Agilent g1607a ce esi ms sprayer kit

CE-TOFMS was performed using an Agilent capillary electrophoresis system equipped with an Agilent 6210 time-of-flight mass spectrometer, Agilent 1100 isocratic high-performance liquid chromatography pump, Agilent G1603A CE-MS adapter kit, and Agilent G1607A CE-ESI-MS sprayer kit (Agilent Technologies, Waldbronn, Germany). The analytical conditions were identical to those used in a previous study [21 (link)].
Raw data were processed using MasterHands, as described previously [21 (link)]. Briefly, among the detected compounds, those annotated in the Human Metabolome Database (ver. 4.0, http://www.hmdb.ca/, accessed on 27 January 2022) or KEGG database (http://www.genome.jp/kegg/, accessed on 27 January 2022) were further analyzed. The relative contents of the annotated compounds were determined by comparing the peaks of compounds with the same MS properties. To compare the relative content of the compounds between the LN and HN groups, the peak areas were normalized to those of the internal standards and sample weights. The abundance of each compound used for comparative analysis was set to zero when the level of the compound was not detected. The file conversion of raw MS data, peak picking, reduction of noise, and alignment of data for multiple samples was conducted as previously described [21 (link)].

CE-time-of-flight mass spectrometry (TOFMS) was carried out using an Agilent CE Capillary Electrophoresis system equipped with an Agilent 6210 Time of Flight mass spectrometer, Agilent 1100 isocratic HPLC pump, Agilent G1603A CE-MS adapter kit, and Agilent G1607A CE-ESI-MS sprayer kit (Agilent Technologies, Waldbronn, Germany). The systems were controlled by Agilent G2201AA ChemStation software version B.03.01 for CE (Agilent Technologies, Waldbronn, Germany). The metabolites were analyzed by using a fused silica capillary (50 μm i.d. × 80 cm total length), with commercial electrophoresis buffer (solution ID: H3301-1001 for cation analysis and H3302-1021 for anion analysis, Human Metabolome Technologies, Inc., Tsuruoka, Japan) as the electrolyte. The sample was injected at a pressure of 50 mbar for 10 s (approximately 10 nL) in cation analysis and 25 s (approximately 25 nL) in anion analysis. The spectrometer was scanned from m/z 50 to 1,000. Other conditions were as described previously [18 (link)-20 (link)].

CE-TOFMS was performed using an Agilent capillary electrophoresis system equipped with an Agilent 6210 time-of-flight mass spectrometer, Agilent 1100 isocratic high-performance liquid chromatography pump, Agilent G1603A CE-MS adapter kit, and Agilent G1607A CE-ESI-MS sprayer kit (Agilent Technologies, Waldbronn, Germany). The analytic conditions were identical to those used in a previous study [99 (link)]. The spectrometer was scanned from m/z 50 to 1000.

The CE-FTMS was performed using an Agilent 7100 CE capillary electrophoresis system equipped with a Q Exactive Plus (Thermo Fisher Scientific Inc., Waltham, MA, USA), Agilent 1260 isocratic HPLC pump, Agilent G1603A CE-MS adapter kit, and Agilent G1607A CE-ESI-MS sprayer kit (Agilent Technologies). The CE-FTMS was conducted using an analyzer by Human Metabolome Technologies Inc. (HMT, Tsuruoka, Japan), according to HMT’s ω Scan package, using a previously described method with some modifications [59 (link)]. In short, 40 µL of saliva samples was added to 10 µL of Milli-Q water containing internal standards (H3304-1002, Human Metabolome Technologies Inc.). The solutions were filtered using a Millipore 5-kDa cutoff filter (ULTRAFREE MC PLHCC, Human Metabolome Technology Inc.) at 9100× g for 60 min at 4 °C. Then, the filtrates were employed for CE-FTMS analysis. PCA was performed using the HMT’s proprietary software, SampleStat.

Fifty μL of plasma were added to 450 μL of methanol with an internal standard (10 μM final concentration solution of methionine sulfone and 10-camphorsulfonic acid, solution ID: H3304-1002, HMT) at 0 °C in order to inactivate enzymes. The extract was further mixed with 500 μL chloroform and 200 μL Milli-Q water. The mixture was centrifuged at 2300 × g for 5 min at 4 °C, and 350 μL of the upper aqueous layer was transferred into an ultrafiltration tube (Ultra-free MC PLHCC, filter-unit 5 kDa, HMT) and filtered to remove proteins. The filtrate was centrifugally concentrated and rehydrated in 50 μL of Milli-Q water for injection into the CE-TOFMS.
CE-TOFMS measurement was carried out using an Agilent CE Capillary Electrophoresis System equipped with an Agilent 6210 time of flight mass spectrometer, Agilent 1100 isocratic HPLC pump, Agilent G1603A CE-MS adapter kit, and Agilent G1607A CE-ESI-MS sprayer kit (Agilent Technologies, Waldbronn, Germany). The systems were controlled by Agilent G2201AA ChemStation software version B.03.01 for CE (Agilent Technologies). Pre-treated samples were applied into the system using fused silica capillaries (50 μm i.d. ×80 cm total length) (Agilent Technologies) to detect hydrosoluble metabolites. The measurement modes for cation and anion metabolites are shown in Table S1.

The metabolites were measured in cation and anion modes by capillary electrophoresis-time-of-flight mass spectrometry (CE-TOF/MS) system (Agilent Technologies Inc., Santa Clara, CA, USA) [29 (link),30 (link),49 (link)]. Peaks were extracted using automatic integration software (MasterHands ver. 2.13.0.8.h; Keio University, Tokyo, Japan). The CE system was equipped with an Agilent 6210 TOF mass spectrometer, Agilent G1603A CE-MS Adapter Kit, Agilent 1100 Isocratic HPLC Pump, and Agilent G1607A CE-ESI-MS Sprayer Kit (Agilent Technologies, Waldbronn, Germany). The system was controlled by Agilent G2201AA ChemStation software version B.03.01 for CE (Agilent Technologies Inc., Santa Clara, CA, USA). A fused silica capillary column with 50 µm i.d × 80 cm total length (Agilent Technologies, Waldbronn, Germany) was used for separation. Analytical conditions were set as follows: run buffer (cation buffer solution (p/n: H3301–1001) and anion run buffer (p/n: I3302–1023), which were also used for rinsing; MS ionization was conducted in positive and negative ion modes; sample injection pressure was 50 mbar for 10 s; CE voltage was 27 kV (cation) and 30 kV (anion); MS capillary voltage was 4000 V (cation) and 3500 V (anion); and the MS scan range was m/z 50–1000.