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๐Ÿงซ Mobile Phase Preparation

Calculate exact volumes of each component for HPLC mobile phase preparation. Add up to 4 components, set the ratio and total volume.

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Preparation Instructions
๐Ÿ’ก Preparation tip: Always prepare mobile phase in a clean volumetric flask. Mix thoroughly before use. Filter through 0.45 ฮผm membrane and degas by sonication or vacuum. For buffer-containing mobile phases, filter the buffer before adding the organic modifier.
โš ๏ธ Verify before use: Volumes assume simple volume-ratio mixing (v/v). Double-check critical mobile phase preparations independently, especially for validated methods.
๐Ÿ“– Example: Mobile phase 65% acetonitrile / 35% water, total 500 mL โ†’ measure 325 mL acetonitrile + 175 mL water. Mix and degas. Report: "Mobile phase: 65:35 (v/v) acetonitrile:water, prepared and degassed prior to use."
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Up to 4 components

Add up to 4 mobile phase components โ€” organic, aqueous, buffer and modifier.

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Ready-to-use instructions

Copy the preparation instructions directly into your lab notebook or report.

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No sign up, no limits. Use for every HPLC mobile phase preparation.

โœ” Formulas last verified against primary references: March 2025  ยท  Report an error
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Preparing and optimising HPLC mobile phases

Mobile phase composition controls HPLC retention, selectivity, and peak shape. Add organic solvent first to prevent salt precipitation, then add aqueous buffer. Degas by sonication or helium sparging. Higher organic content reduces retention in reversed-phase HPLC. Buffer pH controls ionisation of acidic and basic analytes. For LC-MS, use volatile buffers only: ammonium formate or ammonium acetate (0.1–10 mM). Avoid phosphate and borate buffers which damage mass spectrometer sources.