Abstract
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.
Highlights
Drug toxicity is a chief reason for drug withdrawal from the market, and toxicity limits clinical use of many compounds [1,2]
No significant interference was observed at the retention times of the analytes
Minocycline, cefotaxime and netilmicin were m/z 547.1!167.2!468.1, m/z 458.2!441.2!283.3!337.3, m/z 456.0!396.1!167.1!324.1, and m/z 476.2!299.2!191.5!160.1!458.4, respectively and these data agreed with mass fragmentation pathways [34,35,36]
Summary
Drug toxicity is a chief reason for drug withdrawal from the market, and toxicity limits clinical use of many compounds [1,2]. Studies suggest that $802 million and an average of 7 years are needed to develop a new compound and that only 1 in 5,000 candidate compounds enters clinical trials [3]. Accurate toxicity assessment can increase the chances that a drug will reach consumers. Zebrafish (Danio rerio) have been used in genetics, drug discovery, compound optimization, toxicology, and drug safety screening [4,5,6,7]. Zebrafish genes have homologous to human genes of approximate 70%, and partly zebrafish protein sequences, such as app, psen and pen, show over 70% homology to human corresponding proteins [8,9]; and they possess similarities with respect to nervous and cardiovascular systems, liver, PLOS ONE | DOI:10.1371/journal.pone.0124805. Zebrafish genes have homologous to human genes of approximate 70%, and partly zebrafish protein sequences, such as app, psen and pen, show over 70% homology to human corresponding proteins [8,9]; and they possess similarities with respect to nervous and cardiovascular systems, liver, PLOS ONE | DOI:10.1371/journal.pone.0124805 May 4, 2015
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