Tof lc ms

Each sample (20 µl) was applied to Agilent TOF LC/MS and separated by VyDac C18 5-µm-particle-size column (3.2 mm by 150 mm). The column was previously equilibrated with 95% solvent A (water–0.01% formic acid) and 5% solvent B (acetonitrile 0.1% formic acid) with flow rate of 0.5 ml min-1. Following each injection, solvent B was increased by 2.5% per min to 55% over a 20 min period. Finally, solvent B was increased to 95% in 1 min and then returned to starting concentration over the next 1 min. The sodium adducts of each compound provided the most prominent peak and therefore were used for detection purposes whereas the parent ions provided the relative results. Compound obtained in the extracts was quantitated by comparing to molecular weight by mass spectrometry of each purified compound solved in DMSO as well as to elution profiles.

Each preparation (20 µl) was subjected to Agilent TOF LC/MS. Each compound was separated through VyDac C18 5-µm-particle-size column (1.0 mm by 50 mm) using gradient of solvent A (water–0.01% formic acid) and solvent B (acetonitrile–0.1% formic acid). The flow rate was set to 0.5 ml min⁻¹, and the column was equilibrated with 95% solvent A and 5% solvent B. Following each injection, solvent B was increased to 55% over a 20 min period (2.5% increase per min). At 21 min solvent B was increased to 95% in 1 min and then returned to starting conditions over the next 1 min. Each compound was detected by TOF-MS using an Agilent 6230 mass spectrometer in selective ion monitoring mode. The sodium adducts of each compound provided the most prominent peak and therefore were used for detection purposes although the parent ions provided the relative results. The amount of compound obtained in the extracts was determined by comparison to standards of each purified compound dissolved in DMSO. Each compound was confirmed with both elution time over the column and molecular weight by mass spectrometry.