HPLC Separation of 2-Fluorophenylacetic acid, 4-Fluorophenylacetic acid, 3-Fluorophenylacetic acid on Newcrom BH and Primesep SB Columns

Separation type: Liquid Chromatography Mixed-mode

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High Performance Liquid Chromatography (HPLC) Method for Analysis of 2-Fluorophenylacetic acid , 4-Fluorophenylacetic acid,  3-Fluorophenylacetic acid

2-Fluorophenylacetic acid is a popular chiral derivatizing agent used in 19F NMR spectroscopy. 3-Fluorophenylacetic acid is a common precursor for synthesizing various pentaamine and bis-heterocyclic compounds. 4-Fluorophenylacetic acid is a common intermediary compound used to produce fluorinated anesthetics. While these 3 compounds all have the same chemical formula, the position of the Fluoride group varies between them, changing slightly their retention and chromatographic characteristics.
All 3 isomers can be measured at low UV. Using a Primsep SB and a Newcrom BH reverse-phase columns and a mobile phase consisting of acetonitrile (MeCN) and water with either formic acid or acetic acid buffer, these fluorophenylacetic acid isomers can be retained, separated, and UV detected at 264 nm with precise resolution. This method is compatible with Mass Spectrometry.


Condition 1

Column Newcrom BH, 4.6×250 mm, 3 µm, 100A
Mobile Phase MeCN/H2O – 30/70%
Buffer H3PO4 – 0.05%
Flow Rate 1.0 ml/min
Detection UV 210  nm

Condition 2

Column Primesep SB, 4.6×250 mm, 5 µm, 100A
Mobile Phase MeCN/H2O – 40/60%
Buffer H3PO4 – 0.1%
Flow Rate 1.0 ml/min
Detection UV 210  nm


Class of Compounds Acid
Analyzing Compounds 2-Fluorophenylacetic acid (2-FPAA), 4-Fluorophenylacetic acid (4-FPAA), 3-Fluorophenylacetic acid


Application Column

Newcrom BH

The Newcrom columns are a family of reverse-phase-based columns. Newcrom A, AH, B, and BH are all mixed-mode columns with either positive or negative ion-pairing groups attached to either short (25 Å) or long (100 Å) ligand chains. Newcrom R1 is a special reverse-phase column with low silanol activity.

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Primesep SB

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
2-Fluorophenylacetic acid
3-Fluorophenylacetic acid
4-Fluorophenylacetic acid
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.