UHPLC Method Development for Simultaneous Determination of Antihypertensive Combination Agents

Význam tématu

Combination therapy of antihypertensive agents in a single formulation enhances blood pressure control and patient adherence.

Cíle a přehled studie

A robust UHPLC method was developed for rapid simultaneous separation of six antihypertensive drugs using Thermo Fisher Vanquish UHPLC and automated method scouting in Chromeleon CDS 7.2.

Použitá metodika a instrumentace

• UHPLC system: Vanquish System Base, Binary Pump H, Split Sampler HT, Column Compartment H, Diode Array Detector HL.
• Software: Chromeleon CDS 7.2 with Automated Method Scouting and method speed-up tools.
• Column: Accucore Vanquish C18+, 1.5 µm, 2.1×100 mm.
• Mobile phases: A1–water, A2–water/0.1% formic acid, A3–10 mM ammonium acetate pH 4.5; B1–acetonitrile, B2–acetonitrile/0.1% formic acid, B3–methanol.
• Gradient: 5–90% B in 1.25 min; accelerated gradient halved runtime.
• Flow rate: 0.4 mL/min (standard), 0.8 mL/min (accelerated); temperature: 50 °C; injection volume: 1 µL.
• Sample prep: Stock solutions of each API in methanol mixed to standard ratios.

Hlavní výsledky a diskuse

Automated scouting identified the optimal eluent combination (0.1% formic acid in water and acetonitrile) achieving baseline resolution of six compounds within 1.8 min at <700 bar. Doubling flow rate and applying the speed-up tool reduced analysis time by half without loss of resolution. An unknown impurity was detected upon accelerated method. Nanoliter injections (0.01 µL) showed peak area RSD <5% and retention time RSD <0.1%. Calibration across 0.01–1 µL had r² >0.996.

Přínosy a praktické využití metody

The method enables high-throughput screening and quality control with minimal sample and solvent use. Automated scouting and transfer tools simplify method development and ensure reproducible performance.

Budoucí trendy a možnosti využití

Future applications may include coupling to mass spectrometry, adoption of greener solvents, further miniaturization, and integration of AI-driven method development workflows. Expansion to other pharmaceutical combinations and impurity profiling is anticipated.

Závěr

This study demonstrates a fast, reliable UHPLC approach for simultaneous analysis of antihypertensive combinations. The integration of automated scouting and acceleration tools delivers robust separation, high reproducibility, and efficient throughput.

Reference

1. Jones JD, Jackson SH, Agboton C, Martin TS. Pharmacy and Therapeutics. 2011;36(10):634–640.
2. Thermo Fisher Scientific. Vanquish UHPLC System. 2014.
3. Thermo Scientific Technical Note 161: LC Method Development by an Automated Method Scouting System. 2015.
4. Thermo Scientific Technical Note 75: A Universal Tool for Method Transfer From HPLC to UHPLC. 2014.