16a-Hydroxyestradiol Separation on Mixed-Mode HPLC Columns

Separation type: Liquid Chromatography Mixed-mode




 

16a-Hydroxyestradiol is an estrogen very similar to estriol, but with three C13 in the 2,3,4 positions. Estriol is often used as a marker for fetal health since it is produced by the adrenal cortex of the fetus.  16a-Hydroxyestradiol was analyzed for a study focused on emerging contaminants in water by the Federal Institute of Hydrology of Koblenz, Germany. It was retained on both Primesep 100 and Obelisc R HPLC Columns. Obelisc R is a mixed-mode column which uses a long hydrophobic chain and multiple ionic groups to fine tune retention. Primesep 100 is a reverse phase column with embedded acidic ion pairing groups. Method is LC/MS compatible and will be effective for achieving unique retention for many pesticides.



Condition 1

Column Obelisc R, 2.1×150 mm, 5 µm, 100A
Mobile Phase Gradient MeCN – 25-70%, 15 min
Buffer Gradient AmAc pH 3.0- 30-60 mM, 15 min
Flow Rate 0.4 ml/min
Detection UV, 270 nm

 

Condition 2

Column Obelisc R, 2.1×150 mm, 5 µm, 100A
Mobile Phase Gradient MeCN – 10-70%, 15 min
Buffer Gradient AmAc pH 3.0- 20-60 mM, 15 min
Flow Rate 0.4 ml/min
Detection UV, 270 nm

 

Condition 3

Column Primesep 100, 2.1×150 mm, 5 µm, 100A
Mobile Phase Gradient MeCN – 10-70%, 15 min
Buffer Gradient AmAc pH 3.0- 20-60 mM, 15 min
Flow Rate 0.4 ml/min
Detection UV, 270 nm

 

Description

Class of Compounds
 Hormone,  Hydrophobic, Ionizable
Analyzing Compounds 16a-Hydroxyestradiol

 

Application Analytes:
16a-Hydroxyestradiol

Application Detection:
UV Detection
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.

Obelisc R

SIELC has developed the mixed-mode Obelisc™ columns to be the first commercially available columns with Liquid Separation Cell technology (LiSC™). This cost effective duo can replace multiple HPLC columns such as reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion- exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters  - Buffer concentration, Buffer pH, and Organic Modifier Concentration  - users can adjust the column properties with pinpoint precision to separate complex mixtures.

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Primesep 100

The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.

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