RESUMO
Nanomaterials possessing enzyme-like activity have been extensively studied owing to their high stability and tunable catalytic properties. In this work, a simple method has been developed for the synthesis of porous manganese oxide/manganese ferrite (MnOx/MnFe2O4) nanopopcorns (MFNPs) in neutral media. The MFNPs exhibit dual enzymatic activities towards selective oxidation of ketoses followed by H2O2-induced decline of its catalytic activity. MFNPs, with MnFe2O4 as the core material and an outer layer rich in MnOx, were synthesized from ammonium iron(III) citrate and potassium permanganate at 70⯰C for 12â¯h followed by annealing at 300⯰C for 6â¯h. The nanozyme, MFNPs, exhibited oxidase-like activity, which was proved by the oxidation of amplex red (AR) in the presence of dissolved oxygen in the solution, to form fluorescent resorufin. The activity of MFNPs is highly suppressed by H2O2 as a result of its induced dissolution of MnOx. In addition, MFNPs having catalytic activity towards the selective oxidation of ketoses (e.g., fructose) followed by the formation of H2O2. The as-formed H2O2 diminished the catalytic activity of MFNPs for the AR oxidation to form fluorescent resorufin. Upon increasing fructose concentration, the fluorescence of resorufin decreases. Since the MFNPs do not show catalytic activity towards aldose sugars, such as glucose, sucrose, and mannose, the AR/MFNPs probe has high selectivity and sensitivity for detection of fructose with a limit of detection of 32⯵M. Our study shows its great potential for quantitation of fructose in honey samples.
Assuntos
Materiais Biomiméticos/química , Compostos Férricos/química , Cetoses/análise , Compostos de Manganês/química , Nanoestruturas/química , Óxidos/química , Peroxidase/metabolismo , Peróxido de Hidrogênio/farmacologia , Oxidantes/farmacologiaRESUMO
α-Dicarbonyl compounds (α-DCs) are very clinically important as they are considered as advanced glycation end products (AGEs) precursors and biomarkers for many chronic diseases such as diabetes and vascular diseases, in addition to their major role in progression of complications of such diseases. Aromatic aldehydes and ammonium acetate were productively used as a one-pot co-reagents for fluorogenic derivatization of α-DCs yielding fluorescent imidazole derivatives. Among the tried aromatic aldehydes, 4-carbomethoxybenzaldehyde yielded the products with best fluorescent characters. This approach for fluorogenic derivatization of α-DCs overcome the selectivity problem of the most commonly used derivatization reagent for α-DCs, α-diamino compounds, that can react unselectively with α-DCs and aldehydes. Separation of the formed imidazole derivatives of five α-DCs including glucosone, 3-deoxyglucosone, glyoxal, methyl glyoxal and dimethyl glyoxal together with ethylmethylglyoxal as an internal standard was carried out on an octyl column using a mobile phase consisted of methanol-water (15:85, v/v%) containing 0.2% formic acid with time programed flow, followed by fluorescence detection at excitation/emission wavelengths of 310/410â¯nm. The method showed excellent sensitivity for the targeted α-DCs with limits of detections ranging from 0.4 to 5.0â¯nM in human serum. Simple protein precipitation procedure was used for human serum treatment yielding very good recovery (91-105%) for the targeted α-DCs. The developed method was fully validated, then applied to the analysis of the five above mentioned clinically important α-DCs in serum samples of healthy, diabetic, rheumatic and cardiac disorders human volunteers. Due to the excellent analytical features of the developed method, including high selectivity and sensitivity, it was able to detect the pattern of the targeted α-DCs serum levels under the investigated different clinical conditions.
Assuntos
Diabetes Mellitus/sangue , Produtos Finais de Glicação Avançada/sangue , Cardiopatias/sangue , Doenças Reumáticas/sangue , Acetatos/química , Aldeídos/sangue , Aldeídos/química , Aldeídos/metabolismo , Biomarcadores/análise , Biomarcadores/metabolismo , Cromatografia Líquida de Alta Pressão/instrumentação , Cromatografia Líquida de Alta Pressão/métodos , Desoxiglucose/análogos & derivados , Desoxiglucose/análise , Desoxiglucose/química , Desoxiglucose/metabolismo , Diabetes Mellitus/metabolismo , Fluorescência , Produtos Finais de Glicação Avançada/química , Produtos Finais de Glicação Avançada/metabolismo , Voluntários Saudáveis , Cardiopatias/metabolismo , Humanos , Imidazóis/química , Imidazóis/metabolismo , Cetoses/análise , Cetoses/química , Cetoses/metabolismo , Limite de Detecção , Estresse Oxidativo , Doenças Reumáticas/metabolismo , Sensibilidade e EspecificidadeRESUMO
Baby foods are exposed to elevated temperatures during processing treatments such as sterilization or spray drying. These treatments decompose sugars leading to the formation of α-dicarbonyl compounds that are of importance since they have been associated with several metabolic disorders. In this study, an analytical method based on high-performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS) was used to determine α-dicarbonyl compounds in baby foods. The method entailed aqueous extraction of α-dicarbonyl compounds from the samples and derivatization with o-phenylenediamine prior to chromatographic analysis. The results indicated that major degradation product was 3-deoxyglucosone in the samples including cereal-based infant formula, canned fruit and vegetable puree. Its concentration ranged between 3.9 and 827.1 mg/kg in infant formula and between 26.7 and 92.3 mg/kg in fruit puree samples. The concentrations of glucosone, 1-deoxyglucosone, 5-hydroxymethyl-2-furfural, furfural, glyoxal, methylglyoxal, and dimethylglyoxal levels were rather low.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Desoxiglucose/análogos & derivados , Alimentos Infantis/análise , Espectrometria de Massas por Ionização por Electrospray/métodos , Desoxiglucose/análise , Furaldeído/análise , Glioxal/análise , Humanos , Fórmulas Infantis/química , Cetoses/análise , Aldeído Pirúvico/análiseRESUMO
The isomers of monosaccharide always produce multiple chromatographic peaks as volatile derivatives during gas chromatography, which may result in the overlapping of different sugar peaks. Whereas reduction and oximation of sugar carbonyl groups for GC analysis do eliminate many isomer derivatives, the approaches create new problems. One ketose can yield two peaks by oximation, and different aldoses and ketoses can yield the same alditol upon reduction, leading to the inability to detect some important monosaccharides. This paper reports an optimal method that yields a single peak per sugar by acetylation directly. By using a methyl sulfoxide (Me2SO)/1-methylimidazole (1-MeIm) system, the carbohydrates in acetic anhydride (Ac2O) esterification reactions were solubilized, and the oxidation that normally occurs was inhibited. The results demonstrate that acetylated derivatives of 23 saccharides had unique peaks, which indicates aldose, ketose, and alditol can be determined simultaneously by GC-MS.
Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Ginkgo biloba/química , Cetoses/análise , Populus/química , Álcoois Açúcares/análise , Anidridos Acéticos/química , Acetilação , Dimetil Sulfóxido/química , Glucose/análise , Imidazóis/química , Limite de Detecção , Monossacarídeos/químicaRESUMO
Sugar-sweetened carbonated soft drinks (CSDs) are broadly consumed worldwide. The added sugar, particularly high-fructose corn syrup (HFCS), can be an important source of sugar degradation products, such as α-dicarbonyl compounds. This study recorded the α-dicarbonyl profile in CSDs by ultrahigh-performance liquid chromatography with hyphenated diode array-tandem mass spectrometry after derivatization with o-phenylenediamine. Thus, 3-deoxy-D-erythro-hexos-2-ulose (3-DG), D-lyxo-hexos-2-ulose (glucosone), 3-deoxy-D-threo-hexos-2-ulose (3-DGal), 1-deoxy-D-erythro-hexos-2,3-diulose (1-DG), 3,4-dideoxyglucosone-3-ene (3,4-DGE), methylglyoxal, and glyoxal were identified as major α-dicarbonyls and, with the exception of glyoxal, quantified (recovery rates, 85.6-103.1%; RSD, 0.8-3.6%). Total α-dicarbonyl concentration in 25 tested commercial products ranged between 0.3 and 116 µg/mL and was significantly higher in HFCS-sweetened CSDs compared to CSDs sweetened with HFCS and sucrose or with sucrose alone. Predominant was 3-DG (≤87 µg/mL) followed by glucosone (≤21 µg/mL), 3-DGal (≤7.7 µg/mL), 1-DG (≤2.8 µg/mL), methylglyoxal (≤0.62 µg/mL), and 3,4-DGE (≤0.45 µg/mL).
Assuntos
Bebidas Gaseificadas/análise , Frutose/química , Galactose/análogos & derivados , Glioxal/análise , Cetoses/análise , Edulcorantes/química , Cromatografia Líquida de Alta Pressão , Galactose/análise , Reação de Maillard , Espectrometria de Massas em TandemRESUMO
High-fructose corn syrup (HFCS) is a widely used liquid sweetener produced from corn starch by hydrolysis and partial isomerization of glucose to fructose. During these processing steps, sugars can be considerably degraded, leading, for example, to the formation of reactive α-dicarbonyl compounds (α-DCs). The present study performed targeted screening to identify the major α-DCs in HFCS. For this purpose, α-DCs were selectively converted with o-phenylendiamine to the corresponding quinoxaline derivatives, which were analyzed by liquid chromatography with hyphenated diode array-tandem mass spectrometry (LC-DAD-MS/MS) detection. 3-Deoxy-D-erythro-hexos-2-ulose (3-deoxyglucosone), D-lyxo-hexos-2-ulose (glucosone), 3-deoxy-D-threo-hexos-2-ulose (3-deoxygalactosone), 1-deoxy-D-erythro-hexos-2,3-diulose (1-deoxyglucosone), 3,4-dideoxyglucosone-3-ene, methylglyoxal, and glyoxal were identified by enhanced mass spectra as well as MS/MS product ion spectra using the synthesized standards as reference. Addition of diethylene triamine pentaacetic acid and adjustment of the derivatization conditions ensured complete derivatization without de novo formation for all identified α-DCs in HFCS matrix except for glyoxal. Subsequently, a ultra-high performance LC-DAD-MS/MS method was established to quantify 3-deoxyglucosone, glucosone, 3-deoxygalactosone, 1-deoxyglucosone, 3,4-dideoxyglucosone-3-ene, and methylglyoxal in HFCS. Depending on the α-DC compound and concentration, the recovery ranged between 89.2% and 105.8% with a relative standard deviation between 1.9% and 6.5%. Subsequently, the α-DC profiles of 14 commercial HFCS samples were recorded. 3-Deoxyglucosone was identified as the major α-DC with concentrations up to 730 µg/mL HFCS. The total α-DC content ranged from 293 µg/mL to 1,130 µg/mL HFCS. Significantly different α-DC levels were not detected between different HFCS specifications, but between samples of various manufacturers indicating that the α-DC load is influenced by the production procedures.
Assuntos
Análise de Alimentos , Contaminação de Alimentos/análise , Frutose/química , Glioxal/análise , Cetoses/análise , Edulcorantes/química , Zea mays/química , Cromatografia Líquida , Estrutura Molecular , Espectrometria de Massas em TandemRESUMO
Fibrosis and vascular sclerosis are main complications that limit the long-term application of peritoneal dialysis (PD). Low biocompatibility has been largely attributed to the presence of glucose degradation products (GDPs), which are formed during the heat sterilization of PD fluids. GDPs readily modify proteins in the peritoneum, leading to a decline of their biological function. After absorption, GDPs can also promote systemic protein glycation. Additionally, GDPs may augment DNA glycation, a process enhanced in uremia. Apart from their glycating activity, GDPs induce cytotoxicity and interfere with cell signaling in peritoneal mesothelial cells. Targeted screening revealed the nature of the 6 major GDPs with α-dicarbonyl structure as 3-deoxyglucosone, 3-deoxygalactosone, glucosone, glyoxal, methylglyoxal, and 3,4-dideoxyglucosone-3-ene. Valid quantification of these GDPs was achieved by ultrahigh-performance liquid chromatography/diode array detector/tandem mass spectrometry. Identification and quantification of single GDPs allow a structure-dependent risk evaluation. As a consequence, PD fluids and processes can be improved to reduce the GDP burden of patients undergoing PD.
Assuntos
Soluções para Diálise/química , Glucose/química , Diálise Peritoneal/efeitos adversos , Desoxiglucose/análogos & derivados , Desoxiglucose/análise , Soluções para Diálise/efeitos adversos , Galactose/análogos & derivados , Galactose/análise , Glioxal/análise , Temperatura Alta , Humanos , Cetoses/análise , Pironas/análise , Relação Estrutura-AtividadeRESUMO
During heat sterilization of peritoneal dialysis solutions, glucose is partially transformed into glucose degradation products (GDPs), which significantly reduce the biocompatibility of these medicinal products. Targeted α-dicarbonyl screening identified glyoxal, methylglyoxal, 3-deoxyglucosone, 3,4-dideooxyglucosone-3-ene, glucosone, and 3-deoxygalactosone as the major six GDPs with α-dicarbonyl structure. In the present study, an ultra-high-performance liquid chromatography method was developed which allows the separation of all relevant α-dicarbonyl GDPs within a run time of 15 min after derivatization with o-phenylenediamine. Hyphenated diode array detection/tandem mass spectrometry detection provides very robust quantification and, at the same time, unequivocal peak confirmation. Systematic evaluation of the derivatization process resulted in an optimal derivatization period that provided maximal derivatization yield, minimal de novo formation (uncertainty range ±5%), and maximal sample throughput. The limit of detection of the method ranged from 0.13 to 0.19 µM and the limit of quantification from 0.40 to 0.57 µM. Relative standard deviations were below 5%, and recovery rates ranged between 91% and 154%, dependent on the type and concentration of the analyte (in 87 out of 90 samples, recovery rates were 100 ± 15%). The method was then applied for the analysis of commercial peritoneal dialysis fluids (nine different product types, samples from three lots of each).
Assuntos
Desoxiglucose/análise , Soluções para Diálise/química , Contaminação de Equipamentos , Glioxal/análise , Cetoses/análise , Diálise Peritoneal , Espectrometria de Massas em Tandem , Humanos , Estrutura Molecular , Reprodutibilidade dos Testes , EsterilizaçãoRESUMO
UNLABELLED: We investigated the development of antioxidant activity relative to the change of pH, fluorescent intensity, ultraviolet (UV) absorbance (A294), browning (A420), and alpha-dicarbonyl compounds in sugar-amino acid Maillard reaction (MR) model systems comprising fructose, glucose, or ribose each with glycine (Fru-Gly, Glu-Gly, and Rib-Gly) or lysine (Fru-Lys, Glu-Lys, and Rib-Lys), respectively, which were heated at 121 °C for 5 to 90 min. For hexose models, the change in pH was shown to fit a second-order polynomial regression with A294 and A420. Antioxidant activity was significantly and positively correlated with UV absorbance (r = 0.905, P < 0.001) and browning products (r = 0.893, P < 0.001) rather than with fluorescent products or the alpha-dicarbonyl compounds. Type of sugar was most important in evoking a change in UV absorbance, browning, alpha-dicarbonyl compounds, and antioxidant activity of MR products (MRPs). In conclusion, the antioxidant activity of MRPs in six model systems was more closely associated with products derived at the intermediate-to-late stages of the reaction and influenced mostly by the type of sugar. PRACTICAL APPLICATION: We report on the different factors and their interactions that are important for understanding the functional attributes of food components that comprise the generation of Maillard browning products and the associated antioxidant activities generated during high-temperature food processing.
Assuntos
Aminoácidos/química , Antioxidantes/química , Frutose/química , Glucose/química , Reação de Maillard , Modelos Químicos , Ribose/química , Cor , Desoxiaçúcares/análise , Desoxiaçúcares/química , Desoxiglucose/análogos & derivados , Desoxiglucose/análise , Desoxiglucose/química , Frutose/análogos & derivados , Glucose/análogos & derivados , Glicina/análogos & derivados , Glicina/química , Temperatura Alta , Concentração de Íons de Hidrogênio , Cetoses/análise , Cetoses/química , Cinética , Lisina/análogos & derivados , Lisina/química , Pentoses/análise , Pentoses/química , Ribose/análogos & derivados , Espectrometria de Fluorescência , Espectrofotometria UltravioletaRESUMO
In this study, for the first time, the amount of 3-deoxy-D-erythro-hexos-2-ulose (3-DG) in fresh citrus and chestnut honeys was determined. 3-DG was measured as the corresponding quinoxalines after derivatization with orthophenylenediamine using reverse-phase high-performance liquid chromatography (RP-HPLC). Notwithstanding the freshness of the samples, high levels of 3-DG were detected in both honeys. The comparison of 3-DG and 5-hydroxymethylfurfural (HMF) concentrations, which was also quantified by RP-HPLC, showed that citrus honeys had the lowest amount of 3-DG (107 mg/kg) and the highest of HMF (16.7 mg/kg), while chestnut honeys had the opposite (398 and 1.2 mg/kg, respectively). During thermal treatment, different 3-DG and HMF trends were highlighted between the citrus and chestnut honeys; at the end, 3-DG formation was more favored with respect to HMF formation. Moreover, in citrus honeys, a good correlation between 3-DG and HMF levels was observed, which was not found in chestnut honeys, suggesting a role of the high pH values of these honeys on the degradation routes. The kinetic analysis showed the highest k value for 3-DG and HMF formation in chestnut and citrus honeys, respectively. The lowest Ea values related to 3-DG formation and the highest to HMF formation, indicating that the key intermediate 3-DG is easily formed at low temperatures, whilst the formation of HMF requires higher temperatures. For this reason, 3-DG seems to be an aging index rather than a thermal index and its use, at least for honeys at high pH values, together with HMF, could improve their quality assessment.
Assuntos
Mel/análise , Cetoses/análise , Cromatografia Líquida de Alta Pressão , Citrus , Fagaceae , Flores , Furaldeído/análogos & derivados , Furaldeído/análise , Furaldeído/química , Temperatura Alta , Concentração de Íons de Hidrogênio , Cetoses/química , Cinética , Fatores de TempoRESUMO
Glucose degradation products (GDPs) formed during heat sterilization of peritoneal dialysis (PD) fluids exert cytotoxic effects and promote the formation of advanced glycation end-products in the peritoneal cavity. As a result, long-term application of continuous ambulatory peritoneal dialysis is limited. The composition and concentration of GDPs in PD fluids must be known to evaluate their biological effects. The present study describes a targeted screening for novel GDPs in PD fluids. For this purpose, dicarbonyl compounds were converted with o-phenylenediamine to give the respective quinoxaline derivatives, which were selectively monitored by HPLC/diode array detector. Glucosone was thereby identified as a novel major GDP in PD fluids. Product identity was confirmed by LC/MSMS analysis using independently synthesized glucosone as a reference compound. Furthermore, a method was developed to quantify glucosone in PD fluids by HPLC/UV after derivatization with o-phenylenediamine. The method's limit of detection was 0.6 microM and the limit of quantitation 1.1 microM. A linear calibration curve was obtained between 1.1 and 113.9 microM (R(2)=0.9999). Analyzed at three different concentration levels, recovery varied between 95.6% and 102.0%. The coefficient of variation ranged between 0.4% and 4.7%. The method was then applied to the measurement of glucosone in typical PD fluids. Glucosone levels in double chamber bag PD fluids varied between not detectable and 6.7 microM. In single chamber bag fluids, glucosone levels ranged between 28.7 and 40.7 microM.
Assuntos
Cromatografia Líquida de Alta Pressão/instrumentação , Cromatografia Líquida de Alta Pressão/métodos , Soluções para Diálise/química , Glucose/metabolismo , Cetoses/análise , Espectrometria de Massas/instrumentação , Diálise Peritoneal , Padrões de Referência , Reprodutibilidade dos TestesRESUMO
Coffee, a highly processed food, and Maillard mixtures are able to activate nuclear factor kappaB translocation in macrophages via generation of hydrogen peroxide. In this study, a substructure library was prepared and used to identify Maillard products that are responsible for this effect. Three different Maillard reaction products with aminoreductone substructure (C(6)-aminoreductone, C(4)-aminoreductone, and aminohexose reductone) strongly induce nuclear factor kappaB translocation in macrophages. The effect was almost completely blocked by co-incubation with catalase, indicating that cellular activation was mediated by the ability of the test compounds to generate hydrogen peroxide. The cellular effect of a Maillard mixture, which was produced under conditions favoring aminoreductone formation, could be almost completely related to the presence of C(6)-aminoreductone.
Assuntos
Núcleo Celular/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Reação de Maillard , NF-kappa B/metabolismo , Proteínas Nucleares/metabolismo , Substâncias Redutoras/farmacologia , Animais , Catalase/metabolismo , Linhagem Celular , Núcleo Celular/metabolismo , Cromatografia Líquida de Alta Pressão , Manipulação de Alimentos , Peróxido de Hidrogênio/metabolismo , Cetoses/análise , Cetoses/química , Cetoses/farmacologia , Macrófagos/metabolismo , Espectroscopia de Ressonância Magnética , Transporte Proteico/efeitos dos fármacos , Ratos , Substâncias Redutoras/síntese química , Substâncias Redutoras/química , Bibliotecas de Moléculas Pequenas , Espectrofotometria Ultravioleta , Fator de Transcrição RelA/metabolismoRESUMO
A method has been established by which to determine aldoses and ketoses in plant material simultaneously. Monosaccharides were extracted by sonication with 80% ethanol and sugar oximes formed by treatment of the resultant extract with hydroxylamine and pyridine at 90 degrees C. After reaction, one aliquot of the product was derivatised with acetic anhydride at 90 degrees C, whilst a second aliquot was silylated with HMDS and TMCS at 80 degrees C. Both reaction mixtures were analysed by GC-MS in the SIM mode. Quantivation was linear within the range 1-4 microg/mL and the detection limit for monosaccharides was 5-25 ng/mL. The absolute recoveries were between 73.0 and 90.2% and the RSDs were 3.1-10.0%. This method was applied to analyse the free monosaccharides in Lyceum barbarum L.; eight monosaccharides were present in amounts between 0.26 and 368.65 microg/mg.
Assuntos
Carboidratos/análise , Carboidratos/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Cetoses/análise , Frutas/química , Cetoses/química , Lycium/químicaAssuntos
Campylobacter jejuni/química , Helicobacter pylori/química , Cetoses/análise , Polissacarídeos/metabolismo , Pseudomonas aeruginosa/química , Fatores de Virulência/metabolismo , Sequência de Carboidratos , Cetoses/metabolismo , Dados de Sequência Molecular , Polissacarídeos/biossíntese , Difosfato de Uridina/análise , Difosfato de Uridina/metabolismo , Fatores de Virulência/biossínteseRESUMO
Capillary electrophoresis coupled to mass spectrometry (CE-MS) is reported for the first time as an alternative and powerful analytical method for the characterization and monitoring of N-substituted 1-amino-1-deoxyketoses (Amadori compounds). It allows rapid separation and identification of Amadori compounds, while benefiting from the well-known advantages of MS, such as specificity and sensitivity. Amadori compounds of several amino acids, such as glycine, valine, isoleucine, methionine, proline, and phenylalanine, as well as a cysteine-derived compound, were separated and/or discriminated using CE-MS/MS under standard conditions. The technique may also be useful to study the stability and degradation kinetics of other labile charged Maillard intermediates that play an important role in food and medical science.
Assuntos
Aminoácidos/análise , Eletroforese Capilar/métodos , Cetoses/análise , Reação de Maillard , Espectrometria de Massas/métodos , Aminoácidos/química , Aminoácidos/isolamento & purificação , Estabilidade de Medicamentos , Alimentos , Glucose/química , Temperatura Alta , Cetoses/química , Cinética , Preparações Farmacêuticas/análiseRESUMO
While the Maillard reaction of amino acids and proteins as well as its consequences in vivo has been thoroughly investigated, little attention has so far been paid to the glycation of aminophospholipids such as phosphatidylethanolamine (PE) or phosphatidylserine (PS), which are essential for structure and functionality of biological membranes. PE-derived glucosylamines (Schiff-PEs) and aminoketoses (Amadori-PEs) have now for the first time been simultaneously identified and quantified in erythrocytes from diabetics and healthy individuals by liquid chromatography-electrospray mass spectrometry (LC-(ESI)MS). The amounts of glycated PE (gPE) were significantly higher in diabetics (0.18-34.2 mol% Schiff-PE and 0.047-0.375 mol% Amadori-PE) than in controls (0.12-3.99 mol% Schiff-PE and 0.018-0.055 mol% Amadori-PE). A positive correlation between fructosylated hemoglobin (HbA(1c)) and the gPE levels was established. No advanced glycation endproducts (AGEs) like 5-hydroxymethylpyrrole-2-carbaldehyde (pyrrole-PE), carboxymethyl (CM-PE), or carboxyethyl (CE-PE) derivatives were detected. To investigate the influence of gPE on lipid peroxidation of biological membranes, liposomes consisting of soy-PE and synthetically prepared Amadori-PE (16:0-16:0) were incubated for several days and the formation of oxidation products was monitored. It could be shown that Amadori-PE extensively promotes lipid peroxidation even in the absence of transition metal ions like Cu(2+) and Fe(3+). Oxidative damage to membrane lipids therefore is supposed to be at least partially caused by the glycation of aminophospholipids.
Assuntos
Eritrócitos/metabolismo , Peroxidação de Lipídeos/fisiologia , Fosfatidiletanolaminas/metabolismo , Aminas/análise , Aminas/química , Aminas/isolamento & purificação , Cromatografia Líquida , Humanos , Técnicas In Vitro , Cetoses/análise , Cetoses/química , Lipossomos/metabolismo , Oxirredução , Fosfatidiletanolaminas/química , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
A sensitive and selective method for detection and quantification of 1,5-anhydro-D-fructose, microthecin, and 4-deoxy-glycero-hexo-2, 3-diulose using GC-MS in selected ion monitoring mode has been developed. These compounds, which occur in many biological systems, have here been quantified in the red alga Gracilariopsis lemaneiformis. A screening of other algae showed the occurrence of 1, 5-anhydro-D-fructose in several other species of red algae as well as in some green and brown algae.
Assuntos
Eucariotos/química , Frutose/análogos & derivados , Cromatografia Gasosa-Espectrometria de Massas/métodos , Cetoses/análise , Animais , Frutose/análise , Controle de Qualidade , RatosRESUMO
Aldoses, ketoses and uronic acids were derivatized with p-aminobenzoic acid and separated as their borate complexes by capillary zone electrophoresis, using a capillary tube of fused silica containing 150 mM borate buffer, pH 10.0, as carrier. The electrophoretic mobilities of 22 carbohydrates were determined and found to increase with increasing stability of the borate complexes formed. Besides the number of hydroxyl groups and the presence of substituents, complex stability depended most strongly on the configuration of the three vicinal hydroxyl groups at C2, C3 and C4. On-column UV monitoring at 285 nm allowed the detection of glucose with a lower mass detection limit of 15 fmol and a concentration sensitivity of 4 microM. Reproducible quantification of carbohydrates was achieved at least in the concentration range of 0.1-10 mM in reaction solutions by the relative peak area method, using cinnamic acid as internal standard. The method was applied successfully to the determination of the monosaccharide composition of polysaccharides extracted from Radix althaeae.
Assuntos
Aldeídos/análise , Eletroforese/métodos , Cetoses/análise , Ácidos Urônicos/análise , para-Aminobenzoatos , Ácido 4-Aminobenzoico/análise , Aminação , Carboidratos/química , Temperatura Alta , Concentração de Íons de Hidrogênio , Estrutura Molecular , Oxirredução , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
3-Deoxyglucose (3-deoxy-D-erythro-hexos-2-ulose) (3-DG) is a reactive dicarbonyl intermediate involved in the polymerization and browning of proteins by glucose in vitro. Damage to protein by formation of 3-DG in vivo is thought to be limited by enzymes which convert 3-DG to less reactive species, such as 3-deoxyfructose (3-DF). We have developed a sensitive and specific assay for measuring 3-DG and 3-DF in human urine and plasma. In this assay, 3-DG and 3-DF are reduced to 3-deoxy-hexitols (3-DH), using either NaBH4 or NaBD4, and then analyzed by selected ion monitoring gas chromatography-mass spectrometry. Based on comparative analysis of samples reduced with NaBD4 versus NaBD4, 3-DH in urine was derived exclusively (greater than 99%) from 3-DF, while 3-DG accounted for approximately 15% of 3-DH in plasma. The concentrations of 3-DH in fasting human urine and plasma were 5.3 +/- 1.5 micrograms/mg creatinine (n = 18) and 7.2 +/- 1.7 micrograms/dl (n = 18), respectively. The concentrations of 3-DG and 3-DF in plasma (n = 7) were 1.0 +/- 0.2 and 6.7 +/- 1.6 micrograms/dl, respectively. These results suggest that several milligrams of 3-DG are formed in the body per day and detoxified by reduction to 3-DF and support the role of 3-DG as an intermediate in the browning of protein via the Maillard reaction in vivo.