RESUMO
Sphingolipids are a major lipid species found in all eukaryotes. Among structurally complex and diversified lipids, sphingoid bases have been heavily linked to various metabolic diseases. However, most current LC-MS-based methods lack the sensitivity to detect low-abundant sphingoid bases. The 6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) derivatization reagent, which efficiently forms covalent bonds with amino groups, has been widely used for amino acid detection. Nevertheless, the commonly used reverse-phase HPLC method for amino acid analysis is not suitable for amphipathic sphingolipids. To address this issue, we report a robust reverse-phase HPLC-MS/MS method capable of separating and detecting hydrophilic amino acids and sphingoid bases in a single run with high sensitivity. This method is also inclusive of other amino metabolites with an expandable target list. We tested this method under various conditions and samples, demonstrating its high reproducibility and sensitivity. Using this approach, we systematically analyzed human serum samples from healthy individuals, dyslipidemia, and type II diabetes mellitus (T2DM) patients, respectively. Two sphingolipids and five amino acids were identified with significant differences between the control and T2DM groups, highlighting the potential of this method in clinical studies.
RESUMO
1. In order to compare the difference between different derivatisations for amino acids determination of foie gras via, reversed phase high performance liquid chromatography (HPLC), O-phthalaldehyde and 9-fluorenyl-methyl chloroformate (OPA-FMOC group), phenylisothiocyanate (PITC group) and 6-Aminoquinolyl-N-hydrox-ysuccinimidyl Carbamate (AQC group) were applied to derivatisation reagent in this current experiment. The determination results of automatic amino acid analyser were applied, and 17 amino acids were detected by these three derivatisation methods.2. The running times of OPA-FMOC group, PITC group and AQC group were 18, 45 and 35 min, respectively. There was a large difference between the results of OPA-FMOC group and results from the automatic amino acid analyser, although the difference between the results from PITC and the automatic amino acid analyser was minimal.3. In conclusion, the running time of OPA-FMOC group was shorter than that of PITC group and AQC group; the accuracy of the former was better than the OPA-FMOC group and AQC group for the determination of amino acid of foie gras.
RESUMO
Omega-3 fatty acids are in high demand due to their efficacy in treating hypertriglyceridemia and preventing cardiovascular diseases. However, the growth of the industry is hampered by low purity and insufficient productivity. This study aims to develop an efficient RP-MPLC purification method for omega-3 fatty acid ethyl esters with high purity and capacity. The results indicate that the AQ-C18 featuring polar end-capped silanol groups outperformed C18 and others in retention time and impurity separation. By injecting pure fish oil esters with a volume equivalent to a 1.25% bed volume on an AQ-C18 MPLC column using a binary isocratic methanol-water (90:10, v:v) mobile phase at 30 mL/min, optimal omega-3 fatty acid ethyl esters were obtained, with the notable purity of 90.34% and a recovery rate of 74.30%. The total content of EPA and DHA produced increased from 67.91% to 85.27%, meeting the acceptance criteria of no less than 84% set by the 2020 edition of the Pharmacopoeia of the People's Republic of China. In contrast, RP-MPLC significantly enhanced the production efficiency per unit output compared to RP-HPLC. This study demonstrates a pioneering approach to producing omega-3 fatty acid ethyl esters with high purity and of greater quantity using AQ-C18 RP-MPLC, showing this method's significant potential for use in industrial-scale manufacturing.
Assuntos
Cromatografia de Fase Reversa , Ésteres , Ácidos Graxos Ômega-3 , Óleos de Peixe , Ácidos Graxos Ômega-3/química , Ácidos Graxos Ômega-3/isolamento & purificação , Ésteres/química , Ésteres/isolamento & purificação , Óleos de Peixe/química , Cromatografia de Fase Reversa/métodos , Cromatografia Líquida de Alta Pressão/métodos , Ácidos Docosa-Hexaenoicos/química , Ácidos Docosa-Hexaenoicos/isolamento & purificação , Ácido Eicosapentaenoico/química , Ácido Eicosapentaenoico/isolamento & purificaçãoRESUMO
Determination of substrate binding affinity (Kd) is critical to understanding enzyme function. An extensive number of methods have been developed and employed to study ligand/substrate binding, but the best approach depends greatly on the substrate and the enzyme in question. Below we describe how to measure the Kd of BesD, a non-heme iron halogenase, for its native substrate lysine using equilibrium dialysis with subsequent detection with High Performance Liquid Chromatography (HPLC). This method can be performed in anaerobic glove bag settings, requires readily available HPLC instrumentation for subsequent detection, and is adaptable to meet the needs of a variety of substrate affinity measurements.
RESUMO
A simple and reliable HPLC method was developed for quantification of chondroitin sulfate (CS). The procedure is based on precolumn hydrolysis of CS to liberate galactosamine and subsequent derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. Hydrolysis and derivatization conditions were optimized. A linear correlation coefficient of 0.9999 was calculated within the range of 10-1500 µg/mL from the standard curve. The method produces good precision and good accuracy (100.75 % recovery). An advantage over other common methods is its ability to quantify CS of all molecular weights and structures, as evidenced by the determination of CS fractions with narrow molecular weight distributions obtained through depolymerization by different methods, while enzymatic HPLC was proven to be infeasible. Extraction recoveries of CS from monosaccharide mixed samples were > 93 %. The reliability was also validated by a small difference (-1.95 % to 4.12 %) relative to enzymatic HPLC results in analysing representative CS samples of different animal origins and suppliers.
Assuntos
Aminoquinolinas , Carbamatos , Sulfatos de Condroitina , Animais , Peso Molecular , Reprodutibilidade dos Testes , Aminoquinolinas/química , Cromatografia Líquida de Alta Pressão/métodosRESUMO
The performance of an active-quenching single-photon avalanche diode (SPAD) array that is based on the tri-state gates of a field programmable gate array (FPGA) is presented. The array is implemented by stacking a bare 4 × 4 N-on-P SPAD array on a bare FPGA die, and the electrodes of the SPAD pixels and the I/O ports of the FPGA are connected through wire bonding within the same package. The active quenching action on each SPAD pixel is performed by using the properties of the tri-state gates of the FPGA. Digital signal processing, such as pulse counters, data encoders, and command interactions, is also performed by using the same FPGA. The breakdown voltage of the SPAD pixels, with an active area of 60 µm × 60 µm, is 47.2-48.0 V. When the device is reverse biased at a voltage of ~50.4 V, a response delay of ~50 ns, a dead time of 157 ns, a dark count rate of 2.44 kHz, and an afterpulsing probability of 6.9% are obtained. Its peak photon detection probability (PDP) reaches 17.0% at a peak wavelength of 760 nm and remains above 10% at 900 nm. This hybrid integrated SPAD array is reconfigurable and cost effective.
RESUMO
Biogenic amines (BAs) are compounds generated by decarboxylation of their amino acid precursors. Their intake, even at low concentrations, can lead to several types of health problems in sensitive individuals. As they can be easily formed in fermented dairy products, their quantitative determination is very relevant. In the present paper, a method for the quantitative determination of four biogenic amines in different dairy products has been developed, validated and applied to 37 samples of milk, 23 of yogurt, and 14 of kefir. Amines were selectively extracted using solid phase extraction, subsequently derivatizatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate and further determined by High Performance Liquid Chromatography with fluorescence detection. The method's sensitivity was highly satisfactory, with limits of detection lower than 0.2 mg/L. Optimal linearity and repeatability were also achieved. BAs were not detected in most of the milk samples, but they were found frequently at high levels in yogurt and kefir samples, reaching values of up to 79 mg/kg total BAs in kefir samples. Levels measured should not be a cause for concern for the population at large, but should be known by BAs-sensitive individuals.
Assuntos
Produtos Fermentados do Leite , Kefir , Humanos , Aminas Biogênicas , Aminoácidos , Cromatografia Líquida de Alta Pressão , CorantesRESUMO
Raman Spectroscopy is well emerged in the field of Analytical Quality Control (AQC) as a rapid and cost-effective technique useful in many applications. The advantage of Raman spectroscopy is the non-invasiveness of measurements that enablesto analyse samples directly in its container. In this study, the potential of Raman spectroscopy was investigated for analysis of clinical preparations of mAbs. Three commercial formulations of monoclonal antibodies (mAbs) Avastin®, Ontruzant® and Tecentriq® corresponding to Bevacizumab (BVC), Trastuzumab (TRS) and Atezolizumab (ATZ) respectively, were analysed in quartz cuvette in macroscopic analysis and through the wall of perfusion bags in microscopic analysis. The spectra have been compared to those of excipients (trehalose and sucrose) and of γ-Globulin, in order to investigate the origin of Raman bands. As expected, Raman spectra were a combination of bands from monoclonal antibodies and correspoding excipients found in formulas. For quantitative analysis of the solutions, models have been constructed using Partial Least Square Regression (PLSR) with Leave K-Out Cross Validation (LKOCV). The quantification performance was comparable for both macroscopic and microscopic analysis, in terms of error and linearity. The results are thus promising for future AQC in situ, in perfusion bags.
Assuntos
Antineoplásicos Imunológicos , Excipientes , Anticorpos Monoclonais/análise , Bevacizumab , Excipientes/química , Quartzo , Análise Espectral Raman/métodos , Sacarose , Trastuzumab , Trealose , gama-GlobulinasRESUMO
The study of brain diseases has long been of interest to researchers worldwide, and stroke is the third leading cause of death that threatens human health. At the same time, cerebral ischemia-reperfusion injury is closely associated with high rates of disability and mortality. The conditions of the 6-aminoquinolyl N-hydroxysccinimidyl carbamate method for the derivatization of amino acids in the bone marrow fluid and hippocampus of C57BL/6 mice with cerebral ischemia-reperfusion injury were explored and optimized, such as the column temperature, concentration of derivatization reagents and mobile phase concentration. The mobile phase consisted of 20 mm sodium acetate solution (phosphoric acid to adjust pH 5.0) and 60% acetonitrile solution at a flow rate of 1 ml min-1 . The 23 analytes were separated and determined in a gradient elution procedure; the correlation coefficient r was >0.9990 in the range 0.1-8.0 µg ml-1 . The results showed that the content of relevant analytes was significantly changed in the cerebral ischemia-reperfusion injury model, and the method was suitable for the simultaneous determination of 23 amino acids in the bone marrow fluid and hippocampus of C57BL/6 mice.
Assuntos
Medula Óssea , Traumatismo por Reperfusão , Aminoácidos , Aminoquinolinas , Animais , Cromatografia Líquida de Alta Pressão/métodos , Hipocampo , Humanos , Indicadores e Reagentes , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Introduction: Amino acids are critical biomarkers for many inborn errors of metabolism, but amino acid analysis is challenging due to the range of chemical properties inherent in these small molecules. Techniques are available for amino acid analysis, but they can suffer from long run times, laborious derivatization, and/or poor resolution of isobaric compounds. Objective: To develop and validate a method for the quantitation of a non-derivatized free amino acid profile in both plasma and urine samples using mixed-mode chromatography and tandem mass spectrometry. Methods: Chromatographic conditions were optimized to separate leucine, isoleucine, and allo-isoleucine and maintain analytical runtime at less than 15 min. Sample preparation included a quick protein precipitation followed by LC-MS/MS analysis. Matrix effects, interferences, linearity, carryover, acceptable dilution limits, precision, accuracy, and stability were evaluated in both plasma and urine specimen types. Results: A total of 38 amino acids and related compounds were successfully quantitated with this method. In addition, argininosuccinic acid was qualitatively analyzed. A full clinical validation was performed that included method comparison to a reference laboratory for plasma and urine with Deming regression slopes ranging from 0.38 to 1.26. Conclusion: This method represents an alternative to derivatization-based methods, especially in urine samples where interference from metabolites and medications is prevalent.
RESUMO
The majority of human proteins operate as multimeric complexes with defined compositions and distinct architectures. How the assembly of these complexes is surveyed and how defective complexes are recognized is just beginning to emerge. In eukaryotes, over 600 E3 ubiquitin ligases form part of the ubiquitin-proteasome system (UPS) which detects structural characteristics in its target proteins and selectively induces their degradation. The UPS has recently been shown to oversee key quality control steps during the assembly of protein complexes. We review recent findings on how E3 ubiquitin ligases regulate protein complex assembly and highlight unanswered questions relating to their mechanism of action.
Assuntos
Complexo de Endopeptidases do Proteassoma , Ubiquitina , Humanos , Complexo de Endopeptidases do Proteassoma/metabolismo , Controle de Qualidade , Ubiquitina/metabolismo , Ubiquitina-Proteína Ligases/metabolismoRESUMO
In recent years, goat milk powder and camel milk powder have gained popularity among consumers. Due to their potential low allergenicity, these milk powders have become a substitute for breast milk, especially for infants, and for people with lactose intolerance. In this paper, a method was developed for the simultaneous determination of 18 amino acids (AAs), histidine (His), serine (Ser), arginine (Arg), glycine (Gly), aspartic acid (Asp) combined with asparagine (Asn), glutamic (Glu), glutamine (Gln), threonine (Thr), alanine (Ala), proline (Pro), lysine (Lys), tyrosine (Tyr), methionine (Met), valine (Val), isoleucine (Iso), leucine (Leu), and dimer of cysteine (Cys) combined with cysteine (L-Cys-Cys), phenylalanine (Phe), taurine (Tau) in milk, goat milk, and camel milk power. The aim of the research was to compare the three kinds of milk powder from the perspective of the constituent amino acids. Therefore, the amino acid compositions and contents were compared. Thus, 2.0 g of the sample was accurately weighed, added to 16 mL H2O, and mixed thoroughly. Then, 200 mg of the sample was weighed in a glass tube with a stream of nitrogen to displace oxygen. The samples were hydrolyzed in HCl for 24 h at 110 â. Then, the amino acids were pre-column derivatized by 6-aminoquinoline-n-hydroxysuccinimide carbamate (AQC). In precolumn derivatization combined with reverse-phase chromatography, both 2,4-dinitrofluorobenzene (DNFB) and phenylisothiocyanate (PITC) can react with primary amines and secondary amines. However, the derivatization time is approximately 1 h. In contrast, the derivatization time of AQC was greatly shortened. Derivatization led to the conversion of free amino acids into highly stable derivatives, which were separated by ultra performance liquid chromatography (UPLC) with UV detection at 260 nm and quantified by the external standard method. The samples were separated on a BEH C18 column (150 mm×2.1 mm, 1.7 µm) at a flow rate of 0.4 mL/min. The calibration curves showed good linearity, with correlation coefficients greater than 0.999. The limits of detection (LODs) and limits of quantification (LOQs) of the 18 amino acids were 1.3-2.5 (mg/100 g) and 3.9-7.5 (mg/100 g), respectively. Quality control samples of SRM 1849a were used as the reference material. The results were in accordance with the content range. The RSDs ranged from 2.04% to 3.65%. Furthermore, the developed method was successfully applied to determine the types and concentrations of amino acids in 11 samples purchased from local markets in Shanghai and online shops. Abundant amino acids were detected in the three types of milk powder. While all the milk powder samples contained 18 types of amino acids, Tau was not detected in some of the goat and camel milk powder samples. Total essential amino acids (TEAA) in total amino acids (TAA) of milk powder was the highest of all. The TEAA values of TAA in the goat and camel milk powders were similar. The developed method requires only 22 min for the separation of 18 amino acids. This method is suitable for the large-scale analysis of milk powder samples, and it demonstrates high sensitivity and accuracy for the determination and confirmation of the 18 amino acids in different types of milk powders.
Assuntos
Aminoácidos , Análise de Alimentos/métodos , Leite , Aminoácidos/análise , Animais , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Leite/química , PósRESUMO
Proteins are plant cell wall components but they are not included in the definition of dietary fiber. Therefore, dietary fiber preparations have to be corrected for their residual protein contents. This is commonly done by calculating the residual protein concentrations from the nitrogen contents after Kjeldahl digestion. Here, three different methods to determine nitrogen in Kjeldahl digests were compared: conventional titration with hydrochloric acid after steam distillation, a colorimetric assay (24-well microplates and cuvettes), and the determination by using an ammonia electrode. All assays gave similar results but detection using the ammonia electrode was found to be the most time-efficient approach. Also, an amino-acid profiling method, which is not based on commercial kits and which is suitable for routine analysis of dietary fiber preparations, was established. For this purpose, an HPLC-FLD method following amino acid derivatization using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) was optimized for fiber samples. Although all commonly used dietary fiber preparation methods involve the application of proteases the amino acid profiles of fiber samples from different sources were shown to be quite diverse. Considering the amino acid composition of the residual protein in various dietary fiber preparations, residual protein is probably not only based on structural proteins.
RESUMO
ß-N-Methylamino-L-alanine (BMAA) is a non-canonical amino acid implicated as a cause for amyotrophic lateral sclerosis/parkinsonism dementia complex and potentially other neurodegenerative diseases. As interest in this molecule has increased, there has been a proliferation of methods along with a plethora of opinions as to the superiority of some methods over others. We analyzed the literature with reference to BMAA and its naturally occurring isomers, N-(2-aminoethyl) glycine (AEG) and 2,4 diaminobutyric acid (DAB). A comparison of methods, results, and critiques reveal that a single method has been approved by the AOAC but several different methods provide comparable BMAA quantification concentrations in similar tissues. We also describe a productive way to move forward as technology improves and changes.
Assuntos
Diamino Aminoácidos/análise , Neurotoxinas/análise , Diamino Aminoácidos/química , Animais , Cromatografia Líquida , Toxinas de Cianobactérias , Neurotoxinas/química , Reprodutibilidade dos Testes , Espectrometria de Massas em TandemRESUMO
Here we report evidence that new aminoquinoline N-glycan fluorescent labels interfere with the release of core α(1-6) fucose from N-glycans by bovine kidney α-l-fucosidase (BKF). BKF is a commonly employed exoglycosidase for core α(1-6) fucose determination. Molecular simulations of the bound and unbound Fuc-α(1-6)-GlcNAc, where GlcNAc is situated at the reducing end for all N-glycans, suggest that the reduced BKF activity may be due to a nonoptimal fit of the highest populated conformers in the BKF active binding site at room temperature. Population analysis and free energy estimates suggest that an enhanced flexibility of the labeled sugar, which facilitates recognition and binding, can be achievable with extended reaction conditions. We provide these experimental conditions using a sequential exoglycosidase digestion array using high concentrations of BKF.
Assuntos
Aminoquinolinas/farmacologia , Fucose/isolamento & purificação , Polissacarídeos/química , alfa-L-Fucosidase/metabolismo , Animais , Sítios de Ligação , Bovinos , Corantes Fluorescentes , Polissacarídeos/metabolismo , Coloração e RotulagemRESUMO
The study, conducted on Italian preadolscents aged 11 to 13 belonging to the general population, aims to investigate the relationship between the emotional functioning, namely, alexithymia, and the risk of developing behavioral and emotional problems measured using the Strength and Difficulty Questionnaire. The latent class analysis approach allowed to identify two latent variables, accounting for the internalizing (emotional symptoms and difficulties in emotional awareness) and for the externalizing problems (conduct problems and hyperactivity, problematic relationships with peers, poor prosocial behaviors and externally oriented thinking). The two latent variables featured two latent classes: the difficulty in dealing with problems and the strength to face problems that was representative of most of the healthy participants with specific gender differences. Along with the analysis of psychopathological behaviors, the study of resilience and strengths can prove to be a key step in order to develop valuable preventive approaches to tackle psychiatric disorders.
Assuntos
Sintomas Afetivos , Comportamento Problema , Adolescente , Sintomas Afetivos/complicações , Criança , Feminino , Humanos , Itália , Masculino , Modelos Psicológicos , Grupo Associado , Escalas de Graduação Psiquiátrica , PsicologiaRESUMO
To find novel functional beverages from folk teas, 33 species of frequently used non-Camellia tea (plants other than Camellia) were collected and compared with Camellia tea (green tea, pu-erh tea and black tea) for the first time. Data are reported here on the quantities of 20 free amino acids (FAAs) and three purine alkaloids (measured by UHPLC), total polyphenols (measured by Folin-Ciocalteu assay), and antioxidant activity (DPPH). The total amounts of FAAs in non-Camellia tea (0.62-18.99 mg/g) are generally less than that of Camellia tea (16.55-24.99 mg/g). However, for certain FAAs, the quantities were much higher in some non-Camellia teas, such as γ-aminobutyric acid in teas from Ampelopsis grossedentata, Isodon serra and Hibiscus sabdariffa. Interestingly, theanine was detected in tea from Potentilla fruticosa (1.16±0.81 mg/g). Furthermore, the content of polyphenols in teas from A. grossedentata, Acer tataricum subsp. ginnala are significantly higher than those from Camellia tea; teas from I. serra, Pistacia chinensis and A. tataricum subsp. ginnala have remarkable antioxidant activities similar to the activities from green tea (44.23 µg/mL). Purine alkaloids (caffeine, theobromine and theophylline) were not detected in non-Camellia teas. The investigation suggest some non-Camellia teas may be great functional natural products with potential for prevention of chronic diseases and aging, by providing with abundant polyphenols, antioxidants and specific FAAs.
RESUMO
Exposure to ß-N-methylamino-l-alanine (BMAA) might be linked to the incidence of amyotrophic lateral sclerosis, Alzheimer's disease and Parkinson's disease. Analytical chemistry plays a crucial role in determining human BMAA exposure and the associated health risk, but the performance of various analytical methods currently employed is rarely compared. A CYANOCOST initiated workshop was organized aimed at training scientists in BMAA analysis, creating mutual understanding and paving the way towards interlaboratory comparison exercises. During this workshop, we tested different methods (extraction followed by derivatization and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis, or directly followed by LC-MS/MS analysis) for trueness and intermediate precision. We adapted three workup methods for the underivatized analysis of animal, brain and cyanobacterial samples. Based on recovery of the internal standard D3BMAA, the underivatized methods were accurate (mean recovery 80%) and precise (mean relative standard deviation 10%), except for the cyanobacterium Leptolyngbya. However, total BMAA concentrations in the positive controls (cycad seeds) showed higher variation (relative standard deviation 21%-32%), implying that D3BMAA was not a good indicator for the release of BMAA from bound forms. Significant losses occurred during workup for the derivatized method, resulting in low recovery (<10%). Most BMAA was found in a trichloroacetic acid soluble, bound form and we recommend including this fraction during analysis.
Assuntos
Diamino Aminoácidos/análise , Cromatografia Líquida/métodos , Neurotoxinas/análise , Espectrometria de Massas em Tandem/métodos , Diamino Aminoácidos/metabolismo , Animais , Encéfalo/metabolismo , Cianobactérias/metabolismo , Toxinas de Cianobactérias , Daphnia , Neurotoxinas/metabolismo , Reprodutibilidade dos Testes , Ácido Tricloroacético/químicaRESUMO
BACKGROUND: Proteins in human tissues and body fluids continually undergo spontaneous oxidation and glycation reactions forming low levels of oxidation and glycation adduct residues. Proteolysis of oxidised and glycated proteins releases oxidised and glycated amino acids which, if they cannot be repaired, are excreted in urine. SCOPE OF REVIEW: In this review we give a brief background to the classification, formation and processing of oxidised and glycated proteins in the clinical setting. We then describe the application of stable isotopic dilution analysis liquid chromatography-tandem mass spectrometry (LC-MS/MS) for measurement of oxidative and glycation damage to proteins in clinical studies, sources of error in pre-analytic processing, corroboration with other techniques - including how this may be improved - and a systems approach to protein damage analysis for improved surety of analyte estimations. MAJOR CONCLUSIONS: Stable isotopic dilution analysis LC-MS/MS provides a robust reference method for measurement of protein oxidation and glycation adducts. Optimised pre-analytic processing of samples and LC-MS/MS analysis procedures are required to achieve this. GENERAL SIGNIFICANCE: Quantitative measurement of protein oxidation and glycation adducts provides information on level of exposure to potentially damaging protein modifications, protein inactivation in ageing and disease, metabolic control, protein turnover, renal function and other aspects of body function. Reliable and clinically assessable analysis is required for translation of measurement to clinical diagnostic use. Stable isotopic dilution analysis LC-MS/MS provides a "gold standard" approach and reference methodology to which other higher throughput methods such as immunoassay and indirect methods are preferably corroborated by researchers and those commercialising diagnostic kits and reagents. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn.
Assuntos
Produtos Finais de Glicação Avançada/análise , Espectrometria de Massas/métodos , Proteínas/análise , Produtos Finais de Glicação Avançada/química , Humanos , Oxirredução , Proteínas/químicaRESUMO
This paper reports the determination of biogenic amines by high-performance liquid chromatography (HPLC) and fluorescence detection after derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) in balsamic, apple, and red, white, and Sherry wine vinegars. A solid-phase extraction (SPE) with mixed-mode resins method was used before analysis. The method was successfully validated obtaining adequate values of selectivity, response linearity, precision, accuracy, and low detection and quantification limits. The total content of biogenic amines in vinegars ranged from 23.35 to 1445.2 µg/L, being lower than those reported in wines. Putrescine was the amine that showed the highest concentrations in most samples. Methylamine and phenylethylamine were not determined in any vinegar. Balsamic and "Pedro Ximénez" Sherry vinegars reached the highest amounts of biogenic amines, while apple, white and Sherry wine vinegars had the lowest concentrations. Principal component analysis using the biogenic amines as variables, allowed to separate the different kind of vinegars, excepting red vinegars.