Finnigan hplc system

The concentration of DNA was quantified by HPLC-UV analysis of dG in the enzymatic hydrolysate of DNA. The amount of DNA was calculated from the dG content by assuming that 1 mg of DNA contained 3 μmol of nucleotides and that dG accounted for 22% of the total nucleotides in DNA [36 (link)]. The experiment was conducted on a Thermo Finnigan HPLC system with a diode array detector. Gradient elution was performed using a Luna C18 column (4.6×250 mm, 5 μm in particle size, Phenomenex, Torrance, CA) eluted with deionized water (solvent A) and methanol (solvent B) at a flow rate of 0.7 mL/min. The UV signal was monitored at 254 nm. The solvent composition began at 5% B followed by linear increase to 22% B over 15 min. Then the solvent B was raised to 80% over 2 min followed by an isocratic elution for 3 min. Subsequently, the percentage of the solvent B was brought back to 5% over 2 min, followed by an equilibration for 15 min.

The high-performance liquid chromatographic (HPLC) analysis of the anthocyanins from the BC samples was conducted on a Thermo Finnigan HPLC system (Thermo Scientific, Waltham, MS, USA). The separation and identification of the main anthocyanin compounds was employed on a Synergi 4u Fusion-RP 80A (150 × 4.6 mm, 4 μm) column, at the 520 nm wavelength, at an oven temperature of 27 °C. The samples were filtered through a C18 Sep-Pack cartridge-Waters and through a 0.22-μm syringe filters (Bio Basic Canada Inc., Toronto, ON, Canada). The mobile elution phase consisted of two main solvents that were 10% formic acid (A) and 100% methanol (B) with a vertical gradient. The injection volume was 10 µL at a flow rate of 1.0 mL/min. The identification and quantification of the compounds were made based on each compound’s calibration curve, and the results were expressed as mg/g d.w.