All metabolic transformations were analyzed using RP-HPLC with UV-vis detection with 5 mm Suplex pKb-100 analytical column (0.46 cm × 25 cm, C18) (Supelco, Bellefonte, PA, USA) with Waters HPLC system equipped with 1525 binary pump, 7725i Rheodyne injector, 2487 dual λ absorbance detector and 717 plus autosampler controlled with Breeze software (Waters Co., Milford, MA, USA). Aliquots of 200 µL of the prepared samples were analyzed by HPLC-UV-vis at a flow rate of 1 mL/min in 50 mM ammonium formate buffer, pH = 3.2, with a linear gradient from 15% to 80% methanol for 25 min, followed by a linear gradient from 80% to 100% for 3 min. The column was then re-equilibrated at initial conditions for 20 min between runs. The elution of sample components was monitored at 254 nm (testosterone transformation), 330 nm (7-OH-TFC glucuronidation) or 420 nm (C-1305 and C-1311 metabolism).
Breeze software
Manufactured by Waters Corporation
Sourced in United States
Breeze software is a data acquisition and analysis platform developed by Waters Corporation. It serves as a comprehensive software solution for managing and processing data from various analytical instruments, including liquid chromatography and mass spectrometry systems.
Automatically generated - may contain errors
Lab products found in correlation
717 plus autosampler, by Waters Corporation (3 mentions)
1525 binary pump, by Waters Corporation (2 mentions)
1525 hplc system, by Waters Corporation (2 mentions)
2487 dual, by Waters Corporation (1 mentions)
717 plus, by Waters Corporation (1 mentions)
7725i rheodyne injector, by Waters Corporation (1 mentions)
Suplex pkb 100 analytical column, by Merck Group (1 mentions)
C18 column, by Phenomenex (1 mentions)
Standard system, by Waters Corporation (1 mentions)
High pressure isocratic pump, by Waters Corporation (1 mentions)
C18 reversed phase column, by Waters Corporation (1 mentions)
2707 autosampler, by Waters Corporation (1 mentions)
1525 binary hplc pump, by Waters Corporation (1 mentions)
Waters 2414 differential refractive index detector, by Waters Corporation (1 mentions)
Waters 1515 isocratic pump, by Waters Corporation (1 mentions)
Hplc system, by Agilent Technologies (1 mentions)
Soxtec 2055 fat extraction system, by Foss (1 mentions)
Hplc system, by Waters Corporation (1 mentions)
Gemini c18, by Phenomenex (1 mentions)
Waters 2487 dual λ absorbance detector, by Waters Corporation (1 mentions)
22 protocols using breeze software
1
Quantitative HPLC Analysis of Metabolic Transformations
2
HPLC Analysis of Dantrolene Sodium
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The analytical HPLC measurements were performed on a Waters 1525 HPLC System (Waters, Milan, Italy) equipped with a Waters 2587 variable-wavelength UV-Vis detector and a Waters 717 plus autosampler. The chromatographic data were acquired using the Waters Breeze software (version 3.20). Analyses were performed on a Phenomenex C18 column (150 × 4.6 mm i.d., 3 µm particle size; Phenomenex srl, Castel Maggiore, Italy) using a mobile phase of methanol/water (75:25 v/v), with aqueous formic acid 0.1%. The used flow rate was 0.5 mL/min and the injection volume was 20 µL. Wavelength UV-Vis detector was set at 380 nm.
The chemical stability of dantrolene sodium was evaluated in a phosphate buffered saline solution pH 7.4 (10 mM HPO42−/H2PO4−, 100 mM NaCl) at 37 °C. Samples were tested in triplicate starting from three different stocks solution (1 mM), which were prepared separately.
The chemical stability of dantrolene sodium was evaluated in a phosphate buffered saline solution pH 7.4 (10 mM HPO42−/H2PO4−, 100 mM NaCl) at 37 °C. Samples were tested in triplicate starting from three different stocks solution (1 mM), which were prepared separately.
3
HPLC-Based Quantification of Neurotransmitters
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The GABA and glutamate levels were estimated by the method described by Donzanti (Donzanti and Yamamoto, 1988 (link)) with slight modifications, which involved HPLC with an ECD. (A Waters standard system comprising a high-pressure isocratic pump, a 20 microlitres manual sample injector valve, a C18 reversed-phase column, and an ECD used in the study. The mobile phase comprised 100 mM anhydrous disodium hydrogen phosphate, 25 mM EDTA, and 22 % methanol (pH: 6.5). The electrochemical conditions for the experiment were +0.65 V, with sensitivity ranging from 5 to 50 nA. The separation was carried out at a flow rate of 1.2 mL/minutes, and the column temperature maintained at 40 °C. Samples (20 μl) injected manually through a rheodyne valve injector. On the day of the experiment, the frozen brain samples thawed and homogenized in 0.2 M perchloric acid. Then, the samples anteroposterior at 12,000×g for 15 min. The supernatant was derivatized using OPA/β-ME (o-pthalaldehyde/β-mercaptoethanol) and then filtered through the 0.22-mm nylon filters before being injected into the HPLC sample injector). Data were recorded and analyzed using Breeze software version 3.2 purchase from Waters HPLC. The amino acid concentrations calculated from the standard curve generated using a standard in the concentration range of 10–100 ng/ml. The values expressed as a percentage of the normal control group.
4
Optimized HPLC Analysis of Anthocyanins
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For HPLC analysis, anthocyanins were extracted from 1 g of finely ground plant material in 1 mL of 1% (v/v) HCl-methanol for 24 h at 4 °C on a rotating wheel in darkness. Samples were clarified by centrifugation at 13,000× g for 15 min at 4 °C, and then 1.5 mL of the supernatant was transferred to autosampler vials. Analysis was conducted on the HPLC and PDA system, which was equipped with a model 1525 binary solvent delivery system (Waters, Milford, CT, USA), an an on-line degasser (Waters, Milford, CT, USA), and a 2707 autosampler (Waters, Milford, CT, USA). The data acquisition and processing were performed by the Breeze software (Waters, Milford, CT, USA). For all of the samples, anthocyanin separations were carried out on a C18 Diamonsil column (250 mm × 4.6 mm, i.d. 5 μm,) (Dikma, Beijing, China). Solvent A was 10% (v/v) formic acid and solvent B was methanol. The gradient of solvent B was as follows: 0 min, 17%; 1 min, 17%; 9 min, 35%; 20 min, 37%; and 25 min, 100%. The gradient was run at a flow rate of 1 mL/min and a column temperature of 40 °C, with a 5 μL aliquot of sample being injected. Absorbance was measured at 520 nm. Standards were cy3-gal, cy3-glu (Sigma Chemica, St. Louis, MO, USA), cyanidin-3-arabinoside chloride (cy-3-ara) and cyanidin-3-rutinoside chloride (cy-3-rut) (Polyphenols Laboratories, AS, Hanaveien, Bergen, Norway).
5
Quantification of Tocopherols and Phytosterols
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Quantification of free tocopherols and phytosterols was done using a 1250 Series HPLC system (Waters, Milford, CT, USA) equipped with a UV detector (2487, Waters, Milford, CT, USA) and a C18 reversed-phase column (250 × 4.6 mm; 5 μm). The injection volume and column temperature were 20 μL and 30 °C, respectively. Tocopherols were detected at 300 nm wavelength. The mobile phase was a mixture of methanol and high-purity water (98:2, v/v). The flow rate of the mobile phase was set at 1 mL·min−1. Phytosterols were detected at the 210 nm wavelength. The isocratic mobile phase (acetonitrile: high-purity water = 98:2, v/v) was set at a flow rate of 1.5 mL·min−1. Each component was quantified using an external standard method with pure standards of tocopherols and phytosterols. Waters Breeze software (Waters, Milford, CT, USA) was used to calculate the peak areas.
6
HPLC Analysis of Fluorescent Copolymer
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SEC was used to monitor the binding yield of the fluorophore onto the copolymer according to a previously described method32 (link), with a Waters 1515 isocratic HPLC pump (flow rate: 1 mL /min) and a Styragel HR4E Waters column at 30 °C. The eluent was DMF with LiBr (0.05 mol/l). Detection was provided by a Waters 2410 RI and a Waters 2489 UV-visible detector set at 650 nm (AF647 maximum absorption wavelength in water is 649 nm). Analyses were performed by injecting 10 µL of the reaction medium (diluted to 5 mg/mL with DMF). Data acquisition and treatment were carried out using the Breeze software (Waters).
7
HPLC Analysis of Clonidine
8
Polymer Molar Mass Determination by SEC
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A Waters size exclusion chromatography (SEC) instrument (Waters Corporation, Milford, MA, USA) was utilized for the determination of the molar mass (Mw), molar mass distributions, and dispersity index (Mw/Mn) of the synthesized P(SMA-co-OEGMA) copolymer. The chromatography system is equipped with a Waters 1515 isocratic pump (Waters Corporation, Milford, MA, USA), a set of three µ-Styragel mixed pore separation columns (pore size 102–106 Å), and a Waters 2414 differential refractive index detector (equilibrated at 40 °C) (Waters Corporation, Milford, MA, USA). The measurements and data analysis were conducted using the Breeze software (Waters Corporation, Milford, MA, USA). Tetrahydrofuran (THF) containing 5% v/v trimethylamine was the mobile phase, at a flow rate of 1 mL/min and temperature set at 30 °C. The calibration curve was set by utilizing linear polystyrene standards with average molecular mass in the range of 1200–152,000 g·mol−1 and narrow molecular mass distributions. The copolymer was dissolved in the mobile phase and measured at concentration of 1 mg mL−1.
9
Molecular Weight Distribution Analysis
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Referring to the method reported by Jia et al. [27 (link)], the MW distribution was measured using a high-performance liquid chromatography (HPLC) system (Agilent, Santa Clara County, CA, USA) equipped with a TSK gel G2000 column (300 mm × 7.8 mm; Tosoh, Tokyo, Japan). A total of 10 µL of the sample was separated at a flow rate of 0.5 mL min-1 at 30 °C and monitored at 220 nm. Bovine insulin (5733.49 Da), Bacitracin (1422 Da), Gly-Gly-Tyr-Arg (451 Da), and Gly-Gly-Gly (189 Da) were used to plot the standard curve. The data were analyzed using Breeze software (Waters, MA, USA). The peaks of each sample were integrated and divided into 4 ranges (MW > 5000, 3000–5000, 1000–3000, and <1000 Da). The relative content of each molecular weight range was expressed as the corresponding peak area for each region. The logarithm of the relative molecular mass (log MW) was linearly related to the retention time (Rt) with the following equation: Rt = −0.311558(log MW) + 8.51004 (R2 = 0.9867, p < 0.05).
10
Comprehensive Chemical Analysis of Aquatic Samples
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Analysis of diets, whole fish, and feces followed the methodology described by AOAC [16 ]. Dry matter after drying at 105 °C for 24 h; total ash by combustion (550 °C during 6 h) in a muffle furnace (Nabertherm L9/11/B170, Lilienthal, Germany); crude protein (Nx6.25) by a flash combustion technique, followed by a gas chromatographic separation and thermal conductivity detection with a Leco N Analyzer (Model FP-528, Leco Corporation, Benton Harbor, MI, USA); crude lipid by petroleum ether extraction (40–60 °C) using a Soxtec™ 2055 Fat Extraction System (Foss, Hillerød, Denmark), with prior acid hydrolysis with 8.3 M HCl; gross energy in an adiabatic bomb calorimeter (Werke C2000, IKA, Hohenems, Germany). Chromium concentrations in feeds and feces were determined according to Bolin et al. [17 (link)], after perchloric acid digestion. Amino acids were determined after hydrolysis in 6M HCL at 108 °C for 24 h in nitrogen-flushed glass vials. A Waters Pico-Tag reversed-phase HPLC system using norleucine as an internal standard was used. The resulting chromatograms were analyzed with Breeze software (Waters, Milford, MA, USA). The tryptophan in complete feeds was analyzed according to ISO 13904:2016 by HPLC-FD methodology [18 ].
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