsudatos e Transudatos/química , Exsudatos e Transudatos/metabolismo , Guias como Assunto , Humanos , Segurança do Paciente , Derrame Pleural/diagnóstico , Garantia da Qualidade dos Cuidados de Saúde , Sociedades Médicas , Manejo de Espécimes/normas
3.
Forensic Sci Int ; 303: 109959, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31546164

RESUMO

The organ distribution of 3-fluorophenmetrazine (3-FPM), pyrazolam, diclazepam as well as its main metabolites delorazepam, lormetazepam and lorazepam, was investigated. A solid phase extraction (SPE) and a QuEChERS (acronym for quick, easy, cheap, effective, rugged and safe) - approach were used for the extraction of the analytes from human tissues, body fluids and stomach contents. The detection was performed on a liquid chromatography-tandem mass spectrometry system (LCMS/MS). The analytes of interest were detected in all body fluids and tissues. Results showed femoral blood concentrations of 10 µg/L for 3-FPM, 28 µg/L for pyrazolam, 1 µg/L for diclazepam, 100 µg/L for delorazepam, 6 µg/L for lormetazepam, and 22 µg/L for lorazepam. Tissues (muscle, kidney and liver) and bile exhibited higher concentrations of the mentioned analytes than in blood. Additional positive findings in femoral blood were for 2-fluoroamphetamine (2-FA, approx. 89 µg/L), 2-flourometamphetamine (2-FMA, hint), methiopropamine (approx. 2.2 µg/L), amphetamine (approx. 21 µg/L) and caffeine (positive). Delorazepam showed the highest ratio of heart (C) and femoral blood (P) concentration (C/P ratio = 2.5), supported by the concentrations detected in psoas muscle (430 µg/kg) and stomach content (approx. 210 µg/L, absolute 84 µg). The C/P ratio indicates that delorazepam displays susceptibility for post-mortem redistribution (PMR), supported by the findings in muscle tissue. 3-FPM, pyrazolam, diclazepam, lorazepam and lormetazepam did apparently not exhibit any PMR. The cause of death, in conjunction with autopsy findings was concluded as a positional asphyxia promoted by poly-drug intoxication by arising from designer benzodiazepines and the presence of synthetic stimulants.


Assuntos
Benzodiazepinas/farmacocinética , Drogas Desenhadas/farmacocinética , Diazepam/análogos & derivados , Fenmetrazina/análogos & derivados , Mudanças Depois da Morte , Adulto , Benzodiazepinas/análise , Bile/química , Líquidos Corporais/química , Química Encefálica , Drogas Desenhadas/análise , Diazepam/análise , Diazepam/farmacocinética , Toxicologia Forense , Conteúdo Gastrointestinal/química , Humanos , Rim/química , Fígado/química , Lorazepam/análogos & derivados , Lorazepam/análise , Lorazepam/farmacocinética , Pulmão/química , Masculino , Nordazepam/análogos & derivados , Nordazepam/análise , Nordazepam/farmacocinética , Líquido Pericárdico/química , Fenmetrazina/análise , Fenmetrazina/farmacocinética , Músculos Psoas/química , Espectrometria de Massas em Tandem
4.
Forensic Sci Int ; 303: 109957, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31546167

RESUMO

We report the peptide content of decomposition fluid produced under field-based conditions and in the absence of a soil matrix. Sixteen domestic pig (Sus scrofa domesticus) cadavers were used to model human decomposition in trials conducted in the summer and winter months in Western Australia. Physical characteristics were recorded and the peptide components of decomposition fluid were analysed using high performance liquid chromatography-time of flight mass spectrometry. A range of peptides was consistently detected in both summer and winter. Thirty seven peptides were common to both trials; 22 originating from haemoglobin subunit beta, 1 from haemoglobin subunit alpha, 4 from beta-enolase, and 2 from creatine kinase. In agreement with our previous findings, 13 peptides occurred consistently, regardless of trial conditions. Degradation patterns for haemoglobin subunits alpha and beta in summer and winter were similar when expressed in ADD and when adjusted for differences in temperature. The consistent identification of several protein-specific peptides generated during decomposition trials conducted under different temperature and rainfall regimes suggests that quantitative peptide analysis may be useful in estimating time since death.


Assuntos
Líquidos Corporais/química , Peptídeos/análise , Mudanças Depois da Morte , Estações do Ano , Animais , Austrália , Cromatografia Líquida , Patologia Legal , Espectrometria de Massas , Modelos Animais , Proteínas/análise , Suínos
5.
Eur J Pharm Biopharm ; 144: 252-265, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31563633

RESUMO

Nanoscale cerium dioxide (nanoceria) has industrial applications, capitalizing on its catalytic, abrasive, and energy storage properties. It auto-catalytically cycles between Ce3+ and Ce4+, giving it pro-and anti-oxidative properties. The latter mediates beneficial effects in models of diseases that have oxidative stress/inflammation components. Engineered nanoparticles become coated after body fluid exposure, creating a corona, which can greatly influence their fate and effects. Very little has been reported about nanoceria surface changes and biological effects after pulmonary or gastrointestinal fluid exposure. The study objective was to address the hypothesis that simulated biological fluid (SBF) exposure changes nanoceria's surface properties and biological activity. This was investigated by measuring the physicochemical properties of nanoceria with a citric acid coating (size; morphology; crystal structure; surface elemental composition, charge, and functional groups; and weight) before and after exposure to simulated lung, gastric, and intestinal fluids. SBF-exposed nanoceria biological effect was assessed as A549 or Caco-2 cell resazurin metabolism and mitochondrial oxygen consumption rate. SBF exposure resulted in loss or overcoating of nanoceria's surface citrate, greater nanoceria agglomeration, deposition of some SBF components on nanoceria's surface, and small changes in its zeta potential. The engineered nanoceria and SBF-exposed nanoceria produced no statistically significant changes in cell viability or cellular oxygen consumption rates.


Assuntos
Líquidos Corporais/química , Líquidos Corporais/metabolismo , Cério/química , Cério/metabolismo , Nanopartículas/metabolismo , Propriedades de Superfície/efeitos dos fármacos , Células A549 , Células CACO-2 , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Mitocôndrias/efeitos dos fármacos , Nanopartículas/química , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Consumo de Oxigênio/efeitos dos fármacos
6.
Mater Sci Eng C Mater Biol Appl ; 104: 109884, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31500005

RESUMO

In this study, we have successfully doped hydroxyapatite (HA) with zinc (Zn2+), sulphate (SO42-) and fluoride (F-) ions to develop a new composition of bioceramic, Ca10-x Znx(PO4)6-y(SO4)y(OH)2-z-yFz(SO4)y, (x = 0, 0.2, 0.6, 1.0, y = 0, 0.5 and z = 0,1.0 mol), using wet precipitation method. The obtained materials were analysed using XRD, FTIR, FESEM, and XPS techniques to investigate the phase purity, particle morphology and elemental composition, respectively. A model anticancer drug (Doxorubicin, DOX) was loaded onto the surface of the Zn/SO4-FHA materials. About 100% loading of DOX with a controlled release profile was obtained. Degradation of materials in Simulated body fluid (SBF) was greatly improved with the incorporation of Zn2+/SO42- ions in comparison to HA/FHA, which makes it highly bioactive materials. In vitro cell viability and adhesion of Human fetal osteoblast (hFOB) cell were investigated. Cell viability has demonstrated that the hFOB cells proliferated at a high rate on Zn/SO4-FHA materials, confirming the in vitro biocompatibility of the materials. Alkaline phosphatase (ALP) activity and intracellular calcium deposition of hFOB cells seeded on 1ZnSO4-FHA disc surface was statistically higher than observed on pure HA and FHA discs, indicating that hFOB cells differentiated into mature osteoblasts on 1Zn/SO4-FHA disc surfaces. Taken together, our results suggest that HA substituted by (Zn2+, 0.2 mol), (SO42-, 0.5 mol) and (F-, 1 mol) (1Zn/SO4-FHA) material was a promising material for hard tissue scaffolds.


Assuntos
Materiais Biocompatíveis/química , Regeneração Óssea/efeitos dos fármacos , Hidroxiapatitas/química , Hidroxiapatitas/farmacologia , Nanopartículas/química , Sulfatos/química , Zinco/química , Materiais Biocompatíveis/farmacologia , Líquidos Corporais/química , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Durapatita/química , Fluoretos/química , Humanos , Teste de Materiais/métodos , Osteoblastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos
7.
Environ Pollut ; 255(Pt 1): 113171, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31539851

RESUMO

Environmental contaminants, to which humans are widely exposed, cause or worsen several diseases, like cardiovascular diseases and cancers. Among these molecules, polycyclic aromatic hydrocarbons (PAHs) stand out since they are ubiquitous pollutants found in ambient air and diet. Because of their toxic effects, public Health agencies promote development of research studies aiming at increasing the knowledge about PAHs and the discovery of biomarkers of exposure and/or effects. Extracellular vesicles (EVs), including small extracellular vesicles (S-EVs or exosomes) and large extracellular vesicles (L-EVs or microvesicles), are delivery systems for multimolecular messages related to the nature and status of the originating cells. Because they are produced by all cells and detected within body fluids, EV releases could act as cell responses and thereby serve as biomarkers. To test whether EVs can serve as biomarkers of PAHs exposure, we evaluate the effects of these pollutants on EV production using an in vitro approach (human endothelial cell line, HMEC-1) and an in vivo approach (urine samples from PAHs-exposed rats). Our study indicates that, i) PAH exposure increases in vitro the EV production by endothelial cells and in vivo the release of EVs in urine, and that the stimulating effects of PAHs concern both S-EVs and L-EVs; ii) PAH exposure and more particularly exposure to B[a]P, can influence the composition of exosomes produced by endothelial cells; iii) the aryl hydrocarbon receptor, a cytosolic receptor associated to most deleterious effects of PAHs, would be involved in the PAH effects on the release of S-EVs, but not L-EVs. These results suggest that EVs may have utility for monitoring exposure to PAHs, and more particularly to B[a]P, considered as reference PAH, and to detect the related early cellular response prior to end-organ damages.


Assuntos
Células Endoteliais/metabolismo , Poluentes Ambientais/toxicidade , Vesículas Extracelulares/metabolismo , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Urina/química , Animais , Biomarcadores/metabolismo , Líquidos Corporais/química , Linhagem Celular , Exposição Ambiental/efeitos adversos , Monitoramento Ambiental/métodos , Exossomos , Feminino , Humanos , Ratos , Receptores de Hidrocarboneto Arílico/metabolismo
8.
Mater Sci Eng C Mater Biol Appl ; 104: 109974, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31499935

RESUMO

The current work aims at exploring the effects of the microstructure and alloy composition on enhancing the bone osseointegration in Ti-6Al-4V (Ti64) and Ti-6Al-7Nb (Ti67) alloys. This was revealed by investigating the alloy susceptibility to grow hydroxyapatite (HA) on their surfaces after immersion in simulated body fluid (SBF). The specimens were produced by two methods: forging and casting in order to study the influence of the microstructure on the precipitation process. The surface conditions investigated were the polished, alkaline and the hydrothermally treated. It was found that precipitation on both of Ti64 and Ti67 occurs after about 4 weeks and considerably dissolve back with further immersion. Precipitation process is enhanced at some pH levels lower than the neutral level. Forged Ti67 has less reactivity with Hank solution than Ti64 specimens; the reverse is true for cast specimens. In case of the alkaline treated specimens, precipitations on cast specimens were denser than on the forged ones. For the hydrothermally treated specimens, high amounts of Ca and P were observed on cast Ti67 indicating that hydrothermal treatment is considered the best surface modification treatment for alloy Ti67.


Assuntos
Durapatita/química , Titânio/química , Líquidos Corporais/química , Concentração de Íons de Hidrogênio , Osseointegração/efeitos dos fármacos , Propriedades de Superfície
9.
Molecules ; 24(15)2019 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-31382421

RESUMO

A rapid and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination of ochratoxin A (OTA) and its metabolite ochratoxin α (OTα), for the first time, in dairy cow plasma, milk, urine, heart, liver, spleen, lung, and kidney. The established method was extensively validated by determining the linearity (R2 ≥ 0.990), sensitivity (lower limit of quantification, 0.1-0.2 ng mL-1), recovery (75.3-114.1%), precision (RSD ≤ 13.6%), and stability (≥83.0%). Based on the methodological advances, the carry-over of OTA was subsequently studied after oral administration of 30 µg/kg body weight OTA to dairy cows. As revealed, OTA and OTα were detected in urine, with maximal concentrations of 1.8 ng mL-1 and 324.6 ng mL-1, respectively, but not in milk, plasma, or different tissues, verifying the protection effects of rumen flora against OTA exposure for dairy cows. Moreover, 100 fresh milk samples randomly collected from different supermarkets in Shanghai were also analyzed, and no positive samples were found, further proving the correctness of the in vivo biotransformation results. Thus, from the currently available data, regarding OTA contamination issues on dairy cows, no significant health risks were related to OTA exposure due to the consumption of these products.


Assuntos
Cromatografia Líquida , Contaminação de Alimentos/análise , Ocratoxinas/análise , Ocratoxinas/química , Espectrometria de Massas em Tandem , Animais , Líquidos Corporais/química , Bovinos , Cromatografia Líquida/métodos , Leite/química , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem/métodos
10.
Mater Sci Eng C Mater Biol Appl ; 103: 109843, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31349461

RESUMO

A series of germanium (Ge)-containing glasses were synthesized based on a starting glass composition of SiO2-ZnO-CaO-SrO-P2O5. Additions of GeO2 (6 and 12 mol%) were incorporated at the expense of SiO2, which retained the amorphous character, and each glass was processed to present similar particle size and surface area. Glass characterization using x-ray photoelectron spectroscopy (XPS) and magic angle spinning nuclear magnetic resonance (MAS-NMR) determined that the addition of GeO2 increased the fraction of lower Q-speciation and subsequently the concentration of non-bridging oxygens (NBO). Glass Polyalkenoate Cements (GPC) were formulated from each glass with 40, 50 and 60 wt% PAA, and presented time dependent solubility profiles (1, 10, 100, 1000 h) for the release of Si4+ (4-140 mg/l), Ca2+ (1-8 mg/l), Zn2+ (<6 mg/l), Sr2+ (2-37 mg/l), PO43- (2-43 mg/l) and Ge4+ (20-911 mg/l) and attained pH values close to 7.5 after 1000 h. Ge-GPCs containing 40 wt% polyacrylic acid (PAA) presented appropriate working time (Tw) and setting times (Ts), and the corresponding compressive strengths ranged from (14-30 MPa). The Ge-GPCs (40, 50 wt%) presented a linear increase (R2-0.99) with respect to time. Simulated Body Fluid (SBF) testing resulted in the Ge-GPCs encouraging the precipitation of crystalline hydroxyapatite on the GPC surface, more evidently after 100 and 1000 h incubation.


Assuntos
Líquidos Corporais/química , Germânio/química , Cimentos de Ionômeros de Vidro/química , Vidro/química , Humanos , Solubilidade
11.
Environ Pollut ; 252(Pt A): 336-343, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31158662

RESUMO

The health risks and toxicity of heavy metals (HMs) in PM2.5 are not only associated with their total amounts, but also with their species and bioaccessibility. In this study, the speciation (fractions) and bioaccessibility of HMs (Pb, Cd, Cr, Cu and Zn) as well as their correlations in fine particulate matter (PM2.5) samples from four seasons were studied. A sequential extraction procedure was applied to divide the studied HMs into four fractions: acid-soluble fraction (F1), reducible fraction (F2), oxidative fraction (F3) and residual fraction (F4). The simulated body fluids (gastrointestinal and lung phases) were used for in vitro tests in order to evaluate the bioaccessibility of HMs. The distribution of HMs in PM2.5 was season and element dependent. It was found that Zn was the most abundant element among the five measured metals and followed by Pb, Cu, Cr and Cd. The total contents of each HM in different seasons were in the following order: winter > autumn > spring > summer. The studied HMs were mainly concentrated in acid-soluble fraction (F1) with high bioaccessibility (p < 0.05) except for Cr. Zn, Pb and Cu possessed the highest bioaccessibility in summer while Cd and Cr were the highest in winter. In vitro tests indicated that HMs in PM2.5 were much more accessible to gastrointestinal fluids rather than lung phase (Gamble's solution). A significant correlation was found between the results from the optimized BCR sequential extraction and solubility bioaccessibility research consortium (SBRC). The fractions extracted by SBRC were consistent with the first two fractions extracted by the sequential extraction method.


Assuntos
Monitoramento Ambiental/métodos , Trato Gastrointestinal/química , Pulmão/química , Metais Pesados/análise , Material Particulado/análise , Líquidos Corporais/química , China , Cidades , Estações do Ano
12.
Talanta ; 202: 136-144, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31171161

RESUMO

A reliable screening and non invasive method based on the use of microextraction by packed sorbent coupled with desorption electrospray ionization-high resolution mass spectrometry was developed and validated for the detection of new psychoactive substances in oral fluid. The role of different sample substrates in enhancing signal intensity and stability was evaluated by testing the performances of two polylactide-based materials, i.e. non-functionalized and functionalized with carbon nanoparticles, and a silica-based material compared to commercially available polytetrafluorethylene supports. The best results were achieved by using the non-functionalized polylactide substrates to efficiently ionize compounds in positive ionization mode, whereas the silica coating proved to be the best choice for operating in negative ionization mode. LLOQs in the low µg/L, a good precision with CV% always lower than 16% and RR% in the 83(±4)-120(±2)% range, proved the suitability of the developed method for the determination of the analytes in oral fluid. Finally, the method was applied for screening oral fluid samples for the presence of psychoactive substances during private parties, revealing mephedrone in only one sample out of 40 submitted to analysis.


Assuntos
Líquidos Corporais/química , Psicotrópicos/análise , Microscopia de Força Atômica , Espectrometria de Massas por Ionização por Electrospray
13.
Talanta ; 202: 546-553, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31171220

RESUMO

In the field of forensic toxicology, the use of non-destructive and easy-to-use analytical techniques deserves remarkable attention, especially in those situations involving public health and security. In addition, the miniaturization and portability of one-touch devices for the detection of specific threats is required more and more. In this study, a novel on-site MicroNIR/Chemometric platform was developed to perform a real-time prediction of cocaine and its metabolites in non pre-treated oral fluid. Simulated oral fluids were prepared in water in order to calibrate the instrumental response and the matrix effect was consequently evaluated by processing spiked oral fluids collected from volunteers. The procedure was optimized using a proper experimental design taking into account the equilibrium between cocaine and benzoylecgonine in the range 10-100 ng-ml and validated by comparing results with the reference official method (GC-MS). The developed method was statistically able to discriminate oral fluid samples containing cocaine from 10 to 100 ng/ml and demonstrated to be not affected by the variability of the matrix as all the blank samples of different volunteers (smokers and non smokers, assuming caffeine, sugars, chewing-gum or alcohol) as well as spiked oral fluids were correctly predicted by the model. In addition, results from six real samples confirmed the feasibility of the miniaturized platform to provide a correct identification of cocaine abuse and to propose the MicroNIR as innovative personal screening system to prevent accidents and in cases involving workplace surveillance.


Assuntos
Líquidos Corporais/química , Cocaína/análise , Cocaína/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Espectroscopia de Luz Próxima ao Infravermelho
14.
Molecules ; 24(12)2019 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-31234484

RESUMO

Improvement in high-throughput MALDI-TOF MS analysis requires practical and efficient sample preparation protocols for high acquisition rates. The use of hexagonal mesoporous silica (HMS) sorbents in combination with MALDI-TOF MS was explored as a versatile tool for peptidomic profiling of clinical specimens difficult to process, but considered important sources of disease biomarkers: synovial fluid and sputum. A rapid and efficient procedure, based on dispersive solid-phase extraction of peptides using commercially available wormhole mesostructured HMS, was tested for: a) pre-concentration of standard peptides in serially diluted solution up to the sub-nanomolar range; b) peptidome profiling of sputum and synovial fluid. The use of HMS, as dispersed sponges, significantly amplified the peptidic repertoire of sputum and synovial fluid by excluding from the adsorptive process large size proteins, which mask and/or suppress peptidome signals. The protocol proposed, as dispersive solid phase extraction, ensures good analytical performances. Moreover, it is economical and rapid, as it avoids the use of less reproducible and prolonged sample preparation procedures, such as the use of ultrafiltration filter devices. These findings may contribute to defining a high-throughput screening MS-based platform for monitoring key peptidic features of difficult to analyse bodily fluids in a clinical setting.


Assuntos
Peptídeos/análise , Proteômica , Dióxido de Silício/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Biomarcadores , Líquidos Corporais/química , Líquidos Corporais/metabolismo , Humanos , Projetos Piloto , Proteômica/métodos , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Escarro
15.
Anal Chim Acta ; 1075: 1-26, 2019 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-31196414

RESUMO

In recent years, advances in sensitive analytical techniques have encouraged the analysis of various compounds in biological fluids. While blood serum, blood plasma and urine still remain the golden standards in clinical, toxicological and forensic science, analyses of other body fluids, such as breast milk, exhaled breath condensate, sweat, saliva, amniotic fluid, cerebrospinal fluid, or capillary blood in form of dried blood spots are becoming more popular. This review article focuses on capillary electrophoresis and microchip electrophoresis of small ions and molecules (e.g. inorganic cations/anions, basic/acidic drugs, small acids/bases, amino acids, peptides and other low molecular weight analytes) in various less conventional human body fluids and hopes to stimulate further interest in the field.


Assuntos
Secreções Corporais/química , Líquidos Corporais/química , Íons/análise , Compostos Orgânicos/análise , Líquido Amniótico/química , Eletroforese Capilar/métodos , Eletroforese em Microchip/métodos , Humanos , Íons/líquido cefalorraquidiano , Compostos Orgânicos/líquido cefalorraquidiano
16.
Bioanalysis ; 11(10): 913-922, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31218902

RESUMO

Aim: To develop and validate a simple method using LC-MS/MS for determination of dextromethorphan (DXM) and dextrorphan (DT) in human oral fluid. Results: Following protein precipitation, chromatographic separation used a phenyl column with isocratic elution (1 ml/min) of 10 mM ammonium-formate buffer and acetonitrile (65:35; v/v) with 0.1% formic acid. Retention times were 2.6 min for DT and 5 min for DXM. Total run time was 7 min. The intra- and inter-assay deviations (accuracy) for DT (1-100 ng/ml) and DXM (5-1000 ng/ml) ranged from -13.6 to 8.8% and -9.6 to 5.7%, respectively. Precision variations were ≤7.5%. Matrix effect was ≤11.8%. Conclusion: This method may prove helpful for quantification of DT and DXM in oral fluid for either clinical or toxicological purposes.


Assuntos
Líquidos Corporais/química , Cromatografia Líquida/métodos , Testes de Química Clínica/métodos , Dextrometorfano/análise , Dextrorfano/análise , Espectrometria de Massas em Tandem/métodos , Métodos Analíticos de Preparação de Amostras , Calibragem , Humanos , Limite de Detecção , Modelos Lineares , Reprodutibilidade dos Testes
17.
Anal Chim Acta ; 1067: 107-114, 2019 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-31047141

RESUMO

A novel and versatile immunosensing strategy was developed for ultrasensitive and specific detection of proteins by organically integrating interfacial specific target recognition and homogeneous transcription amplification. In principle, classic antigen-antibody sandwich structure on the microplate could realize the specific identification of target protein. Biotinylated DNA probe was subsequently introduced by streptavidin-biotin system as a bridge linking interfacial and homogeneous reaction. The biotinylated DNA initiated exponential transcription amplification in the solution, which converted per target recognition event on the interface to numerous single-stranded RNA products in solution for highly sensitive fluorescence immunosensing. The proposed immunoassay based on interfacial recognition-induced homogeneous exponential transcription (IR-HET) for vascular endothelial growth factor (VEGF) detection showed a good linear range from 0.01 to 1000 pg/mL and the limit of detection as low as 1 fg/mL, which was 3 orders lower than traditional ELISA method. The established strategy was also successfully applied to directly detect VEGF from culture supernatants of tumor cells and clinical body fluid samples, proving very high sensitivity, selectivity and low matrix effect. Therefore, IR-HET-based immunosensing strategy might become a potential powerful tool be applied in ultrasensitive detection of low abundance protein biomarker for clinical early diagnosis, treatment and prognosis.


Assuntos
Biomarcadores Tumorais/análise , Técnicas Biossensoriais , Líquidos Corporais/química , Imunoensaio , Transcrição Genética/genética , Fatores de Crescimento do Endotélio Vascular/análise , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral , Humanos , Técnicas de Amplificação de Ácido Nucleico , Fatores de Crescimento do Endotélio Vascular/genética
18.
Anal Chim Acta ; 1067: 11-30, 2019 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-31047142

RESUMO

The employment of spectroscopically-resolved NMR techniques as analytical probes have previously been both prohibitively expensive and logistically challenging in view of the large sizes of high-field facilities. However, with recent advances in the miniaturisation of magnetic resonance technology, low-field, cryogen-free "benchtop" NMR instruments are seeing wider use. Indeed, these miniaturised spectrometers are utilised in areas ranging from food and agricultural analyses, through to human biofluid assays and disease monitoring. Therefore, it is both intrinsically timely and important to highlight current applications of this analytical strategy, and also provide an outlook for the future, where this approach may be applied to a wider range of analytical problems, both qualitatively and quantitatively.


Assuntos
Líquidos Corporais/química , Espectroscopia de Ressonância Magnética/instrumentação , Espectroscopia de Ressonância Magnética/métodos , Animais , Doença , Medicina Legal , Humanos , Sistemas Automatizados de Assistência Junto ao Leito
19.
J Pharm Biomed Anal ; 172: 183-188, 2019 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-31055183

RESUMO

A novel method using UPLC with tandem mass-spectrometric detection (UPLC-MS/MS) with positive electrospray ionization was developed for the detection of the antiarrhythmic drug, dofetilide, in mouse plasma and urine. Protein precipitation was performed on 10 µL of plasma and 2 µL of urine samples using dofetilide-D4 as an internal standard, and separation of the analyte was accomplished on a C18 analytical column with the flow of 0.40 mL/min. Subsequently, the method was successfully applied to determine the pharmacokinetic parameters of dofetilide following oral and intravenous administration. The calibration curve was linear over the selected concentration range (R2 ≥ 0.99), with a lower limit of quantitation of 5 ng/mL. The intra-day and inter-day precisions, and accuracies obtained from a 5-day validation ranged from 3.00 to 7.10%, 3.80-7.20%, and 93.0-106% for plasma, and 3.50-9.00%, 3.70-10.0%, 87.0-106% for urine, while the recovery of dofetilide was 93.7% and 97.4% in plasma and urine, respectively. The observed pharmacokinetic profiles revealed that absorption is the rate-limiting step in dofetilide distribution and elimination. Pharmacokinetic studies illustrate that the absolute bioavailability of dofetilide in the FVB strain mice is 34.5%. The current developed method allows for accurate and precise quantification of dofetilide in micro-volumes of plasma and urine, and was found to be suitable for supporting in vivo pharmacokinetic studies.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Fenetilaminas/sangue , Fenetilaminas/urina , Plasma/química , Sulfonamidas/sangue , Sulfonamidas/urina , Espectrometria de Massas em Tandem/métodos , Animais , Disponibilidade Biológica , Líquidos Corporais/química , Calibragem , Limite de Detecção , Masculino , Camundongos , Fenetilaminas/farmacocinética , Sulfonamidas/farmacocinética
20.
Molecules ; 24(9)2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-31083395

RESUMO

The United States is currently experiencing an opioid crisis, with more than 47,000 deaths in 2017 due to opioid overdoses. Current approaches for opioid identification and quantification in body fluids include immunoassays and chromatographic methods (e.g., LC-MS, GC-MS), which require expensive instrumentation and extensive sample preparation. Our aim was to develop a portable point-of-care device that can be used for the instant detection of opioids in body fluids. Here, we reported the development of a morphine-sensitive fluorescence-based sensor chip to sensitively detect morphine in the blood using a homogeneous immunoassay without any washing steps. Morphine-sensitive illuminating peptides were identified using a high throughput one-bead one-compound (OBOC) combinatorial peptide library approach. The OBOC libraries contain a large number of random peptides with a molecular rotor dye, malachite green (MG), that are coupled to the amino group on the side chain of lysine at different positions of the peptides. The OBOC libraries were then screened for fluorescent activation under a confocal microscope, using an anti-morphine monoclonal antibody as the screening probe, in the presence and absence of free morphine. Using this novel three-step fluorescent screening assay, we were able to identify the peptide-beads that fluoresce in the presence of an anti-morphine antibody, but lost fluorescence when the free morphine was present. After the positive beads were decoded using automatic Edman microsequencing, the morphine-sensitive illuminating peptides were then synthesized in soluble form, functionalized with an azido group, and immobilized onto microfabricated PEG-array spots on a glass slide. The sensor chip was then evaluated for the detection of morphine in plasma. We demonstrated that this proof-of-concept platform can be used to develop fluorescence-based sensors against morphine. More importantly, this technology can also be applied to the discovery of other novel illuminating peptidic sensors for the detection of illicit drugs and cancer biomarkers in body fluids.


Assuntos
Analgésicos Opioides/análise , Analgésicos Opioides/sangue , Líquidos Corporais/química , Técnicas de Química Combinatória/métodos , Morfina/análise , Morfina/sangue , Peptídeos/química , Cromatografia Líquida , Ensaios de Triagem em Larga Escala , Humanos , Biblioteca de Peptídeos
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