The ABTS+ cation radical method was used to determine the antioxidant activity of the extracts. A 2 mm ABTS+ stock solution containing 3.5 mM potassium sulfate (VI) was prepared by diluting the stock solution eight times in methanol and incubating it overnight at room temperature in the dark to allow radical stabilization. An Acquity UPLC liquid chromatography system (Waters, Milford, MA, USA) with a Waters Acquity PDA detector (Waters, Milford, MA, USA) and an Acquity UPLC® BEH C18 column (150 mm × 2.1 mm, particle size 1.9 μm) (Waters, Dublin, Ireland) were used. The gradient started with A—0.1% aqueous formic acid solution in acetonitrile and B—0.1% aqueous formic acid (10:90, %, v/v to 40:60, %, v/v) for 15 min. The chromatogram was recorded at a 280 nm wavelength. The injection volume of the sample was 10 µL, and the flow rate was 0.4 mL/min. After running through the column, the UPLC elution returned to baseline. The sample was then re-injected with ABTS+. This time, the chromatogram was recorded at a 734 nm wavelength. The antioxidant activity was calculated from the difference in peak areas of both chromatograms and compared with the standard curve for TROLOX. Each commercial compound standard was obtained from Sigma (St. Louis, MO, USA). The obtained results were converted into 1 g of extract [27 (link),28 (link),29 (link)].
Acquity uplc liquid chromatography system
Manufactured by Waters Corporation
Sourced in United States
The ACQUITY UPLC liquid chromatography system is a high-performance liquid chromatography (HPLC) instrument designed for analytical separations. It utilizes ultra-performance liquid chromatography (UPLC) technology to enable rapid and efficient separation of complex samples. The system can be used for a variety of applications in analytical chemistry, pharmaceutical research, and other fields requiring accurate and sensitive determination of chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Acquity pda detector, by Waters Corporation (2 mentions)
Acquity uplc beh c18 column, by Waters Corporation (2 mentions)
Av 400, by Bruker (2 mentions)
Superdex 200 increase 16 60, by GE Healthcare (2 mentions)
Histrap hp column, by GE Healthcare (2 mentions)
High speed refrigerated centrifuge, by Eppendorf (1 mentions)
Milli q gradient a 10 water purification system, by Merck Group (1 mentions)
Thermo syncronis c18 column, by Thermo Fisher Scientific (1 mentions)
Xevo tq s ms ms system, by Waters Corporation (1 mentions)
Synapt g2 qtof high resolutionmass spectrometer, by Waters Corporation (1 mentions)
Acquity uplc system, by Waters Corporation (1 mentions)
Milli q advantage a10 ultra pure water purifier, by Merck Group (1 mentions)
Synapt g2 hdms q tof, by Waters Corporation (1 mentions)
10 protocols using acquity uplc liquid chromatography system
1
ABTS+ Radical Scavenging Assay for Antioxidant Evaluation
2
Analytical Instrumentation for Metabolomic Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The experimental instruments used in this study included the following: Waters ACQUITY UPLC liquid chromatography system, Q-TOF SYNAPT G2 HDMS mass spectrometer, and MassLynx v4.1 chromatographic workstation (Waters, Milford, MA, USA); a high-speed refrigerated centrifuge (Eppendorf, Hamburg, Germany); Vortex Genius 3 vortex oscillator (IKA company, Staufen, Germany); and the Milli-Q Gradient A10 water purification system (Millipore, Billerica, MA, USA).
3
UPLC-QTOF Metabolite Identification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The samples were dried under nitrogen and then reconstituted in 10% acetonitrile in water. Five microliters of these samples were then injected in an Acquity UPLC liquid chromatography system coupled with a Synapt G2 QToF high-resolution mass spectrometers (both from Waters, Milford, MA, USA). The analytes were then separated on a BEH (2.1 × 100 mm) reversed-phase column (Waters) using a linear gradient of acetonitrile in water (5 to 100%). The eluting compounds were analyzed by high-resolution mass spectrometry in both positive and negative ion electrospray modes. Leucine Enkephalin reference standard was used as lock-mass to achieve a mass accuracy below 5 ppm. Metabolites were tentatively identified by interrogating the publicly available HMDB (Human Metabolome Database) and LipidMaps reference databases.
4
Protein Purification and Enzymatic Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All reagents used here were purchased at the highest commercial quality (>95%) and used without further purification unless otherwise specified. Protein purifications were performed on HisTrap HP columns (GE Healthcare) followed by size exclusion chromatography on a Superdex 200 Increase 16/60 in the relevant buffers. In vitro enzymatic assays were performed as described in the Supplementary Methods and analysed using a Waters ACQUITY UPLC liquid chromatography system equipped with an electrospray ionisation (ESI) source. All X-ray diffraction data was obtained at Diamond Light Source in Oxford, UK and the data was processed and refined using PHENIX. Amino acid-DBE substrates were synthesised following a published method and 1H NMR spectra was recorded on a Bruker AV 400 equipped with a BBFO probe. Full experimental details can be found in the Supplementary Methods section of the Supplementary Information .
5
UPLC-MS/MS Quantification of R-/S-HFBA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An ACQUITY UPLC™ liquid chromatography system equipped with a quaternary pump, automatic injector, oven and a Xevo TQS MS/MS system (Waters Corporation, Milford, MA, USA) was used for excretion analyses. Chromatographic separation was performed on a Thermo Syncronis C18 column (50 mm × 2.1 mm, 1.7 μm; Thermo, USA). The analytes were eluted using a gradient method at a flow rate of 0.4 mL min−1. Mobile phase A was 0.1% formic acid in water and mobile phase B was acetonitrile according to the following gradient: 0–1.5 min, 15–46% B; 1.5–2.5 min, 46% B; 2.5–2.6 min, 46–15% B; 2.6–3.5 min, 15% B. The injection volume was 2 μL. The column and automatic injector temperature were set at 35 °C and 4 °C, respectively.
The mass spectrometer was operated in positive electrospray mode using multiple reaction monitoring (MRM). The desolvation temperature and source temperature were 350 °C and 150 °C, respectively. The capillary voltage was 3.0 kV. The desolvation gas flow rate and the cone gas flow rate were 700 L h−1 and 150 L h−1, respectively. The ion transitions (m/z) were 278.0 > 187.9 for R-/S-HFBA, 151.9 > 109.9 for IS. The collision energy were 18 eV for R-/S-HFBA and 14 eV for IS, respectively. The cone voltage were 21 V for R-/S-HFBA and 22 V for IS, respectively. The product ion spectra for R-/S-HFBA and IS were shown in our previous report.23 (link)
The mass spectrometer was operated in positive electrospray mode using multiple reaction monitoring (MRM). The desolvation temperature and source temperature were 350 °C and 150 °C, respectively. The capillary voltage was 3.0 kV. The desolvation gas flow rate and the cone gas flow rate were 700 L h−1 and 150 L h−1, respectively. The ion transitions (m/z) were 278.0 > 187.9 for R-/S-HFBA, 151.9 > 109.9 for IS. The collision energy were 18 eV for R-/S-HFBA and 14 eV for IS, respectively. The cone voltage were 21 V for R-/S-HFBA and 22 V for IS, respectively. The product ion spectra for R-/S-HFBA and IS were shown in our previous report.23 (link)
6
LC-MS/MS Metabolomics Profiling Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All chemicals and reagents used
for sample preparation and liquid
chromatography–mass spectrometry (LC–MS)/MS analysis
were purchased from Aldrich (Milano, Italy). All analyses were carried
out using an ACQUITY UPLC system coupled to a Synapt G2 QToF high-resolution
mass spectrometer (Waters, Milford, MA).
The samples were simply
diluted 10× in acetonitrile/water (50:50). About 5 μL of
these samples were then injected into the Acquity UPLC liquid chromatography
system coupled with the Synapt G2 QToF high-resolution mass spectrometer
(both from Waters, Milford, MA). The analytes were then separated
on a T3 (2.1 × 100 mm) reversed-phase column (Waters) using a
linear gradient of acetonitrile in water (1–100%). The eluting
compounds were analyzed by high-resolution mass spectrometry in both
positive and negative ion electrospray modes. The Leucine Enkephalin
reference standard was used as lock-mass to achieve mass accuracy
below 5 ppm. Metabolites were tentatively identified by interrogating
the publicly available HMDB (Human Metabolome Database), METLIN, and
LipidMaps reference databases.44 (link)−46 (link)
for sample preparation and liquid
chromatography–mass spectrometry (LC–MS)/MS analysis
were purchased from Aldrich (Milano, Italy). All analyses were carried
out using an ACQUITY UPLC system coupled to a Synapt G2 QToF high-resolution
mass spectrometer (Waters, Milford, MA).
The samples were simply
diluted 10× in acetonitrile/water (50:50). About 5 μL of
these samples were then injected into the Acquity UPLC liquid chromatography
system coupled with the Synapt G2 QToF high-resolution mass spectrometer
(both from Waters, Milford, MA). The analytes were then separated
on a T3 (2.1 × 100 mm) reversed-phase column (Waters) using a
linear gradient of acetonitrile in water (1–100%). The eluting
compounds were analyzed by high-resolution mass spectrometry in both
positive and negative ion electrospray modes. The Leucine Enkephalin
reference standard was used as lock-mass to achieve mass accuracy
below 5 ppm. Metabolites were tentatively identified by interrogating
the publicly available HMDB (Human Metabolome Database), METLIN, and
LipidMaps reference databases.44 (link)−46 (link)
7
Fecal Metabolite Extraction and LC-MS Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Feces collected at week 12 (10 mg) were added to methanol and then shaken, sonicated, and
centrifuged. The supernatant (100 µl) was mixed with 900
µl of methanol and vortexed for 30 s. The filtrate was added to the
test bottle. Detection was performed by an AB4000 triple quadrupole mass spectrometer (AB
Sciex, Concord, Canada) coupled to a Waters ACQUITY UPLC liquid chromatography system
(Waters Corporation, Milford, MA, USA).
centrifuged. The supernatant (100 µl) was mixed with 900
µl of methanol and vortexed for 30 s. The filtrate was added to the
test bottle. Detection was performed by an AB4000 triple quadrupole mass spectrometer (AB
Sciex, Concord, Canada) coupled to a Waters ACQUITY UPLC liquid chromatography system
(Waters Corporation, Milford, MA, USA).
8
Daosheng Four Diagnostic Instrument Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DS01-A Daosheng Four Diagnostic instrument (Daosheng, China), Waters Acquity UPLC Liquid Chromatography System, Q-TOF SYNAPT G2 HDMS (Waters Corporation, USA), high-speed refrigeration centrifuge (Termo Scientific, Germany), SpeedVac®SPD131Centrifuge enrichment system (Termo Scientific, Germany), VORTEX GENIUS 3 VORTEX oscillator (IKA, Germany), and Milli-Q Advantage A10 Ultra-pure Water Purifier (Milli-pore, USA).
9
ABTS+ Cation Radical Antioxidant Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ABTS+ cation radicals method was used to determine the antioxidant activity of the extracts. A 2 mm ABTS+ stock solution containing 3.5 mM potassium sulfate (VI) was prepared by diluting the stock solution eight times in methanol and overnight incubation at room temperature in the dark to allow radical stabilization. An Acquity UPLC liquid chromatography system (Waters, Milford, MA, USA) with a Waters Acquity PDA detector (Waters, Milford, MA, USA) and an Acquity UPLC® BEH C18 column (150 mm × 2.1 mm, particle size 1.9 μm) (Waters, Dublin, Ireland) were used. The gradient started with A: 0.1% aqueous formic acid solution in acetonitrile and B: 0.1% aqueous formic acid (10:90, %, v/v to 40:60, %, v/v) for 15 min. The chromatogram was recorded at a 280 nm wavelength. The injection volume of the sample was 10 µL, and the flow rate was 0.4 mL/min. After running through the column, the UPLC elution returned to baseline. The sample was then re-injected with ABTS+. This time the chromatogram was recorded at a 734 nm wavelength. The antioxidant activity was calculated from the difference in peak areas of both chromatograms and compared with the standard curve for TROLOX. Each commercial compound standards was obtained from Sigma (St. Louis, MO, USA). The obtained results were converted into 1 g of extract [53 (link),54 ].
10
Structural Insights into Enzymatic Catalysis
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!