HPLC Method for Separating Corticosteroids such as Prednisone, Prednisolone and Methylprednisolone on Primesep B Column

HPLC Method for Prednisone, Prednisolone, Methylprednisolone on Primesep B by SIELC Technologies

HPLC method for separating corticosteroids such as Prednisone, Prednisolone, and Methylprednisolone on Primesep B Column
HPLC Method for Analysis of Prednisone, Prednisolone, Methylprednisolone on Primesep B Column by SIELC Technologies

 High Performance Liquid Chromatography (HPLC) Method for Analysis of Prednisone, Prednisolone, Methylprednisolone

All three compounds have potent anti-inflammatory effects, making them valuable in treating conditions where inflammation plays a role.


  • Mechanism of Action: Prednisone is a synthetic corticosteroid that mimics the action of cortisol, a natural steroid produced by the adrenal glands. It primarily exerts its effects by binding to glucocorticoid receptors and regulating gene expression.
  • Medical Uses: It is commonly used to treat inflammatory conditions, autoimmune disorders, allergic reactions, and certain cancers.


  • Derived Form: Prednisolone is the active form of prednisone. When prednisone is ingested, the liver converts it into prednisolone.
  • Mechanism of Action: Similar to prednisone, it has anti-inflammatory and immunosuppressive properties.


  • Potency: Methylprednisolone is considered to be more potent than prednisone.
  • Medical Uses: It is used in various conditions, including severe allergic reactions, inflammatory conditions, and as a part of immunosuppressive regimens.

Corticosteroids can be retained, separated, and analyzed using a Primesep B mixed-mode stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water, acetonitrile (MeCN), and sulfuric acid as a buffer. Detection is achieved using UV at 245 nm

ColumnPrimesep B, 4.6 x 150 mm, 5 µm, 100 A
Mobile PhaseMeCN/H2O – 35/35%
BufferH3PO4 -0.1%
Flow Rate1.0 ml/min
DetectionUV 245 nm

Class of Compounds
Analyzing CompoundsPrednisone, Prednisolone, Methylprednisolone

Application Column

Primesep B

Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A

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Application Analytes:

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.