Indole-3-acetic acid

Examples of using this product（for reference only）

LC-MS

For HILIC separation, samples were injected onto an column (2.1 ×100 mm, 1.7 μm; ) at 25 ℃, and using a 21.2 min linear gradient at 0.3 mL/min flow rate and 2 μL injection volume. ESI source conditions were set as follows: curtain gas (CUR) as 30, ion source gas1 (GS1) as 60, and ion source gas2 (GS2); source gas temperature as 600℃; ISVF as 5500 V. The MS/MS acquisition was performed in positive (PI) and negative ion (NI) modes, the mass range was set at 25-1000 m/z, and the accumulation time for product ion scan was set at 0.05 s/spectrum.

The second order spectrum was acquired in information-dependent acquisition (IDA) and high-sensitivity modes. Parameters were set as follows: declustering potential (DP) as±60 V; collision energy as 35 V±15 eV; maximum candidate ions to monitor per cycle as 10; excluding isotopes within 4 Da.

References：

Ma Z, Yang K, Wang J, Ma J, Yao L, Si E, Li B, Ma X, Shang X, Meng Y, Wang H. Exogenous Melatonin Enhances the Low Phosphorus Tolerance of Barley Roots of Different Genotypes. Cells. 2023 May 16;12(10):1397. doi: 10.3390/cells12101397. PMID: 37408231; PMCID: PMC10217165.

LC-MS/MS

LC-MS/MS conditions Analysis was performed using a liquid chromatographytandem mass spectrometer LC–MS/MS equipped with an ESI source. Separation was performed using a C18 column (250 mm × 4.6 mm × 5 μm). The flow rate was 0.4 ml/min, and the column temperature was maintained at 25℃. The mobile phases were formic acid: water (0.1:100, v/v) as mobile phase A, and methanol:acetonitrile (1:1) and 0.1% formic acid as mobile phase B. The binary gradient elution conditions were: (A:B): 0.01 min–5 min, 70:30→5:95; 5–8 min, 5:95; 8.01 min–16 min, 70:30. Detection was performed using multiple reaction monitoring (MRM) in positive mode, and the optimized MRM parameters for each compound are shown in Table 1. The mass condition parameters were set as: nebulizer gas, 3 L/min; drying gas, 10.0 L/min; heating gas, 10.0 L/min; interface temperature, 30℃; collision-induced dissociation (CID) gas, 230 kPa; DL temperature, 250℃; thermal block temperature, 400℃; interface voltage, ?4.5 kV. The autosampler was kept at 4℃.

References：

Xu H, Pan LB, Yu H, Han P, Fu J, Zhang ZW, Hu JC, Yang XY, Keranmu A, Zhang HJ, Bu MM, Jiang JD, Wang Y. Gut microbiota-derived metabolites in inflammatory diseases based on targeted metabolomics. Front Pharmacol. 2022 Sep 27;13:919181. doi: 10.3389/fphar.2022.919181. PMID: 36238574; PMCID: PMC9551995.

Remark：These protocols are for reference only. Solarbio does not independently validate these methods.

Note：

1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.

2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.

3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.