The respective crude metabolites were characterized, using Fourier Transform Infrared (FTIR) and Gas Chromatogram-Mass Spectrometry (GC-MS).
For FTIR analysis, an Infrared spectrometer Varian 660 MidIR Dual MCT/DTGS Bundle with ATR) was used. Prior to the analysis, the samples were first prepared by drying in a desiccator for 24 h. After drying, the samples were directly applied to a diamante crystal of ATR. The resulting spectra were corrected for background air absorbance. The spectra were recorded in a transmittance mode from 4000 to 500/400 cm−1 at a resolution of 4 cm. Infrared spectrum was Fourier transformed and recorded in the absorption mode.
For GC-MS, the samples were analysed using a Varian 3800 gas chromatograph equipped with an Agilent fused silica capillary CP-Sil 5 CB column (30 m × 0.25 mm) and connected to a Varian 4000 mass spectrometer. The carrier gas used was Itroge, at flow rate and split ratio of 1.0 ml/min and 1:10, respectively. The operating temperature was 60 °C for 1 min; rising at 3.0 °C/min to 240 °C and held for 1 min, with the injector and detector also held at 240 °C.
The components were detected and quantified by the mass spectrometer. Through comparison of their retention times and fragmentation pattern of mass with mass spectral libraries (Wiley 9 and NIST 08), the components were identified.
For FTIR analysis, an Infrared spectrometer Varian 660 MidIR Dual MCT/DTGS Bundle with ATR) was used. Prior to the analysis, the samples were first prepared by drying in a desiccator for 24 h. After drying, the samples were directly applied to a diamante crystal of ATR. The resulting spectra were corrected for background air absorbance. The spectra were recorded in a transmittance mode from 4000 to 500/400 cm−1 at a resolution of 4 cm. Infrared spectrum was Fourier transformed and recorded in the absorption mode.
For GC-MS, the samples were analysed using a Varian 3800 gas chromatograph equipped with an Agilent fused silica capillary CP-Sil 5 CB column (30 m × 0.25 mm) and connected to a Varian 4000 mass spectrometer. The carrier gas used was Itroge, at flow rate and split ratio of 1.0 ml/min and 1:10, respectively. The operating temperature was 60 °C for 1 min; rising at 3.0 °C/min to 240 °C and held for 1 min, with the injector and detector also held at 240 °C.
The components were detected and quantified by the mass spectrometer. Through comparison of their retention times and fragmentation pattern of mass with mass spectral libraries (Wiley 9 and NIST 08), the components were identified.