Section of Legal Medicine-Forensic Toxicology and Antidoping Unit, Department of Environmental Medicine and Public Health, Via Falloppio 50, 35121 Padova, Italy.
Forensic Sci Int. 2009 Dec 15;193(1-3):79-83. doi: 10.1016/j.forsciint.2009.09.010. Epub 2009 Oct 20.
Embalming and formalin fixation are common, and yet they can create problems for the forensic scientist if a drug has been the cause of death and if the only available specimens to be analyzed are formalin-fixed tissues. Previous studies have demonstrated that during fixation xenobiotics are extracted into formalin according to tissue and fixing solution characteristics. In some cases formalin can react with the analyte resulting in the production of new chemical entities. Regarding cocaine and its metabolites, Cingolani et al. have reported that formalin-fixation extracts benzoylecgonine (BE) from tissues and that BE is stable in the fixing solution. However, the stability and kinetic properties of cocaine remain so far unexplored. Our data show that in buffered formalin (pH 7.4) cocaine is hydrolyzed to BE in agreement with a pseudo first-order reaction kinetic (half-life time approximately 7 days), whereas in unbuffered formalin (pH approximately 3.5) it is relatively stable over a period of 30 days. The analysis of brain and liver samples at different fixation times indicates that during fixation an extraction process occurs for both analytes and that the extraction is more efficient in the liver than in the brain, probably because of a greater lipophilicity of the brain tissue. In conclusion, our study demonstrates that formalin-fixed tissues and their fixing solutions can be used for cocaine analysis only if a short time period has passed since the fixation beginning. The rapid extraction process of cocaine into formalin and the concomitant hydrolysis to BE occurring in buffered formalin may prevent the identification of cocaine in both tissues and formalin solution already at 15-30 days after fixation. Moreover, the unpredictable extraction rate of both analytes, along with the hydrolysis of cocaine into BE significantly affects tissue concentrations, thus complicating the interpretation of quantitative results.
尸体防腐和福尔马林固定是常见的做法,但如果导致死亡的原因是药物,且唯一可用于分析的样本是福尔马林固定组织,那么这会给法医科学家带来问题。先前的研究表明,在固定过程中,根据组织和固定溶液的特性,外来化合物会被提取到福尔马林溶液中。在某些情况下,福尔马林可能会与分析物发生反应,从而产生新的化学实体。关于可卡因及其代谢物,Cingolani 等人报告称,福尔马林固定提取物从组织中提取出苯甲酰爱康宁(BE),并且 BE 在固定溶液中稳定。然而,迄今为止,可卡因的稳定性和动力学特性仍未得到探索。我们的数据表明,在缓冲福尔马林(pH7.4)中,可卡因按照拟一级反应动力学(半衰期约为 7 天)水解为 BE,而在未缓冲福尔马林(pH 约为 3.5)中,在 30 天的时间内相对稳定。对不同固定时间的脑和肝样本的分析表明,在固定过程中,两种分析物都会发生提取过程,并且在肝脏中的提取效率高于在脑内,这可能是因为脑组织的脂溶性更强。总之,我们的研究表明,只有在固定开始后较短的时间内,才能使用福尔马林固定组织及其固定溶液进行可卡因分析。在缓冲福尔马林溶液中,可卡因迅速被提取到福尔马林溶液中,同时发生水解生成 BE,这可能会导致在固定后 15-30 天,无论是在组织还是福尔马林溶液中都无法识别可卡因。此外,两种分析物的不可预测的提取率以及可卡因水解为 BE 会显著影响组织浓度,从而使定量结果的解释变得复杂。