Nucleosil expert column 100 5 c18

HPLC was performed in modular reversed-phase high-performance liquid chromatography system (RP–HPLC, Schimadzu, Japan) with the use of Nucleosil expert column (EC) 100-5 C18 (Macherey–Nagel, Düren, Germany; column length 250 mm, inner diameter 4 mm, particle size 5 µm, pore size 100 Å). Samples were injected in a volume of 40 µL under isocratic conditions: mobile phase: 15% acetonitrile; flow rate: 0.8 mL/min; column temperature: 30 °C, analysis time: 30 min. Metabolites were detected by an ultraviolet–visible (UV–VIS) detector at 280 and 220 nm and by a fluorescence detector with a xenon lamp at excitation/emission wavelength 280/350 nm and 315/425 nm. The performance of the method was evaluated in terms of accuracy, linearity, imprecision and limit of detection.

Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC)
HPLC was performed in a modular reversed-phase high-performance liquid chromatography system (RP–HPLC, Schimadzu, Japan) with the use of Nucleosil expert column (EC) 100-5 C18 (Macherey–Nagel, Düren, Germany; column length 250 mm, inner diameter 4 mm, particle size 5 µm, pore size 100 Å). Samples were injected in a volume of 40 µL under isocratic conditions: mobile phase: 15% acetonitrile; flow rate: 0.8 mL/min; column temperature: 30 °C, analysis time: 30 min. Metabolites were detected by an ultraviolet–visible (UV–VIS) detector at 280 and 220 nm and by a fluorescence detector with a xenon lamp at excitation/emission wavelength 280/350 nm and 315/425 nm. The performance of the method was evaluated in terms of accuracy, linearity, imprecision, and limit of detection.