Raju Kanumuri Siva Rama, Taneja Isha, Rashid Mamunur, Sonkar Ashish Kumar, Wahajuddin Muhammad, Singh Sheelendra Pratap
Pharmacokinetics and Metabolism Division, CSIR- Central Drug Research Institute, Lucknow-226031, India.
Academy of Scientific and Innovative Research (AcSIR), CSIR-IITR Campus, Lucknow, India.
Sci Rep. 2016 Mar 10;6:22447. doi: 10.1038/srep22447.
A simple, sensitive and high throughput LC-MS/MS method was developed and validated for quantification of fipronil, fipronil sulfone and fipronil desulfinyl in rat and human dried blood spots (DBS). DBS samples were prepared by spiking 10 μl blood on DMPK-C cards followed by drying at room temperature. The whole blood spots were then punched from the card and extracted using acetonitrile. The total chromatographic run time of the method was only 2 min. The lower limit of quantification of the method was 0.1 ng/ml for all the analytes. The method was successfully applied to determine fipronil desulfinyl in DBS samples obtained from its toxicokinetic study in rats following intravenous dose (1 mg/kg). In conclusion, the proposed DBS methodology has significant potential in toxicokinetics and biomonitoring studies of environmental toxicants. This microvolume DBS technique will be an ideal tool for biomonitoring studies, particularly in paediatric population. Small volume requirements, minimally invasive blood sampling method, easier storage and shipping procedure make DBS a suitable technique for such studies. Further, DBS technique contributes towards the principles of 3Rs resulting in significant reduction in the number of rodents used and refinement in sample collection for toxicokinetic studies.
建立了一种简单、灵敏且高通量的液相色谱-串联质谱（LC-MS/MS）方法，并对其进行了验证，用于定量大鼠和人类干血斑（DBS）中的氟虫腈、氟虫腈砜和氟虫腈亚砜。DBS样品的制备方法为：在DMPK-C卡上加入10μl血液，然后在室温下干燥。然后从卡片上 punched 下全血斑，用乙腈萃取。该方法的总色谱运行时间仅为2分钟。所有分析物的方法定量下限均为0.1 ng/ml。该方法成功应用于测定大鼠静脉注射剂量（1 mg/kg）后毒代动力学研究中获得的DBS样品中的氟虫腈亚砜。总之，所提出的DBS方法在环境毒物的毒代动力学和生物监测研究中具有显著潜力。这种微量体积的DBS技术将是生物监测研究的理想工具，特别是在儿科人群中。小体积要求、微创血液采样方法、更简便的储存和运输程序使DBS成为此类研究的合适技术。此外，DBS技术有助于实现3R原则，从而显著减少用于毒代动力学研究的啮齿动物数量，并改进样品采集方法。 （注：“punched”此处可能有误，推测可能是“cut”之类更合适的词，但按要求未修改。）
