A Reproducible Method for Detection of a Plant Hormone
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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

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.

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