Siderophore production by Pseudomonas fluorescence’s LNPF1 (5 × 105 cells/mL) was carried out in SM media in a 5-L flask at 28 °C for 24 h. Following the incubation, centrifugation at 10,000 rpm for 20 min separated siderophore from biomass. The pH of the cell-free supernatant was brought down to 7.0 using 12 M HCl after the successful CAS test. The acidified supernatant was concentrated on a rotary vacuum evaporator (Buchi, R124, Flawil, Switzerland) at 100 rpm and 60 °C (10 times, or to produce approximately 200 mL). After that, purification was applied to the concentrated supernatant [23 (link)].
R 124
Manufactured by Büchi
Sourced in Switzerland
The R-124 is a rotary evaporator designed for efficient solvent removal and sample concentration in laboratory settings. It features a digital display, temperature control, and an adjustable speed drive for consistent and reproducible results. The R-124 is a versatile and reliable piece of equipment for a wide range of applications in analytical, chemical, and pharmaceutical laboratories.
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Lab products found in correlation
10 protocols using r 124
1
Siderophore Production by Pseudomonas fluorescens
2
Nectandra angustifolia Extract: Anti-inflammatory Properties
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As previously described by Ferrini L et al. [19 (link)], aerial parts of Nectandra angustifolia were collected in Corrientes, Argentina (27°50′51.9″ S 58°42′46.3″ W). Briefly, the extract was obtained from air-dried leaves by maceration with ethanol at 95° (48 h). After vacuum filtration, the ethanolic extract was evaporated using a rotary evaporator (Büchi R-124). Until further use, the ethanolic extract (NaE) was kept in desiccators under reduced pressure. Chemical composition and HPLC characterization of the extract has already been published, and it includes quercetin, rutin, quercitrin, quercetin-3-β-D-glucoside, quercetin-3-O-neohesperidoside and natsudaidain-3-glucoside [19 (link),20 (link)]. NaE was administered 30 min prior to the LPS injection in mice and 1 h before LPS stimulation in BV2 microglia cells.
3
Extraction of Bioactive Compounds from CA
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A portion of CA was extracted by non-heating and heating with water or ethanol. For the non-heating extraction, the sample was macerated in the solvents at room temperature for 24 h. For heating extraction, the samples extracted with 50% ethanol and 95% ethanol were refluxing at 50 ± 2 °C for 2 hwhile the samples extracted with distilled water were boiled for 2 h on the electric hot plate. The mixture was filtered using Whatman No.1 filter paper, evaporated under reduced pressure by a rotary evaporator (R-124 Buchi, Switzerland) until dried extracts and kept at 4 °C.
4
Biomass Extraction Protocol
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Cells cultivated in suspension were filtered and the biomasses were pooled and dried at room temperature. Then, dry biomass (100 g) was extracted three times by maceration at room temperature with a mixture of grade-reactive solvent (CH2Cl2:CH3OH 9:1; Merck, Mexico City, Mexico) at a ratio of 1:20 (w/v) at 24 h for each one. The extracts were filtered, pooled, and concentrated to dryness under reduced pressure in a rotaevaporator (Büchi R-124. Postfach, Switzerland). This procedure was performed several times.
5
Extraction of Ethanolic Plant Extracts
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The bark was washed and disinfected with a 2% NaClO solution. It was air-dried at room temperature for seven days. Then, it was submitted to a temperature of 40°C, in an oven with circulating air (Memmert 854, Schwabach, Germany), always verifying humidity loss (three days approximately). The cortex was ground to a fine powder and then weighed.
The crushed and weighed plant material was then subjected to cold maceration for three days in 96% ethanol. Successive concentrations of this extract were made at reduced pressure in a digital rotary evaporator (model R-124, vacuum controller V-800, Buchi) at 40°C to obtain an ethanolic extract, which was stored at 4°C until the phytochemical screening was performed.
The crushed and weighed plant material was then subjected to cold maceration for three days in 96% ethanol. Successive concentrations of this extract were made at reduced pressure in a digital rotary evaporator (model R-124, vacuum controller V-800, Buchi) at 40°C to obtain an ethanolic extract, which was stored at 4°C until the phytochemical screening was performed.
6
Extraction of Fungal Metabolites from Cultures
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The crude extracts of fermented culture broths were prepared [19 , 20 (link)]. Pure culture of MCEF001 growing in PDA unenriched with Chloramphenicol was used to obtain three mycelial culture plugs (0.5 × 0.5 cm2) that were inoculated into 150 ml of sterile Potato Dextrose Broth (PDB) and were incubated at 28 ± 2°C for two weeks on a shaker at 150 rpm.
A double-layered muslin cloth was used to filter the culture broth so as to separate the filtrate from the mycelial mat. The culture filtrate was centrifuged at 4000 rpm at room temperature for 15 min. The mycelia free culture filtrate was then added with 150 ml of ethyl acetate (EA) in a separation funnel and was shaken gently. It was then left stationary for 1 h, and the EA layer was collected. The procedure was repeated twice more, and all three fractions were pooled and concentrated using an angular rotary evaporator (Buchi R-124, Switzerland) (150 rpm at 38°C).
A similar procedure was followed to prepare the EA fractions of the culture broths from MCEF002, MCRF003, and MCRF006.
A double-layered muslin cloth was used to filter the culture broth so as to separate the filtrate from the mycelial mat. The culture filtrate was centrifuged at 4000 rpm at room temperature for 15 min. The mycelia free culture filtrate was then added with 150 ml of ethyl acetate (EA) in a separation funnel and was shaken gently. It was then left stationary for 1 h, and the EA layer was collected. The procedure was repeated twice more, and all three fractions were pooled and concentrated using an angular rotary evaporator (Buchi R-124, Switzerland) (150 rpm at 38°C).
A similar procedure was followed to prepare the EA fractions of the culture broths from MCEF002, MCRF003, and MCRF006.
7
Extraction of Nest Entrance Extracts
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The nest entrances were extracted according to the previous methods with some modification (Kraikongjit et al., 2018) . Briefly, each sample was crushed in small pieces and then macerated in 70% ethanol (one gram per 10 mL of the solvent). The samples were incubated at room temperature for 14 days and followed by vacuum filtration. The filtrated samples were then evaporated to remove all solvent residuals under low pressure and temperature below 40°C using a rotary evaporator (Buchi R-124, Switzerland). The crude extracts from each sample were named as ethanolic Nest Entrance Extracts (eNEEs) and stored at 4°C until further analysis. The eNEEs of each source were renumbered as shown in Table 1.
8
Extraction of Crude Bioactive Compounds
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The production of crude extract was started by cultivating in liquid culture of 500 mL marine NB. The cultures were shaken at 110 rpm for 3 days at room temperature (29±2℃). The harvest was done by centrifugation to separate the cell from the medium at 6,000 rpm for 10 minutes. The medium was mixed with ethyl acetate (EtOAc) and the cell with methanol (MeOH) in a 1:1 (v/v) ratio. Both suspensions were homogenized by shaking for 15 minutes. Then, the organic layer from the water was separated using a separatory funnel (Pyrex). The organic layer was evaporated using a rotary evaporator (Buchi R-124) at 40℃. Further, the mixture of pellets with MeOH was separated using filter paper. The extraction was collected into the clean vial bottle and concentrated using nitrogen gas.
9
Preparation and Encapsulation of Semi-Purified Job's Tear Extract
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Thai Black Loei (TBL) Job's tears were purchased from Loei province, Thailand. DPPC (L-R-Dipalmitoylphosphatidylcholine) was supplied from Nikko Company, Japan. Cholesterol, acridine orange (AO), ethidium bromide (EB), sulforhodamine B (SRB), vitamin C (L-(+)-ascorbic acid), EDTA and dimethyl sulfoxide (DMSO) were from Sigma Chemical Co. (St. Louis, MO). Trypsin was from Fluka (Buchs, Switzerland) and prepared as 0.25% solution in phosphate buffered saline. DMEM was from GIBCO (Invitrogen 95 Corporation, Grand Island, NY) and supplemented with 10% fetal bovine serum, penicillin (100 U/ ml) and streptomycin (100 mg/ml). All other chemicals and reagents were of analytical grade.
Preparation of the semi-purified Job's tear extract Job's tears were ground in a mortar. Hulls (H, 100 g) were separated manually, soaked in 1 l of methanol for 24h and shook at 300 rpm in an orbital shaker at room temperature (25 ± 2 C) for 1 h. The extract was filtered and concentrated under vacuum using a rotary evaporator (R-124 Buchi, Switzerland). For semi-purification, the methanolic extract was dispersed in distilled water and partition extracted by hexane and ethyl acetate (Figure 1). The semi-purified Job's tear extract from ethyl acetate fraction of M-HTBL-N1 (S5) was used to encapsulate in liposomes.
Preparation of the semi-purified Job's tear extract Job's tears were ground in a mortar. Hulls (H, 100 g) were separated manually, soaked in 1 l of methanol for 24h and shook at 300 rpm in an orbital shaker at room temperature (25 ± 2 C) for 1 h. The extract was filtered and concentrated under vacuum using a rotary evaporator (R-124 Buchi, Switzerland). For semi-purification, the methanolic extract was dispersed in distilled water and partition extracted by hexane and ethyl acetate (Figure 1). The semi-purified Job's tear extract from ethyl acetate fraction of M-HTBL-N1 (S5) was used to encapsulate in liposomes.
10
Quantifying Phenolic Compounds in CDRB and SAWE
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The identification and quantification of the CDRB and its SAWE powder were compared to reveal the effects of the extraction process on the phenolic compounds. The extraction method of phenolic compounds described by Shao et al. (2014) (link) was employed, and to that method, slight modifications were made. Concisely, the CDRB and SAWE powder were extracted with 100 mL of 80% methanol with ultrasonication for 30 mins at 40 o C. After centrifugation at 10,000×g for 10 mins (4 o C), the residue was repeatedly extracted and the supernatants were pooled. Then the supernatant was adjusted to a pH value of ~1.5 -2.0 using 2 M HCl acid and was concentrated using a rotary evaporator (Buchi R124, B480, Japan) at 50 o C. Next, the fat in the concentrate was removed by 30 mL of hexane three times and then was extracted with 30 mL of ethyl acetate three times. The ethyl acetate extracts were collected and evaporated by rotary evaporation at 35 o C until dry. The crude free phenolic extract was prepared by adding 5 mL of 80% methanol to dissolve the dried extract.
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