UPLC was performed on a Waters ACQUITY UPLC™ system (Waters Corp., Milford, MA, USA) equipped with a binary solvent delivery system, a UV detector, and an auto-sampler. Chromatographic separation was performed on a Waters ACQUITY BEH C18 column (i.d., 100 mm × 2.1 mm, 1.7 µm particle size; Waters Corp.) and the injection volume was 5 µL. The column temperature was set at 37 °C and the flow rate was 0.3 mL/min. The mobile phase consisted of 0.1% v/v formic acid in water (A) and 0.1% v/v formic acid in acetonitrile (B). The initial condition was 5% B for 1 min and linearly increased to 100% B over 9 min. Total run time was 14 min including re-equilibration of the column to the initial conditions. For MS experiments, the Waters Q–TOF Premier (Micromass MS Technologies, Manchester, UK) was operated in negative ion mode with an m/z range of 100–1000. The source temperature was set at 100 °C, the collision energy was set at 10 eV, the collision gas flow was 0.3 mL/min, and the desolvation gas was set to 650 L/h at a temperature of 300 °C. The capillary voltage and sample cone voltage were set at 2.5 kV and 50 V, respectively. The V mode was used for the mass spectrometer and data were collected in the centroid mode with a scan accumulation of 0.2 s. Leucine encephalin was used as reference lock mass (m/z 554.2615) by independent LockSpray interference.
Waters acquity beh c18 column
Manufactured by Waters Corporation
Sourced in United States, Ireland
The Waters Acquity BEH C18 column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of analytes. The column features a bridged ethylene hybrid (BEH) stationary phase, which provides high efficiency, peak shape, and pressure stability across a wide pH range.
Automatically generated - may contain errors
Lab products found in correlation
Waters acquity uplc system, by Waters Corporation (6 mentions)
Waters acquity uplctm system, by Waters Corporation (1 mentions)
Synapt g2 mass spectrometer, by Waters Corporation (1 mentions)
T6508, by Merck Group (1 mentions)
Dithiothreitol (dtt), by Thermo Fisher Scientific (1 mentions)
Uhplc q exactive mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Avance 3 600 nmr spectrometer, by Bruker (1 mentions)
Acquity uplc 1 class system, by Waters Corporation (1 mentions)
Masslynx v4, by Waters Corporation (1 mentions)
Acquity uplc system, by Waters Corporation (1 mentions)
Targetlynx software, by Waters Corporation (1 mentions)
Q tof premier quadrupole, by Waters Corporation (1 mentions)
Gc tofms, by Leco (1 mentions)
Uplc q tof ms, by Waters Corporation (1 mentions)
Syncronis c18 column, by Thermo Fisher Scientific (1 mentions)
Rtx 5ms column, by Restek (1 mentions)
Waters acquity, by Waters Corporation (1 mentions)
Diode array detector, by Thermo Fisher Scientific (1 mentions)
Ultimate 3000, by Thermo Fisher Scientific (1 mentions)
Ultimate 3000 uhplc system, by Thermo Fisher Scientific (1 mentions)
24 protocols using waters acquity beh c18 column
1
UPLC-MS Analysis of Organic Compounds
2
Quantitative Analysis of Cordycepin in GRC-SC11
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The amount of cordycepin in GRC-SC11 was determined by UPLC quantitative analysis, as described previously [8 (link)]. UPLC-Q-TOF-MS analysis was performed using a Waters Acquity UPLCTM system (Waters Corp., Milford, MA, USA) equipped with a Waters Acquity BEH C18 column (100 mm × 2.1 mm × 1.7 μm particle size; Waters Corp.) and Waters Q-TOF Premier (Micromass MS Technologies, Manchester, UK). The UPLC-Q-TOF-MS analysis was performed following previously described methods [8 (link)].
3
Antibody Peptide Mapping by LC-MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reduced tryptic peptide
mapping was performed by first diluting 50 μg of antibody intermediate
to 10 mg/mL. Samples were denatured and reduced at 37 °C for
30 min in 20 μL of 6.3 M urea, 1 M guanadine HCl, 100 mM Tris
pH 8.0, and 5 mM dithiothreitol (DTT, Thermo Scientific) mixture.
Then, samples were alkylated with 15 mM iodoacetamide (IAM, Thermo
Scientific) for 30 min in the dark at room temperature. Subsequently,
the reaction was diluted with 3 volumes of 100 mM Tris pH 7.5 to allow
for trypsin digestion. Trypsin (V5280, Promega) was added at 1:12.5
protease: protein ratio and incubated at 37 °C for 3–4
h. The reaction was quenched by adding 5 μL of 10% TFA (T6508,
Sigma-Aldrich). The digests were analyzed by LC-MS using a Waters
ACQUITY UPLC system equipped with a Waters ACQUITY BEH C18 column
(1.7 mm, 2.1 × 150 mm), mobile phase A (0.02% TFA in HPLC water),
mobile phase B (0.02% TFA in acetonitrile) over a 90 min gradient,
and a Synapt G2 mass spectrometer (Waters).
mapping was performed by first diluting 50 μg of antibody intermediate
to 10 mg/mL. Samples were denatured and reduced at 37 °C for
30 min in 20 μL of 6.3 M urea, 1 M guanadine HCl, 100 mM Tris
pH 8.0, and 5 mM dithiothreitol (DTT, Thermo Scientific) mixture.
Then, samples were alkylated with 15 mM iodoacetamide (IAM, Thermo
Scientific) for 30 min in the dark at room temperature. Subsequently,
the reaction was diluted with 3 volumes of 100 mM Tris pH 7.5 to allow
for trypsin digestion. Trypsin (V5280, Promega) was added at 1:12.5
protease: protein ratio and incubated at 37 °C for 3–4
h. The reaction was quenched by adding 5 μL of 10% TFA (T6508,
Sigma-Aldrich). The digests were analyzed by LC-MS using a Waters
ACQUITY UPLC system equipped with a Waters ACQUITY BEH C18 column
(1.7 mm, 2.1 × 150 mm), mobile phase A (0.02% TFA in HPLC water),
mobile phase B (0.02% TFA in acetonitrile) over a 90 min gradient,
and a Synapt G2 mass spectrometer (Waters).
4
UHPLC-Q-Exactive Mass Spectrometry Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All biological samples were analyzed using a UHPLC-Q-Exactive mass spectrometer (Thermo Fisher Scientific, MA, USA) equipped with an ESI ion source. Chromatographic separation was performed on a Waters ACQUITY BEH C18 column (2.1 mm × 100 mm × 1.7 μm; Waters Corporation, Milford, MA, USA), and the temperature was set at 25°C. The mobile phase consists of 0.1% formic acid solution (A) and acetonitrile (B), and the linear gradient was set as follows: 0–1 min, 5% B; 1–3 min, 5%–25% B; 3–8 min, 25%–60% B; 8–15 min, 60%–80% B; 15–20 min, 80%–100% B; 20–24 min, 100% B; 24–25 min, 100%–5% B; and 25–30 min, 5% B. The flow rate was 0.3 mL/min and the injection volume was 2 μL.
The optimal operating parameters in negative and positive ion mode were set as follows: electrospray voltage, 4 kV; capillary temperature, 350°C; sheath gas flow rate, 275 kPa; auxiliary gas flow rate, 138 kPa; auxiliary gas temperature, 300°C; and collision energy, 40 eV. Metabolites were detected from m/z 100–1500 using full-scan mass spectrometry with a resolution of 70000. Furthermore, the acquisition mode that relies on the parent ion list- (PIL-) DE was used to obtain the MSn phase of the acquired data set.
The optimal operating parameters in negative and positive ion mode were set as follows: electrospray voltage, 4 kV; capillary temperature, 350°C; sheath gas flow rate, 275 kPa; auxiliary gas flow rate, 138 kPa; auxiliary gas temperature, 300°C; and collision energy, 40 eV. Metabolites were detected from m/z 100–1500 using full-scan mass spectrometry with a resolution of 70000. Furthermore, the acquisition mode that relies on the parent ion list- (PIL-) DE was used to obtain the MSn phase of the acquired data set.
5
Quantitative Analysis of Salidroside in Rhodiola crenulata
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CRV was cut into small pieces and soaked in 40% ethanol at ambient temperature for 2 h for 3 times. The extract was decanted, filtered under vacuum, concentrated in a rotary evaporator, and then lyophilized. The extract was isolated using a Waters Acquity UPLC system (Waters, Prague, Czech Republic) equipped with a photodiode array detector and a 2.1 × 50.0 mm2, 1.7-µm-internal-diameter Waters Acquity BEH C18 column. The mobile phase comprised solvent A, ultrapure water and solvent B, methanol. Elution conditions were 0–5 min of 98–98% A to B; 5–8 min of 98–10% A to B (linear gradient); 8–10 min of 10%–10% A to B; 10–11 min of 10–98% A to B (linear gradient); 11–15 min of 98–98% A to B at a flow rate of 0.8 mL/min. The individual peak area corresponding to salidroside set as the index compound in the UPLC profile was determined at the observed maximal absorbance at 223 nm. A standard calibration curve of salidroside was obtained with a series of standard compound concentrations. Quantification of the index compound in CRV was subsequently performed through UPLC analysis. The peak area of the candidate compound in the chromatogram of CRV was defined, and its content in the sample was obtained on the basis of the quantity calculated from the standard calibration curve.
6
UPLC Identification and Quantification of Phenolic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A hundred mg of each phenolic extract was dissolved in 10 mL 50% (v/v) acetonitrile and filtered using a 0.45 µm syringe filter. The identification and quantification of phenolic compounds was performed using a Thermo® Scientific UPLC (Vanquish, ThermoScientific, Breda, the Netherlands) equipped with a Waters Acquity BEH C18 column (150 × 2.1 mm, 1.7 µm; Waters, France). The mobile phase A contained 0.1% (v/v) formic acid and phase B 100% acetonitrile, an injection volume of 10 μL, and a constant flow rate of 0.35 mL min−1 was used. The gradient (v/v) was obtained, increasing phase B from 4% in 2 min; to 60% in 28 min; to 100% in 5 min; and back to 4% in 5 min. The absorbance was measured at 280 nm due to a strong interference at 210 nm. All samples were analyzed in triplicate.
7
Rapid Purification and Analysis of Allicin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GE (1 mL) was diluted 1:10 with distilled water and loaded on a SepPac SPE tC18 1cc 100 mg cartridge (Waters, Milford, MA, USA), preconditioned with ethanol and distilled water. The SPE column was washed with 2 mL distilled water, before eluting off the GE-fractions with a 2 mL stepwise increased ethanol concentration (10, 20, 40, and 60% ethanol), GE10–GE60. These fractions were directly tested for anti-proliferation activity using the MTT-assay. The 20% fraction was further purified for NMR analysis using a Waters Acquity UPLC system. A Waters Acquity BEH C18 column (2.1 × 100 mm) was used with water and methanol as mobile phases A and B, respectively, both added 0.1% formic acid. The gradient was optimized for a rapid 5-min run, where allicin was eluted at 2.0–2.2 min. This was confirmed by the testing of bioactivity in all 0.5-min fractions. Using the optimized gradient, the bioactivity containing a fraction of 100 injections was collected using a divert valve on the HPLC. This pooled fraction was vacuum-dried and dissolved in D2O prior to NMR analysis. 1H NMR spectra were recorded at room temperature on a Bruker Avance III 600 NMR spectrometer.
8
UPLC Analysis of XLGB Powder
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The XLGB powder (0.2 g) was mixed with 8 mL of 60% methanol, and the mixture was subjected to ultrasonic extraction three times for 30 min and centrifuged at 12000 rpm for 5 min. The supernatant (50 μL) was injected into the UPLC system (Li Z. et al., 2016 (link)).
The measurements were performed using a Waters Acquity UPLC Iclass system (Waters, Milford, MA, United States) with a binary solvent delivery system, an autosampler, and a photodiode-array detection (PDA) system. The separation was carried out on a Waters ACQUITY BEH C18 column (2.1 mm × 100 mm I.D., 1.7 μm, Waters, Milford, MA, United States). The mobile phase consisted of (A) 0.1% formic acid in water and (B) acetonitrile. The UPLC elution conditions were as follows: 2% B (0–1 min), 2–20% B (1–10 min), 20–24% B (10–15 min), 24% B (15–24 min), 24–30% B (24–25 min), 30–80% B (25–40 min), 80–100% B (40–41 min), 100% B (41–42 min). The flow rate was maintained constant at 0.3 ml/min, and the temperature of the column was kept at 25°C (Geng et al., 2014 (link)). The compounds for chromatography were purchased from YuanYe Bio-Technology, Co., Ltd (Shanghai, People’s Republic of China). Other reagents were purchased from Nanjing Chemical Reagent, Co., Ltd (Nanjing, People’s Republic of China). All the reagents were of analytical grade.
The measurements were performed using a Waters Acquity UPLC Iclass system (Waters, Milford, MA, United States) with a binary solvent delivery system, an autosampler, and a photodiode-array detection (PDA) system. The separation was carried out on a Waters ACQUITY BEH C18 column (2.1 mm × 100 mm I.D., 1.7 μm, Waters, Milford, MA, United States). The mobile phase consisted of (A) 0.1% formic acid in water and (B) acetonitrile. The UPLC elution conditions were as follows: 2% B (0–1 min), 2–20% B (1–10 min), 20–24% B (10–15 min), 24% B (15–24 min), 24–30% B (24–25 min), 30–80% B (25–40 min), 80–100% B (40–41 min), 100% B (41–42 min). The flow rate was maintained constant at 0.3 ml/min, and the temperature of the column was kept at 25°C (Geng et al., 2014 (link)). The compounds for chromatography were purchased from YuanYe Bio-Technology, Co., Ltd (Shanghai, People’s Republic of China). Other reagents were purchased from Nanjing Chemical Reagent, Co., Ltd (Nanjing, People’s Republic of China). All the reagents were of analytical grade.
9
UPLC Analysis of Dissolved Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The samples were analyzed with a Waters ACQUITY UPLC system (Waters, Milford, USA), and the system was operated using MassLynx V4.1 software (Waters). A Waters ACQUITY BEH C18 column (1.7 µm, 2.1 mm × 100 mm; Waters) was used for sample separation. The injection volume was 4 µL. The column was maintained at 30°C and the flow rate was set at 0.4 mL/min. The optimal mobile phase consisted of a linear gradient system of (A) acetonitrile and (B) 0.1% formic acid in water: 0–10 min, 2–27% A; 10–18 min, 27–33% A; 18–20 min, 33–35.5% A; 20–22 min, 35.5–38% A; 22–24 min, 38–54% A; 24–26 min, 54–69% A; 26–28 min, 69–71% A; 28–30 min, 71–73% A; 30–32 min, 73–100% A; 32–33 min, 100% A; and 33–35 min, 100–2% A.
The resulting eluates were collected directly into a 96-well plate every minute. After collection, the 96-well plate was placed in a vacuum drying oven and dried under reduced pressure at 50°C. After the solvent was completely evaporated, 100 µL of cell culture medium was added to each well. The complete medium was placed on a shaker for 20 min and then sonicated at 37°C for 30 min to dissolve it completely. Finally, the solution was used in subsequent experiments.
The resulting eluates were collected directly into a 96-well plate every minute. After collection, the 96-well plate was placed in a vacuum drying oven and dried under reduced pressure at 50°C. After the solvent was completely evaporated, 100 µL of cell culture medium was added to each well. The complete medium was placed on a shaker for 20 min and then sonicated at 37°C for 30 min to dissolve it completely. Finally, the solution was used in subsequent experiments.
10
Paraben Retention Enthalpy Determination
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!