The amino acids present in the extract were determined according to Pérez-Palacios et al. (2015) (link) with some modifications. Protein extracts were hydrolysed using 6 M HCl at 105 °C for 12–24 h and deproteinised by acetonitrile, before derivatisation using N-tert-Butyldimethylsilyl-N-methyltrifluoroacetamide with 1% tert-Butyldimethylchlorosilane (MTBSTFA + 1% TBDMSCI) and then analysed by a gas chromatography equipment (7890A, Agilent Technologies) coupled to a mass spectrometer (5975C inert MSD with Triple Axis Detector, Agilent Technologies) equipped with a HP-5MS capillary column. Amino acid standard (AAS18) from Sigma-Aldrich was used for quantitation.
5975c inert msd with triple axis detector
Manufactured by Agilent Technologies
Sourced in United States
The 5975C inert MSD with Triple Axis Detector is a gas chromatography-mass spectrometry (GC-MS) system designed for analytical laboratories. It features an inert ion source and a triple axis detector for enhanced sensitivity and selectivity in compound detection and identification.
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3 protocols using 5975c inert msd with triple axis detector
1
Amino Acid Profiling via GC-MS
2
Quantitative GC-MS Analysis of Organic Compounds
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An Agilent 5975C inert MSD with triple-Axis Detector was used. The MS was operated in the EI mode (70 eV). The GC–MS interface, ion source and quadruple temperatures were set at 266, 230 and 150 °C, respectively. For quantitative determination, the MS was operated in the time-scheduled selected ion monitoring (SIM) mode.
Helium (99.999%) was employed as carrier gas at the constant flow rate of 1.0 mL/min. The gas chromatograph was operated in the splitless mode and the split valve was kept closed for 0.75 min. The injector temperature was kept at 250 °C. The column temperature was raised from 75 °C (hold 3 min) to 120 °C at 25 °C/min, then to 300 °C at 5 °C/ min (hold 11 min). Total time for the GC analysis was 51.8 min. A split/splitless injector operated in the splitless mode was used. The carrier was hydrogen at 9 psi pressure. The flow of carrier gas was applied as 19 ml/min. The injection volume was 2 μl.
Helium (99.999%) was employed as carrier gas at the constant flow rate of 1.0 mL/min. The gas chromatograph was operated in the splitless mode and the split valve was kept closed for 0.75 min. The injector temperature was kept at 250 °C. The column temperature was raised from 75 °C (hold 3 min) to 120 °C at 25 °C/min, then to 300 °C at 5 °C/ min (hold 11 min). Total time for the GC analysis was 51.8 min. A split/splitless injector operated in the splitless mode was used. The carrier was hydrogen at 9 psi pressure. The flow of carrier gas was applied as 19 ml/min. The injection volume was 2 μl.
3
Synthesis and Analysis of Mannosylated Palm Kernel Oil
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All chemicals used in the synthesis of ManPKO, D(+)-mannose monohydrate and boron trifluoride, BF 3 , were purchased from Sigma Aldrich and used without further purification. Palm kernel oil, PKO, was obtained from Golden Jomalina Food Industries Sdn. Bhd. (Malaysia), and the main components were lauric (49%), myristic (16%) and oleic (8%) acids. Dodecanol and cis-9-octadecen-1-ol, used for the synthesis of the pristine mannosides, were purchased from Merck.
Solvents for synthesis were analytical grade and used without further purification.
The mannosides, ManPKO, ManC 12 , and ManC 18:1 , were synthesised using a three-steps method consisting of peracetylation, glycosylation, and finally deacetylation, as shown in Figure 2. Fatty acids of PKO were converted to PKO alcohol by reduction using lithium aluminium The compositions of PKO alcohol were determined using a 450 GC/220 MS (Varian Inc. USA) gas chromatography system equipped with a 5975C inert MSD with triple-axis detector (Agilent Technology, USA). Operating conditions are described in the ESI following previous reports by Oborn [30] .
Solvents for synthesis were analytical grade and used without further purification.
The mannosides, ManPKO, ManC 12 , and ManC 18:1 , were synthesised using a three-steps method consisting of peracetylation, glycosylation, and finally deacetylation, as shown in Figure 2. Fatty acids of PKO were converted to PKO alcohol by reduction using lithium aluminium The compositions of PKO alcohol were determined using a 450 GC/220 MS (Varian Inc. USA) gas chromatography system equipped with a 5975C inert MSD with triple-axis detector (Agilent Technology, USA). Operating conditions are described in the ESI following previous reports by Oborn [30] .
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