To prepare nanoliposomes, cholesterol (Chol), 1,2-dimyristoyl-sn-glycero-3-phosphorylcholine (DMPC), and 1,2-dimyristoyl-sn-glycero-3-phosphorylglycerol (DMPG) (Avanti Polar Lipid; Alabaster, USA) lipids were dissolved in chloroform at the molar ratios of 15:10:75, respectively. Lipid solution was dried to a thin lipid film under reduced pressure using rotary evaporation (Heidolph, Germany). The prepared lipid film was then freeze-dried (VD-800F, Taitech, Japan) overnight to completely remove the solvent. Subsequently, the dried lipids were hydrated with 10 mM HEPES buffer (pH 7.2) containing 5% dextrose, and vortexed and bath-sonicated to disperse completely in the buffer. To obtain small unilamellar vesicles (SUVs) with a uniform size of 100–200 nm, the multilamellar vesicles (MLVs) were sequentially extruded using a mini extruder (Avestin, Canada) with polycarbonate membranes of 600, 400, 200, and 100 nm pore size, respectively. Physical properties of the prepared nanoliposomes, including particle size (diameter, nm), polydispersity index (PDI), and surface charge of the nanoliposomal formulation, were determined using dynamic light scattering (DLS) technique on a Zetasizer (Nano-ZS, Malvern, UK) at the room temperature (RT). The morphology and structure of the manufactured nanoliposomes were also visualized using a Philips CM10 transmission electron microscope (TEM).
Rotary evaporation
Manufactured by Heidolph
Sourced in Germany
The Rotary Evaporation is a laboratory equipment used for the efficient and controlled evaporation of solvents from liquid samples. It operates by applying a combination of gentle heating, reduced pressure, and rotation to facilitate the evaporation process, allowing for the recovery of the solute while leaving the volatile components behind.
Automatically generated - may contain errors
Lab products found in correlation
Mini extruder, by Avestin (2 mentions)
1525 binary pump, by Waters Corporation (1 mentions)
2707 autosampler, by Waters Corporation (1 mentions)
Fraction collector 3, by Waters Corporation (1 mentions)
Xselect csh c18 prep column, by Waters Corporation (1 mentions)
Zetasizer nano z, by Malvern Panalytical (1 mentions)
Liposofast lf 50 extruder, by Avestin (1 mentions)
Freezone 4, by Labconco (1 mentions)
Bath sonicator, by Bandelin (1 mentions)
1n naoh, by Merck Group (1 mentions)
Chloroform methanol, by Merck Group (1 mentions)
10 n hcl, by Merck Group (1 mentions)
No 1 filter paper, by Cytiva (1 mentions)
12 protocols using rotary evaporation
1
Preparation of Lipid-Based Nanoliposomes
2
Isolation of Malonyl Astragalin via Semi-Preparative HPLC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A semi-preparative HPLC system running Empower 2.0 comprised of Waters 1525 Binary pump, Waters 2998 PDA, Waters 2707 autosampler and Waters Fraction Collector III, was used to isolate kaempferol-3-O-(6’’-O-malonyl)-β-D-glucopyranoside (aka malonyl astragalin), for which no commercial standard was available. In brief, a Waters XSelect™ CSH™ C18 prep column (150 × 10 mm, 5 µm particle size), equipped with a Waters XSelect™CSH™C18 prep guard-column (10 × 10 mm, 5 µm particle size), held at 60°C, was used under the following linear gradient at 2 ml/min: solvent A = water; solvent B = acetonitrile; t0 = 95% A; t1min = 95% A, t2min = 70% A; t10min = 67.6% A; t11min = 5% A; t13min = 5% A; t14min = 95% A; t16min = 95% A, with peak detection at 350 nm, triggered to collect by retention time window. A hops leaf methanolic extract (∼10 mg/ml total flavonols) enriched for the peak of interest was used as starting material, with 100 µl on column injected (full loop needle overfill = 300 µl) per chromatographic run, and with corresponding fractions from ∼65 successive runs combined. Acetonitrile was subsequently removed via rotary evaporation (Heidolph Instruments, Schwabach, Germany), and residual water removed via lyophilisation (LabConco Corporation, Kansas City, MO, USA), yielding 19.5 mg of light yellow powder in purity, which was characterized via 2D NMR methods.
3
Extraction and Yield of GST
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
One hundred grams of GST were extracted with 1 L of hot water. The supernatant was filtered, concentrated with rotary evaporation (Heidolph Instruments GmbH & Co., Schwabach, Germany), and lyophilized. The yield of the dried extract was approximately 6.8 g/L.
4
Curcumin Encapsulation in Long-Circulation Liposomes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Curcumin was encapsulated in long circulation liposomes (LCL) at a concentration of 4.7 mg/ml, using the film hydration method with a lipid molar ratio 9.5: 0.5: 1 (DPPC: PEG-2000-DSPE: CHO) as previously described [33 (link),34 ]. Briefly, curcumin and the lipid components were dissolved in ethanol, and the solvent was removed by rotary evaporation (Heidolph, Schwabach, Germany). For size reduction, the liposomal dispersion was sequentially extruded through polycarbonate membranes with a pore size of 100 nm, using a LiposoFast LF-50 extruder (Avestin Europe GmbH, Mannheim, Germany). The liposomal size and polydispersion were determined using dynamic light scattering, and the zeta potential was measured by laser Doppler electrophoresis, using a Zetasizer Nano ZS (Malvern Instruments, Malvern, UK). The proposed formulation had appropriate quality attributes for intravenous administration, including the monodispersion size of 140 nm and zeta potential of −50 mV. To determine whether liposomal encapsulation increased the therapeutic efficacy of curcumin, curcumin solutions of the same concentration were prepared by dilution in ethanol 96% (v/v) and by dilution in saline solution.
5
Acanthocereus tetragonus Phytochemical Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The young stems of Acanthocereus tetragonus were collected near the “Cerro de Horcones” community from the municipality of Alamo-Temapache, Veracruz, Mexico (21.073783, −97.782116) in November 2020. The collected samples were dethorned, and then the cuticle was removed manually. For the cooking process, each sample (2 kg) was sliced and boiled in water (1:3 p/v) at 90 °C for 15 min. Once the cooking process was achieved, the crude and cooked samples were dried in an oven at 40 °C (Gallenkamp, London, UK) for one week in dark conditions and then pulverized in a mortar. The resultant powder was subjected to extraction with methanol (three times, 1:3 p/v) in an ultrasonic bath (30 min each time). The resulting extract was filtered and concentrated by rotary evaporation (Heidolph Instruments, Schwabach, Germany) under reduced pressure at 40 °C. The samples were freeze-dried (FreeZone 4.5; Labconco Corporation, Kansas City, MO, USA), and each freeze-dried sample was resuspended (2.5 mg mL−1) in HPLC-Mass Spectrometry-grade methanol, sonicated for 10 min, filtered, and then used for the phytochemical analysis.
6
Extraction of Jr Bark Compound
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The CAE was prepared by infusion of dried bark of Jr (25 g) with boiling distilled water (500 ml) for 30 minutes. The infusate was filtered and evaporated by rotary evaporation (Heidolph Instruments, Germany) at a temperature of 45°C. The yield of extraction was 11.8 %.
7
Liposomal Encapsulation of SLA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Liposomes containing SLA were prepared using lipid film hydration method. Briefly, two liposomal formulations consisting of (DOTAP/cholesterol) (1:1 molar ratio) as a low Tm formulation and (DSPC/DOTAP/cholesterol) (3:1:1 molar ratio) as a high Tm formulation with DiI or DiD (0.2 mol% of phospholipid as a florescent label) were dissolved in chloroform: methanol (2:1, v/v) in a sterile tube. The solvent was then removed by rotary evaporation (Heidolph, Germany) resulting in deposition of a thin lipid film on the tube’s wall. The lipid film was then freeze-dried (TAITEC, Japan) overnight to ensure complete removal of the solvent. The film was hydrated and dispersed in sterile buffer (10 mM HEPES, 10% sucrose, pH 7.4) containing SLA (1 mg/ml) at 65 °C. The resultant multilamellar dispersions were vortexed and reduced in size and lamellarity by soniaction using bath sonicator (Bandelin, Germany), at 60 °C. Liposomes were then extruded 13 times through 400 and 200 nm polycarbonate filters, respectively, using the Mini Extruder (Avestin, Canada) to make liposomes with the size of around 200 nm. Dialysis against buffer was done using dialysis bags (cut off 300 kD) in order to separate the un-entrapped SLA from liposomal one.
8
Extraction and Saponification of Lipids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissues and organs were minced with a homogenizer (Polytron® PT 1200 with tip PT-DA 1205/2, Kinematica AG, Littau, Switzerland) and extracted twice with 2 mL of chloroform/methanol (1/2, v/v), 2 mL of chloroform/methanol (1/1, v/v), and 2 mL of chloroform/methanol (2/1, v/v) (all purchased from Merck, Darmstadt, Germany). The supernatants of each extract were combined (12 mL), dried by rotary evaporation (Heidolph, Schwabach, Germany) and co-extracted phospholipids and triglycerides, which make up the major lipids in crude lipid extracts, were saponified with 4 mL of 1 N NaOH (Merck) solution for 1 h at 37 °C. Afterwards, the alkaline solution was neutralized dropwise with 400 µL of 10 N HCl (Merck), followed by dialysis against deionized water and drying by rotary evaporation. The extracts were dissolved in defined volumes of chloroform/methanol (2/1, v/v) corresponding to 0.1 mg wet weight per µL or to 0.1 µL of serum per µL dissolvent.
9
Carotenoid Extraction from Red Marine Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Carotenoids were extracted from R. marinus strains TK-1, TK-2, TK-3, SB-71 and ISCaR-493 using two methods. In the first method, aqueous cell suspensions were sonicated in an ice bath for 5 × 2 min, with 1-min rests in-between in order to keep cold sample conditions. After sonication the samples were mixed with ethyl acetate (1:1). In the second method, lyophilized cells were powdered with a glass rod before mixing with dichloromethane (25 mL solvent per g freeze-dried cells). All organic phase extracts were vacuum filtered before being dried by rotary evaporation (Heidolph instruments) at 80 rpm at 40 °C. The extracts were reconstituted in 3 mL of dichloromethane, filtered through a 0.2 μm PTFE syringe filter, flushed with N2 gas and stored at -80 °C until mass spectrometry analysis.
Carotenoids were additionally extracted from strains TK-1, TK-2, TK-3, SB-71 and ISCaR-493, for absorbance spectra analysis, by mixing full loops of cells from agar plates with hexane:acetone (1:1) and sonicating in a bath for 20 min. This was done in triplicates.
Carotenoids were additionally extracted from strains TK-1, TK-2, TK-3, SB-71 and ISCaR-493, for absorbance spectra analysis, by mixing full loops of cells from agar plates with hexane:acetone (1:1) and sonicating in a bath for 20 min. This was done in triplicates.
10
Fractionation and Bioactive Screening of Dried Fruit Powder
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The dried fruit powder was delipidated with hexane and filtered through Whatman No. 1 filter paper. Filtration residues were sequentially fractionated with 100% dichloromethane (DCM), 100% methanol (MeOH), and then 50% MeOH (50 : 50 v/v H2O : MeOH) using an orbital shaker. All filtrates were subjected to rotary evaporation (Heidolph, Germany) and stored in an upright ultralow freezer (Sanyo, Japan) at −40°C for bioactive fraction screening assays. The most potent extracts, as evaluated by performance of bioassays, were subjected to liquid-liquid partitioning by solubilization with water (Fraction C) and then partitioned successively, first with chloroform (Fraction A) and then with ethyl acetate (Fraction B), as illustrated in Figure 1 .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!