Imfeld Anic B, Watterson James H
Department of Forensic Science, Laurentian University, 935 Ramsey Lake Rd, Sudbury, ON, Canada, P3E 2C6.
Int J Legal Med. 2016 Mar;130(2):371-9. doi: 10.1007/s00414-015-1196-0. Epub 2015 May 7.
The semi-quantitative analysis of decomposed bone of rats exposed to colchicine and euthanized following different time intervals postexposure (i.e., dose-death interval, DDI) is described. Rats received colchicine (50 mg/kg, i.p.) and were euthanized 30 min (DDI1; n = 4), 60 min (DDI2; n = 4), or 180 min (DDI3; n = 4) postdose. Drug-free animals (n = 3) served as negative controls. Perimortem heart plasma was collected. Remains were decomposed to skeleton outdoors and then collected and sorted (skull, vertebrae, rib, pelvis, femur, tibia). Bones were dried, pulverized, and prepared by microwave-assisted extraction and microplate solid-phase extraction (MAE-MPSPE), followed by analysis for colchicine, 3-demethylcolchicine (3DMC), and 2-demethylcolchicine (2DMC) by ultra-high-performance liquid chromatography with photodiode array detection (UHPLC-PDA) at 350 nm. Bone type was a main effect (Kruskall-Wallis, p < 0.05) with respect to drug level (expressed as mass-normalized response ratio, RR/m) for each analyte, at each DDI. For all samples, DDI was a main effect (Kruskall-Wallis, p < 0.05) with respect to analyte level, and the ratio of analyte levels (RR3DMC/RRCOLCH, RR2DMC/RRCOLCH, and RR2DMC/RR3DMC). Bone COLCH levels varied by 19-fold, 12-fold, and 60-fold across all bone types in the DDI1, DDI2, and DDI3 groups, respectively. Bone 3DMC levels varied by 12-fold, 11-fold and 17-fold across all bone types in the DDI1, DDI2, and DDI3 groups, respectively. Bone 2DMC levels varied by 20-fold, 14-fold, and 14-fold across all bone types in the DDI1, DDI2, and DDI3 groups, respectively. Values of RR3DMC/RRCOLCH varied by 16-fold, 5-fold, and 5-fold across all bone types in the DDI1, DDI2, and DDI3 groups, respectively. Values of RR2DMC/RRCOLCH varied by 10-fold, 6-fold, and 12-fold across all bone types in the DDI1, DDI2, and DDI3 groups, respectively. Values of RR2DMC/RR3DMC varied by 3-fold, 5-fold, and 2-fold across all bone types in the DDI1, DDI2, and DDI3 groups, respectively. Measured analyte levels in bone correlated poorly with corresponding levels in blood (r = -0.65-+0.31). Measured values of RR2DMC/RRCOLCH and RR2DMC/RR3DMC in bone also correlated poorly with corresponding values in blood. Measured values of RR3DMC/RRCOLCH were well correlated with corresponding blood levels for all bone types except skull (r = 0.91-0.97).
描述了对暴露于秋水仙碱并在暴露后不同时间间隔（即剂量-死亡间隔，DDI）实施安乐死的大鼠的分解骨骼进行的半定量分析。大鼠接受秋水仙碱（50 mg/kg，腹腔注射），并在给药后30分钟（DDI1；n = 4）、60分钟（DDI2；n = 4）或180分钟（DDI3；n = 4）实施安乐死。无药物处理的动物（n = 3）作为阴性对照。采集濒死时的心脏血浆。遗体在户外分解为骨骼，然后收集并分类（颅骨、椎骨、肋骨、骨盆、股骨、胫骨）。将骨骼干燥、粉碎，并通过微波辅助萃取和微孔板固相萃取（MAE-MPSPE）进行制备，随后通过超高效液相色谱-光电二极管阵列检测（UHPLC-PDA）在350 nm波长下分析秋水仙碱、3-去甲基秋水仙碱（3DMC）和2-去甲基秋水仙碱（2DMC）。在每个DDI下，对于每种分析物，骨类型是药物水平（以质量归一化响应比，RR/m表示）的主要影响因素（Kruskal-Wallis检验，p < 0.05）。对于所有样本，DDI是分析物水平以及分析物水平之比（RR3DMC/RRCOLCH、RR2DMC/RRCOLCH和RR2DMC/RR3DMC）的主要影响因素（Kruskal-Wallis检验，p < 0.05）。在DDI1、DDI2和DDI3组中，所有骨类型的骨骼秋水仙碱水平分别相差19倍、12倍和60倍。在DDI1、DDI2和DDI3组中，所有骨类型的骨骼3DMC水平分别相差12倍、11倍和17倍。在DDI1、DDI2和DDI3组中，所有骨类型的骨骼2DMC水平分别相差20倍、14倍和14倍。RR3DMC/RRCOLCH的值在DDI1、DDI2和DDI3组中，所有骨类型分别相差16倍、5倍和5倍。RR2DMC/RRCOLCH的值在DDI1、DDI2和DDI3组中，所有骨类型分别相差10倍、6倍和12倍。RR2DMC/RR3DMC的值在DDI1、DDI2和DDI3组中，所有骨类型分别相差3倍、5倍和2倍。骨骼中测得的分析物水平与血液中的相应水平相关性较差（r = -0.65至+0.31）。骨骼中RR2DMC/RRCOLCH和RR2DMC/RR3DMC的测量值与血液中的相应值相关性也较差。RR3DMC/RRCOLCH的测量值与除颅骨外的所有骨类型的相应血液水平相关性良好（r = 0.91至0.97）。
