Fig. 1.

pHPL is excreted in high amounts by L. infantum promastigotes. Figures A, B and C correspond to HPLC chromatograms. (A) 20 μl of a solution containing 0.5 mM pHPL as standard was run. The peak corresponding to pHPL eluted at 11.14 min. (B) A total volume of 230 ml of complete medium was used to extract the ether soluble fraction as control in order to asses that there are no peaks at the same elution time than the standard in the chromatogram. 20 μl was loaded in the run. (C) A total volume of 230 ml of supernatant obtained from a stationary-phase promastigote culture was used to extract the ether-soluble fraction and analyzed by HPLC. The peak which its retention time is similar to the retention time of the standard was eluted and sent for further analysis by GC–MS. (D) Gas chromatogram of the peak eluted in Fig. 1C. The major peak with an elution time of 13.7 min was isolated and analyzed by mass spectrometry. (E and F) The mass spectrum of the peak (E) was identical to the mass spectrum of the pHPL standard (F).