The main challenge was how to make Bingo Cartridge methods usable as Spring Data Repository methods.
This involved two steps: First the Bingo Cartridge methods were exposed to Hibernate by extending PostgreSQL82Dialect and then were used by creating custom Repository implementations.
Additionally, results for RCP using SEP-PAK cartridge based methods were found to be operator dependent due to the influence of unavoidable variations in elution velocity (Vinberg, 2000; Ponto, 2005; Millar & Hesslewood, 2004).
This should ideally be a single synthetic step in the clinic that requires no purification post labelling, or, if purification is necessary, in which a simple cartridge-based method can be applied.
The final extract was reduced in volume to approximately 1 mL using an automated nitrogen evaporation system (Turbo Vap II, Zymark Inc .. Extracts were then purified by elution through a Florisil solid-phase extraction cartridge utilizing the method reported by [ 28].
We have used a 200-mg Waters Sep-Pak tC18 reversed-phase cartridge with a method optimised for the recovery of oxysterols with a side-chain hydroxyl group (e.g. 22 R-, 24 S-, 25- and 26-HC) and for cholestenoic acids (e.g. 3β-hydroxycholest-5-enoic acid and 3β,7α-dihydroxycholest-5-enoic acid).
Sera of the GEN-treated animals were extracted using the SEP-PAK cartridges solid-phase method according to the standard protocol suggested by the manufacturer (Waters, Eschborn, Germany), and ethanol-redissolved aliquots were subjected to HPLC separation and UV-detection as described above.
U and Th were then separated on a TEVA cartridge (Eichrom) using a method developed in our laboratory (Knight et al. 2014).
The disposable spin cartridge has become the method of choice for hydrophobic fractionation in our laboratory because of its proven functionality, no pressure- build up, low cost, and zero sample-to-sample cross contamination.
To set up 11C MET production at our institution, we used the simplest method, Sep-Pak cartridge without HPLC purification (Gomzina et al. 2011), for our initial attempt.
