Oleic acid

Human embryonic kidney (HEK293) cells were purchased from (ATCC® CRL1573™, USA) and cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F-12) (Sigma-D8437, Germany), supplemented with FBS (Sigma, Germany), and penicillin–streptomycin (Sigma, Germany). HEK293 cells were employed as model cells that normally express both ACE-2 and NRP-1 receptors and TMPRSS2 enzyme and related to high comorbidity in SARS-CoV-2 infection. The purchased metabolites were palmitic acid (Sigma Aldrich, Germany), stearic acid (Sigma Aldrich, Germany), oleic acid (Daejung, Korea), 17-octadecynoic acid (Santa Cruz, USA), 1-nonadecene (TCI, US-Japan), and l-lactic acid (Sigma Aldrich, Germany).

Tolvaptan (purity > 98.5%) was provided by Hwail Pharm (Pyeongtaek, Korea). Triethyl citrate, Peceol^TM (glyceryl monooleate, type 40), Maisin^® CC (glyceryl monolinoleate), Lauroglycol^® 90 (propylene glycol monolaurate type II), Lauroglycol^® FCC (propylene glycol monolaurate type I), Cremophor^® RH40 (PEG-40 hydrogenated castor oil), Capryol^® 90 (propylene glycol monocaprylate type II), Labrafil^® M 2125 CS (laouroyl polyoxyl-6-glecerides), Labrafil^® M 1944 CS (oleoyl polyoxyl-6-glycerides), Labrafac^® lipophile WL 1349 (medium-chain triglyceride), Labrasol^® (caprylocapryl polyoxyl-8-glycerides), and Transcutol^® HP (diethylene glycol monoethyl ether) were obtained from Gattefosse (Saint-Priest, France). Tween (polyethylene glycol sorbitan monolaurate) 20, Tween 40, Tween 80, polyethylene glycol (PEG) 200, PEG 400, castor oil, and oleic acid were purchased from Daejung (Seoul, Korea). Methanol and acetonitrile (HPLC grade) were purchased from Duksan (Seoul, Korea). All other chemicals and reagents were of analytical grade.

Puerarin was purchased from Alibaba Co., Ltd. (Hangzhou, China). Lauric acid, tetrahydrofuran (THF), and acetonitrile were purchased from Junsei (Tokyo, Japan), while myristic acid, palmitic acid, stearic acid, oleic acid, 1,4-dioxane, acetone, tert-butyl alcohol, tert-amyl alcohol, 1,2-dichloroethane, chloroform, and n-heptane were purchased from Dae-Jung (Sigeung-si, Republic of Korea). In addition, palmitic anhydride, vinyl palmitate, ethyl palmitate, butyl palmitate, 1-octanol, and ABTS were obtained from TCI (Tokyo, Japan), and methyl palmitate was purchased from Aladdin (Shanghai, China). Ethyl acetate and dimethyl sulfoxide-d₆ (DMSO-d₆) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Commercially used immobilized enzymes, Novozym 435 (Candida antarctica lipase B immobilized on acrylic resin), Lipozyme TL IM (Thermomyces lanuginosus immobilized on a silica gel carrier), and Lipozyme RM IM (Rhizomucor miehei immobilized on a resin carrier) were obtained from Novozymes (Bagsværd, Demark). The acetic acid, acetonitrile, methanol, and water used for HPLC analysis were purchased from J.T.Baker (Phillipsburg, NJ, USA) in HPLC grade. Formic acid used for LC–MS analysis was purchased from Thermo Fisher Scientific (Waltham, MA, USA).

All reagents and chemicals were procured at the highest purity. M9 minimal salts (5×), 3-Hydroxyoctanoic acid, 3-Hydroxydecanoic acid, and 3-Hydroxydodecanoic acid were purchased from Sigma Aldrich (St. Louis, MO, USA). Chloroform, H₂SO₄, n-hexane, HCl, NH₄Cl, NaOH, methanol, NaCl, sodium acetate, sodium bicarbonate, palmitic acid, oleic acid, stearic acid, linoleic acid, l–cysteine hydrochloride and antifoam were purchased from Daejung Chemical & Metals (Siheung-si, Republic of Korea). LB media, yeast extract, peptone, beef extract, soybean extract and urea were purchased from BD-DIFCO (Detroit, MI, USA).

Lornoxicam was gifted by ATCO Laboratories Ltd., Pakistan. Other chemicals/excipient used were Carbopol 940, oleic acid (OA) and Propylene Glycol (PG) (Dae-Jung, Gyeonggi-do, Korea), Triethanolamine (BDH, Poole, England) and Methanol HPLC grade (TEDIA, Fairfield, USA). The backing membrane (3M-9720), rate-controlling membranes (3M Cotran-9728 and 3M CoTran-9716) and release liner (SCOTCHPAK—9755) were gifted by 3M, St. Paul, USA. Acrylate adhesive Duro–Tak 387/2510 was supplied by Henkel Corporation (Bridgewater, USA).

Naringin (95%) and ethyl acetate (99.8%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Novozym 435 (immobilized Candida antarctica lipase B), Lipozyme TL IM (immobilized Thermomyces lanuginosus), and Lipozyme RM IM (immobilized Rhizomucor miehei) were purchased from Novo Nordisk Bioindustry (Bagsvaerd, Denmark). Dimethyl sulfoxide (DMSO) (99.5%), 1,4-dioxane (99.5%), acetone (99.5%), tert-butanol (99.5%), tert-amyl alcohol (99%), 1,2-dichloroethane (99%), isooctane (98%), oleic acid (99%), and molecular sieves (3-Å) were purchased from Dae-Jung Chemical and Metals Co. Ltd. (Gyunggido, Korea). Acetonitrile (99.5%) and tetrahydrofuran (THF) (99.5%) were purchased from Junsei Chemical Co. Ltd. (Tokyo, Japan). Methanol, water, and acetic acid (HPLC grade) were purchased from J.T. Baker (Phillipsburg, NJ, USA).

Cesium nitrate [CsNO₃] (Sigma-Aldrich, Seoul, South Korea; high purity grade), ammonium metavanadate [NH₄VO₃] (Sigma-Aldrich, high purity grade), 1-octadecene [CH₃(CH₂)₁₅CH=CH₂] (Sigma-Aldrich; technical grade, 90%), oleylamine [CH₃(CH₂)₇CH=CH(CH₂)₇CH₂NH₂] (Sigma-Aldrich; technical grade, 70%), oleic acid [CH₃(CH₂)₇CH=CH(CH₂)₇COOH] (Daejung, Sinan, South Korea; extra pure), ammonium hydroxide solution [NH₄OH] (Sigma-Aldrich; ~25% NH₃ basis), and acetone [CH₃COCH₃] were of analytical grade. Triple-distilled de-ionized (DI) water was used to dissolve the CsNO₃. All the above-mentioned reagents were used without additional purification.