Lars Holm Rasmussen1, Henrik AErenlund Pedersen2. 1. Department of Technology, Metropolitan University College, Copenhagen, Denmark. 2. Natural History Museum of Denmark, University of Copenhagen, Copenhagen, Denmark.
Abstract
INTRODUCTION: Ptaquiloside (PTA) is a noxious carcinogen found widespread in Bracken (Pteridium sp.) but with scattered and unresolved distribution outside the genus. The carcinogen causes Bovine Enzootic Haematuria among cattle all-over the World and is under suspicion of causing human cancers. OBJECTIVE: To set-up a methodology for large-scale qualitative studies on the distribution of PTA in ferns using already available herbarium specimens as source. METHODOLOGY: PTA and the main degradation product pterosin B (PtB) were quantified in aqueous frond extracts by HPLC-DAD. PTA was quantified after forced reaction into PtB. Optimal reaction conditions were tested using hydrochloric acid, sulphuric acid, trifluoroacetic acid (TFA), acetic acid and temperature as the experimental variables. A pair-wise test for PTA degradation in plant-press were used to explore the effect of this preservation regime. A selection of up to 50 years old Bracken herbarium specimens were tested for PTA and PtB. The methodology was applied on 21 fern species from Denmark. RESULTS: An optimised TFA-based method results in 30.7% higher conversion and a 1:1 reaction between PTA and PtB. Full three-dimensional resolution of the analyte was obtained. Preservation of fronds in a plant press increase formation of PtB. Hence, the method is only suitable for qualitative studies. Presence of PTA and PtB were found in samples up to 50 years old. Among 21 ferns tested, the compounds were only found in Pteridium aquilinum. CONCLUSION: Herbarium specimens up to 50 years old can be used for explorative risk assessment of ferns using HPLC-DAD for quantification and identification.
INTRODUCTION:Ptaquiloside (PTA) is a noxious carcinogen found widespread in Bracken (Pteridium sp.) but with scattered and unresolved distribution outside the genus. The carcinogen causes BovineEnzootic Haematuria among cattle all-over the World and is under suspicion of causing humancancers. OBJECTIVE: To set-up a methodology for large-scale qualitative studies on the distribution of PTA in ferns using already available herbarium specimens as source. METHODOLOGY:PTA and the main degradation product pterosin B (PtB) were quantified in aqueous frond extracts by HPLC-DAD. PTA was quantified after forced reaction into PtB. Optimal reaction conditions were tested using hydrochloric acid, sulphuric acid, trifluoroacetic acid (TFA), acetic acid and temperature as the experimental variables. A pair-wise test for PTA degradation in plant-press were used to explore the effect of this preservation regime. A selection of up to 50 years old Bracken herbarium specimens were tested for PTA and PtB. The methodology was applied on 21 fern species from Denmark. RESULTS: An optimised TFA-based method results in 30.7% higher conversion and a 1:1 reaction between PTA and PtB. Full three-dimensional resolution of the analyte was obtained. Preservation of fronds in a plant press increase formation of PtB. Hence, the method is only suitable for qualitative studies. Presence of PTA and PtB were found in samples up to 50 years old. Among 21 ferns tested, the compounds were only found in Pteridium aquilinum. CONCLUSION: Herbarium specimens up to 50 years old can be used for explorative risk assessment of ferns using HPLC-DAD for quantification and identification.
Authors: Paulo César Dos Reis Aranha; Lars Holm Rasmussen; Godelind Alma Wolf-Jäckel; Henrik Michael Elvang Jensen; Hans Christian Bruun Hansen; Christian Friis Journal: PLoS One Date: 2019-06-21 Impact factor: 3.240