Anhydrous sodium sulfate was purchased from Merck KGaA (Darmstadt, Germany). Limonene (≥99.0%), γ-terpinene (≥98.5%), and LPSs from Escherichia coli O55:B5 were purchased from Sigma–Aldrich (St Louis, MO, USA). Authentic standards for the identification of volatile aroma components were obtained from Sigma–Aldrich and Tokyo Chemical Industry (Tokyo, Japan). RPMI-1640, sodium nitrite, and squalane were obtained from Wako Pure Chemical Industries (Osaka, Japan). Dimethyl sulfoxide was purchased from Nacalai Tesque, Inc. (Kyoto, Japan) and fetal bovine serum (FBS) was obtained from Thermo Fisher Scientific (Waltham, MA, USA). Griess reagent comprised of 2.5% sulfanilamide (Sigma–Aldrich), 0.05% N-(1- naphthyl) ethylenediamine (Sigma–Aldrich), and 2.5% phosphoric acid (Wako Pure Chemical Industries). CellTiter 96™ Aqueous One Solution Cell Proliferation Assay reagent containing 3-(4,5-dimethylthiazol-2-yl)-5-(3carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) was obtained from Promega Corporation (Madison, WI, USA).
Squalane
Manufactured by Fujifilm
Sourced in Japan
Squalane is a natural, saturated hydrocarbon found in the skin's sebum. It functions as a skin conditioning agent, providing moisturization and enhancing the skin's barrier function.
Automatically generated - may contain errors
Lab products found in correlation
Anhydrous sodium sulfate, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Limonene, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Nacalai Tesque (1 mentions)
γ terpinene, by Merck Group (1 mentions)
Phosphoric acid, by Fujifilm (1 mentions)
Sulfanilamide, by Merck Group (1 mentions)
N 1 naphthyl ethylenediamine, by Merck Group (1 mentions)
Rpmi 1640, by Fujifilm (1 mentions)
Sodium nitrite, by Fujifilm (1 mentions)
Celltiter 96 aqueous one solution cell proliferation assay reagent, by Promega (1 mentions)
Lpss from escherichia coli o55 b5, by Merck Group (1 mentions)
4 5 dimethylthiazol 2 yl 5 3carboxymethoxyphenyl 2 4 sulfophenyl 2h tetrazolium, by Promega (1 mentions)
Gcms qp2010se system, by Shimadzu (1 mentions)
3 protocols using squalane
1
Cytotoxicity and Nitric Oxide Assay
2
Porcine Eye Topical Solution Preparation
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TSs were prepared with saline, CO (Sioe Pharmaceutical Co., Ltd., Hyogo, Japan), SH (Artz Dispo 25 mg, Seikagaku Corp., Tokyo, Japan) and DHS (squalane, FUJIFILM Wako Pure Chemical Corp.,
Osaka, Japan). TSs examined in this study were as follows: 1) saline, 2) saline/0.5% (V/V) SH/1% (V/V) CO or DHS solutions and 3) saline/0.1%, 0.25% or 0.3% (V/V) SH/1%, 2.5% or 5% (V/V) DHS
solutions. To prepare respective TSs, individual reagents were simply mixed without additives. Each prepared TS was stored in a sterilized eye dropper, which was shaken well just before
application to the porcine eyes.
Osaka, Japan). TSs examined in this study were as follows: 1) saline, 2) saline/0.5% (V/V) SH/1% (V/V) CO or DHS solutions and 3) saline/0.1%, 0.25% or 0.3% (V/V) SH/1%, 2.5% or 5% (V/V) DHS
solutions. To prepare respective TSs, individual reagents were simply mixed without additives. Each prepared TS was stored in a sterilized eye dropper, which was shaken well just before
application to the porcine eyes.
3
Quantification of Squalene in A549 Cells
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A549 cells were seeded in 60-mm dishes. After incubation with various concentrations of squalene (0–300 μM) for 3 h, the cells were washed in cold PBS and harvested. For extraction of squalene, 0.8 mL of chloroform/methanol (2:1 v/v) and squalane (Wako Pure Chemical Industries, Ltd., Osaka, Japan), as an internal standard, was added to the cell pellets, which were then sonicated. After centrifugation (10,000 rpm, 5 min, 4°C), the supernatants were evaporated and dissolved in hexane for GC-MS analysis.
GC-MS analysis was performed using a Shimadzu GCMS-QP2010 SE system (Shimadzu, Kyoto, Japan). Separation was carried out on a fused silica DB-23 capillary GC column (30 m × 0.25 mm I.D., 0.25 μm film; Agilent Technologies, Inc., Santa Clara, CA, USA). Helium gas (99.999%) was used as the carrier gas at a constant flow rate of 1.45 mL/min, an injection volume of 1 μL at a split ratio of 15:0, an injector temperature of 250°C, and an ion-source temperature of 250°C. The oven temperature was programmed at 110°C with an increase of 10°C/min up to 200°C followed by 5°C/min up to 250°C, and ending with a 5 min isothermal at 250°C. The total GC running time was 24 min. Data acquisition and processing were performed using GCMSsolution Ver. 2.7 (Shimadzu, Kyoto, Japan).
GC-MS analysis was performed using a Shimadzu GCMS-QP2010 SE system (Shimadzu, Kyoto, Japan). Separation was carried out on a fused silica DB-23 capillary GC column (30 m × 0.25 mm I.D., 0.25 μm film; Agilent Technologies, Inc., Santa Clara, CA, USA). Helium gas (99.999%) was used as the carrier gas at a constant flow rate of 1.45 mL/min, an injection volume of 1 μL at a split ratio of 15:0, an injector temperature of 250°C, and an ion-source temperature of 250°C. The oven temperature was programmed at 110°C with an increase of 10°C/min up to 200°C followed by 5°C/min up to 250°C, and ending with a 5 min isothermal at 250°C. The total GC running time was 24 min. Data acquisition and processing were performed using GCMSsolution Ver. 2.7 (Shimadzu, Kyoto, Japan).
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