2 propanol

Resveratrol (99.0% purity, trans-form) was purchased from Kukjeon Pharm Co., Ltd. (Gyeonggi, Korea). Carbamazepine (99.0% purity) and sodium lauryl sulfate (SLS) were purchased from Sigma-Aldrich Co. (St Louis, MO, USA). Diethylene glycol monoethyl ether (DEGEE, Transcutol^® HP, Gattefossè, Saint-Priest, France) was obtained from Masung & Co., Ltd. (Seoul, Korea). Polyvinylpyrrolidone (PVP K12, K17, K25, and K30) and polyvinylpyrrolidone vinyl acetate VA64 (PVP VA64) were obtained from BASF Co., Ltd. (Kollidon^®, Ludwigshafen, Germany). Hydroxypropyl cellulose polymers (HPC-L, HPC-SL, and HPC-SSL) were purchased from Nippon Soda Co., Ltd. (Tokyo, Japan). Hydroxypropyl methyl cellulose (HPMC, Hypromellose 3 cp, 6 cp, and 15 cp) was obtained from Shin-Etsu Chemical Co., Ltd. (Tokyo, Japan). Acetone (99.8% purity), 1-butanol (99.5% purity), and ethyl acetate (99.5% purity) were purchased from Daejung Chemical & Metals Co., Ltd. (Gyeonggi, Korea). Dichloromethane was purchased from Junsei Chemical Co., Ltd. (Tokyo, Japan). High-performance liquid chromatography (HPLC)-grade methanol, ethanol, acetonitrile, 1-propanol, and 2-propanol were purchased from Honeywell Burdick & Jackson (Morristown, NJ, USA).

Pentaerythritol, methacryloyl chloride, triethylamine, tetrahydrofuran (THF; extra dry), and acetone (extra dry) were purchased from Acros Organics. CL was obtained from Tokyo Chemical Industry. 3,6-Dimethyl-1,4-dioxane-2,5-dione (DLLA) was purchased from Huizhou Foryou Medical Devices Co. Ltd. or Acros Organics. Tin(II)-2-ethylhexanoate [Sn(Oct)₂], 2,2-diethyl-1,3-propanediol, sodium bicarbonate (NaHCO₃), sodium chloride (NaCl), (+)-α-tocopherol (vitamin E), NVP, BAPO, 1-(phenyldiazenyl)naphthalen-2-ol (Sudan I), borane-tert-butylamine complex [(CH₃)₃CNH₂ BH₃], and lithium bromide (LiBr) were purchased from Sigma-Aldrich. Hydrogen tetrachloroaurate(III) trihydrate (HAuCl₄) was obtained from abcr. Hexane, dichloromethane (DCM), and dimethylformamide (DMF) were purchased from Thermo Fisher Scientific. Dimethyl sulfoxide-d6 (DMSO-d6) was obtained from ReseaChem. Methanol and 2-propanol were provided by VWR Chemicals. Midazolam was purchased from Sintetica. Alfaxalon was purchased from Jurox UK. Isoflurane was purchased from Provet AG. PBS was obtained from Thermo Fisher Scientific. All chemicals were used as received.

p-HPEA, 3,4-DHPEA, verbascoside, rosmarinic acid, and α-tocopherol were purchased from Merk (Milan, Italy). The secoiridoid derivatives (i.e., oleacein (3,4-DHPEA-EDA) and oleocanthal (p-HPEA-EDA)) were obtained as reported in the previous work of Taticchi et al. [39 (link)]. Then, 2,4,6-tripyridyls-triazine (TPTZ), 6-hydroxy-2,5,7,8-tetramethyl-chroman-2-carboxylic acid (Trolox), 2,2-diphenyl-1-picrilidrazil (DPPH˙), and butylated hydroxytoluene (BHT) were purchased from Merk (Milan, Italy). Formic acid, n-hexane, phosphoric acid, hydrochloric acid, methanol, ethanol, 2-propanol, and acetonitrile were supplied by VWR (Milan, Italy). HPLC-grade methanol and water for HPLC-MS were supplied by Carlo Erba (Milan, Italy). Finally, 4-methyl-2-pentanol, as an internal standard for pesto volatile compounds, was purchased from Sigma-Aldrich (Milan, Italy).

Reagents were of analytical purity. Media components (peptone, glucose monohydrate, sodium nitrate, potassium hydrogenphosphate, magnesium sulfate, potassium chloride, ferrous sulfate) were from Reanal (Budapest, Hungary). Allyl isothiocyanate, allyl cyanide, 2-phenylethyl isothiocyanate and phenylpropionitrile and glutathione (reduced form) were from Sigma Aldrich (MO, USA). Pure glucosinolates were from Phytoplan (Germany). LC-MS grade acetonitrile, water and formic acid were purchased from Fisher Scientific (Belgium). Solvents (1-propanol, 2-propanol) and media additives (streptomycin, chloramphenicol and dichloran) were from VWR. Double distilled water was used throughout the study.

The reagents that were used for the synthesis of the SBA-15 were: triblock copolymer Pluronic P123 (PEO₂₀PPO₇₀PEO₂₀ average Mn ~5800, Sigma-Aldrich, Saint Louis, MO, USA) as template, tetraethylorthosilicate (TEOS 98%, Aldrich) as silicon source, zirconium propoxide (Zr(OCH₂CH₂CH₃)₄, 70 wt% in 1-propanol, Aldrich) as zirconium source, and hydrochloric acid (HCl 37%, VWR, Radnor, PA, USA). The pore expander was of 1,3,5-trimethylbenzene (TMB 98%, Aldrich), while the fluoride source was ammonium fluoride (NH₄F 99.5%, Aldrich).
The chemicals used in the MPV reaction were: furfural (Sigma-Aldrich, 99%), 2-propanol (VWR, HPLC grade, 99.9%), 2-butanol (Sigma-Aldrich, 99.5%), and cyclohexanol (Sigma-Aldrich, 99%), used as sacrificing alcohols, and o-xylene (Sigma-Aldrich, 99.9%) was employed as the internal standard. The gases that were employed in this research were He (Air Liquide, Paris, France, 99.99%), H₂ (Air Liquide, 99.999%), and N₂ (Air Liquide, 99.9999%).

Naproxen was obtained from Tokyo Chemical Industry Co (Japan). Other excipients used to prepare the liqui-pellet included microcrystalline cellulose (Avicel PH-101), (FMC corp., UK), hydrophilic fumed silica with specific surface area of 300 m²/g (Aerosil 300) (Evonik Industries AG, Hanau, Germany), sodium starch glycolate Type A (Primojel) (DFE Pharma, Goch, Germany), croscarmellose (Primellose) (DFE Pharma., Goch, Germany), 2-propanol (VWR Chemicals, Fontenay Sous Bois France), polysorbate 80 (Tween 80) and PEG with molecular weight of 1500 (Acros Organics, UK). All other reagents and solvent were of analytical grades.

10 µm tissue sections were mounted onto spatially barcoded glass slides with poly-T reverse transcription primers, one section per array. The prepared slides were warmed to 37 °C, following which the sections were fixed using 4% formaldehyde solution (P087.1, Carl Roth, Karlsruhe, Germany) for 10 min, which was then rinsed using PBS. The fixed sections were covered with 2-Propanol (20842312, VWR International, Radnor, PA, USA). Post evaporation for 40 s, sections were incubated in Mayer’s Hematoxylin (1092490500, VWR International, Radnor, PA, USA) for 7 min, Dako bluing buffer (CS70230-2, Agilent Technologies, Santa Clara, CA, USA) for 90 s, and finally in Eosin Y (E4382, MilliporeSigma, St. Louis, MO, USA) for 1 min. The glass slides were then washed with RNase/DNase-free water and incubated at 37 °C for 5 min or until dry. Before imaging, the glass slides were mounted with 87% glycerol (A3739, AppliChem, Darmstadt, Germany) and covered with coverslips (01-2450/1, R. Langenbrinck, Emmendingen, Germany). Brightfield imaging was performed at 10x magnification with an inverted microscope (Axio Imager 2, Zeiss, Jena, Germany), post-processed using ImageJ software. Post Imaging, coverslips, and glycerol were removed by washing the glass slides in RNase/DNase-free water, after which the slides were washed using 80% ethanol to remove final traces of glycerol.