Phosphatidylethanol in Human Organs and Blood: A Study on Autopsy Material and Influences by Storage Conditions

Abstract

The rat experiments indicated that the very high PEth concentrations found in the organs of the alcoholics were probably largely formed while the organs were frozen at -20 degrees C. Our data suggest that tissue material from bodies that were exposed to ethanol must be stored properly to obtain reliable results from subsequent analysis for PEth. Tissue should not be frozen at -20 degrees C but instead stored refrigerated until extraction, preferably within hours of autopsy, or frozen in liquid nitrogen and stored at -80 degrees C. Blood samples that contain ethanol can be stored refrigerated for up to 72 hr or frozen in liquid nitrogen and stored at -80 degrees C without affecting PEth levels.

Quantification of phosphatidylethanol 16:0/18:1, 18:1/18:1, and 16:0/16:0 in venous blood and venous and capillary dried blood spots from patients in alcohol withdrawal and control volunteers

“…They are biomarkers of alcohol consumption [2] present in blood, mainly located in erythrocytes [3], and in different organs [4]. Up to forty-eight different PEths have been detected in blood collected in autopsy cases of heavy drinkers [5].…”

“…Some publications reported other types of sample preparation, such as protein precipitation with methanol [18] or protein precipitation followed by an online-solid phase extraction [19]. A number of detection methods is based on HPLC with normal phase columns coupled to light-scattering detection (ELSD); chromatography has been carried out with hexane and propanol-based gradients containing acetic acid and triethylamine [3,4,11,12]. Quantification limits (LLOQ) obtained with these methods ranged between 100-500 ng/mL [4,11], analysing 250 to 300 µL of whole blood.…”

“…A number of detection methods is based on HPLC with normal phase columns coupled to light-scattering detection (ELSD); chromatography has been carried out with hexane and propanol-based gradients containing acetic acid and triethylamine [3,4,11,12]. Quantification limits (LLOQ) obtained with these methods ranged between 100-500 ng/mL [4,11], analysing 250 to 300 µL of whole blood. PEths have also been analysed with non-aqueous capillary electrophoresis (CE) coupled to UV [13] detection.…”

Phosphatidylethanol Accumulation Promotes Intestinal Hyperplasia by Inducing ZONAB-Mediated Cell Density Increase in Response to Chronic Ethanol Exposure

“…Furthermore, ethanol could also undergo chemical coupling to membrane phospholipids via the action of phospholipase D (PLD; refs. 15,16), resulting in the conversion of phosphatidylcholine into phosphatidylethanol (17)(18)(19). In the presence of high ethanol concentrations, this transphosphatidylation reaction dominates over the hydrolytic reaction done by PLD in the absence of alcohol, which generates phosphatidic acid, and the resulting loss of phosphatidic acid can interfere with signal transduction in hepatocytes and glia (20)(21)(22).…”

“…In the presence of high ethanol concentrations, this transphosphatidylation reaction dominates over the hydrolytic reaction done by PLD in the absence of alcohol, which generates phosphatidic acid, and the resulting loss of phosphatidic acid can interfere with signal transduction in hepatocytes and glia (20)(21)(22). However, lower concentrations of ethanol are found in the intestine, where the accumulation of phosphatidylethanol has previously been detected following chronic ethanol exposure (17,18,23).…”