Venous blood samples (1.5–2.0 mL) were centrifuged for 1 min at 12000 × g to separate plasma and red blood cells. Plasma was deproteinized by adding perchloric acid (HClO4; final concentration 0.4N) and centrifuging for 5 min at 12000 × g. The supernatant was neutralized with KOH (pH 7.0–7.5) and filtered twice (Millex GS 0.22 μm, Millipore, Billerica, MA). Aliquots (0.1 mL) of whole blood, plasma and the final supernatant were counted in a gamma counter. Count data for plasma and whole blood aliquots were decay corrected and used to calculate the relative concentrations of [18F]2 in plasma and whole blood (Cp/Cwb). The final supernatant was analyzed by HPLC (Synergi 10 μm Hydro-RP column, 4.6 × 250 mm, 60 mM sodium phosphate buffer, pH 5.4 with 8% ethanol, flow rate 1.0 mL/min) with an in-line radiation detector. Under the HPLC conditions used [18F]2 had a retention time Rt = 12.7 min, while the main polar radiometabolite formed had Rt = 9.3 min. Peak area analysis of the radiation detection curve was used to estimate the percentage intact parent tracer fraction (fintact) for each plasma sample.
Millex gs 0.22 μm
Manufactured by Merck Group
Sourced in Germany
The Millex GS 0.22 μm is a laboratory filter designed for the filtration of aqueous solutions and other liquids. It features a 0.22 μm pore size membrane that effectively removes particulate matter and microorganisms from the sample.
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Lab products found in correlation
4 protocols using millex gs 0.22 μm
1
Radiotracer Quantification in Plasma
2
Synthesis of 11C-Choline Radiotracer
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With minor modifications, 11C-choline was produced according to the method described by Hara et al. [37 (link)]. In brief, 11C-CO2 was converted to 11C-CH3I by the catalytic gas-phase iodination reaction via 11C-CH4 (GE MeI MicroLab). 11C-CH3I, swept with a helium flow at 50 ml/min, was passed through a Light-CM cartridge loaded with N,N-dimethylethanolamine (25 μl). The column was washed with 10 ml ethanol followed by 10 ml water before eluting the product with isotonic saline (2-5 ml of 0.9% NaCl) through a Millipore filter (Millex GS, 0.22 μm) into a sterile vial. Quality control was performed using HPLC (LiChrosorb RP18, 250×4.6 mm; 1 mM sodium naphthalene sulfonic acid, 50 mM M H3PO4, 1.5 ml/min; k=3.7).
3
Solubilization of Compound 4R
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The solubility of 4R at 12 mg/mL was tested using 5 different vehicle mixtures. The one with the best solubility at 12 mg/mL was chosen. 4R was prepared at room temperature by adding 10% DMSO and 90% polyethylene glycol 400 (PEG400). The 4R formulation was observed to be a clear colorless solution. The solution was sterile filtered using a Millipore Millex-GS 0.22 μm syringe filter. Dose formulations were prepared on Day 1 prior to dose administration and were maintained at room temperature until administration to the animals.
4
Synthesis of [68Ga]Ga-NeoB Radiotracer
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The whole radiotracer production was performed inside a hot cell isolator (cleanroom class A, ITD, Dresden-Rossendorf, Germany) under GMP conditions 5 mL 0.1 M suprapur HCl were automatically eluted through the 68Ge/68Ga-generator and through a sterile filter (Millex-GS 0.22 μm, SLGSV255F, Millipore) directly into the reaction vial containing the NeoB precursor (50 ± 5 μg, 31.7 ± 0.6 nmol), resulting in a total reaction mixture volume of 3 mL containing 1100 ± 100 MBq 68Ga. Subsequently, the kit labeling buffer (0.50–0.55 mL 1 M formic acid with gentisic acid, pH 5) was added. Silicon-coated cannulas (0.6 × 60 mm, Sterican, B. Braun) were used throughout the whole synthesis process. Radiolabeling was performed at pH 3.6–4.0 for 7–10 min at 89 °C inside the reaction vial in a heating block (95 °C). The resulting solution contained 712 ± 73 MBq [68Ga]Ga-NeoB in a radiochemical purity (RCP) of 96–99%, as confirmed by radio-thin-layer chromatography (radio-TLC) and radio-HPLC (tR = 9.7 min).
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