RESUMEN
Developing synthetic materials, with enzyme-like molecular recognition capabilities, as functional receptors in electronic or electrochemical devices for the timely diagnosis of major diseases is a great challenge. Herein, we present the development of Fe/MIPpy nanozymes, characterized as enzyme-like artificial receptors, for the precise and non-invasive monitoring of cancer biomarkers in aqueous solutions and human saliva. Through the integration of PVA-stabilized FeFe2O4 nanocrystals in a molecularly imprinted conducting polypyrrole matrix, the Fe/MIPpy nanozymes demonstrate 424 nA cm-2 nM-1 sensitivity and 220 pM detection limit. Charge-transfer mechanisms, Fe/MIPpy-spermine interactions, and the principle of spermine recognition are investigated by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The disposable Fe/MIPpy sensor operates wirelessly and offers rapid and remote quantification of spermine, making it a promising material for the development of cost-effective tools for non-invasive cancer diagnosis and prognosis.
Asunto(s)
Técnicas Electroquímicas , Polímeros , Pirroles , Espermina , Humanos , Espermina/química , Espermina/análisis , Pirroles/química , Polímeros/química , Neoplasias/diagnóstico , Catálisis , Compuestos Férricos/química , Pronóstico , Propiedades de Superficie , Tamaño de la PartículaRESUMEN
Small fruits such as strawberries, are a good source of natural antioxidants. In recent decades, many efforts have been made to increase the shelf life of strawberries and maintain its nutritional value in post-harvest conditions. In the present study, the effects of spermine (Spm) and spermidine (Spd) (0, 1.0 and 1.5 mM) on the post-harvest life and quality of strawberry fruits during the 3rd, 6th, and 12th days of storage, were investigated. Applications of Spm and Spd decreased the rate of weight loss, fruit decay, soluble solids content, fruit juice pH and taste index during the storage period in compared to the control. However, titratable acids and vitamin C contents, tissue stiffness, phenolic compounds and antioxidant activity increased in compared to the control. These growth regulators prevented the aging and loss of bioactive compounds of the fruit by increasing the antioxidant activity and preventing the destruction of the fruit tissue. Among the studied treatments, applications of 1.5 mM of Spm and Spd were the most effective treatments to enhance the storage life and quality characters of strawberry fruits.
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Fragaria , Espermidina , Espermidina/farmacología , Espermidina/análisis , Espermina/farmacología , Espermina/análisis , Antioxidantes/farmacología , Antioxidantes/análisis , FrutasRESUMEN
(1) Background: Spermidine is a biogenic polyamine that plays a crucial role in mammalian metabolism. As spermidine levels decline with age, spermidine supplementation is suggested to prevent or delay age-related diseases. However, valid pharmacokinetic data regarding spermidine remains lacking. Therefore, for the first time, the present study investigated the pharmacokinetics of oral spermidine supplementation. (2) Methods: This study was designed as a randomized, placebo-controlled, triple-blinded, two-armed crossover trial with two 5-day intervention phases separated by a washout phase of 9 days. In 12 healthy volunteers, 15 mg/d of spermidine was administered orally, and blood and saliva samples were taken. Spermidine, spermine, and putrescine were quantified by liquid chromatography-mass spectrometry (LC-MS/MS). The plasma metabolome was investigated using nuclear magnetic resonance (NMR) metabolomics. (3) Results: Compared with a placebo, spermidine supplementation significantly increased spermine levels in the plasma, but it did not affect spermidine or putrescine levels. No effect on salivary polyamine concentrations was observed. (4) Conclusions: This study's results suggest that dietary spermidine is presystemically converted into spermine, which then enters systemic circulation. Presumably, the in vitro and clinical effects of spermidine are at least in part attributable to its metabolite, spermine. It is rather unlikely that spermidine supplements with doses <15 mg/d exert any short-term effects.
Asunto(s)
Espermidina , Espermina , Animales , Adulto , Humanos , Espermidina/análisis , Espermina/análisis , Putrescina/metabolismo , Cromatografía Liquida , Saliva/química , Espectrometría de Masas en Tándem , Poliaminas/metabolismo , Plasma/química , Suplementos Dietéticos/análisis , Mamíferos/metabolismoRESUMEN
The evaluation of the biogenic amines (BAs) profile of different types of craft beers is herein presented. A previously developed and validated analytical method based on ion-pair chromatography coupled with potentiometric detection was used to determine the presence of 10 BAs. Good analytical features were obtained for all amines regarding linearity (R2 values from 0.9873 ± 0.0015 to 0.9973 ± 0.0015), intra- and inter-day precision (RSD lower than 6.9% and 9.7% for beer samples, respectively), and accuracy (recovery between 83.2-108.9%). Detection and quantification limits range from 9.3 to 60.5 and from 31.1 to 202.3 µg L-1, respectively. The validated method was applied to the analysis of four ale beers and one lager craft beer. Ethylamine, spermidine, spermine, and tyramine were detected in all analyzed samples while methylamine and phenylethylamine were not detected. Overall, pale ale beers had a significantly higher total content of BAs than those found in wheat pale and dark samples. A general least square regression model showed a good correlation between the total content of BAs and the brewing process, especially for Plato degree, mashing, and fermentation temperatures. Knowledge about the type of ingredients and manufacturing processes that contribute to higher concentrations of these compounds is crucial to ensuring consumer safety.
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Cerveza , Aminas Biogénicas , Cerveza/análisis , Aminas Biogénicas/análisis , Cromatografía Líquida de Alta Presión/métodos , Espermidina/análisis , Espermina/análisisRESUMEN
The polyamine content of human breast milk, which is the first exogenous source of polyamines for the newborn, can be affected by several factors associated with the mother, the infant, or breastfeeding itself. The aim of this study was to evaluate the influence of different breastfeeding factors on the polyamines found in human milk. For this study, a cohort of 83 mothers was considered for up to 4 months, and a subgroup of 33 mothers were followed during the first six months of breastfeeding. Two breast milk samples were collected at each sampling point (foremilk and hindmilk) and the polyamine content was determined by UHPLC-FL. Polyamine levels varied considerably between the mothers and tended to decrease over time. Putrescine was the minor polyamine, whereas spermidine and spermine contents were very similar. The concentrations of the three polyamines were significantly higher in hindmilk than foremilk (p < 0.001). Spermidine and spermine levels decreased significantly through the lactation progress (p < 0.05). Finally, slightly higher levels of polyamines were observed in the milk of mothers providing partial, rather than full, breastfeeding, although the differences were not significant. The polyamine content in human milk was found to change during a single feed (foremilk versus hindmilk) and as lactation progressed, mainly in response to the specific circumstances of the newborn.
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Lactancia Materna/métodos , Leche Humana/química , Poliaminas/análisis , Adolescente , Adulto , Factores de Edad , Peso al Nacer , Índice de Masa Corporal , Cromatografía Líquida de Alta Presión , Estudios de Cohortes , Parto Obstétrico/métodos , Femenino , Humanos , Lactante , Recién Nacido , México , Madres , Poliaminas/química , Putrescina/análisis , Espermidina/análisis , Espermina/análisis , Adulto JovenRESUMEN
We developed a facile detection method of spermine based on the fluorescence (FL) quenching of the ciprofloxacin-Tb3+ complex, which shows astrong green emission. Ciprofloxacin (CP) makes efficient bondings to Tb3+ ion as a linker molecule through carboxylic and ketone groups to form a kind of lanthanide coordination polymer. The addition of spermine that competes with Tb3+ ions for the interaction with CP due to its positive charge brings about weakened coordination linkage of CP and Tb3+. The probe exhibited high sensitivity, selectivity, and good linearity in the range of 2-180 µM with a low limit of detection of 0.17 µM. Moreover, we applied this method on the paper strip test (PST), along with the integration of a smartphone and Arduino-based device. The practical reliability of the developed probe was evaluated on human serum samples with acceptable analytical results.
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Ciprofloxacina/química , Colorantes Fluorescentes/química , Elementos de la Serie de los Lantanoides/química , Polímeros/química , Espectrometría de Fluorescencia/métodos , Espermina/análisis , Terbio/química , Cationes/química , Humanos , Reproducibilidad de los Resultados , Espectrometría de Fluorescencia/instrumentaciónRESUMEN
A method for the determination of 8 biogenic amines in aquatic products and their derived products was established by HPLC-MS/MS without derivatization. The samples were extracted by 5% perchloric acid solution. N-hexane was used to clean the extract. The analytes were separated by a column of ACQUITY UPLC HSS T3 (100 mm × 2.1 mm, 1.8 µm), and gradient eluted with a mixed solution of (0.5% formic acid) and acetonitrile. Good linearity was obtained with correlation coefficients (R2) >0.99. This method achieved higher sensitivity (from 0.1 mg/kg for tyramine, 2-phenylethylamine and tryptamine to 1.0 mg/kg for spermidine, spermine, cadaverin, histamine and putrescine). The average recoveries were demonstrated in the range of 70.9%-113.1%, with relative standard deviations (RSDs) from 0.33% to 10.81%. This method was suitable for the detection of BAs in aquatic products and their products.
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Aminas Biogénicas/análisis , Cromatografía Líquida de Alta Presión/métodos , Alimentos Marinos/análisis , Espectrometría de Masas en Tándem/métodos , Cadaverina/análisis , Histamina/análisis , Fenetilaminas/análisis , Putrescina/análisis , Espermidina/análisis , Espermina/análisis , Triptaminas/análisis , Tiramina/análisisRESUMEN
AIM: To assess the applicability of the novel technique based on the detection of spermine in solutions by spectrocolorimetric method using gold and silver colloidal nanoparticles. MATERIALS AND METHODS: Colloidal solution of gold nanoparticles were synthesized by chemical reduction of tetrachlorauric acid with trisodium citrate. Colloidal solution of silver nanoparticles was obtained by chemical reduction of silver nitrate with tryptophan. The absorption spectra of gold/silver metal colloids and their mixtures with polyamines were recorded. RESULTS: The increase of spermine concentration in solution caused the change in the intensity of the band of localized surface plasmon resonance that was not affected by the excess of spermidine. The color shift in colloidal gold due to its aggregation with spermine was registered spectrophotometrically. CONCLUSION: The principal possibility of selective quantification of spermine in the presence of spermidine in extremely high concentration using colloidal gold has been shown. This method can be used to assay selectively spermine in biological fluids.
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Oro , Nanopartículas del Metal , Plata , Espermina/análisis , Humanos , Espectrofotometría/métodos , Espermidina/análisisRESUMEN
During maize production, drought throughout the flowering stage usually induces seed abortion and yield losses. The influence of postpollination drought stress on seed abortion and its underlying mechanisms are not well characterized. By intervening in the competition for assimilates between kernel siblings under different degrees of postpollination drought stresses accompanied by synchronous pollination (SP) and incomplete pollination (ICP) approaches, the mechanisms of postpollination abortion were investigated at physiological and molecular levels. Upon SP treatment, up to 15% of the fertilized apical kernels were aborted in the drought-exacerbated competition for assimilates. The aborted kernels exhibited weak sucrose hydrolysis and starch synthesis but promoted the synthesis of trehalose-6-phosphate and ethylene. In ICP where basal pollination was prevented, apical kernel growth was restored with reinstated sucrose metabolism and starch synthesis and promoted sucrose and hexose levels under drought stress. In addition, the equilibrium between ethylene and polyamine in response to the drought and pollination treatments was associated with the abortion process. We conclude that competition for assimilates drives postpollination kernel abortion, whereas differences in sugar metabolism and the equilibrium between ethylene and polyamines may be relevant to the "live or die" choice of kernel siblings during this competition.
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Grano Comestible/fisiología , Zea mays/fisiología , Carbohidratos/análisis , Deshidratación , Grano Comestible/química , Grano Comestible/crecimiento & desarrollo , Etilenos/metabolismo , Fotosíntesis/fisiología , Hojas de la Planta/fisiología , Polinización/fisiología , Putrescina/análisis , Espermidina/análisis , Espermina/análisis , Agua/metabolismo , Zea mays/crecimiento & desarrolloRESUMEN
BACKGROUND: Nitric oxide (NO) and abscisic acid (ABA) are important regulators of plant response to cold stress, and they interact in response to cold signals. The primary goal of this study was to determine the roles of exogenous NO and ABA on the synthesis of endogenous NO and ABA in cold-stored peach fruit. RESULTS: Exogenous NO and ABA maintained a relatively high content of NO, increased nitrate reductase (NR) activity, and inhibited the activity of NO synthase (NOS)-like and the levels of polyamine biosynthesis in peaches during cold storage. Treatments of potassium 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO), NO, N-nitro-l-Arg-methyl ester (L -NAME), and sodium tungstate did not influence ABA content. Exogenous ABA increased the content of carotenoids and the activities of aldehyde oxidase (AO), 9-cis-epoxycarotenoid dioxygenase (NCED), and zeaxanthin epoxidase (ZEP) of ABA synthesis in peaches during cold storage, and upregulated the gene expression of PpAO1, PpNCED1, PpNCED2, and PpZEP. The production of endogenous NO was differentially inhibited by NO scavengers, ABA inhibitors, and NR inhibitors, but not affected by NOS-like inhibitors during cold storage. CONCLUSION: Exogenous NO and ABA can induce endogenous NO synthesis in cold-stored peaches by the nitrate reductase pathway, and ABA can mediate endogenous ABA synthesis by the autocatalytic reaction. NO does not regulate ABA synthesis. © 2019 Society of Chemical Industry.
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Ácido Abscísico/metabolismo , Óxido Nítrico/metabolismo , Prunus persica/metabolismo , Adenosilmetionina Descarboxilasa/metabolismo , Aldehído Oxidasa , Arginina/análisis , Carboxiliasas/metabolismo , Dioxigenasas , Almacenamiento de Alimentos , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Nitratos/análisis , Óxido Nítrico Sintasa/metabolismo , Nitritos/análisis , Oxidorreductasas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Espermidina/análisis , Espermina/análisisRESUMEN
AIM: To analyze the performance of biosensor based on nanoparticles of zinc oxide for the detection of spermine and spermidine in solution and in cell culture. MATERIALS AND METHODS: Zinc oxide nanoparticles were used for preparing biosensor containing antibodies to spermine and spermidine. Polyamine concentration is solutions of spermine and spermidine as well as in lyophilisate of MCF-7 cells was measured by luminescence of the samples excited by laser beam at 380 nm. RESULTS: The minimum concentration for the detection of polyamines in model solutions is 10 ng/ml, and maximum one is 100 ng/ml. A higher level of luminescence intensity of nanoparticles was found during analysis the polyamines in MCF-7 lyophilisate allowing for detecting polyamines at concentrations from 100 cells/ml to 100,000 cells/ml. CONCLUSIONS: The proposed biosensor system for determining the level of biogenic polyamines in cell lyophilisate using the optical properties of zinc oxide nanoparticles is promising for further improvement of the methodology and its implementation for detection and measurement of polyamines in biological systems.
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Anticuerpos Inmovilizados/química , Técnicas Biosensibles/métodos , Nanopartículas/química , Poliaminas/análisis , Óxido de Zinc/química , Humanos , Células MCF-7 , Espermidina/análisis , Espermina/análisisRESUMEN
The sustainable development of point-of-care testing (POCT) for spermine detection is important to check for food spoilage, early diagnosis of various malignancies and diminished anticonvulsant drug carbamazepine response in chronic epilepsy. Herein, the synthesis, characterization and spectroscopic properties of perylene diimide EA-PDIâ©Cu2+ complex based nanoparticles towards spermine were studied in detail. This EA-PDIâ©Cu2+ complex can be used for the ultrasensitive detection of spermine as low as 86.3 nM (UV-vis) and 90 pM (fluorescence) in aqueous medium, in urine and blood serum samples (recovery 99 ± 3) and in the solid state (0.1 µg L-1), and EA-PDI shows minimal cytotoxicity to cells and can easily enter into Human Osteosarcoma MG-63 cells for bio-imaging of Cu2+ and spermine. This EA-PDIâ©Cu2+ complex can be established as a cost-effective method to develop a diagnostic kit for POCT of spermine in terms of a solution-based test kit for real time detection of spermine in vapor and solution form released from fermented food samples.
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Cobre/química , Colorantes Fluorescentes/química , Contaminación de Alimentos/análisis , Imidas/química , Nanopartículas/química , Perileno/análogos & derivados , Pruebas en el Punto de Atención , Juego de Reactivos para Diagnóstico , Espermina/análisis , Colorantes Fluorescentes/síntesis química , Humanos , Tamaño de la Partícula , Perileno/química , Propiedades de SuperficieRESUMEN
Spermine contamination ranks as one of the food safety issues, it will cause some adverse reactions if the intake of spermine is excessive in human body. So it is of great significance to establish fast and efficient analysis method to detect spermine in foods. In this study, the spermine aptamers with high affinity and specificity were obtained by the capture systematic evolution of ligands by exponential enrichment (Capture-SELEX) technique. Forty-one aptamer sequences were obtained by cloning and sequencing, and were divided into eight families based on homology and secondary structure analysis. The affinity and specificity of candidate aptamers was analyzed by isothermal titration calorimetry (ITC) and fluorescence assay. The aptamers named APJ-6 was picked out as the optimal aptamer that recognizes spermine specifically with the Kd value of 9.648⯱â¯0.896â¯nM. In order to verify the practicability of the selected aptamers, the sensitive aptamer-based fluorescene assay was designed. Under optimized conditions, this aptasensor exhibited a low detection limit of 0.052â¯nM, as well as a linear within the range of 0.1-20â¯nM. Besides, it has been further applied for the determination of spermine in pork samples and the recoveries ranged from 86.45% to 98.15%, showing its great potential for sensitive analysis in food safety control.
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Aptámeros de Nucleótidos/química , ADN de Cadena Simple/química , Espermina/análisis , Animales , Contaminación de Alimentos/análisis , Límite de Detección , Carne de Cerdo/análisis , Técnica SELEX de Producción de Aptámeros/métodos , PorcinosRESUMEN
Bioactive amines are found in food and can be relevant for the assessment of fruits shelf life and nutritional quality. The pulp and peel of 20 banana and plantain were analyzed and the bioactive amine content varied according to the genotype, ripening stage, fruit tissue and thermal processing. In most of the analyzed genotypes, tyramine, histamine, dopamine, serotonin, spermidine, and spermine were decreased during the ripening process in the pulps. By contrast, there was an increase in putrescine level. In many genotypes of plantains, the serotonin and dopamine contents in pulp decreased until stage 5 and increased at stage 7. Peels contain higher levels of serotonin, dopamine, histamine and tyramine than pulps. Additionally, thermal processing affects the content of amines present in fruit. Boiling with the peel should be preferred in domestic preparations, regardless of the genotype used.
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Aminas/análisis , Frutas/metabolismo , Musa/metabolismo , Plantago/metabolismo , Aminas Biogénicas/análisis , Dopamina/análisis , Frutas/química , Genotipo , Histamina/análisis , Musa/química , Musa/genética , Plantago/química , Plantago/genética , Putrescina/análisis , Serotonina/análisis , Espermidina/análisis , Espermina/análisis , Temperatura , Tiramina/análisisRESUMEN
Non-covalent interaction in the binary systems of polyamines (putrescine, spermidine, spermine) with citric acid and complex formation in the binary as well as ternary systems of lanthanide(III) ions, citric acid and polyamine have been investigated. The studies were performed in aqueous solution. The overall stability constants of the complexes were determined using the potentiometric method with computer analysis of the data. Only mononuclear type of complexes were found in the ternary systems and polyamines were located in the outer as well as inner coordination sphere. Non-covalent interaction between biogenic amines and citric acid in the binary and ternary systems were confirmed on the basis of the equilibrium constants analysis and spectroscopic studies.
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Ácido Cítrico/química , Complejos de Coordinación/química , Elementos de la Serie de los Lantanoides/química , Sustancias Luminiscentes/química , Poliaminas/química , Concentración de Iones de Hidrógeno , Luminiscencia , Mediciones Luminiscentes , Poliaminas/análisis , Putrescina/análisis , Putrescina/química , Espermidina/análisis , Espermidina/química , Espermina/análisis , Espermina/químicaRESUMEN
Twenty-two compounds belonging to several classes of polyamine analogs have been examined for their ability to inhibit the growth of the human malaria parasite Plasmodium falciparum in vitro and in vivo. Four lead compounds from the thiourea sub-series and one compound from the urea-based analogs were found to be potent inhibitors of both chloroquine-resistant (Dd2) and chloroquine-sensitive (3D7) strains of Plasmodium with IC50 values ranging from 150 to 460 nM. In addition, the compound RHW, N1,N7-bis (3-(cyclohexylmethylamino) propyl) heptane-1,7-diamine tetrabromide was found to inhibit Dd2 with an IC50 of 200 nM. When RHW was administered to P. yoelii-infected mice at 35 mg/kg for 4 days, it significantly reduced parasitemia. RHW was also assayed in combination with the ornithine decarboxylase inhibitor difluoromethylornithine, and the two drugs were found not to have synergistic antimalarial activity. Furthermore, these inhibitors led to decreased cellular spermidine and spermine levels in P. falciparum, suggesting that they exert their antimalarial activities by inhibition of spermidine synthase.
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Antimaláricos/farmacología , Plasmodium falciparum/efectos de los fármacos , Poliaminas/farmacología , Espermidina/análisis , Espermina/análisis , Animales , Antimaláricos/administración & dosificación , Modelos Animales de Enfermedad , Sinergismo Farmacológico , Concentración 50 Inhibidora , Malaria/tratamiento farmacológico , Ratones , Carga de Parásitos , Parasitemia , Pruebas de Sensibilidad Parasitaria , Plasmodium falciparum/química , Plasmodium falciparum/crecimiento & desarrollo , Plasmodium yoelii/efectos de los fármacos , Poliaminas/administración & dosificaciónRESUMEN
Shuidouchi is a traditional Chinese fermented soybean product and its quality is largely affected by the microbes involved in the fermentation. In this study, eleven Shuidouchi samples were collected from southwest China and the microbial diversity and its correlations with chemical characteristics were investigated. Bacterial community was detected using 16S rRNA sequencing, along with bacterial and fungal viable plate counts. Biogenic amines and other chemical characteristics were determined by HPLC and corresponding chemical reaction methods. Among eleven Shuidouchi samples, 21 phyla and 356 genera were identified. Firmicutes, Bacteroidetes and Proteobacteria were the predominant phyla while Bacillus, Bacteroides and Lactobacillus were the main genera. The average cell number of bacteria, lactic acid bacteria and fungi were 1.6â¯×â¯106, 5.9â¯×â¯104 and 7.6â¯×â¯103â¯CFU/g, respectively. HPLC results showed that the mean concentration of tryptamine, ß-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine were 23.11, 3.66, 12.21, 7.12, 8.13, 22.98, 24.72, and 39.00â¯mg/kg, respectively. The average content of other characteristics including amino acid nitrogen, titratable acidity, and reducing sugar were 2.08, 3.44, and 25.78â¯g/kg, respectively. Shuidouchi samples were slightly acidic or neutral. Fibrinolytic enzyme activity was detected only in one sample. Among top 52 identified genera, 9 genera showed positive correlations with the chemical characteristics of Shuidouchi while 15 genera were negatively associated. Our results indicated that Shuidouchi contained rich microbial resources and were edible safety based on the tested indexes. The associations identified between microbes and chemical characteristics could be further utilized in the food fermentation industry.
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Biodiversidad , Alimentos Fermentados/análisis , Alimentos Fermentados/microbiología , Microbiología de Alimentos , Glycine max/química , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Aminas Biogénicas/análisis , Cadaverina/análisis , China , Cromatografía Líquida de Alta Presión , Recuento de Colonia Microbiana , Fermentación , Hongos/clasificación , Hongos/genética , Histamina/análisis , Fenetilaminas/análisis , Putrescina/análisis , ARN Ribosómico 16S/genética , Espermidina/análisis , Espermina/análisis , Triptaminas/análisis , Tiramina/análisisRESUMEN
A new method for determination of underivatized biogenic amines in cheese based on ion exchange chromatography coupled with tandem mass spectrometric detection was proposed. The method was applied to the analysis of 10 biogenic amines (trimethylamine, putrescine, cadaverine, histamine, 2-phenylethylamine, spermine, spermidine, tryptamine, agmatine, and tyramine) in different types of cheese. The amines were extracted only with water without any additional derivatization step or sample cleanup. This is a great advantage in terms of simplicity of sample pretreatment procedure compared with other currently existing methods in the literature. Biogenic amines were separated using cation exchange column, under gradient elution conditions by mixing formic acid (1.00 M) and deionized water. Detection was achieved using tandem MS/MS, with the instrument set into multiple reaction monitoring mode to ensure high specificity. The detection and quantification limits were in the ranges of 12-46 µg/L and 40-153 µg/L, respectively. The exceptions were spermidine and spermine, with detection limits of 0.8 and 5.4 mg/L, respectively. The linearity for most of the biogenic amines was from 10 µg/L up to 10 mg/L. The best recoveries were observed for trimethylamine, tyramine, and cadaverine, and were 89, 94, and 102%, respectively. The results showed that this method can be used for routine determination of biogenic amines in different types of cheeses as well other food matrices. It must be stressed that the proposed method is capable of determining 10 biogenic amines, including tyramine, which is reported to cause food intoxication commonly associated with cheeses.
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Aminas Biogénicas/análisis , Queso/análisis , Cromatografía por Intercambio Iónico/métodos , Análisis de los Alimentos/métodos , Espectrometría de Masas en Tándem/métodos , Cadaverina/análisis , Límite de Detección , Metilaminas/análisis , Espermidina/análisis , Espermina/análisis , Tiramina/análisisRESUMEN
We developed a competitive fluorescent molecularly imprinted polymer (MIP) assay to detect biogenic amines in fish samples. MIPs synthesized by precipitation polymerization using histamine as template were used in a batch binding assay analogous to competitive fluoroimmunoassays. Introducing a complex sample matrix, such as fish extract, into the assay changes the environment and the binding conditions, therefore the importance of the sample preparation is extensively discussed. Several extraction and purification methods for fish were comprehensively studied, and an optimal clean-up procedure for fish samples using liquid-liquid extraction was developed. The feasibility of the competitive MIP assay was shown in the purified fish extract over a broad histamine range (1 - 430µM). The MIP had the highest affinity towards histamine, but recognized also the structurally similar biogenic amines tyramine and tryptamine, as well as spermine and spermidine, providing simultaneous analysis and assessment of the total amount of biogenic amines.
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Aminas Biogénicas/análisis , Peces , Fluoroinmunoensayo/métodos , Impresión Molecular , Polímeros/química , Animales , Aminas Biogénicas/química , Aminas Biogénicas/aislamiento & purificación , Reproducibilidad de los Resultados , Alimentos Marinos/análisis , Espermidina/análisis , Espermidina/química , Espermidina/aislamiento & purificación , Espermina/análisis , Espermina/química , Espermina/aislamiento & purificación , Triptaminas/análisis , Triptaminas/química , Triptaminas/aislamiento & purificación , Tiramina/análisis , Tiramina/química , Tiramina/aislamiento & purificaciónRESUMEN
Thermospermine, a structural isomer of spermine, is widely spread in the plant kingdom and has recently been shown to play a key role in the repression of xylem differentiation in vascular plants. However, a standard high-performance liquid chromatography (HPLC) protocol for detecting polyamines as their dansyl derivative cannot distinguish themospermine from spermine. These isomers become separated from each other after benzoylation. In this chapter, we describe a simple protocol for extraction, benzoylation, and HPLC detection of thermospermine and spermine with other polyamines from plant material.