Oiestad Elisabeth Leere, Johansen Unni, Stokke Opdal Mimi, Bergan Stein, Christophersen Asbjørg Solberg
Norwegian Institute of Public Health, Division of Forensic Toxicology and Drug Abuse, P.O. Box 4404 Nydalen, NO-0403 Oslo, Norway.
J Anal Toxicol. 2009 Sep;33(7):372-8. doi: 10.1093/jat/33.7.372.
A liquid chromatography-tandem mass spectrometry (LC-MS-MS) method has been developed and validated for the determination of digoxin and digitoxin in whole blood samples in autopsy cases. Samples were prepared by liquid-liquid extraction (LLE) with ethyl acetate/heptane/dichloromethane (3:1:1). LC separation was achieved using an Atlantis dC(18)-column (2.1 x 50 mm, 3 microm). The time between injections was 11 min. Mass detection was performed by positive ion mode electrospray LC-MS-MS on the ammonium adducts with two transitions for each analyte and one for the internal standard (digoxin-d(3)). Within-day precision was between 8.3 and 10.8%, between-day precision was between 8.7 and 14.2% and accuracy (bias) was between -17.3 and 11.5%. LOQ was 0.1 nmol/L (0.08 ng/mL), with an accuracy and precision of 19% and -17% (digoxin) and 18% and 3% (digitoxin). Matrix effects ranged from 104 to 117%. Good qualitative correlation with previous findings was achieved for 38 autopsy cases. The median (range, number of cases) B-digitoxin and B-digoxin found with the LC-MS-MS method in this very limited material were 9.3 (3.4-23.8, n = 24) and 5.6 (3.4-26.5, n = 4) nmol/L, respectively.
已开发并验证了一种液相色谱 - 串联质谱（LC-MS-MS）方法，用于测定尸检病例全血样本中的地高辛和洋地黄毒苷。样品通过用乙酸乙酯/庚烷/二氯甲烷（3:1:1）进行液 - 液萃取（LLE）制备。使用Atlantis dC(18)柱（2.1×50 mm，3微米）实现LC分离。进样间隔时间为11分钟。通过正离子模式电喷雾LC-MS-MS对铵加合物进行质量检测，每种分析物有两个跃迁，内标（地高辛 - d(3)）有一个跃迁。日内精密度在8.3%至10.8%之间，日间精密度在8.7%至14.2%之间，准确度（偏差）在 - 17.3%至11.5%之间。定量限为0.1 nmol/L（0.08 ng/mL），地高辛的准确度和精密度分别为19%和 - 17%，洋地黄毒苷为18%和3%。基质效应范围为104%至117%。对38例尸检病例与先前结果实现了良好的定性相关性。在这种非常有限的材料中，用LC-MS-MS方法测得的B - 洋地黄毒苷和B - 地高辛的中位数（范围，病例数）分别为9.3（3.4 - 23.8，n = 24）和5.6（3.4 - 26.5，n = 4）nmol/L。
