The quinones from the dimeric RC–LH1 samples were extracted using 1:1 methanol:chloroform (v:v) containing 0.02% (w/v) FeCl3, and were injected into a Shimadzu 20A HPLC system employing a Shim-pack GIST C-18 reversed-phase column (4.6 mm × 250 mm). The column was pre-equilibrated and was then eluted with 8:2 methanol:isopropanol (v:v) at a flow rate of 1 mL min−1 for 1 h at 40 °C. The elution was analyzed by an LC-20AT detector (Shimadzu), monitored at 275 nm. Ubiquinone-10 (UQ-10) was identified by comparing the retention time of the quinones from the RC–LH1 complexes with that of the UQ-10 standard (TargetMol, China, Cat No.: T2796).
20a hplc system
Manufactured by Shimadzu
Sourced in Japan
The Shimadzu 20A HPLC system is a high-performance liquid chromatography (HPLC) instrument designed for analytical applications. It features a dual-plunger pump, a built-in degasser, and a UV-Vis detector. The 20A HPLC system is capable of performing isocratic and gradient separations with a wide range of mobile phases.
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Lab products found in correlation
4000qtrap mass spectrometer, by Shimadzu (2 mentions)
Lc 20aht, by Shimadzu (2 mentions)
Lc solutions analyst software, by Shimadzu (2 mentions)
Dgu 20a5 degasser, by Shimadzu (2 mentions)
Atlantis c18 column, by Waters Corporation (1 mentions)
Ltq orbitrap xl system, by Thermo Fisher Scientific (1 mentions)
Capcell pak mgii column, by Shiseido (1 mentions)
Lc 20ad, by Shimadzu (1 mentions)
Inertsil ods 3 column, by GL Sciences (1 mentions)
C18 column, by Merck Group (1 mentions)
Lc 20at pump, by Shimadzu (1 mentions)
Spd m20a photodiode array, by Shimadzu (1 mentions)
Cosmosil 5c18 ar 2 column, by Nacalai Tesque (1 mentions)
14 protocols using 20a hplc system
1
Quinone Extraction and HPLC Analysis
2
Quantitative Analysis of Serum S1P
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Briefly, S1P was separated from serum with an ACE reverse phase C18 HPLC column. Formic acid 0.5% in 5mM NH4Ac (A) and formic acid 0.5% in acetonitrile/water (9/1) (B) were used as the mobile phase. Lipids were detected with positive multiple reaction monitor (MRM) scanning at 330.20/264.40 m/z for S1P using a Sciex API-4000 MS/MS combined with a Shimazu 20A HPLC system. The internal standard for this method was Carbutamide. The quantification limit was 1 ng/ml. The calibration range was 1–2000ng/ml. The accuracy of the standards and control samples ranged from 88.7% to 120%. The information of patients was described in Table S1 and S2 .
3
Quantitative Profiling of Free Fatty Acids in B-ALL Cells
4
HPLC-MS/MS Analysis of Target Chemicals
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Target chemicals were analyzed using a 20A HPLC system (Shimadzu, Tokyo, Japan) coupled with a Q-Trap 5500 tandem mass spectrometer (MS/MS; Applied Biosystems, Foster City, CA, USA). Chromatographic separation of the analytes was performed on an Atlantis C18 column (1.7 μm, 3.0 × 100 mm, Waters, Dublin, Ireland), accompanied by mobile phases of water and methanol. The gradient elution conditions and the mass spectrometric information of the target chemicals are shown in Tables S1 and S2 , respectively. The injection volume was 5 µL and the column temperature was set at 40 °C. The flow rate was set at 0.3 mL/min. The mass spectrometer was performed in negative electron spray ionization (ESI) mode. The source temperature was set as 550 °C and the ionization voltage was −4500 V.
5
Quantitative Profiling of Free Fatty Acids in B-ALL Cells
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DKO and TKO B-ALL cells were plated at 500,000 cells/mL in regular or low glucose media. After 24 hours, 3 million cells per sample were centrifuged at 1,000 rpm for 5 minutes and washed once with 1x PBS. Supernatants were removed and cell pellets were shipped on dry ice to the Metabolomics Facility at Washington University (P30 DK020579) for analysis. Cell suspensions (1 × 107 cells/mL) were prepared by vortexing cell pellets with H2O. Protein precipitation was performed to extract free fatty acids (FFA) from 50 μL of cell suspension. The d4-FFA (16:0) was used as an internal standard, which was added to the samples before extraction. Analysis of FFAs was performed in positive MRM mode on a 4000QTRAP mass spectrometer coupled to a Shimadzu 20A HPLC system. Data processing was conducted with Analyst 1.6.3. Quality control (QC) samples were prepared by pooling aliquots of the samples and were used to monitor instrument performance. The QC samples were injected between every five experimental samples. Only the lipid species with coefficient of variance less than 15% in QC injections were reported. The relative quantification of lipids was provided as the peak area ratios of the analytes to the corresponding internal standards.
6
Quantification of Epyrifenacil and S-3100-CA in PXB Mouse Liver
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At necropsy, samples of approximately 100 mg of PXB mouse liver were collected and added to 300 µL of methanol, respectively. After extraction using a ball mill and then centrifugation at 16,000 × g for 5 min, the supernatant was collected and subjected to LC/MS/MS analysis. The analysis was performed on a Shimadzu 20A HPLC system (Shimadzu Corporation, Kyoto, Japan) equipped with a CAPCELL PAK MGII column (50 mm × 2.0 mm, particle size 3 µm, SHISEIDO CO., LTD., Tokyo, Japan). The conditions were as follows: mobile phase, 0.1% formic acid in water (A) and acetonitrile (B); flow rate, 1.0 mL/min; gradient conditions, 30% B at 0 min, 76% B at 4.0 min, 100% B at 4.0-4.5 min, and 30% B at 4.5-5.0 min; injection volume, 5 µL; and column oven temperature, room temperature. An LTQ Orbitrap XL system (Thermo Fisher Scientific Inc., Waltham, MA, USA) was used to perform the MS/MS analysis in positive ESI mode. The selective reaction monitoring transition was m/z 518/473 for epyrifenacil and m/z 490/473 for S-3100-CA. The amounts of the analytes in the samples were calculated with the linear calibration line including the analytical standards.
7
HPLC Separation of Compounds
8
HPLC Analysis of Compounds
9
HPLC Analysis of Indole Compounds
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Three strains with the highest production of indole compounds were subject to a quantitative evaluation for IAA production by high resolution liquid chromatography (HPLC) according to Hoffman et al. (2013) . Samples were analyzed in a 20A HPLC system (Shimadzu, Kyoto, Japan), equipped with an SPD-M20A diode array detector fitted with an Inertsil ODS-3 column (GL Sciences, Tokyo, Japan) with a particle size of 5 μm and dimension of 4.6 × 250 mm with injection volumes of 10 μL.
10
Identification of Phenolic Compounds by HPLC-MS/MS
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A Shimadzu HPLC 20A system (Shimadzu, Tokyo, Japan) was used in conjunction with an Applied Biosystems Q-Trap 3200 LC-MS/MS (3200 Q TRAP. Mundelein, IL, USA) system to identify phenolic chemicals. At a mass range of 150–800 amu, mass spectrum studies were conducted in the negative ionization mode. For the chromatographic analysis, a 250 × 4.6 mm, 5 µm ODS analytical column was employed at 40 °C. UV Chromatograms were taken at 280 and 320 nm. CH3OH:H2O:CH2O2 (10:89:1, v/v/v) (solvent A) and CH3OH:H2O:CH2O2 (89:10:1, v/v/v) (solvent B) were used for the gradient analysis at a flow rate of 1 mL/min. The content of B was increased from 15% to 100% over 40 min.
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