RESUMO
BACKGROUND: Respiratory Distress Syndrome (RDS) is a prematurity-related breathing disorder caused by a quantitative deficiency of pulmonary surfactant. Surfactant replacement therapy is effective for RDS newborns, although treatment failure has been reported. The aim of this study is to trace exogenous surfactant by 13C variation and estimate the amount reaching the lungs at different doses of the drug. METHODS: Forty-four surfactant-depleted rabbits were obtained by serial bronchoalveolar lavages (BALs), that were merged into a pool (BAL pool) for each animal. Rabbits were in nasal continuous positive airway pressure and treated with 0, 25, 50, 100 or 200 mg/kg of poractant alfa by InSurE. After 90 min, rabbits were depleted again and a new pool (BAL end experiment) was collected. Disaturated-phosphatidylcholine (DSPC) was measured by gas chromatography. DSPC-Palmitic acid (PA) 13C/12C was analyzed by isotope ratio mass spectrometry. One-way non-parametric ANOVA and post-hoc Dunn's multiple comparison were used to assess differences among experimental groups. RESULTS: Based on DSPC-PA 13C/12C in BAL pool and BAL end experiment, the estimated amount of exogenous surfactant ranged from 61 to 87% in dose-dependent way (p < 0.0001) in animals treated with 25 up to 200 mg/kg. Surfactant administration stimulated endogenous surfactant secretion. The percentage of drug recovered from lungs did not depend on the administered dose and accounted for 31% [24-40] of dose. CONCLUSIONS: We reported a risk-free method to trace exogenous surfactant in vivo. It could be a valuable tool for assessing, alongside the physiological response, the delivery efficiency of surfactant administration techniques.
Assuntos
Produtos Biológicos/metabolismo , Isótopos de Carbono/metabolismo , Pulmão/metabolismo , Fosfolipídeos/metabolismo , Surfactantes Pulmonares/metabolismo , Animais , Produtos Biológicos/administração & dosagem , Isótopos de Carbono/administração & dosagem , Relação Dose-Resposta a Droga , Pulmão/efeitos dos fármacos , Masculino , Fosfolipídeos/administração & dosagem , Surfactantes Pulmonares/administração & dosagem , Coelhos , Tensoativos/administração & dosagem , Tensoativos/metabolismoRESUMO
This study reports the development of an all-in-one elemental analyser isotope ratio mass spectrometry (EA-IRMS) system modified for simultaneous analysis of dissolved organic carbon (DOC) concentration and its stable carbon isotope footprint (δ13CDOC) in aqueous samples. The method involves a quantitative oxidation of DOC in a 200 µL liquid sample to CO2, after sample acidification and stripping by nitrogen. The detection limit of the method for DOC quantification was 0.2 mg C/L with an analytical precision of 12 %. Uncertainty of stable isotope determinations was 2 % at 0.2 mg DOC/L, while decreasing to 0.3 % at 20 mg DOC/L. Quantitative oxidation of DOC in aqueous samples was validated by using ring test water samples and Deep Sea reference seawater. The method performances of isotope analysis were evaluated by analysing different isotopic standard solutions. The applicability of the method was tested through the analysis of different environmental types of water, showing that δ13CDOC ranged from - 23.30 to -31.85 , allowing to characterize samples of different environmental origin. The developed method offers several advantages including rapidity, use of small sample volumes and minimal sample pre-treatment, making it a valuable tool for routine DOC concentration measurements paired with isotopic characterization.
Assuntos
Carbono , Matéria Orgânica Dissolvida , Carbono/análise , Isótopos de Carbono/análise , Espectrometria de Massas/métodos , ÁguaRESUMO
Plastic waste is a growing global environmental problem, particularly in the marine ecosystems, in consideration of its persistence. The monitoring of the plastic waste has become a global issue, as reported by several surveillance guidelines proposed by Regional Sea Conventions (OSPAR, UNEP) and appointed by the EU Marine Strategy Framework Directive. Policy responses to plastic waste vary at many levels, ranging from beach clean-up to bans on the commercialization of plastic bags and to Regional Plans for waste management and recycling. Moreover, in recent years, the production of plant-derived biodegradable plastic polymers has assumed increasing importance. This study reports the first preliminary characterization of carbon stable isotopes (δ13C) of different plastic polymers (petroleum- and plant-derived) in order to increase the dataset of isotopic values as a tool for further investigation in different fields of polymers research as well as in the marine environment surveillance. The δ13C values determined in different packaging for food uses reflect the plant origin of "BIO" materials, whereas the recycled plastic materials displayed a δ13C signatures between plant- and petroleum-derived polymers source. In a preliminary estimation, the different colours of plastic did not affect the variability of δ13C values, whereas the abiotic and biotic degradation processes that occurred in the plastic materials collected on beaches and in seawater, showed less negative δ13C values. A preliminary experimental field test confirmed these results. The advantages offered by isotope ratio mass spectrometry with respect to other analytical methods used to characterize the composition of plastic polymers are: high sensitivity, small amount of material required, rapidity of analysis, low cost and no limitation in black/dark samples compared with spectroscopic analysis.
Assuntos
Biopolímeros/análise , Monitoramento Ambiental/métodos , Petróleo/análise , Plásticos/análise , Resíduos Sólidos/análise , Poluentes Químicos da Água/análise , Isótopos de Carbono/análise , Ecossistema , Látex/análise , Espectrometria de Massas/métodosRESUMO
The aim of the study was to determine surfactant palmitate disaturated-phosphatidylcholine (DSPC-PA) synthesis in vivo in humans by the incorporation of deuterium from total body water into DSPC-PA under steady state condition. We studied three newborns and one infant (body weight (BW) 4.6 +/- 2.9 kg, gestational age 37.5 +/- 2 weeks, age 9 +/- 9 days) and four preterm newborns (BW 1.3 +/- 0.6 kg, gestational age 30.3 +/- 2.5 weeks, postnatal age 8.8 +/- 9.2 h). All infants were mechanically ventilated during the study and the four preterm infants received exogenous surfactant at the start of the study. We administered 0.44 g (2)H(2)O/kg BW as a bolus intravenously, followed by 0.0125 g (2)H(2)O/kg BW every 6 h to maintain deuterium enrichment at plateau over 2 days. Urine samples and tracheal aspirates (TA) were obtained prior to dosing and every 6 h thereafter. Isotopic enrichment curves of DSPC-PA from sequential TA and urine deuterium enrichments were analyzed by Gas Chromatography-Isotope Ratio-Mass Spectrometry (GC-IRMS) and normalized for Vienna Standard Mean Ocean Water. Enrichment data were used to measure DSPC-PA fractional synthesis rate (FSR) from the linear portion of the DSPC-PA enrichment rise over time, relative to plateau enrichment of urine deuterium. Secretion time (ST) was defined as the time lag between the start of the study and the appearance of DSPC-PA deuterium enrichment in TA. Data were given as mean +/- SD. All study infants reached deuterium-steady state in urine. DSPC-PA FSR was 6.5 +/- 2.8%/day (range 2.6-10.2). FSR for infants who did not receive exogenous surfactant was 5.7 +/- 3.5%/day (range 2.6-9.9%/day) and 7.3 +/- 2.1%/day (range 5.1-10.2%/day) in the preterms, whereas DSPC-PA ST was 10 +/- 10 h and 31 +/- 10 h respectively. Surfactant DSPC-PA synthesis can be measured in humans by the incorporation of deuterium from body water. This study is a simpler and less invasive method compared to previously published methods on surfactant kinetics by means of stable isotopes.
Assuntos
Água Corporal/química , Deutério/metabolismo , Fosfatidilcolinas/biossíntese , Surfactantes Pulmonares/metabolismo , Deutério/administração & dosagem , Deutério/análise , Deutério/farmacocinética , Feminino , Humanos , Recém-Nascido , Cinética , Masculino , Fosfatidilcolinas/análise , Respiração ArtificialRESUMO
Deuterium (2H) in water and urine can be measured by off-line and, more recently, on-line techniques using isotope ratio mass spectrometry (IRMS). We describe a new simple on-line pyrolysis method for the analysis of 2H/1H in water and urine samples by continuous flow IRMS, normally used for 2H/1H measurements in organic compounds. A deactivated column connected the split injector to a high-temperature conversion reactor (TC HD), and 0.5 microL of sample was injected. Accuracy and precision were determined with Vienna Standard Mean Ocean Water (VSMOW), Standard Light Antarctic Precipitation (SLAP), and Greenland Ice Sheet Precipitation (GISP). The range of linearity was measured with a calibration curve of enriched water from 0 up to 0.1 atom percent excess (APE) (i.e. -72 up to 6323 delta per mil (deltaD per thousand)) with a precision of <5 per thousand and accuracy ranging between 1 and 55 per thousand. Blinded reanalysis of urine samples by an equilibration device (Gas Bench) and by a dedicated pyrolysis system (TC/EA) was performed and results compared by the Bland-Altman test. Enrichments ranged between 600 and 2400 per thousand deltaD(VSMOW) with a precision of +/-5 per thousand. Urine enrichments described by our method were strongly correlated with values obtained by Gas Bench and TC/EA (p < 0.0001). There was a significant memory effect that was reduced by injecting the sample 15 times and discarding the first 10 injections, together with accurate furnace conditioning and appropriate cleaning of the syringe. Data indicate that the method is accurate, and that it can be used for water and urine deuterium determination when a Gas Bench or TC/EA instrument is not available and the amount of sample is limited.