S. aureus (25923) was purchased from ATCC (Manassas, VA, USA) and P. putida F1 (6899) from DSMZ (Braunschweig, Germany). The bacteria were grown in BHI agar for 24 h at 37 °C. A few isolated colonies were suspended overnight in 12.5 mL mineral medium (MM) supplied with 1% glucose as the carbon source [38 (link)], followed by incubation at 25 °C and low agitation at 70 rpm, in order to reach the log phase at the beginning of the experiment. The culture was centrifuged (Avanti J-E centrifuge, Beckman Coulter, Brea, CA, USA) at 4000 g for 10 min at 4 °C, and the sediment was washed with ultrapure (UP) water with a resistance of 18.4 MΩ-cm at 25 °C (Synergy UV water purification system, Merck, Darmstadt, Germany). The bacterial sediment was suspended in different concentrations of PBS and UP water, to a final optical density of 0.01 at 600 nm, using a spectrophotometer (Genesys 10S UV-VIS, Thermo Scientific, Waltham, MA, USA).
Synergy uv water purification system
Manufactured by Merck Group
Sourced in United States, Germany, Italy, Canada
The Synergy UV water purification system is a laboratory equipment designed to produce high-quality purified water. The core function of this system is to remove contaminants and impurities from water using ultraviolet light technology.
Automatically generated - may contain errors
Lab products found in correlation
Formic acid, by Thermo Fisher Scientific (3 mentions)
Sodium azide, by Merck Group (3 mentions)
Isopropanol lc ms grade, by Merck Group (2 mentions)
Lc ms grade acetonitrile, by Avantor (2 mentions)
Cannabidiol d3 cbd d3, by Merck Group (2 mentions)
Delta 9 tetrahydrocannabinol d3 thc d3, by Merck Group (2 mentions)
Milli q water, by Merck Group (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Lc ms grade methanol, by Thermo Fisher Scientific (2 mentions)
Acetonitrile, by Merck Group (2 mentions)
Avanti j e centrifuge, by Beckman Coulter (1 mentions)
S aureus, by American Type Culture Collection (1 mentions)
Genesys 10s uv vis, by Thermo Fisher Scientific (1 mentions)
Hplc grade methanol, by Thermo Fisher Scientific (1 mentions)
Sorbitane monostearate span 60, by Merck Group (1 mentions)
Rhodamine b, by Merck Group (1 mentions)
Ammonium persulfate, by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Sodium phosphate dibasic, by Merck Group (1 mentions)
Acrylic acid, by Merck Group (1 mentions)
25 protocols using synergy uv water purification system
1
Cultivation and Preparation of Bacterial Strains
2
Hydrogel Synthesis and Drug Encapsulation
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Acrylic acid (anhydrous 99%), N,N′-methylenebisacrylamide (99%), N,N,N′,N′-tetramethylethylenediamine (TEMED) (reagent plus 99%), ammonium persulfate (≥98%), sorbitane monostearate (Span 60®), sodium chloride, sodium phosphate monobasic (ReagentPlus® ≥ 99%), sodium phosphate dibasic (ReagentPlus® ≥ 99%), amitriptyline hydrochloride (AMT) (≥98% TLC powder), and rhodamine B were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cyclohexane (≥99.5%) was from Merk (Darmstadt, Germany), sodium hydroxide was from Riedel-de Haën (Sigma-Aldrich, St. Louis, MO, USA) and methanol (HPLC grade) was from Fisher Chemical (Hampton, VA, USA). All chemicals were used as received. Wherever we mentioned water in this work it means Ultrapure (Type 1) water from Synergy® UV Water Purification System (Merck, Darmstadt, Germany).
3
Honey Sugars Profiling by HPLC
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Sugar profiles of 11 honey samples were analyzed by HPLC using the Shimadzu chromatographic system (Kyoto, Japan) with the RID-10A refractive index detector. The mobile phase (Milli-Q water obtained using the Elix® Essential 3 Water Purification System with Synergy® UV Water Purification System, Merck Millipore, Darmstadt, Germany) was run at a flow rate of 0.6 mL/min at 75 °C through the REZEX RPM-Monosaccharide Pb2+ column (300 × 7.8 mm, Phenomenex, Torrence, USA). The column was calibrated using sixteen carbohydrate standards, including mono-, di- and trisaccharides. Standard solutions of mono-, di- and trisaccharides: glucose, fructose, galactose, rhamnose, xylose, mannose, sucrose, turanose, maltose, celobiose, fucose, trehalose, melibiose, erlose, melezitose and raffinose (Sigma-Aldrich, Saint Louis, MO, USA) were used for interpretation and quantification of sugars in the honey samples. Sugar concentrations were expressed in g/100 g honey.
4
Simultaneous HPLC Determination of Metformin and Rosuvastatin
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Metformin hydrochloride of pharmaceutical purity grade was purchased from Wanbury Limited (BSEL Tech Park, Maharashtra, India). Rosuvastatin calcium of pharmaceutical purity grade was obtained from MSN Laboratories Private Limited (Rudraram, Telengana, India). The internal standard, N-despropyl ropinirole was purchased from Vitalife Chemipharma Pvt. Ltd. (Mumbai, Maharashtra, India). HPLC grade solvents were used and purchased from E. Merck (Darmstadt, Germany). A Synergy UV water purification system (Merck Millipore, Danvers, MA, USA) was used to provide water HPLC grade. Hydrophilic polytetrafluorethylene membrane syringe filters (PTFE, 13 mm, pore size 0.45 μm) were purchased from Merck Millipore (Danvers, MA, USA). Pooled drug-free human plasma was obtained from National Red Cross General Hospital, Athens, Greece.
5
Functionalization of Porous Silicon Microcavities for Protein Immobilization
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The PYP solution preparation followed the process as previously described in numerous studies [14 (link),15 (link),16 (link),17 (link),18 (link)]; however, the functionalization protocol for the protein-solid interface was adapted to the PSi substrate.
To promote microcavity infiltration with the protein sample, previously fabricated porous silicon substrates with microcavities were exposed to oxygen plasma at 400 mtorr pressure (@29.6 W RF power) for 60 s (PDC-002 Expanded Plasma Cleaner, Harrick Plasma, Ithaca, NY, USA) hydrophilizing their surfaces. Then, the PYP solution was pipetted on the substrates to fill these cavities. The protein films were left drying for 24 h on the substrates. Finally, the extra volumes of the PYP coatings on the slides were washed mildly using deionized water (MilliQ water, Synergy® UV Water Purification System, Merck-Millipore, Burlington, MA, USA), and the PYP functionalized porous silicon microcavity samples became ready-to-use for the measurements.
To promote microcavity infiltration with the protein sample, previously fabricated porous silicon substrates with microcavities were exposed to oxygen plasma at 400 mtorr pressure (@29.6 W RF power) for 60 s (PDC-002 Expanded Plasma Cleaner, Harrick Plasma, Ithaca, NY, USA) hydrophilizing their surfaces. Then, the PYP solution was pipetted on the substrates to fill these cavities. The protein films were left drying for 24 h on the substrates. Finally, the extra volumes of the PYP coatings on the slides were washed mildly using deionized water (MilliQ water, Synergy® UV Water Purification System, Merck-Millipore, Burlington, MA, USA), and the PYP functionalized porous silicon microcavity samples became ready-to-use for the measurements.
6
Quantitative Analysis of Cannabinoids
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Olive oil (pharmaceutical grade), CBD, cannabinol (CBN), CBDA, cannabidiol-d3 (CBD-d3), THC, (-)-delta-9-tetrahydrocannabinol-d3 (THC-d3), and isopropanol LC-MS grade were purchased from Sigma-Aldrich (Milan, Italy). THCA was purchased from LGC (Milan, Italy). Acetonitrile LC-MS grade was purchased from VWR (Milan, Italy).
HPLC-grade water was produced with Elix-coupled with Synergy-UV water purification system (Merck Millipore, Milan, Italy).
HPLC-grade water was produced with Elix-coupled with Synergy-UV water purification system (Merck Millipore, Milan, Italy).
7
Optimization of Macroporous Resin Purification
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EOULs were collected from the Cili Du-zhong Forestry Centre (Zhangjiajie, China). Fresh leaves were dehydration dried at 60 °C by electro-thermostatic blast oven (Shanghai Jing Hong Laboratory Instrument Co., Ltd. Shanghai, China), and pulverized before storage at 4 °C. Chromatographic grade methanol was provided by Merch (Darmstadt, Germany). Deionized water was purified using a Merck Millipore Synergy UV water purification system (Billerica, MA, USA). All other chemicals used were of analytical grade. Macroporous resins (HPD-100, HPD-300, HPD-600, D-3250, X-5, D-140, NKA-9, NKA-II, D-101, AB-8, S-8 and Polyamide) were supplied by Zhengzhou Qinshi Technology Co., Ltd. (Zhengzhou, China). The chemical and physical properties of the tested resins were provided by the manufacturer and shown in the Table 1 . Macropous resins were pretreated according to the manufacturer’s recommendation. Prior to use, the resins were soaked with ethanol for 24 h and washed several times with deionized water. Subsequently, the resins were soaked in 1.0 M NaOH for 4 h, then washed before undergoing another soak in 1.0 M HCl for 4 h. Finally, the resins were thoroughly washed with deionized water before use.
8
HPLC-Based Enzymatic Reaction Protocol
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All chemicals and solvents of HPLC (high performance liquid chromatography) grade were obtained from commercial suppliers and used as received. High purity water used both in reactions and chromatographic analysis was obtained using a Merck Millipore Synergy UV water purification system. Colorless Corning CoStar 0.65 ml centrifuge tubes were used for enzymatic reactions and sample preparation (dilution and centrifugation), while colorless 2 ml glass vials with PTFE-lined screw-cap septa and 0.2 ml glass inserts were used for short-term sample storage and injection on HPLC equipment. A stock solution of the enzyme CGTase derived from Bacillus macerans was received as a kind gift from Amano Enzyme, Inc., Nagoya, Japan. The stock solution was stored at 5°C and used as received.
9
Cannabinoid Capsule Formulation Protocol
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The other materials used for producing the galenic preparation described below (olive oil, distilled water, micronized silica anhydride, type 0 rigid capsules), were purchased from a pharmaceutical supplies company (Farmalabor s.r.l, Canosa di Puglia, Bari, Italy) and complied with the relevant monograph of the European Pharmacopoeia (Eur.Ph). The preparation of the capsules was performed using a 100-hole manual capsule filling machine (Farmalabor, Optima Aluminium®) and a precision pipette (Gilson, Microman®).
Regarding quantitative analysis, olive oil (pharmaceutical grade), CBD, cannabinol (CBN), CBDA, cannabidiol-d3 (CBD-d3), THC, (-)-delta9-tetrahydrocannabinol-d3 (THC-d3) and isopropanol LC-MS grade were purchased from Sigma–Aldrich (Milan, Italy). THCA was purchased from LGC (Milan, Italy). Acetonitrile LC-MS grade was purchased from VWR (Milan, Italy). HPLC grade water was produced with Elix-coupled with Synergy-UV water purification system (Merck Millipore, Milan, Italy).
Regarding quantitative analysis, olive oil (pharmaceutical grade), CBD, cannabinol (CBN), CBDA, cannabidiol-d3 (CBD-d3), THC, (-)-delta9-tetrahydrocannabinol-d3 (THC-d3) and isopropanol LC-MS grade were purchased from Sigma–Aldrich (Milan, Italy). THCA was purchased from LGC (Milan, Italy). Acetonitrile LC-MS grade was purchased from VWR (Milan, Italy). HPLC grade water was produced with Elix-coupled with Synergy-UV water purification system (Merck Millipore, Milan, Italy).
10
Quantitative Metabolite Extraction and Analysis
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The infusion was lyophilized (Alpha-Christ, Christ, Osterode am Harz, Germany) and stored at −20 °C. Samples (30 mg) and 50 µL of 0.2 mg/mL ribitol (Sigma Aldrich, St. Louis, MO, USA) in methanol (Acros organics, Morris, NJ, USA) as internal standard, were dissolved in 0.2 mL DDW (Synergy® UV Water Purification System, Merck, Darmstadt, Germany). A SEP-PAK C18 cartridge (Waters, Milford, MS, USA) was pre-washed with 1 mL methanol, followed by 1 mL H2O. The sample was loaded on the cartridge and eluted with 1.5 mL DDW. The aqueous phase was collected. Next, the cartridge was washed with 1.5 mL methanol, and the methanolic phase was collected. A 0.5 mL sample of each phase was lyophilized and then derivatized, according to an established protocol [19 (link)].
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