Determination of gibberellins in fermentation broth produced by Fusarium verticilliodes MTCC 156 by high-performance liquid chromatography tandem mass spectrometry

Abstract

A method for the detection of gibberellins produced by Fusarium verticilliodes is described using HPLCMS/MS (HPLC tandem MS). A Hypersil (5 μm) octadecylsilane column with methanol/water as eluent in the ratio 3:1 at a flow rate of 0.5 ml/min was used. In the HPLCMS, GA₃ (gibberellic acid; m/z 346.3) eluted at retention time t_r=3.08 min, with the corresponding mass chromatogram having peaks at m/z 346.7 and 328.8 corresponding to the M⁺ and M⁺-H₂O ions respectively. The ethyl acetate extract from the broth, subjected to HPLCMS analysis under similar conditions, showed a constituent with t_r=2.13 min, the mass chromatogram of which exhibited peaks at m/z 348.9 and 331.9 corresponding to the MH⁺ and MH⁺ -H₂O ions respectively. Comparison of the MS and MS/MS results (direct infusion) of an authentic sample of GA₃ and the ethyl acetate extract from the broth revealed the formation of reduced GA₃ in the broth. The present study, utilizing HPLCMS/MS, describes an improved methodology for the unambiguous determination and estimation of gibberellins from fermentation broth.