Agilent 5973c network mass selective detector

The extracted organic compounds were separated using an Agilent 7890A Series Gas Chromatograph interfaced to an Agilent 5973c Network Mass Selective Detector and an Agilent 7683 Series Injector (Agilent Technologies, Santa Clara, CA, USA). A 5 µL sample was injected with a split ratio of 1:5 by 0.3% standard deviation into an HP-5MS column (30 m × 0.25 mm, 0.25 µm film thickness; Agilent Technologies, USA) using helium as the carrier gas at 1 mL/min. The ion source was maintained at 250 °C. The GC oven was programmed with a temperature gradient starting at 100 °C (for 3 min), which was gradually increased to 300 °C (for 5 min) at 8 °C/min.
MS was carried out in the electron-impact mode at an ionizing potential of 70 eV by selected ion monitoring (SIM). The presence of CoPPIX-specific m/z 621 ion was monitored.
The qualitative and quantitative analyses of CoPPIX were performed using synthetic CoPPIX as a standard (Figure S1). Its concentration was determined based on the standard curve (0.08, 0.8, 1.6, 16, and 32 µM (50, 500, 1000, 10,000, and 20,000 µg/L)). A concentration curve was plotted with the peak areas corresponding to the concentration of CoPPIX tested (Figure S2). The analysis was performed in triplicate. Error bars represented the classical standard deviation for 3 replicates. The statistical significance of the obtained results was tested using the Student’s t-test.

The separation of organic compounds was performed using an Agilent 7890A Series Gas Chromatograph (GC) interfaced to an Agilent 5973c Network Mass Selective Detector and an Agilent 7683 Series Injector (Agilent Technologies, Santa Clara, CA, USA). A 5 µL sample was injected with split 1:5 (sample; carrier gas) by 0.3% SD to a HP-5MS column (30 m × 0.25 mm I.D., 0.25 µm film thickness, Agilent Technologies, Santa Clara, CA, USA) using He as the carrier gas at 1 mL min⁻¹. The ion source was maintained at 250 °C; the GC oven was programmed with a temperature gradient starting at 100 °C (for 3 min), and this was gradually increased to 300 °C (for 5 min) at 8 °C min⁻¹. Mass spectrometry analysis was carried out in the electron impact mode at an ionizing potential of 70 eV. Mass spectra were recorded from m/z 40 to 800 (0–30 min).

Gaseous compounds were separated using an Agilent 7890A Series Gas Chromatograph interfaced to an Agilent 5973c Network Mass Selective Detector (Agilent Technologies, Santa Clara, CA, USA). A 5 cm³ gas sample was injected into an HP-1MS column (30 m × 0.25 mm I.D., 0.25 µm film thickness; Agilent Technologies, Santa Clara, CA, USA) with a splitless injector using helium as the carrier gas (1 mL/min). The ion source was maintained at a temperature of 250 °C. The GC oven was programmed with a temperature gradient starting at 35 °C (for 5 min) and gradually increasing to 100 °C at 12 °C/min.
MS analysis was carried out in the electron-impact mode at an ionizing potential of 70 eV. The mass spectra of the samples were recorded at the mass-to-charge (m/z) ratio of 0–150 (0–10.5 min).
Selected compounds were identified using Agilent Technologies Enhanced ChemStation (G1701EA ver. E. 02.00.493) and Wiley Registry of Mass Spectral Data (version 3.2; Palisade Corporation, 8th Edition with Structures (Copyright 1988–2000), and John Wiley and Sons, Inc. (Copyright 2000)) using a 3% cutoff threshold.