Extraction of Indole 3 carboxylic acid from Strata-X-A on Gemini 5u, C18, 150x4.6mm Column

Sample Preparation Details

Part No:8B-S123-TAK
Solid Phase Extraction (SPE) procedure:Note: The solvent volumes shown below are for a 30 mg bed mass. The solvent volumes will need to be adjusted for a smaller or larger bed mass.
Load:500 ul H Plasma (Na2 EDTA) spiked with Indole 3 carboxylic acid at 5 ug/mL, diluted with 500uL water, acidified with 2% H3PO4.
Dry:2 mins at 10" of Hg
Final Prep and Analysis: Elution solvent was applied in 2 aliquots of 500uL followed by evaporation to dryness at 45 deg C under N2 and reconstitute with 500 uL initial mobile phase (20mm KH2PO4(PH 2.5)/ ACN=75/25) spiked with 50ug/mL Salicylic acid (IS)
Inject: 50 µL on HPLC / UV-Vis Abs.-Variable Wave.(UV) @ 220 nm (ambient)


Name Spiked
Log P pKa % Rec %RSC (n=0)
1 Indole-3-carboxylic acid 91 3.1
2 Salicylic acid 50 

Application Detail (App ID: 19274)

Part No:00F-4435-E0
Dimensions:150 x 4.6 mm ID
Elution Type:Isocratic
Elution A:20 mM KH2PO4(pH 2.5)
Elution B:Acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 75 25

Flow Rate:1 mL/min
Col. Temp: ambient
Detection: UV-Vis Abs.-Variable Wave.(UV) @ 220 nm (22°C)
Note:SecurityGuard™ Guard Cartridge System extends column lifetime.
- SecurityGuard Cartridges, Gemini C18 4 x 3.0mm ID, 10/Pk Part No.: AJ0-7597
- Holder Part No.: KJ0-4282

Analyte Details

Analyte Detail not available

Indole-3-carboxylic acid

Salicylic acid