HPLC Determination of Organic Acids

info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Scope:

This method is intended for the analysis of organic acid concentrations. Samples are
analyzed by High Performance Liquid Chromatography (HPLC) using ultraviolet (UV)
detection at 210 nanometers.

Safety:

Please consult the appropriate Material Safety Data Sheets for the below reagents to
obtain more detailed safety information.

Apparatus:

Liquid Chromatograph Waters Breeze HPLC

Detector Waters 2998 Photodiode Array Detector
Analytical Column Allure Organic Acids C18:5 micron (300 x 4.6 mm)
Software Breeze 2.0

Reagents:

100 mM Phosphate Buffer HPLC Grade

Water HPLC Grade
-----------------------------------------------------------------------------
Mobile Phase 100 mM phosphate buffer (pH 2.5)
Dilute 3.695 g of monosodium phosphate and 9.812 g of
disodium phosphate to 900 mL with HPLC water and mix
thoroughly. Adjust pH to 2.5 with 25% phosphoric acid.
Dilute to 1 L with HPLC water.

Standard 2.0 mg/mL in mobile phase

Sample 20.0 mg/ml in mobile phase

HPLC Chromatographic Parameters:

Column Temperature 40˚C

Flow Rate 1.0 mL/minute
Solvents Isocratic: Pump A: 100% Mobile Phase
Wavelength 210 nm
Injection Volume 10 uL

This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied.
This information is offered solely for your investigation, verification, and consideration.
Version6/01-28-19/WI#188
 HPLC Determination of Organic Acids
info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Procedure:
After running standards of organic acids under the above parameters, prep and run
sample in the same way. Create a calibration curve of standards using Breeze 2.0
software, then integrate/calibrate the standard curve to the sample injection to
quantitate the levels of organic acids.

Reviewed 01/28/2019 – Jillian Barrow, Manager of Analytical Chemistry -

Approved 01/28/2019– Erica Segura, Director of R&D -

This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied.
This information is offered solely for your investigation, verification, and consideration.
Version6/01-28-19/WI#188