Indole-3-Butyric Acid Analyzed with HPLC - AppNote
October 28, 2021
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A Reproducible Method for Detection of a Plant Hormone
Click HERE for Column Ordering Information.
Method Conditions
Column: Cogent Phenyl Hydride™, 4μm, 100Å
Dimensions: 4.6mm x 75mm
Mobile Phase:
--A: DI Water with 0.1% Formic Acid (v/v)
--B: Acetonitrile with 0.1% Formic Acid (v/v)
Gradient:
Injection vol.: 1μL
Flow rate: 1.0mL / minute
Detection: UV @ 280nm
Sample Preparation: Indole-3-Butyric Acid prepared as 1.0mg / mL standard solution in (50:50) Acetonitrile / DI Water
Notes: Indole-3-Butyric Acid a substance that is closely related in structure and function to a natural growth regulator found in plants. Indole-3-butyric acid is used on many crops and ornamentals to promote growth and development of roots, flowers and fruits, and to increase crop yields.
Click HERE for Column Ordering Information.
A rapid, sensitive, and Reproducible Method has been developed for Analysis of Indole-3-Butyric Acid. The data below, (overlay of 10 chromatograms ) illustrates how the compound can be adequately Retained and detected using a simple Gradient in Reversed Phase HPLC. The Method demonstrates good Peak Shape and run-to-run Precision with RSD values less than 0.3%.
A Phenyl ring in the Column Stationary Phase provides beneficial π-π Interaction with the Analyte making possible the use of a very simple, Mass Spec friendly Mobile Phase with Formic Acid as an additive.
10 Injections of Indole-3-Butyric Acid Overlay
Method Conditions
Column: Cogent Phenyl Hydride™, 4μm, 100Å
Dimensions: 4.6mm x 75mm
Mobile Phase:
--A: DI Water with 0.1% Formic Acid (v/v)
--B: Acetonitrile with 0.1% Formic Acid (v/v)
Gradient:
| Time (minutes) | %B |
| 0 | 25 |
| 1 | 25 |
| 5 | 85 |
| 6 | 85 |
| 7 | 25 |
| 10 | 25 |
Flow rate: 1.0mL / minute
Detection: UV @ 280nm
Sample Preparation: Indole-3-Butyric Acid prepared as 1.0mg / mL standard solution in (50:50) Acetonitrile / DI Water
Notes: Indole-3-Butyric Acid a substance that is closely related in structure and function to a natural growth regulator found in plants. Indole-3-butyric acid is used on many crops and ornamentals to promote growth and development of roots, flowers and fruits, and to increase crop yields.