The fresh leaves of S. lavanduloides (1 Kg) were dried at room temperature and stored away from light. The dry material (100 g) was crushed and macerated for 24 h sequentially (in triplicate) with two liters of ascending polarity solvents, i.e., n-hexane, ethyl acetate, and dichloromethane (Merck, Darmstadt, Germany). Each extract was filtered and concentrated under reduced pressure using a rota-evaporator (Heidolph at 50 °C) to obtain extracts Sl-Hex, Sl-AcOEt, and Sl-D, respectively.
Rota evaporator
Manufactured by Heidolph
Sourced in Germany
The Rota evaporator is a laboratory equipment used for the separation and concentration of liquid samples through evaporation. It efficiently removes solvents from a wide range of liquid samples, allowing for the recovery of solid or semi-solid residues.
Automatically generated - may contain errors
Lab products found in correlation
N hexane, by Merck Group (1 mentions)
Ethyl acetate, by Merck Group (1 mentions)
Dichloromethane, by Merck Group (1 mentions)
N n dimethylaniline, by Merck Group (1 mentions)
N methylaniline, by Merck Group (1 mentions)
Silica gel 60 f254, by Merck Group (1 mentions)
Vibra cell, by Sonics (1 mentions)
Diax 900, by Heidolph (1 mentions)
9 protocols using rota evaporator
1
Extraction of S. lavanduloides Leaf Compounds
2
Biodegradation Potential of Chlorpyrifos
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The degradation potential of HM01 was initially characterized for its pH (3–10) and temperature (25–45 °C) requirement, tolerance to substrate concentration (CP, 20–1000 mg L−1). The degradation study was performed at 37 °C (except for temperature profile) under shaking condition (150 rpm) in mMSM medium containing CP (100 mg L−1, except for substrate concentration) as a sole source of carbon. The substrate usage profile of HM01 was also studied with stereo-chemically different 13 OPs compounds, 4 substituted mononuclear aromatic compounds and a metabolic intermediate of CP degradation, 3,5,6-trichloro-2-pyridinol (organo-heterocyclic compound), in mMSM medium supplemented with 100 mg L−1 of each compound separately, following plate assay method.
To study the rate of degradation of CP, residual pesticide and degraded intermediates from the entire medium (100 mL) was solvent extracted using ethyl acetate (100 mL) (at an interval of 2 h till 30 h) and concentrated using rota-evaporator (Heidolph Instruments GmbH & Co. KG, Germany) following standard procedure. The CP degradation was measured using HPLC as mention in “Statistical analysis ” section. The bacterial growth was measured at 600 nm.
To study the rate of degradation of CP, residual pesticide and degraded intermediates from the entire medium (100 mL) was solvent extracted using ethyl acetate (100 mL) (at an interval of 2 h till 30 h) and concentrated using rota-evaporator (Heidolph Instruments GmbH & Co. KG, Germany) following standard procedure. The CP degradation was measured using HPLC as mention in “
3
Aromatic Amine Monitoring via GC-FID
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In order to monitor the volatile intermediates and by-products (mainly, the most concerning aromatic amines), the samples were injected in a Shimadzu GC-2010 gas chromatograph provided with flame ionization detector (FID) and split injection mode; 5.0 mL of sample were analyzed in this case. Three successive extractions were conducted with 3.0 mL portions of ethyl acetate (Panreac, 99.5%); the sample was concentrated in a Heidolph rotaevaporator at 40°C up to 2.0 mL, and then measured. Separations were made under a temperature program by using an PTA-5 Fused Silica capillary column (30 m × 0.53 mm × 1.5 μm from SUPELCO). The GC conditions of separation were: Detector and injector temperature: 220°C, carrier gas He, make up gas N2 (40 mL/min), oven temperature program: 130–150°C under a ramp of 1.0°C/min, then 150–180°C under a ramp of 10°C/min, and finally 1 min at 180°C. Following standards were used to calculate response factors: Phenylamine (Mallinckrodt, 99.99%), N-methylaniline (Sigma-Aldrich, ≥99%), N,N-dimethylaniline (Sigma-Aldrich, 99.57%), N,N-dimethyl-p-phenylenediamine (Alfa Aesar, 96%), 3-Dimethylaminophenol (Alfa Aesar, 97%).
4
Fungal Pigment Extraction by Flask Fermentation
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G. triuniae culture was subjected to flask scale fermentation in a total of 6 L (four flasks containing 1.5 L of media) of natural potato dextrose broth (PD broth). Each flask was inoculated with 20–25 mycelial disks (6 mm diameter) of G. triuniae from 3-weeks-old PDA culture plate using a cork borer and incubated at 25 °C with 100 rpm for 4–6 weeks. After incubation, the coloured culture broth was filtered through pre-weighed blotting paper, and culture filtrate was collected in a separate flask. Later, the pigments from the culture filtrates were extracted thrice with an equal volume of Hexane. With the help of a separating funnel, the Hexane part was separated from the culture filtrate. The separated hexane part was evaporated to dryness under reduced pressure in a rota evaporator (Heidolph, Schwabach, Germany). The resulting concentrated hexane extract was used for further experiments. Finally, biomass collected in a pre-weighed blotting paper was dried at 105 °C for 12–15 h and weighed to measure the yield of biomass concentration [35 (link)].
5
Gravimetric Lipid Extraction from Algal Biomass
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The lipid extraction procedure was adapted from [23 (link)] and modified according to laboratories conditions and calculated gravimetrically. The dried algal (100 mg) biomass was taken and 2 mL of chloroform, 4 mL of methanol and 1 mL of distilled water in the ratio of 1:2:0.5 were added to the flask [24 (link)]. The mixture was continuously stirred for 1 h. After 1 h of extraction, 3 mL of chloroform and 3 mL of methanol were added to the mixture. The mixture was centrifuged at 4602× g for 10 min, and after centrifugation, the organic phase was taken and stored in a new tube. Then, 0.5 mL of KOH (5%) and NaOH 0.5 mL (0.76%) were added to the tube and allowed to have phase separation, and then the lower phase was taken. The extraction procedure was repeated until the remaining biomass color was lost. The solvent was evaporated by using a Rota evaporator, from Heidolph, India. The total lipid content was gravimetrically quantified [24 (link)].
6
Identification of Brown Pigment Metabolites
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The brown pigment degradation products were identified by hydrolysis of the pigment as described by Ellis and Griffiths 197441 (link) with slight modifications. Five milligrams of the dried pure brown pigment was added to 0.5 g of KOH in a 5 ml tightly sealed screw cap glass tube. The mixture was brought to boil on hot water bath (1 h) and the dark color residue thus formed was allowed to cool. To the dried residue 1.5 ml, distilled water was added and acidified (pH 2) with concentrated HCl and metabolites were extracted with diethyl ether and dried under rota evaporator (Heidolph, Germany) finally dissolved in HPLC grade methanol. To detect the indoles, the hydrolyzed fraction was run on TLC (Merk, Silica gel 60 F254, 20×10 cm, 0.2 mm,) using a mixture of Chloroform: Methanol: Glacial acetic acid (9: 0.95: 0.05 v/v) as a solvent system and TLC plate was developed using indole-specific TLC reagent prepared as described by Ehmann et al.42 (link). Alkaline hydrogen peroxide oxidation of brown pigment was carriedout according to Ito et al. 2011 and LCMS analysis was performed as described in HRLC-MS.
7
Extraction and Fractionation of Bauhinia suaveolens Leaves
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Leaves of B. suaveolens were first washed to remove adhered dirt and then air-dried under shade. The coarse powder of the air-dried plant was prepared using a grinder and passed through sieve no. 16. Standard procedures were followed for the extraction and fractionation of the plant material employing bottled solvents. About 52 g of coarse plant material was extracted first with petroleum ether for defatting and then extracted with ethanol by continuous hot percolation in Soxhlet apparatus. Further, the ethanolic extract of the plant was polarity-based fractionated using chloroform, ethyl acetate, and n-butanol. Extract and fractions were filtered and the solvent was evaporated to dryness under reduced pressure in a rota evaporator at 40ºC (Heidolph, Germany). All the dried extracts were kept in well-closed containers under refrigeration (2-4ºC) until used for biological testing.
[ 28,29 ]
[ 28,29 ]
8
Zotepine-Loaded Solid Lipid Nanoparticles
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Zotepine loaded solid lipid nanoparticles (ZT-SLNs) were prepared using homogenization-probe sonication method, based on film hydration method [22] . Required amounts of ZT, solid lipid, and soy lecithin were dissolved in 10 mL mixture of chloroform and methanol (1:1) in a round bottom flask. Organic solvents were removed using rota evaporator (Heidolph, Germany). Drug embedded lipid layer was molten by heating at 5 °C above the melting point of the solid lipid. Aqueous phase was prepared by dissolving Poloxamer-188 in double distilled water and heated to the same temperature of oil phase. Hot aqueous phase was added to oil phase. Homogenization was carried out at 12,000 rpm for 5 min to form pre-emulsion. The obtained emulsion was ultra-sonicated using Probe soniactor with 12 T probe tip (Bandelin, Germany) for 20 min. ZT-SLNs were obtained upon cooling to room temperature. The compositions of the SLNs are given in Table 1.
9
Preparation and Characterization of Atorvastatin Solid Lipid Nanoparticles
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OM-loaded SLNs were prepared by hot homogenization followed by the ultrasonication [16, 17] . OM, lipid, and egg lecithin were dissolved in 5 mL mixture of chloroform and methanol (1:1). Organic solvents were completely removed using a rota evaporator (Heidolph, Germany). The drug embedded lipid layer was molten by heating at 5 C above melting point of the lipid. An aqueous phase was prepared by dissolving Poloxamer 188 in double distilled water and heated to same temperature (based on lipid melting point) of oil phase. Hot aqueous phase was added to the oil phase, and homogenization was carried out (at 12,000 rpm) using homogenizer (Diax900, Heidolph, Germany) for 4 min. The coarse hot oil in water emulsion so obtained was ultrasonicated using a 12 T probe sonicator (Vibracell, Sonics, Newtown, CT) for 20 min. OM-loaded solid lipid nanoparticles were obtained by allowing hot nanoemulsion to cool to room temperature. The compositions of various formulations are shown in Table 1.
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