Abstract：
Phosphatidylethanol (PEth) measurement in whole blood samples is established as a specific alcohol biomarker with clinical and forensic applications. Establishment of dried blood spots (DBSs) as a specimen for PEth determination offers several advantages and was the focus of this work. A liquid chromatography tandem mass spectrometry method using a 96-well format for sample preparation was developed and validated. PEth was extracted from DBSs by using isopropanol containing PEth-d5 as internal standard. The blood sampling used a commercial volumetric DBS device having a phosholipase D inhibitor incorporated to stop continuous PEth formation. The method quantified PEth in the range of 0.05-10 μmol/L, with a bias and imprecision of less than 15%. In a clinical study (n = 25) using fingerprick blood, the volumetric device offered more precise quantifications (CV 4.6%) compared with the Whatman 903 Protein Saver card device (CV 16.6%). In another clinical study (n = 48), the use of dried venous and capillary blood, and liquid venous blood was compared under real-life conditions with samples sent by postal service. The capillary and venous DBS samples gave identical results while the liquid blood gave slightly higher values. Calculation of elimination half-life (PEth 16:0/18:1) in 31 cases based on two consecutive samples with 2-9 days in between gave results (mean 6.2 days) that agree with literature but several cases with values over 10 days. In conclusion, this study demonstrates that volumetric DBS is a valid specimen for determination of PEth blood concentrations, offering several advantages.
摘要：
全血样品中的磷脂酰乙醇（PEth）测量被确立为具有临床和法医应用的特定酒精生物标志物。建立干血斑（DBS）作为PEth测定的样本具有几个优点，并且是这项工作的重点。开发并验证了使用96孔格式进行样品制备的液相色谱串联质谱方法。通过使用含有PEth-d5的异丙醇作为内标从DBS中提取PEth。血液取样使用商业体积DBS装置,该装置具有掺入的磷酸脂酶D抑制剂以停止连续的PEth形成。该方法定量PEth在0.05-10μmol/L范围内，偏差和不精确度小于15%。在一项使用手指刺血的临床研究（n=25）中，与Whatman903ProteinSaver卡装置(CV16.6%)相比,容量装置提供了更精确的定量(CV4.6%)。在另一项临床研究中（n=48），使用干燥的静脉和毛细血管血，在现实生活中，液体静脉血与邮政部门发送的样本进行了比较。毛细管和静脉DBS样品给出相同的结果,而液体血液给出稍高的值。在31例病例中，基于两个连续样品，中间有2-9天，计算消除半衰期（PEth16：0/18：1），得出的结果（平均6.2天）与文献一致，但有几例病例的值超过10天。总之,这项研究表明，容量DBS是测定PEth血液浓度的有效标本，提供几个优点。
