1200 isocratic pump

Pieces of whole liver were minced, weighed into individual wells, then homogenization buffer [63 (link)] was added to those corresponding wells. Control tissue homogenate was made by adding homogenization buffer at a 9 to 1 ratio to weighed amount of mouse untreated liver. Aliquots and appropriate amounts of calibration standards were added into wells. Internal standard and approximately 0.25cm³ granite beads were added and then extracted via a liquid-liquid extraction with ammonium hydroxide and phenol: chloroform: isoamyl alcohol (25:24:1). The aqueous layer was then further processed via solid phase extraction utilizing a Phenyl plate. Eluates had a final pass through a protein precipitation plate before dry down under nitrogen at 50°C. Dried samples were reconstituted in water containing 100 μM EDTA. These samples were injected into an Agilent LCMS instrument consisting of a 1260 binary pump, a 1200 isocratic pump, a column oven, an auto sampler, and a 6130 single quadrupole mass spectrometer for analysis (Agilent, Wilmington, DE, USA).

Two different commercially available PCLs were used as bulk materials. (Table 1). Molecular weight was characterized at the Technical University Braunschweig. Gel permeation chromatography (GPC) measurements were performed on a PSS SECcurity2 instrument equipped with a SECcurity2 vacuum degasser, a SECcurity2 TCC6500 column oven (all PSS GmbH, Mainz, Germany), a 1200 isocratic pump (Agilent Technologies Germany GmbH & Co. KG, Waldbronn, Germany), a SECcurity2 1200 refractive index detector (PSS GmbH, Mainz, Germany) and a manual injection valve (Rheodyne^®, IDEX Health & Science LLC, Oak Harbor, WA, USA). Both PCL species were recorded in THF at 40 °C on a PSS triple SDV column setup (1 × pre-column, 2 × main columns, each 10 µm particle and pore size) with a concentration and flow rate of 1 g/L and 1 mL/min, respectively.