TY - JOUR
T1 - An improved method for rapidly quantifying fatty acid ethyl esters in meconium suitable for prenatal alcohol screening
AU - Hutson, Janine R.
AU - Rao, Chitra
AU - Fulga, Netta
AU - Aleksa, Katarina
AU - Koren, Gideon
N1 - Funding Information:
JRH is supported by a CIHR MD/PhD Studentship , the McLaughlin Centre for Molecular Medicine , and a U of T Open Fellowship . GK is the holder of the Ivey Chair in Molecular Toxicology, The Department of Medicine, University of Western Ontario.
PY - 2011/3
Y1 - 2011/3
N2 - Fatty acid ethyl esters (FAEEs) are nonoxidative metabolites of ethanol, and elevated levels of FAEE in meconium are a useful biomarker for heavy prenatal alcohol exposure. FAEE in meconium has been recommended as useful and cost-effective for universal screening for prenatal alcohol exposure. To support an efficient universal screening program, an analytical method to detect and quantify FAEE in meconium needs to be accurate, inexpensive, and rapid. The purpose of this study was to develop an analytical method that would satisfy these criteria and to validate this method using established laboratory guidelines. A method was developed and validated to detect and quantify four FAEEs (ethyl palmitate, ethyl linoleate, ethyl oleate, and ethyl stearate) from 0.5. g of meconium using d5-ethyl esters as internal standards. The sample undergoes liquid-liquid extraction with heptane:acetone, the heptane layer is isolated and evaporated, and then, the resulting residue undergoes headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. The detection limits of the four FAEEs ranged from 0.020 to 0.042. nmol/g and are 6- to 25-fold lower than the individual FAEE threshold concentrations (0.5. nmol/g). This method also has good precision with the coefficient of variation ranging from 2.6 to 19.4% for concentrations of individual FAEE between 0.5 and 2.62. nmol/g meconium (n=4). Calculated concentrations of FAEE that underwent extraction from meconium were 100-101% of the expected concentration, demonstrating the accuracy of the method. The peak shape and retention time of each FAEE were unaffected by the presence of the matrix, and there is no carryover at clinically relevant concentrations. This method was also able to produce clean chromatograms from meconium samples that could not be quantified using a previous method because of high chromatographic background. This method provides an optimal approach to detecting and quantifying FAEE in meconium that could be used in a universal screening program for prenatal alcohol exposure.
AB - Fatty acid ethyl esters (FAEEs) are nonoxidative metabolites of ethanol, and elevated levels of FAEE in meconium are a useful biomarker for heavy prenatal alcohol exposure. FAEE in meconium has been recommended as useful and cost-effective for universal screening for prenatal alcohol exposure. To support an efficient universal screening program, an analytical method to detect and quantify FAEE in meconium needs to be accurate, inexpensive, and rapid. The purpose of this study was to develop an analytical method that would satisfy these criteria and to validate this method using established laboratory guidelines. A method was developed and validated to detect and quantify four FAEEs (ethyl palmitate, ethyl linoleate, ethyl oleate, and ethyl stearate) from 0.5. g of meconium using d5-ethyl esters as internal standards. The sample undergoes liquid-liquid extraction with heptane:acetone, the heptane layer is isolated and evaporated, and then, the resulting residue undergoes headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. The detection limits of the four FAEEs ranged from 0.020 to 0.042. nmol/g and are 6- to 25-fold lower than the individual FAEE threshold concentrations (0.5. nmol/g). This method also has good precision with the coefficient of variation ranging from 2.6 to 19.4% for concentrations of individual FAEE between 0.5 and 2.62. nmol/g meconium (n=4). Calculated concentrations of FAEE that underwent extraction from meconium were 100-101% of the expected concentration, demonstrating the accuracy of the method. The peak shape and retention time of each FAEE were unaffected by the presence of the matrix, and there is no carryover at clinically relevant concentrations. This method was also able to produce clean chromatograms from meconium samples that could not be quantified using a previous method because of high chromatographic background. This method provides an optimal approach to detecting and quantifying FAEE in meconium that could be used in a universal screening program for prenatal alcohol exposure.
KW - Alcohol
KW - Biomarker
KW - Fatty acid ethyl esters
KW - Fetal alcohol spectrum disorder
KW - Meconium
KW - Prenatal screening
UR - http://www.scopus.com/inward/record.url?scp=79751526650&partnerID=8YFLogxK
U2 - 10.1016/j.alcohol.2010.07.005
DO - 10.1016/j.alcohol.2010.07.005
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C2 - 20705417
AN - SCOPUS:79751526650
SN - 0741-8329
VL - 45
SP - 193
EP - 199
JO - Alcohol
JF - Alcohol
IS - 2
ER -