Indole-3-Butyric Acid Analyzed with HPLC – AppNote – MicroSolv Technology Corporation

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ARTICLES, APPNOTES, FAQ (9292)
- AppNotes Library (1888)
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October 28, 2021
Tanya

AppNotes Cogent Phenyl Hydride / AppNotes Library / AppNotes Listed by Column / AppNotes Pharmaceuticals / AppNotes Reversed Phase Methods / ARTICLES, APPNOTES, FAQ

A Reproducible Method for Detection of a Plant Hormone
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

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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