Analysis of chlorogenic acids in Helianthus tuberosus Linn leaves using high performance liquid chromatography-mass spectrometry

To identify chlorogenic acids in Helianthus tuberosus Linn leaves, a method of high performance liquid chromatography-ultraviolet-mass spectrometry (HPLC-UV-MS) was developed. HPLC analysis was performed on an Inertsil ODS-3 column (250mm x 4. 6 mm 5 mu m). The mobile phase consisted of 1% acetic acid (A) and methanol (B). A gradient program was adopted as follows: 0 - 10 min, 20% B to 35% B; 10 - 25 min, 35% B to 50% B; 25 - 35 min, 50% B to 80% B. The flow rate was set at 1. 0 mL/min and the column temperature was 35 degrees C. HPLC chromatogram was extracted at 327 nm. The mass spectrometer used was a TSQ triple quadrupole MS equipped with an electrospray ionization (ESI) interface. Initially, the mass spectrometer was programmed to perform full scan ranging from m/z 250 - 1 200. For MS/MS, negative ion monitoring mode was used and the collision energy was set at 10 - 45 eV. By analyzing UV characteristics and MS fragmentation patterns, 7 chlorogenic acids were assigned to be three categories, i.e., three caffeoylquinic acids, one feruloylquinic acid and three dicaffeoylquinic acids. As the results of the method development efforts, an effective and fast method for the qualitative identification of the chlorogenic acids in Helianthus tuberosus Linn leaves was established.