Forensic Toxicology Research Laboratory, Laurentian University, 935 Ramsey Lake Rd., Sudbury, Ontario, Canada.
Forensic Sci Int. 2011 Apr 15;207(1-3):40-5. doi: 10.1016/j.forsciint.2010.08.021. Epub 2010 Sep 29.
The effect of dose-death interval and tissue distribution on the detection of meperidine in selected skeletal tissues was examined using a rapid microwave-assisted extraction (MAE) methodology. Rats (n=14) were dosed with 0 (n=2) or 30 mg/kg (n=12) meperidine (i.p.). Drug-positive rats were sacrificed with CO(2) after 20, 30, 90 and 150 min (n=3 per group). Heart blood was collected immediately after death. Tibiae were excised and frozen for further analysis. The remaining carcasses were allowed to decompose outside in secured cages to the point of complete skeletonization in a rural Northern Ontario location during the late summer months. Vertebrae and pelvi were collected for each animal. Tibial marrow was homogenized in 3 mL PB6 (phosphate buffer, 0.1M, pH 6). Fresh tibiae, and decomposed vertebrae and pelvi were cleaned in PB8.5 (phosphate buffer, 0.1M, pH 8.5) and sonicated to remove remaining soft tissue. Samples of dried, ground bone (0.5-1g) suspended in 2 mL PB6 were then irradiated in a domestic microwave oven (1100 W) at atmospheric pressure for 15 min. Samples of vertebral bone (1g) were also extracted by passive incubation in methanol (3 mL, 50°C, 72 h). All supernatants then underwent solid-phase extraction and analysis by GC/MS, using electron impact ionization in the Selected Ion Monitoring (SIM) mode. Mean GC/MS responses for each tissue type were negatively correlated with dose-death interval, with correlation coefficients ranging from -0.32 to -0.87. Analysis of variance showed dose-death interval to be a main effect (p<0.05) with respect to GC/MS response for blood, marrow, tibial epiphyses prepared by MAE, and vertebral bone prepared by passive extraction, but not for tibial diaphyses, pelvi or vertebrae prepared by MAE. Overall, MAE is advantageous as a rapid extraction tool for screening purposes in skeletal tissues, but assignment of significance to quantitative expressions of skeletal drug concentrations is complex and should be approached with caution.
使用快速微波辅助提取 (MAE) 方法研究了剂量-死亡间隔和组织分布对选定骨骼组织中哌替啶检测的影响。将大鼠（n=14）用 0（n=2）或 30 mg/kg（n=12）哌替啶（ip）给药。在 20、30、90 和 150 分钟后，用 CO2 处死药物阳性大鼠（n=3/组）。死后立即采集心血。取出胫骨并冷冻用于进一步分析。其余尸体在安大略省北部农村的安全笼中分解，直到夏末完全骨骼化。为每个动物收集椎骨和骨盆。将胫骨骨髓在 3 mL PB6（磷酸盐缓冲液，0.1M，pH 6）中匀浆。新鲜胫骨、分解的椎骨和骨盆用 PB8.5（磷酸盐缓冲液，0.1M，pH 8.5）清洗，并用超声处理以去除剩余的软组织。将悬浮在 2 mL PB6 中的干燥、研磨骨（0.5-1g）的样品置于家用微波炉（1100 W）中，在大气压下辐照 15 分钟。还通过在甲醇（3 mL，50°C，72 h）中被动孵育提取 1g 椎骨样品。所有上清液然后通过固相萃取和 GC/MS 分析，在选择离子监测（SIM）模式下使用电子轰击电离。每种组织类型的 GC/MS 响应平均值与剂量-死亡间隔呈负相关，相关系数范围为-0.32 至-0.87。方差分析表明，剂量-死亡间隔是血液、骨髓、MAE 制备的胫骨骨骺和被动提取制备的椎骨 GC/MS 响应的主要影响因素（p<0.05），但对胫骨骨干、骨盆或 MAE 制备的椎骨没有影响。总体而言，MAE 作为骨骼组织筛选目的的快速提取工具具有优势，但对骨骼药物浓度的定量表达进行赋值是复杂的，应该谨慎处理。
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