Simultaneous determination of ethanol's four types of non-oxidative metabolites in human whole blood by liquid chromatography tandem mass spectrometry.

The importance of ethanol non-oxidative metabolites as the specific biomarkers of alcohol consumption in clinical and forensic settings is increasingly acknowledged. Simultaneous determination of these metabolites can provide a wealth of information like drinking habit and history, but it was difficult to achieve because of their wide range of polarity. This work describes development and validation of a simple liquid chromatography tandem mass spectrometry (LC-MS/MS) assay for 4 types of ethanol non-oxidative metabolites (ethyl glucuronide, ethyl sulfate, fatty acid ethyl esters and phosphatidylethanols) in 50 µL of human whole blood. Pretreatment method, column and MS conditions were optimized. For the first time, the four types of ethanol non-oxidative metabolites with enormous discrepancies of property were simultaneously extracted and analyzed in one run within 40 min. The limits of detections (LODs) were among 0.1-10 ng/mL, and good linearity was obtained. Deviations in precision and accuracy were all lower than 15% at three QC levels. This method was then applied to two forensic samples, resulting in information on drinking habits and drinking time which were very useful for the interpretation of the blood alcohol results.

Elevated plasma aldosterone is an independent risk factor for erectile dysfunction in men.

INTRODUCTION: Erectile dysfunction (ED) and cardiovascular disease (CVD) share a great number of common risk factors. There is growing evidence that aldosterone, an independent CVD risk factor, is associated with ED. AIMS: The purpose of this study was to determine the relationship between plasma aldosterone and erectile dysfunction. METHODS: This study recruited 287 participants, ranging from 18 to 84 years old; 217 were suffering from ED, diagnosed by the International Index of Erectile Function 5 (IIEF-5) scores. Based on IIEF-5 scores, patients were divided into one control group and three ED groups (mild ED; moderate ED; severe ED). MAIN OUTCOME MEASURES: The differences in principal characteristics, blood routine, sexual hormone, adrenal hormone, thyroid hormone, renal function, liver function and blood lipid were compared between ED and control groups. RESULTS: Our study demonstrated that the difference of mean plasma aldosterone levels between ED group and the control group was statistically significant (P < 0.05). Stepwise logistic regression analysis of all the possible factors support the role of aldosterone as an independent risk factor for ED (OR 1.011; 95 % CI 1.003-1.018; P = 0.004). Similar statistical methods were applied to the comparison between moderate to severe ED group and control to mild ED group (OR 1.017; 95 % CI 1.009-1.024; P < 0.001). ROC curve and the area under the curve (0.718; 95 % CI 0.643-0.794; P < 0.001) were performed to assess the diagnostic effect and to compare the severity of risk with the known independent risk factors, such as age and cholesterol (0.704; 95 % CI 0.631-0.778; P < 0.001). When using a 374 pg/mL cut-off value from Youden index, the OR of ED group versus controls is 3.106 (95 % CI 1.458-6.617), while the OR of moderate to severe ED versus control and mild ED is 5.480 (95 % CI 3.108-9.662). CONCLUSIONS: We determined that elevated plasma aldosterone concentration is an independent risk factor for ED. Our findings also indicate that the aldosterone, a well-recognized contributor to vascular injury, might be a potential bond between ED and CVD.

Assessment of the Role Played by N-propanol Found in Postmortem Blood in the Discrimination Between Antemortem Consumption and Postmortem Formation of Ethanol Using Rats.

This study disproves the reliability of n-propanol as a biomarker to establish whether the ethanol found in postmortem blood is derived from antemortem ingestion or postmortem putrefactive processes. Two groups of rats were given ethanol or normal saline solution, respectively, and sacrificed 1.5 h later. After putrefaction, blood and, in a few cases, urine samples from the rats were analyzed for ethanol and n-propanol by head-space gas chromatography equipped with flame ionization detection. Although the concentration ratios of ethanol/n-propanol in the postmortem blood collected from the bodies without prior alcohol consumption were expected to be <20 (as per limited case reports and previous in vitro studies), in samples from several rats that were on saline solution, this ratio was found to exceed 20. In conclusion, the concentration ratio of ethanol/n-propanol in postmortem blood does not allow for the discernment between antemortem ingestion and the postmortem synthesis of ethanol.

Effects of triglycerides levels in human whole blood on the extraction of 19 commonly used drugs using liquid-liquid extraction and gas chromatography-mass spectrometry.

Liquid-liquid extraction (LLE) is the most commonly sample preparation procedure used by forensic toxicologists in China for screening drugs in whole human blood. It extracts numerous substances from blood including targeted drugs and interfering substances, specifically triglycerides (TG). With increasing prevalence of hyperlipidemia, the influences of TG on LLE and on subsequent analysis with gas chromatography-mass spectrometry (GC-MS) may become a major issue for forensic laboratories. This study aims to elucidate the influences of TG on LLE and to provide possible solutions to this problem. Nineteen commonly encountered drugs in forensic cases were spiked to human whole blood with different TG concentrations. Diethyl ether, ethyl acetate/hexane mixed solutions, chlorobutane and several other frequently used solvents were tested for the extraction of drugs from spiked whole blood. The supernatant organic layer was evaporated to dryness and reconstituted with methanol. The resultant products were analyzed by GC-MS, and the extraction recovery was calculated. LLE with diethyl ether, ethyl acetate/hexane (9:1) and chlorobutane all possessed effective and reliable extraction recoveries for blood sample with low TG concentrations (0.63-6.85 mmol/L). At high TG concentrations, diethyl ether produced a highly turbid substance that could not be further analyzed using GC-MS. Extraction recoveries drastically dropped for ethyl acetate/hexane (9:1) mixture at high TG concentrations, while chlorobutane experienced minimal drops in extraction recoveries. In conclusion, TG levels in whole blood noticeably influence drug recovery to variable extents depending on the LLE solvent. Chlorobutane showed minimal influences from TG content in whole blood and thus is the recommended LLE solvent for forensic drug extraction.

Characteristics of 1226 alcohol-positive drivers involved in nonfatal traffic crashes in Shanghai, China.

OBJECTIVE: The purpose of our study was to better characterize and evaluate drunk driving for governmental reference in order to further reduce alcohol-impaired driving. METHOD: This article reports the characteristics of 1226 alcohol-positive drivers with blood alcohol concentrations (BACs) at or over the legal limit of 0.20 mg/mL involved in nonfatal traffic accidents in Shanghai, China, from 2008 to 2011. The mean BAC, age, and gender of these drivers are discussed as well as the vehicle types and times of day when the crashes occurred. RESULTS: The mean BAC was 1.41 mg/mL and the mean age was 38 years old, and the vast majority of drivers were male (96.9%). The mean BAC of male drivers (1.42 mg/mL) was higher than that of female drivers (1.20 mg/mL). The mean age of male drivers (38) was also higher than that of female drivers (33). Distributions of vehicle types involved were studied. Cars had the highest percentage of occurrences (56.4%), followed by motorcycles (32.8%), electric bicycles (6.8%), trucks (1.5%), and bicycles (0.8%). It was found that these alcohol-related traffic crashes most often occurred between 7:00 p.m. and 10:59 p.m., representing 48.1 percent (n = 590) of the 1226 cases. Single-vehicle crashes were overrepresented (70.3%) in all cases. The mean BAC in multivehicle crashes (1.69 mg/mL) was higher than that in single-vehicle crashes (1.30 mg/mL). CONCLUSIONS: The results indicated a notable need for more governmental attention that would prevent accidents caused by driving under the influence of alcohol.