Developing a Separation for Eleven Boronic Acids Using Maxpeak™ Premier Column Technology on an Arc™ HPLC System

Significance of the Topic

The ability to separate and quantify boronic acids by HPLC is critical for synthetic chemistry workflows, including Suzuki–Miyaura coupling and diol tagging. Reliable analysis supports reaction monitoring, quality control, and compound library development.

Objectives and Overview

This study aimed to establish a robust HPLC method for simultaneous separation of eleven structurally related boronic acids. A systematic screening protocol was applied on an Arc HPLC System using MaxPeak Premier column technology to achieve baseline resolution in a single run.

Methodology and Instrumentation

A tiered screening approach comprised two main stages:

1. pH Assessment – Initial evaluation of retention at low pH (2% formic acid) and high pH (200 mM ammonium hydroxide) on a C18 stationary phase to gauge analyte ionization effects.
2. Stationary Phase Screening – Comparison of five MaxPeak Premier columns (BEH C18, BEH Phenyl, BEH C8, Premier CSH C18, Premier HSS T3) under both methanol and acetonitrile as organic modifiers.

Gradient conditions maintained 5% buffer throughout with a linear increase to 95% organic over 16.4 minutes, followed by re-equilibration. Data acquisition was performed with Empower 3 software.

Used Instrumentation

• Arc HPLC System with single column heater
• 2998 Photodiode Array Detector (UV detection at 254 nm)
• Empower 3 Chromatography Data System
• Columns evaluated (4.6 × 100 mm, 3.5 μm): XBridge Premier BEH C18, BEH Phenyl, BEH C8; XSelect Premier CSH C18; XSelect Premier HSS T3

Main Results and Discussion

Low-pH conditions generally provided stronger retention than high pH, where partial deprotonation reduced analyte retention. Stationary phase screening revealed distinct selectivity patterns:

• Phenyl and C8 phases exhibited elution order shifts for certain isomers due to π–π and hydrophobic interactions.
• CSH C18 showed unique ionic interactions, altering retention of amine-containing analytes.
• HSS T3 delivered the most promising separation for all eleven boronic acids under acetonitrile conditions despite limited early retention.

Gradient optimization by starting at 100% aqueous and preserving slope increased retention factors (15-fold for early eluters) and improved USP resolution from 2.09 to 3.43 for the first peaks, achieving baseline resolution in under 11 minutes.

Benefits and Practical Applications

• Baseline separation of eleven boronic acids in a single sub-11-minute run.
• Systematic screening reduces development time and empirical guesswork.
• MaxPeak Premier HPS technology prevents non-specific adsorption, ensuring reproducible chromatography.
• Method is directly applicable to reaction monitoring, QA/QC, and compound tagging workflows.

Future Trends and Opportunities

Continued integration of hybrid column surfaces and advanced screening algorithms will further accelerate method development. Expansion to UHPLC platforms, coupling with MS detection, and AI-driven optimization promise even faster, more selective analyses of boronic acids and related chemistries.

Conclusion

A streamlined, reliable HPLC method for eleven boronic acids was established using a systematic screening protocol on an Arc HPLC System with MaxPeak Premier columns. The final HSS T3–based method delivers rapid, baseline resolution and is highly suitable for diverse analytical applications.

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