Fiber-in-tube SPME-CapLC-MS/MS method to determine Aß peptides in cerebrospinal fluid obtained from Alzheimer's patients.

Mass spectrometry is characterized by its high sensitivity, ability to measure very low analyte concentrations, specificity to distinguish between closely related compounds, availability to generate high-throughput methods for screening, and high multiplexing capacity. This technique has been used as a platform to analyze fluid biomarkers for Alzheimer's disease. However, more effective sample preparation procedures, preferably antibody-independent, and more automated mass spectrometry platforms with improved sensitivity, chromatographic separation, and high throughput are needed for this purpose. This short communication discusses the development of a fiber-in-tube SPME-CapLC-MS/MS method to determine Aß peptides in cerebrospinal fluid obtained from Alzheimer's disease patients. To obtain the fiber-in-tube SPME capillary, we longitudinally packed 22 nitinol fibers coated with a zwitterionic polymeric ionic liquid into the same length of the PEEK tube. In addition, this communication compares this fiber-in-tube SPME method with the conventional HPLC scale (HPLC-MS/MS) and when directly coupled to CapESI-MS/MS without chromatographic separation, and, as a case study, discusses the benefits and challenges inherent in miniaturizing the flow scale of the sample preparation technique (fiber-in-tube SPME) to the CapLC-MS/MS system. Fiber-in-tube SPME-CapLC-MS/MS provided LLOQ ranging from 0.09 to 0.10 ng mL-1, accuracy ranging from 91 to 117 % (recovery), and reproducibility of less than 18 % (RSD). Analysis of the cerebrospinal fluid samples obtained from Alzheimer's disease patients evidenced that the method is robust. At the capillary scale (10 µL min-1), this innovative method presented higher analytical sensitivity than the conventional HPLC-MS/MS scale. Although fiber-in-tube SPME directly coupled to CapESI-MS/MS offers advantages in terms of high throughput, the sample was dispersed and non-quantitatively desorbed from the capillary at low flow rate. These results highlighted that chromatographic separation is important to decrease the matrix effect and to achieve higher detectability, which is indispensable for bioanalysis.

The role of vitamin B12 in viral infections: a comprehensive review of its relationship with the muscle-gut-brain axis and implications for SARS-CoV-2 infection.

This comprehensive review establishes the role of vitamin B12 as adjunct therapy for viral infections in the treatment and persistent symptoms of COVID-19, focusing on symptoms related to the muscle-gut-brain axis. Vitamin B12 can help balance immune responses to better fight viral infections. Furthermore, data from randomized clinical trials and meta-analysis indicate that vitamin B12 in the forms of methylcobalamin and cyanocobalamin may increase serum vitamin B12 levels, and resulted in decreased serum methylmalonic acid and homocysteine concentrations, and decreased pain intensity, memory loss, and impaired concentration. Among studies, there is much variation in vitamin B12 doses, chemical forms, supplementation time, and administration routes. Larger randomized clinical trials of vitamin B12 supplementation and analysis of markers such as total vitamin B12, holotranscobalamin, total homocysteine and methylmalonic acid, total folic acid, and, if possible, polymorphisms and methylation of genes need to be conducted with people with and without COVID-19 or who have had COVID-19 to facilitate the proper vitamin B12 form to be administered in individual treatment.

A Dual Ligand Sol⁻Gel Organic-Silica Hybrid Monolithic Capillary for In-Tube SPME-MS/MS to Determine Amino Acids in Plasma Samples.

This work describes the direct coupling of the in-tube solid-phase microextraction (in-tube SPME) technique to a tandem mass spectrometry system (MS/MS) to determine amino acids (AA) and neurotransmitters (NT) (alanine, serine, isoleucine, leucine, aspartic acid, glutamic acid, lysine, methionine, tyrosine, and tryptophan) in plasma samples from schizophrenic patients. An innovative organic-silica hybrid monolithic capillary with bifunctional groups (amino and cyano) was developed and evaluated as an extraction device for in-tube SPME. The morphological and structural aspects of the monolithic phase were evaluated by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), nitrogen sorption experiments, X-ray diffraction (XRD) analyses, and adsorption experiments. In-tube SPME-MS/MS conditions were established to remove matrix, enrich analytes (monolithic capillary) and improve the sensitivity of the MS/MS system. The proposed method was linear from 45 to 360 ng mL-1 for alanine, from 15 to 300 ng mL-1 for leucine and isoleucine, from 12 to 102 ng mL-1 for methionine, from 10 to 102 ng mL-1 for tyrosine, from 9 to 96 ng mL-1 for tryptophan, from 12 to 210 ng mL-1 for serine, from 12 to 90 ng mL-1 for glutamic acid, from 12 to 102 ng mL-1 for lysine, and from 6 to 36 ng mL-1 for aspartic acid. The precision of intra-assays and inter-assays presented CV values ranged from 1.6% to 14.0%. The accuracy of intra-assays and inter-assays presented RSE values from -11.0% to 13.8%, with the exception of the lower limit of quantification (LLOQ) values. The in-tube SPME-MS/MS method was successfully applied to determine the target AA and NT in plasma samples from schizophrenic patients.

Association between polymorphisms in genes encoding estrogen receptors (ESR1 and ESR2) and excreted bisphenol A levels after orthodontic bracket bonding: a preliminary study.

BACKGROUND: Bisphenol A (BPA) is released from orthodontic composites used for bracket bonding. Genetic variations could modify the metabolism of this chemical within the organism. Considering that free BPA binds to estrogen receptors causing harmful effects to health, the present in vivo study aimed to evaluate the association between genetic polymorphisms in genes encoding estrogen receptors (ESR1 and ESR2) and excreted BPA levels in orthodontic patients. METHODS: Quantification of BPA levels in the urine of 16 patients was performed in a gas chromatograph mass spectrometer before (T0), at 24 h (T1), and 1 week (T2) after bracket bonding. DNA was extracted from saliva, and one genetic polymorphism in ESR1 (rs2234693) and two in ESR2 (rs4986938 and rs1256049) were analyzed by real-time PCR. Increases in BPA levels in the urine at T1 and T2 were grouped according to the genotype, and mean differences were compared by unpaired T test or Mann-Whitney test according to the normality of the data. The established alpha was 5%. RESULTS: BPA levels increased significantly at T1 and T2. There were no statistically significant differences in the increases in BPA levels according to the genotype for any genetic polymorphism (P > 0.05), at neither 24 h nor 1 week after bracket bonding. CONCLUSIONS: The results suggested that there are no association between excreted BPA levels after bracket bonding and the evaluated genetic polymorphisms in ESR1 and ESR2. Further research should be performed in order to confirm these results.

The legacy of organochlorine pesticide usage in a tropical semi-arid region (Jaguaribe River, Ceará, Brazil): Implications of the influence of sediment parameters on occurrence, distribution and fate.

Between the 1940s and 1990s, immeasurable amounts of organochlorine pesticides (OCPs) were used in endemic disease control campaigns and agriculture in the tropical semi-arid regions of Brazil. The present study evaluated the legacy of banned OCP usage, considering the levels, ecological risk and dependence on sediment physicochemical properties for the fate and distribution in the Jaguaribe River. The sum concentration of OCPs (ΣOCPs) ranged from 5.09 to 154.43 ng·g(-1), comparable to the levels found in other tropical and subtropical regions that have traditionally used OCPs. The environmental and geographical distribution pattern of p,p-DDT, p,p-DDD and p,p-DDE shows that the estuarine zone contained more than 3.5 times the levels observed in the fluvial region, indicating that the estuary of the Jaguaribe River is a sink. The temporal pattern indicates application of dichloro-diphenyl-trichloroethanes (DDTs) in the past; however, there is evidence of recent input of these pesticides. High ecological risk was observed for levels of γ-hexachlorocyclohexanes (γ-HCH) and heptachlor, and moderate ecological risk was observed for levels of DDTs in sediments from the Jaguaribe River. The heptachlor, γ-HCH and hexachlorobenzene (HCB) concentrations depend on the organic and inorganic fractions of sediment from the Jaguaribe River, whereas the p,p-DDE, p,p-DDD, p,p-DDT and α-endosulfan concentrations depend solely on the organic fraction of the sediment.

Hybrid silica monolith for microextraction by packed sorbent to determine drugs from plasma samples by liquid chromatography-tandem mass spectrometry.

The present study (1) reports on the synthesis of two hybrid silica monoliths functionalized with aminopropyl or cyanopropyl groups by the sol-gel process; (2) evaluates these monoliths as selective stationary phase for microextraction by packed sorbent (MEPS) to determine drugs in plasma samples via liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the multiple reactions monitoring (MRM) mode; and (3) discusses important factors related to the optimization of MEPS efficiency as well as the carryover effect. The prepared hybrid silica monoliths consisted of a uniform, porous, and continuous silica monolithic network. The structure of the aminopropyl hybrid silica monolith was more compact than the structure of the cyanopropyl hybrid silica monolith. The Fourier-transform infrared spectroscopy (FTIR) spectra of the hybrid silica monoliths displayed readily identifiable peaks, characteristic of the cyanopropyl and aminopropyl groups. Compared with the aminopropyl hybrid silica phase, the cyanopropyl hybrid silica phase exhibited higher binding capacity for most of the target drugs. The developed method afforded adequate linearity at concentrations ranging from the lower limit of quantification (0.05-1.00 ng mL(-1)) to the upper limit of quantification (40-10,500 ng mL(-1)); the coefficients of determination (r(2)) were higher than 0.9955. The precision of the method presented coefficients of variation (CV) lower than 14%; the relative standard error (RSE) of the accuracy ranged from -12% to 14%. The developed method allowed for simultaneous analysis of five antipsychotics (olanzapine, quetiapine, clozapine, haloperidol, and chlorpromazine) in combination with seven antidepressants (mirtazapine, paroxetine, citalopram, sertraline, imipramine, clomipramine, fluoxetine), two anticonvulsants (carbamazepine and lamotrigine), and two anxiolytics (diazepam and clonazepam) in plasma samples from schizophrenic patients, which should be valuable for therapeutic drug monitoring purposes.

Determination of pesticides in lettuce using solid-liquid extraction with low temperature partitioning.

This work describes the optimization and validation of a method employing solid-liquid extraction with low temperature partitioning (SLE/LTP) together with analysis by gas chromatography with electron capture detection (GC/ECD) for the determination of nine pesticides (chlorothalonil, methyl parathion, procymidone, endosulfan, iprodione, λ-cyhalothrin, permethrin, cypermethrin, and deltamethrin) in lettuce. The method was found to be selective, accurate, and precise, with means recovery values in the range of 72.3-103.2%, coefficients of variation ⩽ 12%, and detection limits in the range 0.4-37 µg kg(-1). The matrix components significantly influence the chromatographic response of the analytes (above 10%). The optimized and validated method was applied to determine the residual concentrations of the fungicides iprodione and procymidone that had been applied to field crops of lettuce. The maximum residual concentrations of the pesticides in the lettuce samples were 13.6 ± 0.4 mg kg(-1) (iprodione) and 1.00 ± 0.01 mg kg(-1) (procymidone), on the day after application of the products.

Rimonabant effects on anxiety induced by simulated public speaking in healthy humans: a preliminary report.

OBJECTIVE: We investigated the hypothesis that rimonabant, a cannabinoid antagonist/inverse agonist, would increase anxiety in healthy subjects during a simulation of the public speaking test. METHODS: Participants were randomly allocated to receive oral placebo or 90 mg rimonabant in a double-blind design. Subjective effects were measured by Visual Analogue Mood Scale. Physiological parameters, namely arterial blood pressure and heart rate, also were monitored. RESULTS: Twelve participants received oral placebo and 12 received 90 mg rimonabant. Rimonabant increased self-reported anxiety levels during the anticipatory speech and performance phase compared with placebo. Interestingly, rimonabant did not modulate anxiety prestress and was not associated with sedation, cognitive impairment, discomfort, or blood pressure changes. CONCLUSIONS: Cannabinoid-1 antagonism magnifies the responses to an anxiogenic stimulus without interfering with the prestress phase. These data suggest that the endocannabinoid system may work on-demand to counteract the consequences of anxiogenic stimuli in healthy humans.

Simultaneous plasma lamotrigine analysis with carbamazepine, carbamazepine 10,11 epoxide, primidone, phenytoin, phenobarbital, and PEMA by micellar electrokinetic capillary chromatography (MECC).

The determination of lamotrigine (LTG) simultaneously with carbamazepine (CBZ), carbamazepine 10,11 epoxide (CBZ-E), primidone (PRM), phenytoin (PHT), phenobarbital (PB), and 2-phenyl-2-ethyl-malonamide (PEMA) in human plasma was developed using micellar electrokinetic capillary chromatography (MECC) with a diode-array detector. The reproducibility of both separation and quantitation with MECC analysis were appropriate for the intra- and interassay coefficients. The evaluated drugs concentration intervals of LTG, 0.5-10.0 micro g/mL; CBZ, 1.0-16.0 micro g/mL; PEMA, 1.0-20.0 micro g/mL; PB, 1.0-60.0 micro g/mL; PRM, 1.0-20.0 micro g/mL; PHT, 0.7-40.0 micro g/mL; and CBZ-E, 1.0-14.0 micro g/mL were linear with correlation coefficients higher than 0.987 and coefficients of the variation of the points of the calibration curve lower than 10%. The limit of quantitation of the investigated drugs in plasma varied from 0.5 to 1.0 micro g/mL, depending upon the drug. The MECC technique was sensitive enough to work with microsamples into the subtherapeutic, therapeutic, and toxic concentrations, as well as showed to be simple and efficient when applied to monitoring therapeutic drugs in patients treated with a combination of lamotrigine and other antiepileptic drugs such as hepatic enzyme-inducing agents.

Simultaneous Determination of six Antiepileptic Drugs in Plasma Samples by High-Performance Liquid Chromatography / Simultaneous Determination of six Antiepileptic Drugs in Plasma Samples by High-Performance Liquid Chromatography

Determinação Simultânea de Seis Antiepilépticos em Amostras de Plasma por Cromatografia a Líquido de Alta Eficiência. Um método simples, rápido e sensível foi proposto para a determinação simultânea de carbamazepina, fenitoína, fenobarbital, peimidona e seus principais metabólitos em amostras plasma por cromatografia a líquido de alta eficiência. Acetonitrila foi adicionada às aoatras de plasma para precipitar as proteínas. Após a centrifugação, 100µL do sobrenadante foram transferidos para um tubo de ensaio cônico e evaporado à secura com N2. O extrato foi reconstituído com 100µL de água e 10µL foram utilizados para análises cromatográficas. A separação dos fármacos foi realizada em coluna analítica C18 (25cm x 4,6 mm D.I.x 0,5 µm partícula) com a fase móvel tampão fosfato pH 4,8 - acetonitrila - metanol (55:25:20 v/v/v). A detecção foi realizada com detector ultravioleta a 210nm