Benzyl itc

Lepidium graminifolium L. samples were obtained from wild-growing plants collected from two locations: Rab Island (43°30ʹ41.8ʹʹ N, 16°28ʹ02.7ʹʹ E; October, 2016) and Split (44°43ʹ53.8ʹʹ N, 14°50ʹ27.9ʹʹ E; June 2021). The botanical identity of the plant material was confirmed by a local botanist, Dr. Mirko Ruščić, from the Faculty of Natural Sciences, University of Split, Croatia and stored under voucher numbers HCPMFST 2030 (old v.n. DBLG001) for Rab sample and HCPMFST 4350 for Split sample. Myrosinase, benzyl ITC and sinigrin were obtained from Sigma Aldrich (St. Louis, MO, USA). All other chemicals and reagents were of analytical grade. Cancer cell lines (human bladder cancer cell line UM-UC-3 and human glioblastoma cell line LN229) were cultured in a humidified atmosphere with 5% CO₂ at 37 °C, in Dulbecco’s modified Eagle medium (DMEM, EuroClone, Milan, Italy) containing 4.5 g/L glucose, 10% fetal bovine serum (FBS) and 1% antibiotics (penicillin streptomycin, EuroClone, Milan, Italy).

The following ITCs were used during the course of this study: 2-propenyl-ITC (allyl-ITC; CAS 57-06-7; Sigma-Aldrich (St. Louis, MO, USA) 377430; 95%), phenyl-ITC (CAS 103-72-0; Sigma-Aldrich 139742; 98%), benzyl-ITC (CAS 622-78-6; Sigma-Aldrich 252492; 98%), 2-phenylethyl-ITC (phenethyl-ITC; CAS 2257-09-2; Sigma-Aldrich 253731; 99%), 4-phenylbutyl-ITC (CAS 61499-10-3; Maybridge, Altrincham, UK, TL00409EA; 95%).

Isobutyl ITC 97%, 4-pentenyl ITC 95% and sec-butyl ITC 97% were purchased from ABCR (Karlsruhe, Germany), 3-sec-butyl-2-methoxypyrazine from Acros Organics (Geel, Belgium), (3S,3aS,7aR)-wine lactone >95% from Aromalab (Freising, Germany), 2-(methylthio)ethyl ITC >96% from FCH Group (Chernigiv, Ukraine), butanoic acid ≥99% from Fluka (Steinheim, Germany), (Z)-3-hexenal 50% from SAFC (Steinheim, Germany) and 3-methylbutyl ITC >98% from Santa Cruz Biotechnology (Heidelberg, Germany). Acetic acid >99%, AITC 95%, p-anisaldehyde 98%, benzyl ITC 98%, 3-hydroxy-4,5-dimethyl-2(5H)-furanone (sotolone) ≥97%, 3-isobutyl-2-methoxypyrazine ≥99%, 3-isopropyl-2-methoxypyrazine 99%, 2-methylbutanoic acid 98%, 3-methylbutanoic acid 99%, skatole 98%, 3-(methylthio)propanal 98%, 3-(methylthio)propyl ITC 98%, 1-octen-3-one 50%, 2-phenylacetaldehyde 90% and PEITC >99% were purchased from Sigma-Aldrich (Steinheim, Germany) and 3-butenyl ITC 95% from TCI Europe (Eschborn, Germany). CETP was synthesized as described in our previous study (Kroener and Buettner, 2017 (link)).
Dichloromethane (DCM), which was freshly distilled for purification prior to use, and anhydrous sodium sulfate were purchased from VWR International GmbH (Darmstadt, Germany).