Tracegold tg 5ms column

The alkaloid extracts were dissolved in dichloromethane and filtered with a 0.45-µm regenerated cellulose syringe filter before gas chromatography–mass spectrometry (GC-MS) analysis, following the conditions described by Magalhães et al. (2017) (link) with modifications. The chromatographic analysis of the extracts was performed in a Thermo Fisher Scientific (Waltham, MA, USA) Trace 1300, ISQ single-quadrupole mass spectrometer equipped with a TraceGOLD TG-5MS column (30 m × 0.25 mm; 0.25 µm) from Thermo Fisher Scientific. The oven temperature was programmed as follows: 150°C for 1 min; followed by an increase at 5°C min⁻¹ until the temperature reaches 235°C, it is then held for 15 min; and then the temperature is increased at 10°C min⁻¹ until 280°C is reached, then it is held for 10 min. The injection volume was 1 µL and a split ratio of 1:5 was used. The identification of the compounds was performed by the analysis of standards or by comparison with the National Institute of Standards and Technology (NIST)’s database (2001) .

A Trace 1310 GC system equipped with a TraceGOLD TG-5MS column (30 m, 0.25 mm, 0.25 µm), coupled to a Q-Exactive GC Orbitrap HRMS detector, all from Thermo Fisher Scientific (Waltham, MA, USA), were used for GC-HRMS analysis.
A Conterm laboratory oven, from J.P. Selecta (Barcelona, Spain); an Advanced IR vortex mixer, from VELP Scientifica (Usmate Velate, Italy); and an Allegra X-15R centrifuge, from Beckman Coulter (Indianapolis, IN, USA), were also used.

Derivatized samples were injected in the gas chromatography–mass spectrometry (GC-MS) system, and an alkane standard mixture (C10–C40 all even) was injected at the start and end of the sample analysis for retention index (RI) calculation. For the analysis of metabolites extracted from plasma samples, 1 μl of derivatized samples was injected into the splitless mode using a Triplus 100 autosampler (Thermo Scientific) in the Trace 1300 gas chromatograph equipped with a TSQ 8000 mass spectrometer. Metabolites were separated on a TraceGOLD TG-5MS column (Thermo Scientific) with a diameter of 0.25 mm, a thickness of 0.25 µm, and a length of 30 m. Ultra-high purity grade helium and argon were used as the carrier gas and collision gas, respectively, with a flow rate of 1 ml min⁻¹. The injector port temperature was set at 200°C, whereas the transfer line and ion source temperature were set at 250°C. The GC program was started with an initial oven temperature of 50°C and held for 1 min; then, the temperature was increased to 100°C at a rate of 6°C min⁻¹, ramped up to 200°C at 4°C min⁻¹, and finally to 280 C at the rate of 20°C min⁻¹ which was kept constant for 3 min. All the samples were run on full scan mode ranging from m/z 60 to 650 for the metabolite data acquisition, and raw data obtained were collected for further analysis.