Anhydrous magnesium sulfate

PEG 8 kDa and dextran 500 kDa polymers were purchased from Alfa Aesar. Fluorescein isothiocyanate (FITC)‐dextran 500 kDa conjugate was from Sigma‐Aldrich. The following reagents were purchased from Sigma‐Aldrich and used as they received: sodium chloride (NaCl, VWR, 100%), copper (II) sulfate pentahydrate (CuSO₄ 5H₂O, 99%), anhydrous magnesium sulfate (MgSO₄, 99.5%), N‐hydroxysulfosuccinimide sodium salt (sulfo‐NHS, 98%), 2‐(N‐morpholino) ethanesulfonic acid hydrate (MES, 99.5%), N,N‐diisopropylethylamine (DIPEA, 99%) and mineral oil. Sodium ascorbate (NaAsc, 99%) was obtained from Alfa Aesar. Methoxypolyethylene glycol (mPEG2000) and N‐(3‐Dimethylaminopropyl)‐N′‐ethylcarbodiimide hydrochloride (EDC, 98%) were purchased from TCI. Rhodamine cadaverine was purchased from VWR. Cuprisorb was purchased from Seachem. 5‐(6)‐carboxy‐x‐rhodamine N‐succinimidyl ester (C₃₇H₃₃N₃O₇, Chemodex, 95%) was bought from Adipogen. Phosphate buffer solution (10×, pH 7.4) was purchased from Euromedex. Dialysis was conducted using a Spectra/Por6 dialysis membrane. The following solvent was used without further purification: acetonitrile (ACN, VWR chemicals, HPLC grade, 99.9%), diethyl ether (Et₂O, VWR chemicals, 97%) and dimethyl sulfoxide (DMSO, Sigma‐Aldrich, 99.9%). Water with a resistivity of 18.2 MΩ cm⁻¹ is prepared using a Millipore Milli‐Q system.

To extract essential oils, 300 g of cut pieces of fresh plant materials was mixed with 600 mL of distilled water and the mixture subjected to hydrodistillation for 4-5 hours using glass distillation apparatus. The distillate (oil/water mixture) was collected and the essential oils were extracted from the distillate with HPLC grade hexane (VWR International, Sweden). Anhydrous magnesium sulfate (VWR International, Sweden) was added to the hexane extract to remove any trace of water. After filtration, hexane was evaporated with a rotary evaporator (Buchi Rotavapor R210, Switzerland). The essentials oils obtained were then weighed and the yields calculated as percentage of fresh starting plant materials.

To analyze fat oxidation, fat was extracted from the pastes following a modification of the procedure proposed by Folch et al. (1957) (link). Briefly, 70 g of paste was mixed with 150 mL chloroform/methanol mixture (2:1 v/v, VWR) solution, shaken for 5 min, and then filtered. The solid phase was extracted two more times with 50 mL of the chloroform/methanol mixture. The obtained filtrates were pooled and washed twice with 50 mL of a 0.88% sodium chloride solution. After phase separation, the organic phase was dried using anhydrous magnesium sulfate (VWR) after which the solvent was removed using a rotary evaporator (Büchi). The fat extracts were stored at −18°C until further analysis.
The fat obtained by the Folch method was used to determine the peroxide and p-anisidine values. Peroxide values, expressed as units of meq. O₂/kg fat sample, were determined using the modified method of Wu and Mao (2008) (link) as described by Lenaerts et al. (2018) (link). p-anisidine values were determined using the method described by Tenyang et al. (2017) (link) and Lenaerts et al. (2018) (link).