Hjelt Institute, Department of Forensic Medicine, University of Helsinki, Helsinki, Finland.
Forensic Sci Int. 2013 Dec 10;233(1-3):328-32. doi: 10.1016/j.forsciint.2013.10.009. Epub 2013 Oct 14.
Differentiation of insulin analogues is required in forensic and clinical toxicology as well as in sports doping control. Immunoassay results provide only weak evidence for exogenous administration of insulin, as concentrations cannot be reliably interpreted and specific information on the insulin species remains unknown. In post-mortem blood, insulin degrades rapidly. In this study, improved methodology consisting of precipitation of proteins, immunoaffinity purification and liquid chromatography coupled to high resolution/high accuracy mass spectrometry were applied to post-mortem vitreous humour. Ten successive cases with a post-mortem interval from four to ten days were investigated for insulin analogues. The cause of death in these cases was connected with diabetes and its complications, as well as with chronic cardiovascular disease, alcoholism and cancer. In all cases, the manner of death was natural (disease). Insulin was positively detected in post-mortem vitreous humour in three cases out of ten by mass spectrometry. In two cases, the method revealed the long-acting insulin glargine (Lantus) metabolite M2 (DesB31-32 Lantus), and human insulin was detected in one case. The findings were in agreement with the documented history of insulin medication. No other obvious reason could be found for the failure of detecting insulins in the other cases than insulin degradation during the lengthy post-mortem interval. Vitreous humour is still a most prospective specimen for detection of insulin analogues post-mortem.
在法医毒理学和临床毒理学以及运动兴奋剂控制中，需要区分胰岛素类似物。免疫测定结果仅为外源性给予胰岛素的弱证据，因为无法可靠地解释浓度，并且对胰岛素种类的具体信息仍然未知。在死后血液中，胰岛素迅速降解。在这项研究中，应用改进的方法学，包括蛋白质沉淀、免疫亲和纯化和液相色谱与高分辨率/高精度质谱联用，用于死后玻璃体液。对连续 10 例死后间隔 4 至 10 天的案例进行了胰岛素类似物的研究。这些案例的死因与糖尿病及其并发症、慢性心血管疾病、酗酒和癌症有关。在所有情况下，死亡方式均为自然（疾病）。通过质谱法在 10 例中的 3 例中检测到了死后玻璃体液中的胰岛素。在 2 例中，该方法揭示了长效胰岛素 glargine（Lantus）代谢物 M2（DesB31-32 Lantus），在 1 例中检测到了人胰岛素。这些发现与胰岛素用药的记录病史相符。在其他病例中，除了在漫长的死后间隔期间胰岛素降解外，未能检测到胰岛素的其他明显原因。玻璃体液仍然是死后检测胰岛素类似物最有前途的标本。
