Zhang Fan, Qin Wei, Zhang Jing-Pu, Hu Chang-Qin
National Institutes for Food and Drug Control, Graduate School of Peking Union Medical College, Beijing, China.
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Graduate School of Peking Union Medical College, Beijing, China.
PLoS One. 2015 May 4;10(5):e0124805. doi: 10.1371/journal.pone.0124805. eCollection 2015.
Evaluation of drug toxicity is necessary for drug safety, but in vivo drug absorption is varied; therefore, a rapid, sensitive and reliable method for measuring drugs is needed. Zebrafish are acceptable drug toxicity screening models; we used these animals with a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method in a multiple reaction monitoring mode to quantify drug uptake in zebrafish to better estimate drug toxicity. Analytes were recovered from zebrafish homogenate by collecting supernatant. Measurements were confirmed for drugs in the range of 10-1,000 ng/mL. Four antibiotics with different polarities were tested to explore any correlation of drug polarity, absorption, and toxicity. Zebrafish at 3 days post-fertilization (dpf) absorbed more drug than those at 6 h post-fertilization (hpf), and different developmental periods appeared to be differentially sensitive to the same compound. By observing abnormal embryos and LD50 values, zebrafish embryos at 6 hpf were considered to be suitable for evaluating embryotoxicity. Also, larvae at 3 dpf were adapted to measure acute drug toxicity in adult mammals. Thus, we can exploit zebrafish to study drug toxicity and can reliably quantify drug uptake with LC-MS/MS. This approach will be helpful for future studies of toxicology in zebrafish.
评估药物毒性对于药物安全性而言是必要的，但体内药物吸收存在差异；因此，需要一种快速、灵敏且可靠的药物检测方法。斑马鱼是可接受的药物毒性筛选模型；我们使用这些动物，采用液相色谱 - 串联质谱（LC-MS/MS）方法，在多反应监测模式下对斑马鱼体内的药物摄取进行定量，以更好地评估药物毒性。通过收集上清液从斑马鱼匀浆中回收分析物。对浓度范围在10 - 1000 ng/mL的药物进行了测量确认。测试了四种不同极性的抗生素，以探究药物极性、吸收和毒性之间的任何相关性。受精后3天（dpf）的斑马鱼比受精后6小时（hpf）的斑马鱼吸收更多药物，并且不同发育阶段对同一化合物的敏感性似乎存在差异。通过观察异常胚胎和半数致死剂量（LD50）值，受精后6 hpf的斑马鱼胚胎被认为适合评估胚胎毒性。此外，3 dpf的幼虫适合用于测量成年哺乳动物的急性药物毒性。因此，我们可以利用斑马鱼来研究药物毒性，并能够通过LC-MS/MS可靠地定量药物摄取。这种方法将有助于未来斑马鱼毒理学的研究。
