Return to study ST003190 main page
MB Sample ID: SA347866
| Local Sample ID: | WT-IP8-FigS2b |
| Subject ID: | SU003309 |
| Subject Type: | Plant |
| Subject Species: | Arabidopsis thaliana |
| Taxonomy ID: | 3702 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN005238 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Agilent 1290 Infinity |
| Column | Agilent InfinityLab Poroshell 120 EC-C18 (100 x 3mm,2.7um) |
| MS Type | Other |
| MS instrument type | HPLC-MS/MS |
| MS instrument name | Agilent 5973 |
| Ion Mode | POSITIVE |
| Units | mg/g |
Chromatography:
| Chromatography ID: | CH003965 |
| Chromatography Summary: | InsPs were detected using Hydrophilic Interaction High Performance Liquid Chromatography-Tandem Mass Spectrometry on an Agilent 1290 infinity HPLC system coupled to an Agilent 6460 triple Quad LC-MS/MS using InfinityLab Poroshell 120 HILIC-Z (2.1 × 100) column (Agilent Technologies, USA). Nitrogen was used as the sheath gas and drying gas. The nebulizer pressure was set to 45 psi and the flow rate of drying gas was 5 L / min. The flow rate and temperature of the sheath gas were 11 L / min and 350℃, respectively. Chromatographic separation was carried out on a HPLC column (100 × 2.1 mm, 2.7 µm). The column temperature was 35℃. The mobile phases consisted of (A) ammonium acetate in distilled water (pH 10.0) and (B) Acetonitrile. The gradient program was as follows: 0-10 min, 90 % → 55 % of B; 10-12 min, 55 % → 90 % of B; 12-20 min, 90 % of B. The flow rate was set at 0.3 ml / min, and the injection volume was 10 μl. Mass spectrometric detection was completed by use of an electrospray ionization (ESI) source in negative ion multiple-reaction monitoring (MRM) mode. InsPs were identified based on comparison to known InsPs species. The mass spectrometry parameters corresponding to different InsPs show as below: InsP3 (MRM: 419 -> 321, 419 -> 337, Acquisition time is 6-7 min); InsP4 (MRM: 499 -> 401, 499 -> 417, Acquisition time is 6-7 min); InsP5 (MRM: 579 -> 480.9, 579 -> 382.8, Acquisition time is 6-7 min); InsP6 (MRM: 659 -> 560.8, 659 -> 577, Acquisition time is 6-7 min); InsP7 (MRM: 739 -> 575, 739 -> 657, Acquisition time is 5-6 min); InsP8 (MRM: 819 -> 737, 819 -> 655, Acquisition time is 4-5 min). According to the regression equation calculated from the standard sample, substitute the response value of the sample into the equation to convert the corresponding concentration. For InsP3/InsP4/InsP5/InsP6/InsP7/InsP8 detection in seeds and seedlings, 10 g of 12-day-old seedlings or 2.4 g of dry seeds were used for InsPs enrichment with 300 mg of TiO2 beads for each sample. The enriched substances were analyzed by HPLC-MS/MS. The purchased InsP3 (1,4,5-InsP3, MedChemExpress), InsP4 (1,3,4,5-InsP4, MedChemExpress), InsP5 and InsP6, InsP7 (5-InsP7) and InsP8 (1,5-InsP8) from Lei lab25 were used as standard samples for generating the calibration curves. |
| Instrument Name: | Agilent 1290 Infinity |
| Column Name: | Agilent InfinityLab Poroshell 120 EC-C18 (100 x 3mm,2.7um) |
| Column Temperature: | 35℃ |
| Flow Gradient: | 0-10 min, 90 % → 55 % of B; 10-12 min, 55 % → 90 % of B; 12-20 min, 90 % of B |
| Flow Rate: | 0.3 mL/min |
| Solvent A: | 100% Distilled Water; 10% ammonium acetate (pH 10.0) |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
