In vivo pharmacokinetic, biodistribution, and elimination profiles of CLP after intravenous injection
This protocol is extracted from research article:
A self-illuminating nanoparticle for inflammation imaging and cancer therapy
Sci Adv, Jan 9, 2019; DOI: 10.1126/sciadv.aat2953

BALB/c mice (6 to 8 weeks) were intravenously injected with 0.1 ml of saline containing CLP nanoparticles at 25 mg/kg. At predetermined time periods, whole blood and major organs were collected after mice were euthanized. The plasma was obtained by centrifugation at 2000g for 10 min. After the proteins were precipitated via methanol, the plasma CLP concentrations were quantified by high-performance liquid chromatography (HPLC; Prominence-i LC-2030C, Shimadzu), using a Shim-pack GIST C18 column (250 mm by 4.6 mm; particle size, 5 μm) and an RF-20A fluorescence detector. The excitation wavelength was set at 401 nm, while the emission wavelength was set at 660 nm. The mobile phase consisted of water and methanol at a volume ratio of 10:90, with a flow rate of 1.0 ml/min. The column temperature was 40°C. For different organ tissues, they were homogenized in PBS and centrifuged, and the supernatant was collected. The CLP concentrations were also determined by HPLC.

In another study, excretion of CLP was examined in BALB/c mice that were housed individually in metabolic cages. Briefly, after CLP nanoparticles were administered via intravenous injection at 25 mg/kg, urine and feces were separately collected at predefined time points. Urine samples were precipitated via methanol, while feces were extracted with methanol. The CLP concentrations were quantified by HPLC.

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