Vaiano Fabio, Favretto Donata, Palumbo Diego, Cooper Gail, Mactier Helen, Busardò Francesco P, Mari Francesco, Bertol Elisabetta
Department of Health Science, Forensic Toxicology Division, University of Florence, Largo Brambilla, 50134, Florence, Italy.
Legal Medicine and Toxicology, Department SCTV, University-Hospital of Padova, Via Falloppio 50, Padova, 35121, Italy.
Anal Bioanal Chem. 2016 Apr;408(10):2587-94. doi: 10.1007/s00216-016-9364-8. Epub 2016 Feb 12.
Fatty acid ethyl esters (FAEEs) and ethyl-glucuronide (EtG) in meconium have been widely studied as biomarkers of maternal alcohol consumption during pregnancy. Many analytical approaches have been proposed for their analysis, mostly consisting of separated extraction procedures requiring the use of two meconium aliquots. This study aimed to validate a new analytical procedure for the simultaneous extraction of FAEEs and EtG from a meconium aliquot through a single solid-phase extraction (SPE) applied to 242 anonymized samples of meconium. Targeted FAEEs were: ethyl-myristate (Myr), ethyl-palmitate (Pal), ethyl-oleate (Ole) and ethyl-stearate (Ste). Two hundred milligrams of meconium was sonicated with acetonitrile, and a single SPE performed by means of aminopropyl columns. FAEEs were eluted with hexane, followed by EtG elution with water. Both the mixtures were dried, recovered, and analyzed by liquid chromatography-tandem mass spectrometry using C8 (FAEEs) and C18 (EtG) columns. Transitions were: m/z 257 → 57,88, Myr; m/z 262 → 57,88, Myr-d5; m/z 285 → 57, 72, Pal; m/z 290 → 57,258, Pal-d5; m/z 311 → 72,114, Ole; m/z 316 → 72,265, Ole-d5; m/z 257 → 57,72 Ste; m/z 318 → 57,286, Ste-d5; m/z 221 → 75,85, EtG; m/z 226 → 75,85, EtG-d5. Lower limit of quantification range was 10-15 ng/g for FAEEs and 10 ng/g for EtG. Linearity was evaluated for different concentration ranges; the mean coefficients of determination (R (2)) were above 0.9961. Precision and accuracy for FAEEs and EtG were consistently ≤20 % and ±20 %, respectively. Ion suppression was observed for all the analytes. Matrix effect did not significantly affect the analyses. Recovery efficiency was 93 % for EtG and 75-85 % for FAEEs.
胎粪中的脂肪酸乙酯(FAEEs)和乙基葡萄糖醛酸苷(EtG)已被广泛研究作为孕期母亲饮酒的生物标志物。针对它们的分析已经提出了许多方法,大多由需要使用两份胎粪等分试样的分离提取程序组成。本研究旨在验证一种新的分析程序,通过应用于242份匿名胎粪样本的单一固相萃取(SPE),从一份胎粪等分试样中同时提取FAEEs和EtG。目标FAEEs为:肉豆蔻酸乙酯(Myr)、棕榈酸乙酯(Pal)、油酸乙酯(Ole)和硬脂酸乙酯(Ste)。将200毫克胎粪用乙腈超声处理,并通过氨丙基柱进行单一SPE。FAEEs用己烷洗脱,随后EtG用水洗脱。两种混合物均经干燥、回收,并使用C8(用于FAEEs)和C18(用于EtG)柱通过液相色谱 - 串联质谱法进行分析。跃迁为:m/z 257 → 57,88,Myr;m/z 262 → 57,88,Myr - d5;m/z 285 → 57, 72,Pal;m/z 290 → 57,258,Pal - d5;m/z 311 → 72,114,Ole;m/z 316 → 72,265,Ole - d5;m/z 257 → 57,72 Ste;m/z 318 → 57,286,Ste - d5;m/z 221 → 75,85,EtG;m/z 226 → 75,85,EtG - d5。FAEEs的定量下限范围为10 - 15 ng/g,EtG为10 ng/g。在不同浓度范围内评估线性;平均测定系数(R(2))均高于0.9961。FAEEs和EtG的精密度和准确度分别始终≤20%和±20%。观察到所有分析物均存在离子抑制。基质效应未对分析产生显著影响。EtG的回收率为93%,FAEEs为75 - 85%。