Optical rotations were measured on a JASCO P-1020 digital polarimeter (JASCO, Tokyo, Japan). The NMR spectra were recorded on a Bruker AVANCE 400 MHz spectrometer (Bruker, Ettlingen, Germany). HRESI-MS were obtained in the positive or negative ion mode with an AB SCIEX Trip TOFTM 5600+ (AB Sciex, Framingham, MA, USA). HPLC was carried out on a waters Breeze system (Waters, Milford, MA, USA) with an ODS column (Sunfire ODS, 4.6 × 250 mm, 5 μm,), Semipreparative HPLC was carried out on a waters Breeze system (Waters, Milford, MA, USA) with an ODS column (Sunfire ODS, 10 × 250 mm, 5 μm, 3 mL/min). MPLC was performed on a LC3000 series (Tong Heng Innovation Technology, Beijing, China) with a Flash C18 cartridge (50 μm, 40 g, YMC, Kyoto, Japan); all solvents were HPLC grade. Column chromatography was performed with silica (200–300 mesh, Qingdao Marine Chemical Inc, Qingdao, China) and Sephadex LH-20 (Amersham Biosciences Inc, Uppsala, Sweden), respectively. Thin layer chromatography (TLC) was carried out with glass precoated silica gel GF254 plates. The reagents for analysis were purchased from Xilong Scientific Co., Ltd. (Gongdong, China).
Waters breeze system
Manufactured by Waters Corporation
Sourced in United States
The Waters Breeze system is a high-performance liquid chromatography (HPLC) instrument designed for analytical applications. It provides reliable and consistent separation and detection of chemical compounds. The system includes components for solvent delivery, sample introduction, and data acquisition and processing.
Automatically generated - may contain errors
Lab products found in correlation
Sunfire ods, by Waters Corporation (1 mentions)
Flash c18 cartridge, by YMC (1 mentions)
Avance 400 mhz spectrometer, by Bruker (1 mentions)
P 1020, by Jasco (1 mentions)
Esa coulochem 3 detector, by Thermo Fisher Scientific (1 mentions)
Waters 1525 binary pump, by Waters Corporation (1 mentions)
Waters 717 plus, by Waters Corporation (1 mentions)
Bacitracin, by Merck Group (1 mentions)
Cytochrome c, by Merck Group (1 mentions)
Glutathione (gsh), by Merck Group (1 mentions)
Aprotinin, by Merck Group (1 mentions)
1525 binary hplc pump, by Waters Corporation (1 mentions)
Waters 2487 dual wavelength absorbance detector, by Waters Corporation (1 mentions)
5 protocols using waters breeze system
1
Spectroscopic Analysis of Organic Compounds
2
Quantification of Fermentation Metabolites
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Concentrations of cellobiose, succinate, formate and acetate in culture samples were analyzed with HPLC. Samples taken at different time points were stored at −20 °C. Before HPLC analysis, the samples were thawed, filtered through a 0.2-μm nylon filter and acidified with 2 M sulfuric acid. Twenty microliters each of the acidified samples were then applied to an Aminex HPX-87H, 300 × 7.8 mm column (Bio-Rad, Hercules, CA) on a Hitachi LaChrom Elite System (Hitachi High Technologies America, Inc., San Jose, CA) or a Waters Breeze system (Waters Corp., Milford, MA). Analysis was performed at a flow rate of 0.5 ml/min in 5 mM H2SO4 for 35 min as previously described39 (link). Soluble fermentation products were identified by comparison with retention times and peak areas of corresponding standards.
3
Dopamine Quantification in Mouse Brain
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Mice were sacrificed, their brains removed and placed on ice. Dorsal striatum and nucleus accumbens were dissected and weighed, then flash-frozen and stored at −80 °C. Tissue samples were ultrasonicated in 0.1 M perchloric acid and stored at −80 °C until extraction. Upon thawing, the samples were homogenized in 0.1 M perchloric acid and centrifuged at 25,000 g for 12 min. Dopamine levels were measured by HPLC with electrochemical detection. The analytical column was a SunFire C18 5 lm (4.6–150.0 mm) from Waters (Milford, MA, USA). The mobile phase was 0.01 M sodium dihydrogen phosphate, 0.01 M citric acid, 1.2 mM sodium EDTA, 1.2 mM sodium 1- heptane sulfonic acid, 10% methanol, pH 3.5; the flow rate was 1.0 mL/min and the column temperature was 34 °C. The installation consisted of a Waters 717 Plus automated injection system, a Waters 1525 Binary pump, and an ESA Coulochem III detector (Dionex, Sunnyvale, CA, USA). Waters Breeze system was used for analysis.
4
Molecular Weight Distribution Analysis
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Relative molecular weight distributions of hydrolysates were determined by size-exclusion chromatography using a Waters Breeze System (Waters Co., Ltd. Milford, MA, USA). Hydrolysates were separated on a TSK gel 2000 SWXL (300 × 7.8 mm, Tosho Co., Ltd. Numama, Zushishi, Kanagawaken, Japan), pre-equilibrated with aqueous acetonitrile (55:45, v/v) containing trifluoroacetic acid (0.1 mL/100 mL), and eluted with the same solvent at a flow rate of 0.5 mL/min. Ultraviolet (UV) light absorbance was monitored at 220 nm. A calibration curve was prepared using the following external standards: Cytochrome C (12,500 Da), aprotinin (6511 Da), bacitracin (1422 Da), and glutathione (189 Da), all of which were obtained from Sigma Aldrich (St. Louis, MO, USA).
5
HPLC Analysis of Cellular Nucleotide Levels
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ATP, ADP and AMP levels were measured by HPLC. All steps were carried out on ice or at 4 °C. For the nucleotides extraction, cells were rinsed with PBS, followed by the addition of 500 μl ice-cold 0.6 M HClO 4 to each dish. The dishes were scraped and the lysates were centrifuged for 10 min at 14,000×g. The resulting pellet was re-suspended in 500 μl of 1 M NaOH for posterior protein quantification, while the supernatant was neutralized with 3 M KOH/1.5 M Tris, and centrifuged again for 10 min at 14,000×g. The supernatant was collected and immediately analyzed by reverse-phase high performance liquid chromatography (HPLC), at a wavelength of 254 nm. The chromatographic apparatus was a Waters ® Breeze™ system, consisting of a 1525 Binary HPLC Pump and a Waters ® 2487 Dual Wavelength Absorbance Detector. The column was a LiChrospher ® 100 RP-18 (5 μm) LiChroCART ® 125-4 (Merck) and the analysis was performed using the software Waters ® Breeze™ HPLC software. An isocratic elution with 100 mM phosphate buffer (KH 2 PO 4 ), pH 6.5 and 1.2% (v/v) methanol was performed with a flow rate of 1 ml/min. To determine the nucleotide concentrations, standard curves of ATP, ADP and AMP were previously run. The required time for each analysis was 5.5 min. The adenylate energy charge (EC) was then calculated as EC = (ATP +½ADP)/(ATP + ADP + AMP).
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