Quantitation of ethyl glucuronide in serum & urine by gas chromatography - mass spectrometry.

Background & objectives: Alcohol misuse has now become a serious public health problem and early intervention is important in minimizing the harm. Biochemical markers of recent and high levels of alcohol consumption can play an important role in providing feedback regarding the health consequences of alcohol misuse. Existing markers are not sensitive to recent consumption and in detecting early relapse. Ethyl glucuronide (EtG), a phase-II metabolite of ethanol is a promising marker of recent alcohol use and can be detected in body fluids. In this study an analytical technique for quantitation of EtG in body fluids using solid-phase extraction (SPE) and gas chromatography (GC) with mass spectrometric detection (MS) was developed and validated. Methods: De-proteinization of serum and urine samples was done with perchloric acid and hydrochloric acid, respectively. Serum samples were passed through phospholipids removal cartridges for further clean up. EtG was isolated using amino propyl solid phase extraction columns. Chromatographic separation was achieved by gas chromatography with mass spectrometry. Results: Limit of detection and limit of quantitation were 50 and 150 ng/ml for urine and 80 and 210 ng/ml for serum, respectively. Signal to noise ratio was 3:1, mean absolute recovery was 80-85 per cent. Significant correlation was obtained between breath alcohol and serum EtG levels (r=0.853) and urine EtG and time since last abuse (r = -0.903) in clinical samples. Interpretation & conclusions: In the absence of other standardized techniques to quantitate EtG in biological samples, this gc -ms method was found to have high throughput and was sensitive and specific.

Determination of ethyl glucuronide in blood by GC-MS/MS / 法医学杂志

OBJECTIVE@#To develop a method for determining ethyl glucuronide(EtG) in human blood with gas chromatograph-tandem mass spectrometry (GC-MS/MS).@*METHODS@#Human blood protein was precipitated with acetonitrile. The supernatant was transferred and air flow dried after centrifugated. The residue was derived with N, O-bis (trimethylsilyl)trifluoroacetamide (BSTFA), and analyzed with GC-MS/MS.@*RESULTS@#The detection limit of EtG in blood was 0.05 microg/mL. Calibration curve covered a span from 0.1-10 microg/mL with a good linear relationship (r = 0.999 9). The method showed a excellent performance when was used to authentic blood sample analysis for EtG.@*CONCLUSION@#The method is suitable for blood EtG analysis.

Glomerulopressin activity in peripheral blood of newly diagnosed type I (insulin-dependent) diabetic patients and in normal subjects treated with glucagon

Se estudió en dos ensayos biológicos la actividad de glomerulopresina en sangre periférica de voluntarios normales, pacientes diabéticos Tipo I (insulino-dependientes) de reciente diagnóstico (DID), y en voluntaríos normales tratados con glucagon. Los dos bioensayos fueron: a) variación en la tensión tónica contráctil del fundus de estómago de rata (TTC), y b) aumento de la presión ureteral del sapo que es considerada como índice de la presión glomerular (delta IPG). El ultrafiltrado obtenido de cuatro voluntarios normales tuvo una pequeña actividad en el TTC, y en otros dos sujetos no se observó actividad. El ultrafiltrado de cinco de estos sujetos no tuvo actividad en el delta IPG. El ultrafiltrado de los pacientes DID fue activo en los bioensayos. En tres de los ultrafiltrados de los normales tratados con glucagon el ultrafiltrado fue muy activo en el TTC, y en otros tres sujetos tratados con glucagon el ultrafiltrado fue poco activo en el TTC. En cinco de los voluntarios tratados con glucagon el ultrafiltrado fue muy activo en el ensayo del sapo. Estas observaciones sugieren que la actividad de glomerulopresina está aumentada en la sangre periférica de los pacientes DID de reciente diagnóstico y en sujetos normales tratados con glucagon