A novel method for analyzing solanesyl esters in tobacco leaves using atmospheric pressure chemical ionization/mass...

A direct and simple method for analyzing solanesyl esters found in tobacco leaves was developed. Sample preparation was performed by accelerated solvent extractor 200 (ASE200) using n-hexane followed by evaporating solution in vacuo and dissolving residue with acetone. The separation of analytes was conducted through high-performance liquid chromatography (HPLC) equipped with an SIL-C18/5C column and the non-aqueous reversed phase chromatography (NARP) technique using acetone and acetonitrile as the mobile phase with a linear gradient. Atmospheric pressure chemical ionization/mass spectrometer (APCI/MS) in positive mode was used to detect solanesyl esters in the following conditions: capillary voltage 4000 V, corona current 10 microA, drying gas flow 5 mL/min, fragmentor voltage 200 V, nebulizer pressure 60 psi, and vaporizer temperature 500 degrees C. Each solanesyl ester was identified by the comparison of analyte with synthesized solanesyl esters. Quantification was conducted by selected ion monitoring (SIM) mode in order to detect the specific product ion (613.6 m/z) fragmented from solanesyl ester. The calibration curve was made in the range of 0.1-40 microg/mL with a regression coefficient over 0.999 on almost all solanesyl esters. The limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.01 to 0.05 microg/mL and from 0.03 to 0.15 microg/mL, respectively, on the SIM mode of MS for quantification. Recovery (%) ranged from about 80 to 120%. The direct quantification using the developed method succeeded in showing a different amount and composition of solanesyl esters among various tobacco leaves.