Watterson James H, Vandenboer Trevor C
Forensic Toxicology Research Laboratory, Department of Forensic Science, Laurentian University, 935 Ramsey Lake Rd., Sudbury, Ontario, Canada P3E 2C6.
J Anal Toxicol. 2008 Oct;32(8):631-8. doi: 10.1093/jat/32.8.631.
Ketamine exposure was detected in skeletal tissues by ELISA and liquid chromatography-tandem mass spectrometry (LC-MS-MS). Rats (n = 9) received ketamine hydrochloride acutely (75 mg/kg, i.p.) and were euthanized within 15, 30, or 90 min. Drug-free control animals (n = 3) were also euthanized. Extracted femora were separated into epiphyseal and diaphyseal fragments, with marrow isolated from the medullary cavity. Bone was ground and incubated in methanol. Extracts were dried and reconstituted in phosphate buffer (0.1 M, pH 7.3), and marrow was homogenized in alkaline solution. Both then underwent solid-phase extraction. Extracts were assayed by ELISA, with data expressed in terms of relative decrease in absorbance (%DA, drug-positive tissues vs. matrix-matched drug-free controls) and binary classification test sensitivity (S). Generally, %DA decreased in the order of marrow > epiphyseal bone > diaphyseal bone, and was negatively correlated with dose-death interval (DDI). Measured S values were 100% in ELISA analysis of extracts of all tissue types. Sensitivity values were computed from LC-MS-MS data using a 5 ng/mL cutoff. Sensitivity values for ketamine detection were 100%, 0-100% and 0%, at the 15, 30, and 90 min DDI, respectively, and sensitivity values for norketamine detection were 0-66%, 0-66%, and 0% at the 15, 30, and 90 min DDI, respectively. These results suggest that the tissue type sampled and DDI may influence the sensitivity of detection of ketamine exposure in skeletal tissues.
通过酶联免疫吸附测定法（ELISA）和液相色谱 - 串联质谱法（LC-MS-MS）在骨骼组织中检测到氯胺酮暴露。大鼠（n = 9）急性接受盐酸氯胺酮（75 mg/kg，腹腔注射），并在15、30或90分钟内安乐死。无药物的对照动物（n = 3）也被安乐死。取出的股骨被分离为骨骺和骨干碎片，并从骨髓腔中分离出骨髓。将骨头研磨并在甲醇中孵育。提取物干燥后用磷酸盐缓冲液（0.1 M，pH 7.3）复溶，骨髓在碱性溶液中匀浆。然后两者都进行固相萃取。通过ELISA对提取物进行检测，数据以吸光度的相对下降率（%DA，药物阳性组织与基质匹配的无药物对照）和二元分类测试灵敏度（S）表示。一般来说，%DA按骨髓>骨骺骨>骨干骨的顺序降低，并且与剂量 - 死亡间隔（DDI）呈负相关。在ELISA分析所有组织类型的提取物中，测得的S值均为100%。使用5 ng/mL的截断值从LC-MS-MS数据计算灵敏度值。在15、分别为30和90分钟的DDI时，氯胺酮检测的灵敏度值分别为100%、0 - 100%和0%，而去甲氯胺酮检测的灵敏度值在15、30和90分钟的DDI时分别为0 - 66%、0 -66%和0%。这些结果表明，所采样的组织类型和DDI可能会影响骨骼组织中氯胺酮暴露检测的灵敏度。
