Similar protocols

Pipeline publication

[…] natant was removed and freeze-dried. The oximation reaction was performed by adding 100 μL of the methoxyamine solution to the dried sample and incubating the mixture in a 37 °C water bath for 90 min. Subsequently, 80 μL of MSTFA was added to the sample for silylation reaction and incubated in a 37 °C water bath for 60 min. The sample was analyzed on a Shimadzu single quadrupole GCMS-QP2010 plus (Kyoto, Japan). An Agilent DB-5 MS fused silica capillary column (30 m × 0.25 mm × 0.25 μm) was used for separation. Selected ion monitoring (SIM) mode was chosen for the instrumental analysis. Peak deconvolution and the identification and selection of the characteristic ions were performed by using AMDIS, Leco ChromaTOF and in-house software. The spit ratio was 10:1. The carrier gas was helium, and the flow rate was 1.2 mL/min. The injector, interface and ion source temperatures were set to 280 °C, 280 °C, and 240 °C, respectively. The detector voltage was 0.92 kV. The ionization mode was electron impact (EI) at 70 eV., For LC-MS, peak alignment was achieved with XCMS. For CE-MS, peak deconvolution, detection and area integration were performed by using Agilent Masshunter qualitative and quantitative analysis software. For GC-MS, the molecular features were obtained from the acquired SIM tables. Peak tables with mass, retention time and abundance were established for the three methods. PCA was performed to investigate the changes among the different developmental stages by using SIMCA-P software (Version 11.5, Umetrics AB, Umeå, Sweden). Nonparametric tests were conducted to identify the senescence-related metabolites with Multiexperiment Viewer (MeV) (Version 4.9, Dana-Farber Cancer Institute, Boston, MA, USA). The significance level was less than 0 […]

Pipeline specifications

Software tools AMDIS, ChromaTOF, XCMS
Organisms Nicotiana tabacum
Chemicals Amino Acids, Carbohydrates, Polyamines, Sterols, Steroids, Sugar Alcohols, Amino Acids, Dicarboxylic, Excitatory Amino Acids, Amino Acids, Acidic, Heterocyclic Compounds, 2-Ring, Hydrocarbons, Cyclic