Riluzole concentrations were quantified using an liquid chromatography‐tandem mass spectrometry assay method previously developed and validated.10 (link) Riluzole and its radiolabeled internal standard (IS) [13C,15N2] riluzole were isolated from plasma by liquid‐liquid extraction using ethyl acetate. Riluzole and IS elution at 1.9 minutes was achieved using 0.4 mL/min isocratic flow through a Waters AQUITY UPLC BEH C18 column (2.1 × 50 mm, 1.7‐μm particle size) with the mobile phase composed of 20% ACN and 80% MeOH:water (70:30, v/v) containing 0.1% formic acid. Riluzole (m/z 235 → 166) and IS (m/z 238 → 169) were detected by electrospray ionization using multiple reaction monitoring in a positive mode on a QTRAP 3200 System (AB SCIEX, Framingham, Massachusetts). The assay had linearity established between 0.5 (the lower limit of quantitation) and 800 ng/mL and intraday and interday accuracy and precision of riluzole assay within 10%, meeting the requirements of FDA guidelines.
Aquity uplc beh c18 column
Manufactured by Waters Corporation
Sourced in United States
The Aquity UPLC BEH C18 column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. It features a 1.7 μm particle size and a bonded C18 stationary phase, which provides efficient separation and high resolution. The column is compatible with ultra-high-pressure liquid chromatography (UPLC) systems and is suitable for a variety of applications, including pharmaceutical, environmental, and food analysis.
Automatically generated - may contain errors
Lab products found in correlation
Spd m20a, by Shimadzu (6 mentions)
Nexpera x2 uplc chromatograph, by Shimadzu (2 mentions)
Xevo tq s cronos, by Waters Corporation (2 mentions)
Enzymate beh pepsin column, by Waters Corporation (1 mentions)
Acquity uplc m class, by Waters Corporation (1 mentions)
Synapt g2 s mass spectrometer, by Waters Corporation (1 mentions)
Hdx manager, by Waters Corporation (1 mentions)
Proteinlynx global server plgs, by Waters Corporation (1 mentions)
Targetlynx software, by Waters Corporation (1 mentions)
Acquity uplc 1 class system, by Waters Corporation (1 mentions)
Xevo tq s mass spectrometer, by Waters Corporation (1 mentions)
Arachidonic acid d8, by Cayman Chemical (1 mentions)
Xcalibur 2, by Thermo Fisher Scientific (1 mentions)
Orbitrap elite, by Thermo Fisher Scientific (1 mentions)
15 hydroxyeicosatetraenoic acid hete d8, by Cayman Chemical (1 mentions)
Nexpera x2 uhplc chromatograph, by Shimadzu (1 mentions)
Labsolutions software, by Shimadzu (1 mentions)
Acquity uplc system, by Waters Corporation (1 mentions)
Acquity uplc pda detector, by Waters Corporation (1 mentions)
Ella instrument, by R&D Systems (1 mentions)
17 protocols using aquity uplc beh c18 column
1
Quantification of Riluzole in Plasma
2
UPLC-based Quantification of Bioactive Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Individual compounds were identified and quantified by Ultra-Performance Liquid Chromatography (UPLC) as described and validated in [81 (link),82 (link)]. A Shimadzu Nexpera X2 UPLC chromatograph equipped with a Diode Array Detector (DAD) (Shimadzu, SPD-M20A, Columbia, MA, USA) was used. Separation was performed at 40 °C on a reversed-phase Aquity UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 µm particle size; from Waters, Milford, MA, USA) equipped with a pre-column of the same material. Samples were eluted with two HPLC grade solvents, water/formic acid (0.1%) and 100% acetonitrile, at a flow rate of 0.4 mL/min. Biocompounds were identified by comparing their UV spectra and retention times with those of the corresponding standards. Calibration curves were drawn for a range of concentrations between 250 and 2.5 mg/mL per compound analyzed (vanillic acid, chlorogenic acid, catechin, epicatechin, p-coumaric acid, ellagic acid, naringin, hesperidin, resveratrol, ferulic acid, quercetin, 3,4-dihydroxybenzoic, taxifolin, aloin and kaempferol (R2 > 0.99)). Compounds were quantified and identified at different wavelengths (209–370 nm). Results were expressed in milligrams per liter (mg/L).
3
Automated Hydrogen-Deuterium Exchange Mass Spectrometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H/DX-MS experiments were performed on a fully automated system equipped with a Leap robot (HTS PAL; Leap Technologies, NC), a Waters ACQUITY M-Class UPLC, a H/DX manager (Waters Corp., Milford, MA) and a Synapt G2-S mass spectrometer (Waters Corp., Milford, MA), as described elsewhere (Zhang et al., 2014 (link)). The protein samples were diluted in a ratio of 1:20 with deuterium oxide containing PBS buffer (pH 7.4) and incubated for 0 s, 10 s, 1 min, 10 min, 30 min or 2 hr. The exchange was stopped by diluting the labeled protein 1:1 in quenching buffer (200 mM Na2HPO4 × 2 H2O, 200 mM NaH2PO4 × 2H2O, 250 mM Tris (2-carboxyethyl)phosphine, 3 M GdmCl, pH 2.2) at 1°C. Digestion was performed on-line using an immobilized Waters Enzymate BEH Pepsin Column (2.1 × 30 mm) at 20°C. Peptides were trapped and separated at 0°C on a Waters AQUITY UPLC BEH C18 column (1.7 µm, 1.0 × 100 mm) by a H2O to acetonitrile gradient with both eluents containing 0.1% formic acid (v/v). Eluting peptides were subjected to the Synapt TOF mass spectrometer by electrospray ionization. Samples were pipetted by a LEAP autosampler (HTS PAL; Leap Technologies, NC). Data analysis was conducted with the Waters Protein Lynx Global Server PLGs (version 3.0.3) and DynamX (Version 3.0) software package.
4
Quantitative Analysis of Brain Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Analysis of the brain samples was performed using an Acquity UPLC I-Class system coupled to a Xevo TQ-S (triple quadrupole MS/MS) mass spectrometer (Waters, Ireland). The derivatized analytes were separated on an Aquity UPLC BEH C18 column (1.7 μm, 2.1 × 50 mm) (Waters, Wexford, Ireland) maintained at 60 °C in a column oven. The mobile phase consisted of (A) 0.1% formic acid in water and (B) 0.1% formic acid in methanol and was delivered as a gradient at a flow rate of 600 μL/min. The gradient was as follows: 0.0 min, 35% B; 0.5 min, 35% B; 4.0 min, 75% B; 5.0 min, 99% B; 6.25 min, 99% B; 6.3 min, 35% B and 7.0 min, 35% B.
The instrument was operated in electrospray positive mode and the ionization parameters were as follows: capillary voltage 1.5 kV, cone voltage 20 V, source offset 60 (Arbitrary unit), source temperature 150 °C, desolvation temperature 450 °C and desolvation gas flow 800 L/h. Targeted detection and quantification were achieved using multiple reaction monitoring (MRM) with the transitions optimized manually to achieve maximum sensitivity (Table 4 ). The samples were evaluated using TargetLynx software (Waters, Ireland).
The instrument was operated in electrospray positive mode and the ionization parameters were as follows: capillary voltage 1.5 kV, cone voltage 20 V, source offset 60 (Arbitrary unit), source temperature 150 °C, desolvation temperature 450 °C and desolvation gas flow 800 L/h. Targeted detection and quantification were achieved using multiple reaction monitoring (MRM) with the transitions optimized manually to achieve maximum sensitivity (
5
Quantitative LC-MS/MS Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The LC-MS/MS system was composed of the chromatographic system, consisting of two Accela pumps (ACQUITY UPLC I-CLASS BSM), an autosampler (ACQUITY UPLC I-CLASS SM-FIN) and a column oven (ACQUITY UPLC I-CLASS CH-A), and a Xevo TQ-S mass spectrometer equipped with heated electrospray ionization (Waters, USA). The LC-MS/MS was performed using the MassLynx software (version 4.1). Briefly, 2 μL pretreated sample was injected into an Aquity UPLC® BEH C18 column (2.1 mm × 50 mm, dp = 1.7 μm, Waters, USA), which was protected by an In-line Filter Assembly (ACQUITY GUARD FILTER, USA). H1650R desktop high-speed refrigerated centrifuge was purchased from Shanghai Lu Xiang Yi Centrifuge Instrument Co., Ltd. (China). BT125D electronic balance was provided by Sartorius Lab Instruments GmbH & Co. KG (Germany). The G560E vortex mixer was obtained from Scientific Industries (USA). The XINW-M48 high-throughput tissue homogenizer was supplied from Shanghai Xin Weng Scientific Instrument Co., Ltd. (China).
6
Lipidomic Analysis of Fatty Acid Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lipidomic analysis was performed by LC–MS system as reported with modifications5 (link). In brief, samples were obtained by using centrifugal columns (Monospin C18-AX, GL Sciences, Tokyo, Japan) after the addition of deuterium-labeled internal standard [15-hydroxyeicosatetraenoic acid (HETE)-d8, arachidonic acid-d8, and leukotriene D4-d4; Cayman Chemical, Ann Arbor, MI, USA]. Fatty acid metabolites were analyzed by using a UPLC system (Aquity; Waters, Milford, MA, USA) coupled with a mass spectrometer (Orbitrap Elite; ThermoFisher Scientific, Waltham, MA, USA). UPLC separation was performed with a 1.7-mm, 1.0 × 150 mm Aquity UPLC BEH C18 column (Waters). Mass spectrometric analysis for quantification was based on the ion-trap MS2 detection method in anion mode. Data analysis was performed by using the software Xcalibur 2.2 (ThermoFisher Scientific).
7
Comprehensive UPLC Phenolic Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Individual phenolic compounds were identified and quantified by ultra-performance liquid chromatography (UPLC) as defined and validated by Ferreira-Santos et al. [24 (link)]. A Shimadzu Nexpera X2 UPLC chromatograph equipped with a diode array detector (DAD) (Shimadzu, SPD-M20A, Columbia, MD, USA) was used. Separation was performed at 40 °C on a reversed-phase Aquity UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 µm) from Waters (Milford, MA, USA) and eluted with water/formic acid (0.1%) and 100% acetonitrile at a flow rate of 0.4 mL/min. The biocompounds were identified and quantified by comparison with their UV spectra (wavelengths 209–370 nm) and retention times with that of corresponding standards. Calibration curves were performed for each compound at concentrations between 250–2.5 mg/L (vanillic acid, chlorogenic acid, catechin, epicatechin, p-coumaric acid, ellagic acid, naringenin, hesperidin, resveratrol, ferulic acid, quercetin, 3,4-dihydroxybenzoic, taxifolin, aloin, and kaempferol (R2 > 0.99)). Results were expressed in milligrams per liter (mg/L).
8
Phenolic Compounds Analysis in OP Waste Extracts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The phenolic compounds present in the extracts obtained from OP waste were analyzed by ultra-high-pressure liquid chromatography (UHPLC) according to Jesus et al. [26 (link)]. Briefly, a Shimadzu Nexpera X2 UHPLC chromatograph equipped with a Shimadzu SPD-M20 A diode array detector and a reversed-phase Aquity UPLC BEH C18 column by Waters (2.1 mm × 100 mm, 1.7 µm particle size) at 40 °C were used to separate and identify the compounds. The eluent involved an aqueous 0.1% formic acid solution as the solvent A, and acetonitrile as the solvent B under the following gradient profile: 95% A and 5% B (0 to 5.5 min), a linear increase to 60% B (5.5 to 17 min), a linear increase to 100% B (17 to 18.5 min) and finally, 95% A and 5% B (18.5 to 30.0 min) for the column equilibration. The flow rate of the mobile phase and the injection volume of the samples were maintained to 0.4 mL/min and 1 µL, respectively. Different HPLC grade phenolic compounds were used to prepare standard curves to identify and quantify the compounds found in the extracts. The responses of the UV detector were integrated using the LabSolutions software (Shimadzu, Kyoto, Japan).
9
UPLC Analysis of Phenolic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Identification and quantification analysis of phenolic presents in PBE were performed as described previously [5 (link)] using a Shimatzu Nexpera X2 UPLC chromatograph equipped with Diode Array Detector (DAD) (Shimadzu, SPD-M20A, Columbia, MA, USA). Separation was performed on a reversed-phase Aquity UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 µm particle size; from Waters, Milford, MA, USA) and a pre-column of the same material at 40 °C. The HPLC grade solvents used were water/formic acid (0.1%) and acetonitrile as eluents and the flow rate was 0.4 mL/min. Phenolic compounds were identified by comparing their UV spectra and retention times with that of corresponding standards. Quantification was carried out using calibration curves for each compound analyzed using concentrations between 250–2.5 mg/mL (250, 125, 100, 50, 25, 10, 5, 2.5 mg/mL). In all cases, the coefficient of linear correlation was R2 > 0.99. Compounds were quantified and identified at different wavelengths (209–370 nm).
10
UPLC Analysis of Phenolic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples were analyzed by Shimatzu N expera X2 UPLC chromatograph equipped with Diode Array Detector (DAD) (Shimadzu, SPD-M20A) according to the method described by (Ferreira-Santos et al., 2019) (link). Separation was performed on a reversed phase Aquity UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 μm particle size; from Waters) and a precolumn of the same material at 40 • C. The flow rate was 0.4 mL/ min. HPLC grade solvents water/formic acid 0.1% (A) and acetonitrile (B) were used. The elution gradient for solvent B was as follows: from 0.0 to 5.5 min eluent B at 5%, from 5.5 to 17 min linearly increasing from 5 to 60%, from 17.0 to 18.5 min a linearly increasing from 60 to 100%; the column was equilibrated at 5% from 18.5 to 30.0 min. Phenolic compounds were identified by comparing their UV spectra and retention times with that of corresponding standards. All standards used were of analytical grade (purity level above 94%) and procured from Sigma Aldrich (St. Louis, USA). Quantification was carried out using calibration curves for each compound analyzed using concentrations between 250 and 2.5 mg/mL. In all cases, the coefficient of linear correlation was R 2 greater than 0.99. Compounds were quantified and identified at different wavelengths (250-370 nm). All analyses were made in triplicate.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!