tIn order to improve the economic sustainability of the Jatropha-biofuel chain, seed cake detoxificationand utilization of ‘non toxic’ Jatropa curcas accessions are the main activities pursued with the aim ofusing J. curcas seed cake in animal feed. Given this growing interest, a robust and reliable method forphorbol esters (PEs) determination is necessary. HPLC-UV is a well-established method to detect andquantify the PEs content in Jatropha seeds and related products, but it seems to be unsuitable for morecomplex matrices like Jatropha leaves and animal tissues, due to the presence of interfering compounds.The objective of this work was to develop and optimize a LC–MS/MS method for the quantitative determi-nation of PEs in seeds and leaves of J. curcas L. plants from Ghana and Mexico and in liver (as an organ withthe function of accumulation) from goats fed with PEs in their diet. The HPLC-UV analysis evidenced fivechromatographic peaks in the toxic seed kernels corresponding to the factors C1, C2, C3, C6 and C4–C5,respectively, with a PEs concentration of about 5100 microg/g (as TPA equivalent). No PEs related peaks weredetected in Mexican kernel seeds while in the case of leaves and liver the analysis was hampered bythe presence of interfering compounds. The toxic kernel seed extract was used as a standard solutionfor the PEs quantitation in leaves and liver samples by LC–MS/MS, with the standard addition method.The most intense MRM transitions used to quantify and qualify the PEs were: 675 → 311, 693 → 311, and293 → 265 m/z. The LC–MS/MS method with a LOD and a LOQ of 0.07 and 0.21 microg/g, respectively, resultedin more sensitivity and selectivity than the HPLC-UV method. All three MRM transitions were present inGhanaian toxic kernel seed, while no peaks were present in the supposed non-toxic Mexican kernel seed.PEs concentration in the leaves of toxic Ghanaian accession resulted in about 1/10 of that in the kernel, while no PEs peaks were found in the J. curcas leaves from Mexico and in liver samples.

Determination of phorbol esters in seeds and leaves of Jatropha curcas and in animal tissue by high-performance liquid chromatography tandem mass spectrometry

BALDINI, Mario;FERFUIA, Claudio;BORTOLOMEAZZI, Renzo;VERARDO, Giancarlo;PIASENTIER, Edi;FRANCESCHI, Loretta
2014-01-01

Abstract

tIn order to improve the economic sustainability of the Jatropha-biofuel chain, seed cake detoxificationand utilization of ‘non toxic’ Jatropa curcas accessions are the main activities pursued with the aim ofusing J. curcas seed cake in animal feed. Given this growing interest, a robust and reliable method forphorbol esters (PEs) determination is necessary. HPLC-UV is a well-established method to detect andquantify the PEs content in Jatropha seeds and related products, but it seems to be unsuitable for morecomplex matrices like Jatropha leaves and animal tissues, due to the presence of interfering compounds.The objective of this work was to develop and optimize a LC–MS/MS method for the quantitative determi-nation of PEs in seeds and leaves of J. curcas L. plants from Ghana and Mexico and in liver (as an organ withthe function of accumulation) from goats fed with PEs in their diet. The HPLC-UV analysis evidenced fivechromatographic peaks in the toxic seed kernels corresponding to the factors C1, C2, C3, C6 and C4–C5,respectively, with a PEs concentration of about 5100 microg/g (as TPA equivalent). No PEs related peaks weredetected in Mexican kernel seeds while in the case of leaves and liver the analysis was hampered bythe presence of interfering compounds. The toxic kernel seed extract was used as a standard solutionfor the PEs quantitation in leaves and liver samples by LC–MS/MS, with the standard addition method.The most intense MRM transitions used to quantify and qualify the PEs were: 675 → 311, 693 → 311, and293 → 265 m/z. The LC–MS/MS method with a LOD and a LOQ of 0.07 and 0.21 microg/g, respectively, resultedin more sensitivity and selectivity than the HPLC-UV method. All three MRM transitions were present inGhanaian toxic kernel seed, while no peaks were present in the supposed non-toxic Mexican kernel seed.PEs concentration in the leaves of toxic Ghanaian accession resulted in about 1/10 of that in the kernel, while no PEs peaks were found in the J. curcas leaves from Mexico and in liver samples.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11390/975947
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