Liquid chromatography time-of-flight mass spectrometry (LC-TOFMS) was carried out using an Agilent 1200 HPLC pump with an Agilent 6210 time-of-flight mass spectrometer (Agilent Technologies). LC-TOFMS was conducted using an analyzer by HMT Inc., according to HMT’s LC package, by performing previously described methods with some modifications [60 (link),61 (link)]. Briefly, 80 µL of saliva samples was mixed with 240 µL of methanol containing internal standards (H3304-1002, HMT Inc., Tsuruoka, Japan). The mixtures were centrifuged at 2300× g at 4 °C for 5 min, and the supernatants were collected and evaporated. Then, the supernatants, resuspended in 160 µL of 50% isopropanol (v/v), were applied for LC-TOFMS analysis. PCA was performed using SampleStat software.
Agilent 1200 hplc pump
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 1200 HPLC pump is a high-performance liquid chromatography (HPLC) pump designed for reliable and precise solvent delivery. It features a dual-piston design with active inlet valve control for accurate and consistent flow rates.
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Lab products found in correlation
4 protocols using agilent 1200 hplc pump
1
Saliva Metabolite Profiling by LC-TOFMS
2
Nano-scale Protein Purification by LC-MS
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Protein
samples were pressure-loaded onto a fused silica capillary
tubing (75 μm i.d., with a 5 μm pulled tip) packed with
12 cm of PSDVB resins (PolyRP, 5 μm, 1000 Å, Sepax Technologies).
The Agilent 1200 HPLC pump (Palo Alto, CA) was operated with a flow
split to elute proteins at a constant flow rate of 300 nL/min using
the flowing gradient: 5% buffer B (80% acetonitrile, 20% water, 0.1%
formic acid) at start, 20% buffer B at 5 min, 65% buffer B at 50 min,
100% buffer B at 55 min, and 5% buffer B at 60 min.
samples were pressure-loaded onto a fused silica capillary
tubing (75 μm i.d., with a 5 μm pulled tip) packed with
12 cm of PSDVB resins (PolyRP, 5 μm, 1000 Å, Sepax Technologies).
The Agilent 1200 HPLC pump (Palo Alto, CA) was operated with a flow
split to elute proteins at a constant flow rate of 300 nL/min using
the flowing gradient: 5% buffer B (80% acetonitrile, 20% water, 0.1%
formic acid) at start, 20% buffer B at 5 min, 65% buffer B at 50 min,
100% buffer B at 55 min, and 5% buffer B at 60 min.
3
Plasma Metabolomics Using LC-TOFMS
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80μL of plasma was added to 240μL of 1% formic acid/acetonitrile containing the internal standard (same as CE-FTMS) at 0°C. The mixture was centrifuged at 2,300× g, 4°C for 5 min and then filtered using a hybrid SPE phospholipid cartridge (Hybrid SPE - Phospholipid 30 mg/mL, SUPELCO) to remove phospholipids. The filtrate was then evaporated to dryness under nitrogen, dissolved in 80μL of 50% isopropanol (v/v), and conducted a metabolomic analysis using liquid chromatography time-of-flight mass spectrometry (LC-TOFMS) based on the methods previously described (HMT’s LC package) [24, 25 (link)]. Briefly, LC-TOFMS analysis was performed using an Agilent 1200 HPLC pump, an ODS column (2 mm×50 mm, two μm i.d.), and an Agilent 6210 time-of-flight mass spectrometer (Agilent Technologies, USA). The system was controlled by MassHunter (Agilent Technologies, USA), and the spectrometer was scanned from m/z 50 to 1,000.
4
LC-MS/MS Based Metabolite Quantification
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The LC-MS/MS system consisted of a Q-Trap 5500 triple quadrupole/ion trap mass spectrometer (ABSCIEX™, USA) equipped with a Turbo Spray ion source operated at 350 °C. The pMRM–IDA-EPI (Predicted Multiple reaction monitoring mode – Information dependent acquisition - Enhanced production scan) method used pMRM (predicted MRM) as a survey scan. The information-dependent acquisition (IDA) method was employed to trigger the enhanced product ion (EPI) scans by analyzing MRM signals. A total of 66 MRM transitions were used as the survey experiment in positive mode. The system included an Agilent 1200 HPLC pump, degasser, auto sampler and column heater (Agilent, USA). The in house developed and validated HPLC separation was performed using a 50 × 2.1 mm, Luna 5 μm NH2 100 Å column (Agilent, USA) operated at a flow rate of 1000 μl/min. A mobile phase system consisted of (A) water and (B) acetonitrile and was used with the following gradient: 30% B for 0.75 min, 30 to 90% B from 0.75 to 7 min, and finally 30% B isocratic from 7 to 12 min.
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