Exactive orbitrap high performance benchtop lc ms

Analytical HPLC was performed on a Shimadzu LC-10Avp series HPLC system consisting of an autosampler, high-pressure pumps, column oven and PDA. HPLC conditions: C18 column (Eurospher 100-5, 250 × 4.6 mm) and gradient elution (MeCN/0.1 % (v/v) TFA 0.5/99.5 in 30 min to MeCN/0.1 % (v/v) TFA 100/0, MeCN 100 % for 10 min), flow rate 1 mL min⁻¹. Preparative HPLC was performed on a Shimadzu LC-8a series HPLC system with PDA. LC-MS measurements were performed using an Exactive Orbitrap High Performance Benchtop LC-MS with an electrospray ion source and an Accela HPLC system (Thermo Fisher Scientific, Bremen). HPLC conditions: C18 column (Betasil C18 3 µm 150 × 2.1 mm) and gradient elution (MeCN/0.1 % (v/v) HCOOH (H₂O) 5/95 for 1 min, going up to 98/2 in 15 min, then 98/2 for another 3 min; flow rate 0.2 mL min⁻¹). NMR spectra were recorded on a Bruker AVANCE III 600 MHz instrument equipped with a Bruker cryo platform. The residual solvent signals were used as an internal reference.

Analytical HPLC was performed on a Shimadzu LC-10Avp series HPLC system consisting of an autosampler, high-pressure pumps, column oven and photodiode array detector. HPLC conditions were as follows: C18 column (Eurospher 100-5, 250 × 4.6 mm) and gradient elution (MeCN/0.1% (v/v) trifluoroacetic acid (TFA) 0.5/99.5 in 30 min to MeCN/0.1% (v/v) TFA 100/0, MeCN 100% for 10 min), flow rate 1 ml min⁻¹. Preparative HPLC was performed on a Shimadzu LC-8a series HPLC system with photodiode array detector. LC-MS measurements were performed using an Exactive Orbitrap High Performance Benchtop LC-MS with an electrospray ion source and an Accela HPLC system (Thermo Fisher Scientific, Bremen). HPLC conditions were as follows: C18 column (Betasil C18 3 μm 150 × 2.1 mm) and gradient elution (MeCN/0.1% (v/v) HCOOH (H₂O) 5/95 for 1 min, going up to 98/2 in 15 min, then 98/2 for another 3 min; flow rate 0.2 ml min⁻¹). For tandem mass spectrometry measurements, a Q Exactive Orbitrap mass spectrometer with an electrospray ion source (Thermo Fisher Scientific) was used. NMR spectra were recorded on a Bruker AVANCE III 600 MHz instrument equipped with a Bruker cryo platform. Spectra were normalized to the residual solvent signals. The infrared spectra were recorded on a JASCO FT/IR-4100 type A.

Exactive Orbitrap High Performance Benchtop LC-MS (Thermo Fisher Scientific) with an electron spray ion source and an Accela HPLC System, C18 column (Betasil C18, 150 × 2.1 mm, Thermo Fisher Scientific), solvents: acetonitrile and distilled water (both supplemented with 0.1% formic acid), flow rate: 0.2 mL/min; program: hold 1 min at 5% acetonitrile, 1–16 min 5–99% acetonitrile, hold 15 min 99% acetonitrile, 19–20 min 99% to 5% acetonitrile, hold 11 min at 5% acetonitrile. The metabolic profiles of the acquired fractions were monitored with HR-ESI-LC/MS.