Fast Separation of Cannabinoids on Mixed-Mode HPLC Column Cannsep B

With legalization of marijuana in much of the United States, more and more attention is paid to developing quick, robust and reliable method for quantitation of cannabinoids in marijuana plants and related edible products. The main challenge lies in sample preparation and quantitative analysis of four major cannabinoids: cannabidiol, cannabinol, tetrahydrocannabinol and tetrahydrocannabinolic acid. All compounds are hydrophobic with only THCA (THC-A) and cannabidiolic acid (CBDA) being an acid. Hydrophobic interaction is the main interaction in the separation of cannabinoids on reverse phase columns. On our mixed mode columns acidic molecules retained by ion-exchange mechanism  in addition to hydrophobic. Method can be used for analysis and prep separation of cannabinoids in marijuana plants, seeds and other cannabis-based products. Extraction of cannabinoids is the main procedure in sample preparation. Extraction can be done with organic solvents like chloroform, methanol, ethanol, acetonitrile.  Cannabinoids can be monitored by UV and LC/MS. Mixed-mode columns containing polar embedded groups provide different selectivity than regular reversed-phase columns.



Column Cannsep B, 4.6×150 mm, 5 µm, 100A
Mobile Phase Gradient MeCN 40-60% , 20 min
Buffer H2SO4 – 0.1%
Flow Rate 1.0 ml/min
Detection UV, 210 nm



Class of Compounds
Drug,  Hydrophilic, Ionizable, Supplements
Analyzing Compounds Cannabidivarin (CBDV), Tetrahydrocannabivarin (THCV), Cannabidiol (CBD) Delta-9-Tetrahydrocannabinol (THC), Cannabigerol (CBG), Cannabichromene (CBC), Tetrahydrocannabinolic acid (THCA-A)


Application Column

Cannsep B

The Cannsep family of columns is a specially developed line of reverse-phase columns designed to retain and separate most of the compounds responsible for cannabis's physiological properties. With three options available - Cannsep A, Cannsep B, and Cannsep C - customers can resolve all the various cannabinoids, each providing unique and significantly orthogonal selectivity.

Select options
Application Analytes:
Cannabichromene (CBC)
Cannabidiol (CBD)
Cannabidivarin (CBDV)
Cannabigerol (CBG)
Cannabinol (CBN)
Tetrahydrocannabinol (THC)
Tetrahydrocannabinolic acid (THCA-A)

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 so we can ensure you get optimal results for your compound/s of interest.