What are some method development starting points for Atlantis PREMIER BEH C18 AX mixed-mode, reverse-phase anion exchange columns? - WKB108534

ENVIRONMENT

• High Performance Liquid Chromatography (HPLC)
• Ultra High Performance Liquid Chromatography (UHPLC)
• High Performance Liquid Chromatography (HPLC)
• Method development
• Atlantis PREMIER BEH C18 AX mixed-mode, reverse-phase, anion exchange columns
• Polar compound retention
• VanGuard

ANSWER

Methods Development  - Good places to Start

• Buffer Concentration
  • Ammonium formate is a good choice for pH 2.7 – 4.7 and ammonium acetate for pH 3.8 – 5.8 (both are MS compatible)
  • Start with 10 – 20 mM buffer
    • Lower Concentration
      • Retention increases for anions, decreases for cations
      • Tailing for anions increases
    • Higher Concentration
      • Retention decreases for anions, increases for cations
      • Improved peak shape for anions
• Buffer pH
  • Starting at low pH (3 - 4) tends to give sufficient retention for acids with the best peak shapes
  • At higher pH (>6) the surface charge of the stationary phase gradually decreases
    • Lower retention for anions, higher retention for cations

• Organic Content
  • Increase the % organic for hydrophobic analytes

ADDITIONAL INFORMATION

Atlantis Columns Web Page

Atlantis PREMIER Columns Infographic

Atlantis Premier BEH C18 AX Columns Care and Use Manual