Table 2. Comparison of LC-MS analysis of the F16 sub-fractions obtained from the RP-HPLC and their effect on the A549 cell viability.
| Fraction | |||||||||
| control | F3 | F4 | F5 | F6 | F7 | F8 | F9 | F10 | |
| Metabolites | Relative LC-MS peak intensity | ||||||||
| Tryptoquivaline F | nd | 42.0 | 100 | 6.7 | nd | nd | nd | nd | |
| Fumiquinazoline C | nd | 31.3 | 100 | 8.3 | 4.2 | nd | nd | nd | |
| Fumagillin | nd | 97.5 | 100 | 93.1 | nd | nd | nd | ||
| Trypacidin | nd | 100 | 8.8 | 3.0 | nd | nd | |||
| Monomethylsulochrin | nd | 100 | 40.9 | 2.3 | nd | nd | |||
| Questin | nd | 100 | 54.7 | 1.6 | nd | nd | |||
| Verruculogen | nd | nd | 100 | 58.9 | nd | nd | |||
| Helvolic acid | nd | nd | nd | 100 | nd | nd | |||
| Fumitremorgin B | nd | nd | nd | 100 | nd | nd | |||
| Emodin | nd | nd | nd | nd | nd | nd | |||
| Fumigaclavine A | nd | nd | nd | nd | nd | nd | |||
| Unknown (m/z 453)* | nd | nd | nd | 60.3 | 100 | 48.1 | |||
| cell viability (%) | 92.5 | 80.7 | 79.0 | 77.2 | 35.5 | 48.7 | 47.0 | 47.1 | 70.4 |
Sub-fractionation of F16 fraction using RP-HPLC. F3 to F7 were each collected for 2 min, F8 for 5 min, F9 for 10 min and F10 for 6 min. Each fraction was evaporated and the dried material was dissolved in 50 µl PBS-DMSO 12%. The final concentration of DMSO in the assay medium was 1.2%. The cell viability was measured by the MTT assay described in the Materials and Methods after exposing A549 cells to the metabolites for 48 h. The effect of 1.2% DMSO was used as negative control. For each metabolite, the figures are arbitrary units indicating its elution profile, 100 corresponding to the maximum normalized level measured by the LC-MS sensor. nd means not detectable.
*m/z in positive mode electrospray ionization.