Cell viability was determined using the Cell Counting Kit-8 (CCK-8; Dojindo, Kumamoto, Japan) as described by the manufacturer. Briefly, 2 × 103 cells per well were plated in 96-well culture plates and incubated in 100 µl of complete growth medium. The synergistic effects of sesamin on human lung cancer A549 cell line viability were assessed by treating cells with 10, 20, 40, 60, 80, and 100 µM sesamin (purity > 98%; Chengdu Must Bio, Chengdu, China) dissolved in chloroform (Amresco, Solon, OH, USA) for 12, 24, 36, and 48 h; cells treated with vehicle (0.1% chloroform) were used as controls. After treatment, 10 µl CCK-8 solution was added to each well and plates were incubated for an additional 4 h at 37°C. The percentage of viable cells was determined by measuring optical density in triplicate wells at 450 nm using a microplate reader (Molecular Devices, Sunnyvale, CA, USA).
Chloroform
Manufactured by Avantor
Sourced in United States, Poland, Germany, United Kingdom, France, Belgium, Italy, Portugal, Sweden, Austria, Singapore, Netherlands, Canada, Switzerland, India
Chloroform is a colorless, dense liquid with a characteristic, sweet odor. It is a widely used solvent in various industrial and laboratory applications, primarily due to its ability to dissolve a wide range of organic compounds. Chloroform has a high boiling point and low flammability, making it a suitable choice for specific chemical processes and analyses.
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Lab products found in correlation
Methanol, by Avantor (82 mentions)
Ethanol, by Avantor (29 mentions)
Acetonitrile, by Avantor (25 mentions)
Ethyl acetate, by Avantor (19 mentions)
Sodium hydroxide (naoh), by Avantor (18 mentions)
Sodium chloride (nacl), by Avantor (17 mentions)
Diethyl ether, by Avantor (16 mentions)
N hexane, by Avantor (15 mentions)
Acetone, by Avantor (15 mentions)
Hydrochloric acid (hcl), by Avantor (15 mentions)
Formic acid, by Merck Group (14 mentions)
Isopropanol, by Avantor (14 mentions)
Methanol, by Merck Group (13 mentions)
Acetic acid, by Avantor (12 mentions)
Formic acid, by Avantor (12 mentions)
Hexane, by Avantor (11 mentions)
Toluene, by Avantor (11 mentions)
Acetonitrile, by Merck Group (10 mentions)
Sodium chloride (nacl), by Merck Group (10 mentions)
Hydrochloric acid (hcl), by Merck Group (9 mentions)
310 protocols using chloroform
1
Evaluating Sesamin's Effects on Lung Cancer Cell Viability
2
RNA Extraction from Adipose Tissue and Cell Culture
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For adipose tissue, 100 mg of frozen tissue was homogenized in TRI Reagent® (#T9424 Sigma-Aldrich, UK); for cell culture, cells were lysed in TRI Reagent. Total RNA was then extracted with chloroform (#J67241.AP, VWR International Ltd., UK) (0.2 mL, TRI Reagent®:chloroform 5:1 v/v) and isopropanol (0.5 mL, TRI Reagent®: Isopropanol 2:1 v/v). Samples were measured on a spectrophotometer at 260 nm.
3
Electrospinning of Porous and Smooth PCL Fibers
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PCL (poly(caprolactone) Mn 80,000, Mw/Mn < 2) (Sigma Aldrich, Germany) pellets were weighed and dissolved in 90:10 (v/v) chloroform (VWR, Germany) and DMSO (Dimethyl sulfoxide) (VWR, Germany) to obtain 12.5 wt/v% solution for porous surface fibers. Similarly, to obtain smooth surface fibers, PCL pellets were dissolved in 50:50 (v/v) chloroform and methanol (VWR, Germany) resulting in a 7.5 wt/v% solution. The electrospinning set-up consisted of an earthed spinneret as a syringe with a flat-tipped 27 gauge needle filled with the respective polymer solution. As a collector an aluminium foil of 20 cm × 20 cm was connected to a high voltage supply.
A high enough electric field to overcome the droplet surface tension was used to generate a taylor cone that elongated to form fibers and collected as non-woven mesh on the aluminium collector.
The spinning parameters were optimized to 21 kV, 17 cm (distance between collector and spinneret), and 0.75 mL/h to obtain homogenous non-woven fibrous meshes with pores on the fiber surface. Similarly, the spinning parameters to obtain homogenous non-woven meshes with smooth fibres were optimized to 21 kV, 10 cm and 1 mL/h. The time of electrospinning was constant at 2 min for 10 µm thin meshes in both the cases.
A high enough electric field to overcome the droplet surface tension was used to generate a taylor cone that elongated to form fibers and collected as non-woven mesh on the aluminium collector.
The spinning parameters were optimized to 21 kV, 17 cm (distance between collector and spinneret), and 0.75 mL/h to obtain homogenous non-woven fibrous meshes with pores on the fiber surface. Similarly, the spinning parameters to obtain homogenous non-woven meshes with smooth fibres were optimized to 21 kV, 10 cm and 1 mL/h. The time of electrospinning was constant at 2 min for 10 µm thin meshes in both the cases.
4
Liposome Preparation with Modifications
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Liposomes were prepared according to the method described by Csiszár et al. [14 (link)] with few modifications. In brief, lipid components like neutral lipid, positively charged lipid and the fluorescent compounds were mixed in chloroform (VWR, Darmstadt, Germany) at a ratio of 1/1/0.01–0.1 mol/mol. All liposomal compositions tested here are summarized in Table S1 (Supplemental) . chloroform was evaporated under a vacuum for 0.5 h. Then, lipids were dispersed in 20 mM 2-(4-(2-hydroxyethyl)-1-piperazinyl)-ethansulfonic acid (HEPES) buffer (VWR, Darmstadt, Germany) at a total lipid concentration of 2 mg/mL at pH 7.4 (π = 70 mOsm). The solution was vortexed for 1–2 min to produce multilamellar liposomes. After homogenization in an ultrasonic bath (Sonocool, Bandelin electronic GmbH, Berlin, Germany) for 20 min at 5 °C, mainly unilamellar vesicles were formed.
5
Formulation and Characterization of Microparticles
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Sodium chloride and chloroform were obtained from VWR (Radnor, PA, USA), whereas 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and trifluoroacetic acid were acquired from Roth (Karlsruhe, Germany). Eudragit ® S100 was purchased from Evonik Industries (Darmstadt, Germany), and PEG (12 kDa) was obtained from FLUKA, part of Fischer Scientific (Hampton, NH, USA). Chitocoat ® for chitosan coating of microspheres was obtained from Freund Corporation (Tokyo, Japan). Lyso-PC (purity: 80 %) and phosphatidylcholine (purity: 99.1 %), both from soybean oil, were kindly donated from Lipoid (Steinhausen, Schwitzerland). Chlorophyll Cl, erythrosine and tartrazine were from Caldic (Rotterdam, Netherlands).
chloroform and Sodium chloride were purchased from VWR (Radnor, PA, USA). All other chemicals were bought from Sigma-Aldrich (St. Louis, MO, USA). Ultrapure water was provided from an Ultra Clear water system produced by Siemens (Munich, Germany).
chloroform and Sodium chloride were purchased from VWR (Radnor, PA, USA). All other chemicals were bought from Sigma-Aldrich (St. Louis, MO, USA). Ultrapure water was provided from an Ultra Clear water system produced by Siemens (Munich, Germany).
6
Langmuir trough solvent preparation
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The chloroform (≥98.5%) was supplied by Avantor Performance Materials Poland S.A. (Gliwice, Poland) and was used as a spreading solvent for the used substances. The concentration of the tested solutions was 1 mg cm−3. To prepare a kaempferol or myricetin solution, initially, the substance was dissolved in a small amount of ethanol (≥99.8%, Avantor Performance Materials Poland S.A., Gliwice, Poland), and then chloroform was added. The ultrapure water subphase solution was used as the subphase. High-purity methanol (≥99.8%, Avantor Performance Materials Poland S.A., Gliwice, Poland) was used to clean the Langmuir trough.
7
Langmuir Monolayer Characterization of Flavonoids
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Chloroform ((≥98.5%), obtained from Avantor Performance Materials, Poland S.A.) was used as a solvent to prepare the solutions. (Gliwice, Poland). Solutions with concentrations of 1 mg cm−3 were used.
For the preparation of the kaempferol or myricetin solutions, the substance was first dissolved in a small amount of ethanol (≥99.8%) from Avantor Performance Materials Poland S.A., Gliwice, and then Chloroform was added. The preparation of the lipoic acid solution did not require the addition of ethanol. Ultrapure water was used as a subphase. After each measurement, the Langmuir bath was cleaned, and high-purity methanol ((≥99.8%) from Avantor Performance Materials Poland S.A., Gliwice, Poland) was used to clean it.
For the preparation of the kaempferol or myricetin solutions, the substance was first dissolved in a small amount of ethanol (≥99.8%) from Avantor Performance Materials Poland S.A., Gliwice, and then Chloroform was added. The preparation of the lipoic acid solution did not require the addition of ethanol. Ultrapure water was used as a subphase. After each measurement, the Langmuir bath was cleaned, and high-purity methanol ((≥99.8%) from Avantor Performance Materials Poland S.A., Gliwice, Poland) was used to clean it.
8
Phytochemical Screening and Extraction
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Chemicals used in this study were methanol (Fisher scientific), ethanol (Fisher scientific), n-hexane (Avantor performance materials,chloroform (Avantor performance materials), acetone (Central drug house Pvt. Ltd., petroleum ether (Fisher Scientific), Imipramine hydrochloride (Antidep 25 mg Torrent Laboratories), Isotretinoin (Acneone 20 mg Genesis Biotech), Diacerein (Orcerin 50 mg Macleods Pharmaceuticals), Levodopa (Bidopal 500mg Tablet Glaxo SmithKline), Haloperidol (Serenace Injection RPG Life Sciences), Ferric citrate (Citraphos Pure & cure Healthcare) were utilized as solvents and compounds in the studies for extraction & experimental work. For phytochemical screening, additional reagents including hydrochloric acid (Qualigens), ammonia (Avantor performance materials), sodium hydroxide (Avantor performance materials), chloroform (Avantor performance materials), etc. were utilized. Chemicals were obtained from the store house of the Hygia institute of pharmaceutical education & research Lucknow.
9
Lipid Extraction and GC Analysis
10
Wax Ester Characterization by HPLC
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