Gc 2010 system

Compound identification was carried out on a GCMS-QP2010 system (Shimadzu, Kyoto, Japan) equipped with a split–splitless injector. Data files were collected and elaborated by using Shimadzu “GCMS solution” software (version 4.45) (Kyoto, Japan).
Compound quantification was performed on a GC2010 system (Shimadzu, Kyoto, Japan) equipped with a split–splitless injector. Data files were collected and elaborated by using Shimadzu LabSolutions software (version 5.92) (Kyoto, Japan).

The activity of PvEH1 or its mutant as well as the conversion ratio (c) of rac-mCSO defined as the ratio of its depleted concentration to initial one was assayed by high-performance liquid chromatography (HPLC), using an e2695 apparatus with an XBridge BEH C18 column (Waters, Milford, MA). The mobile phase of methanol/H₂O (7:3, v/v) was used at 0.8 mL/min, and continuously monitored using a Waters 2489 UV–Vis detector at 220 nm. The generated diols (R)- and (S)-mCPED were analyzed by HPLC with a Chiralcel OD-H column (Daicel, Osaka, Japan). The n-hexane/isopropanol (9:1, v/v) was used as a mobile phase. Because (R)- and (S)-mCSO can not be separated by OD-H, they were assayed by chiral gas chromatography (GC), using a GC-2010 system (Shimadzu, Tokyo, Japan) with a CP-Chirasil-DEX CB column (Agilent, Santa Clara, CA) and a flame ionization detector. The injector and detector temperatures were 220 °C, while the column temperature was programmed from 110 to 190 °C at 10 °C/min. The nitrogen gas carrier was used at 3.0 mL/min.

Isolated rBGL2ur was subjected to carbohydrate profiling using gas chromatography (GC) analysis to determinate whether rBGL2ur possesses the TP antigen determinant sugar moiety, 3-O-methyl-mannose. Monosaccharides were detected and quantified by gas liquid chromatography-flame ionization detector (GLC-FID) on a Shimadzu GC-2010 system after conversion to alditol acetate derivatives, as previously described [20 (link)]. A Crossbond 50% cyanopropylmethyl/50% phenylmethyl polysiloxane column was used (15 m × 0.25 mm with 0.25 μm film thickness, RTX-225, Restek, Bellefonte, PA, USA). The GLC conditions were as follows: injector at 220°C, detector at 240°C, and a temperature program of 215°C for 2 min, then 4°C/min up to 230°C before holding for 11.25 min, run at constant linear velocity of 33.4 cm/sec and split ratio of 50:1. Pure 3-O-methyl-mannose (Omicron Biochemicals, South Bend, IN, USA) was used as a reference.

Achiral GC-FID analysis of the derivatised hydroxylated fatty acids was performed with a Shimadzu type GC-2014 equipped with a CP-Sil5 CB column (50 m × 0.53 mm × 1.0 μm) using N₂ as carrier gas. The following conditions were used for the achiral separation using direct injection: injector 340 °C, detector (FID) 360 °C, column flow rate 20.0 ml/min, temperature program: start at 130 °C, hold time 4 min, rate 15 °C/min to 330 °C hold time 5 min.
Gas chromatography-mass spectrometry of derivatised hydroxylated fatty acids was performed with the Shimadzu GC-2010 system which is connected to the GCMS-QP2010s mass detector from Shimadzu. The column CP-Sil5 CB (25 m × 0.25 mm × 0.4 μm) was used. Injections were performed with the autoinjector AOC-20i from Shimadzu. The injector temperature was kept at 250 °C. The injector was used in split-mode with a split ratio of 30:1 at a pressure of 51.2 kPa. The temperature program for fatty acids 1 and 3-15: start at 130 °C, hold time 4 min, rate 15 °C/min to 330 °C hold time 5 min; temperature program for fatty acid 2: start at 130 °C, hold time 4 min, rate 5 °C/min to 325 °C hold time 7 min. Structure determination was based on the comparison of monomer peaks using external standards.

A GC with MS detection was used to determine the specific migration of individual substances. Gas chromatographic separation was performed using a Shimadzu GC2010 system equipped with a Rxi-5Sil MS capillary column (30 m × 250 µm × 0.25 µm). Helium was used as carrier gas in linear velocity flow control mode with a constant flow of 40 cm s⁻¹. Aliquots of 1 µL were injected using a high-pressure splitless injection with 250 kPa for 0.8 min and a column flow of 4.74 mL min⁻¹; a gas saver was switched on in a ratio of 1:5 after 2 min. The septum purge flow was 3 mL min⁻¹, the injection temperature 270 °C and sampling time 1 min. The initial oven temperature was 70 °C (1 min) and was raised at 6 °C min⁻¹ to 340 °C (5 min). Detection was done with GCMS-QP2010 PLUS mass selective detector. Ions were generated with electron ionization (70 eV), the detector voltage was relative to tune, mass spectrometer scanned from m/z 35 to 500 with a scan rate of 1666 amu s⁻¹ and an event time of 0.3 s. The software used was “GC-MS Solution” version 4.42.