Flm 3000 flow manager module

All the chemicals and reagents
used for high-performance liquid chromatography separation coupled
to electrospray-ion trap mass spectrometry (HPLC-ESI-IT-MS) analysis
were purchased from Sigma Aldrich (St. Louis, MO, USA). HPLC-ESI-MS
analyses were performed with a Surveyor HPLC system connected to an
LCQ Advantage mass spectrometer (Thermo Fisher Scientific San Jose,
CA, USA). The chromatographic column was a Vydac C8 reverse phase
(Hesperia, CA, USA) (150 × 2.1 mm, particle diameter 5 μm).
HPLC-high-resolution ESI-MS and MS/MS experiments were carried out
using an Ultimate 3000 Micro HPLC apparatus (Dionex, Sunnyvale, CA,
USA) equipped with a FLM-3000-Flow manager module and coupled to an
LTQ-Orbitrap Elite apparatus (Thermo Fisher). The column was a Zorbax
300SB-C8 (3.5 μm particle diameter; 1.0 × 150 mm).

All general chemicals and reagents were of analytical grade and were purchased from Merck (Damstadt, Germany) and J.T.Baker (Deventer, The Netherlands).
Low-resolution HPLC-ESI-MS measurements were carried out by a Surveyor HPLC system (Thermo Fisher, San Jose, CA, USA) connected by a T splitter to a photodiode-array detector and an LCQ Deca XP Plus mass spectrometer (Thermo Fisher). The mass spectrometer was equipped with an ESI source. High-resolution HPLC-ESI-MS/MS experiments were carried out by an Ultimate 3000 Micro HPLC apparatus (Dionex, Sunnyvale, CA, USA) equipped with a FLM-3000-Flow manager module and coupled to an LTQ Orbitrap XL apparatus (Thermo Fisher) (12 (link)).

High-resolution HPLC-ESI-MS/MS experiments were carried out using an Ultimate 3000 Micro HPLC apparatus (Dionex, Sunnyvale, CA, USA) equipped with a FLM-3000-Flow manager module and coupled to an LTQ Orbitrap XL apparatus (Thermo Fisher Scientific, Waltham, MA). The chromatographic column was a Zorbax 300 SB-C8 (3.5 µm particle diameter; column dimension 1 mm i.d. × 15 cm). High-resolution HPLC-ESI-MS experiments were performed by using the following eluents: (A) 0.1% formic acid in water and (B) 0.1% formic acid in acetonitrile-water 80/20 (v/v). The applied gradient was 0–4 min 5% B, 4–34 min from 5 to 50% B (linear), 34–54 min from 50 to 90% B (linear), at a flow rate of 80 µL/min. High-resolution positive MS/MS spectra were collected in data-dependent acquisition mode; the three most intense multiply charged ions were selected in a time window of 3 milliseconds and fragmented by using collision induced dissociation (35% normalized collision energy) and spectra were recorded. Tuning parameters were: capillary temperature 250°C, source voltage 4 kV, capillary voltage 48 V, tube lens voltage 170 V.