Eba 20 centrifuge

The rats were randomized, weighed, and tail marked into two groups, namely a capmatinib-administered group and a placebo group (n = 6). The capmatinib was freshly dissolved in a 0.5% carboxymethylcellulose (CMC) solution prepared in distilled water while the placebo solution consisted of 0.5% CMC alone. The fasting rats (12 h, water ad libitum) were administered either capmatinib at 10 mg/kg dose or an equivalent amount of 0.5% CMC solution by oral gavage using a stainless-steel oral gavage needle (Harvard Apparatus, Holliston, USA).
A volume of 200 µL of the blood samples was obtained from the retro-orbital plexus using clean capillary tubes and collected into Minicollect EDTA tubes. Blood sampling was performed under light isoflurane anesthesia (Hikma Pharmaceuticals, Amman, Jordan) (5% induction and 2.5% maintenance) carried by oxygen (dual-flow oxygen concentrator, China) using a low-flow anesthesia system (SomnoSuite, Kent Scientific, Torrington, CT, USA). An initial blood sample, a zero-time sample, was collected from each animal. The blood samples were then collected at specific time intervals at 0.25, 0.50, 1, 2, 4, 8, and 24 h post-drug administration. The collected blood tubes were centrifuged for 10 min at 6000 rpm using a Hettich EBA 20 centrifuge, Tuttlingen, Germany. The clear plasma samples were then collected and stored at −80 °C until HPLC analysis.

All experiments were performed using a 1200 high-performance liquid chromatograph combined with a 6410 triple quadrupole mass spectrometer from Agilent Technologies (Santa Clara, CA, USA). The system was equipped with an electrospray ionization (ESI) operating in positive mode. An InfinityLab Poroshell 120 EC-C18 column (4.6 $\times$ 100 mm $\times$ 2.7 µm) also from Agilent was used for chromatographic separation. Data were collected and processed using the MassHunter Quantitative Analysis (QqQ) software, using the multiple reaction monitoring (MRM) mode. SigmaPlot 13.1 (Systat Software, San Jose, CA, USA) and MetaboAnalyst 5.0 software were used for the statistical analysis.
An EBA 20 centrifuge (Hettich, Tuttlingen, Germany), an XcelVap air-drying system (Horizon Technology, Salem, MA, USA) and an LLG-uniTEXER vortex (Heathrow Scientific, IL, USA) were used during sample treatment. Tissue samples were ground with an IKA A11 basic mixer (Wilmington, USA). Nylon syringe filters of 0.45 μm $\times$ 25 mm and 0.2 μm $\times$ 13 mm from Agilent were used to filter the sample extracts.