Easy Method Transfer from UHPLC to HPLC

Introduction

The scalability of the 1.7, 2.6, and 5μm Kinetex® core-shell particles is illustrated in this study. This flexibility makes Kinetex the only core-shell particles in the market that are optimized for different system platforms and are transferable from UHPLC systems to conventional HPLC systems to preparative applications.

TN-1135 APPLICATIONS

With the performance advancements in Ultra High Performance Liquid Chromatography (UHPLC) and High Performance Liquid Chromatography (HPLC) during the last decade, the differences in performance between various instruments has become more evident and new requirements have been identified. The Kinetex 1.7 μm, 2.6 μm and 5 μm core-shell particles are fully scalable across the particle size range by precisely maintaining their core-to-shell ratio. Each particle size in the Kinetex core-shell family is specifically engineered and optimized for UHPLC and/or conventional HPLC platforms.The sub-2μm core-shell particles, designed to be used for the UHPLC systems, are capable of delivering extremely high resolving power that exceeds the performance of fully porous sub-2μm silica sorbents. The 2.6μm core-shell particles are designed as a bridge between the UHPLC and HPLC platforms, and are able to achieve UHPLC level performance on optimized conventional HPLC systems. The newly introduced 5μm Kinetex core-shell media is particularly useful as a “plug-and-play” replacement that delivers immediate improvement for existing separation methods that employ 3μm and 5μm fully porous sorbents. Kinetex 5 μm core-shell particles are also available in the award-winning AXIATM preparative column format and is the first core-shell sorbent available on the market for small-scale preparative applications. Therefore, the family of Kinetex coreshell sorbents, available in various bonded phases, particle sizes, and column formats, is designed to be a complete, effective and scalable set of solutions for any separation challenge, on any system platform.

Material and Methods

Columns

1. Kinetex 1.7 μm C18, 50 x 2.1 mm
2. Kinetex 2.6 μm C18, 50 x 2.1 mm
3. Kinetex 5μm C18, 50 x 2.1 mm

Sample Preparation:
The sample of steroid mixtures is made neat in MeOH at 500 μg/mL for each analyte. Components: (1) Estriol (2) Hydrocortisone (3) Corticosterone (4) Cortisone Acetate (5) β-estradiol (6) 21-Hydroxyprogesterone (7) Estrone (8) Deoxycorticosterone (9) Progesterone. HPLC Conditions:
The mobile phase consisted of deionized water (mobile phase A) and acetonitrile (mobile phase B) in gradient conditions (see Table 1). The method is run at 30 °C and the detection is done using a UV detector at 220 nm. Volume of 0.5 μL prepared sample is injected.

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