HPLC Separation of Alizarin and Alizarin Red S on Newcrom BH Column

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Alizarin and Alizarin Red S are both dyes with applications in locating calcium deposits in tissues. They differ in Alizarin Red S having an additional sulfonic acid group. They can be separated in HPLC using a Newcrom BH mixed-mode column using a mobile phase of methanol (MeOH) and water and sulfuric acid (H2SO4) as a buffer. UV detection at 250nm.


Column Newcom BH, 4.6×50 mm, 5 µm, 100A
Mobile Phase MeOH/H2O – 75/25%
Buffer H2SO4 – 0.5%
Flow Rate 1.0 ml/min
Detection UV 250 nm


Class of Compounds Acid, Dyes
Analyzing Compounds Alizarin, Alizarin Red S


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.

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 columns are standard reverse phase columns with octyldecyl silane chains (C18) attached to porous silica. We have also designed a line of Newcrom R1 columns with a new outer design. This column and corresponding adapter entirely eliminate the need for any high-pressure fittings or tubing as well as minimizing all possible dead volumes. Furthermore, if a leak ever occurs in the high-pressure column inlet, the mobile phase is contained within the column adapter (no external leakage).

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Application Analytes:
Alizarin Red S