PerkinElmer
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Application Brief

Optimizing the Separation of Four Tocopherols and Four Tocotrienols via RP-HPLC

Introduction

The knowledge about tocochromanol-related compounds, especially tocotrienols, tocodienols,tocomonoenols, and others, is still limited due to several challenges faced in analytical chemistry. These challenges include separation resolution, co-elution, price and absence of standards, and low analyte concentration in plant material. Application of different column stationary phase chemistries can assist in the challenges faced in compound separation.

Superficially porous particle (SPP) columns can also be used to improve separation of these compounds. SPP particles are made of a solid, non-porous core surrounded by a shell of a porous silica material, resulting in a shorter diffusion path in comparison with fully porous based columns. With a shorter diffusion path within the SPP particle itself, coupled with a uniform packed bed and ultra-inert silica surface, reductions in run times can be observed. Such phases benefit from increased efficiency, with separation resembling that of a UHPLC column. They can be used on standard HPLC instrumentation, without concerns regarding high backpressures, which often compromise column longevity.

In this application brief, five different Quasar SPP column phases were screened for the separation of four tocopherol and four tocotrienol homologs, with focus on resolving β and γ isomers.