Pu1580 unit

SCE was carried out in a laboratory-scale supercritical fluid extraction system (Jasco Europe Srl, Cremella, Italy). The system was operated in a semi-continuous mode by pumping scCO₂ with a Jasco-PU1580 unit, through a stainless-steel column, 10 mm i.d. and 200 mm length, equipped with 10 µ porous stainless steel 316 L sintered filter disks. The column was filled with the dehydrated samples, heated using a Jasco CO-4061 oven to the chosen temperature, and scCO₂ was pumped to the target pressure. After a pre-equilibrating time of 30 min, SCE was performed.
The dried leaves (7.0 g) were transferred in the CO₂ extractor added with 5 mL ethanol and extracted for 1 h at 150 bar and 55 °C. The dried fruits (7.0 g) were transferred to the CO₂ extractor and extracted for 1 h at 250 bar and 80 °C. The extraction mixtures (F_S and L_S) were immediately analyzed by ¹H and ¹³C NMR and HPLC analysis or stored at 4 °C until the analyses were performed.

Different high performance liquid chromatography (HPLC) methods were applied using a Jasco PU 1580 unit (Jasco Germany GmbH, Pfungstadt, Germany) with degasser, quaternary pump, autosampler and UV/Vis (Jasco UV 2075 Plus) as well as fluorescence detector (Jasco FP 1520) to examine the production of the Fusarium mycotoxins deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZON) and alpha- and beta-zearalenol (α- and β-ZOL) as well as the Alternaria mycotoxins alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT), and tenuazonic acid (TeA). The wheat grains overgrown with bacteria and fungi were ground with a mortar and then extracted to detect the mycotoxins. Detailed extraction and HPLC detection methods for Fusarium toxins are described by Müller et al. (2010) (link) and for Alternaria toxins by Müller and Korn (2013) (link) and Kahl et al. (2016) (link). Photodiode array detection was performed to control toxin identity. Standard substances of DON, NIV, and ZON were obtained from Sigma-Aldrich Chemie GmbH (Taufkirchen, Germany), AOH, AME, ALT and TeA from Romer Labs Diagnostic GmbH (Tulln, Austria). The toxin detection limits in these grains were 10 ng DON, 20 ng NIV, 1 ng ZON, 2 ng α- and β-ZOL, 50 ng AOH, AME, or ALT, and 100 ng TeA per g of fresh substrate mass. All toxin concentrations were calculated on the dry mass of the substrate (ng g^-1 DM).