Agilent hp 1100 series

To assess malondialdehydes (MDA) concentrations, breast and thigh muscle samples were homogenized in a standard buffer solution, centrifuged at 700 × g, and the clear supernatant layer was collected (23 (link)). The samples were analyzed using the Agilent HPLC apparatus (Agilent HP 1100 series, USA). Supelcosil C18 was used as the analytical column (particle size of 5 m and a pore size of 8 nm). The detection wavelength was 250 nm and the flow rate was 1.5 ml/min.

All the isotope-labelled pHLIP (NH₂-GGEQNPIYWARYADWLFTTPLLLLDLALLVDADEGT-CO₂H, with labelled sites shown in bold) were synthesized manually using the routine 9-fluorenylmethyloxycarbonyl (Fmoc) protecting group chemistry with preloaded Fmoc-Thr(tBu)-Wang resin (0.3 mmol g⁻¹ substitution, AAPPTec Inc., Louisville, KY). The isotope-labelled amino acids were purchased from Cambridge Isotope Laboratory, Inc. (Tewksbury, MA), and Fmoc protection of the free amino group was accomplished using literature approaches52 (link). The crude peptides were purified using high-performance liquid chromatography (Agilent HP 1100 series, Agilent Technologies Inc., Santa Clare, CA) installed with a Zorbax C18 reversed-phase column (Agilent Technologies Inc., Santa Clare, CA). A linear gradient with water and acetonitrile was utilized for the purification. The final products were verified with matrix-assisted laser desorption/ionization–time of flight mass spectrometry (Mass Spectrometer Facility, University of Illinois).