Learn how to select, store, and handle HPLC-grade water to improve chromatographic performance. Discover best practices that ensure cleaner baselines, longer column life, and more reliable HPLC results.
HPLC water is not just “clean” water; it is specifically purified to meet the stringent demands of chromatographic separation. It is one of the most vital pillars of RP (reverse phase HPLC). Its purity, storage, and handling directly affect the reliability, reproducibility, and sensitivity of your results.
In this post, we’ll explore how to select the right HPLC water, how to store it properly, and why its quality matters so much in chromatographic performance.
HPLC Water is used as a primary solvent in reversed-phase HPLC, the most common chromatographic technique. However, water from typical laboratory sources-such as distilled or deionised water, is often not pure enough for HPLC. Impurities like organic compounds, particulate matter, dissolved gases, or metal ions can lead to:
| Contaminant Type | Potential Issues |
| Particulates | Clogged columns or filters |
| Organic compounds | Elevated baseline noise |
| Ions and metals | Peak distortion or suppression |
| Microorganisms | Biofilm formation, especially in aqueous buffers |
HPLC-grade water is usually classified by its low levels of:
When buying or producing HPLC-grade water, ensure it is:
Some labs use on-site purification systems, while others prefer prepackaged water from reputable manufacturers. Whichever route you take, consistency and traceability are key.
Even the purest water can become a liability if stored improperly. Water is a universal solvent and can easily absorb atmospheric gases, dust, and microbial contaminants.
Impurities in water can contribute to:
Even seemingly minor contaminants can have a disproportionate effect when analysing trace levels of analytes or working with highly sensitive detectors.
Investing in high-quality HPLC water and following strict storage protocols isn’t just good practice’s essential. The cleaner your water, the cleaner your data.
Whether you’re troubleshooting a noisy baseline or setting up a new HPLC system, start with the foundation: your solvents. Pure water, properly handled, is one of the simplest yet most effective ways to ensure consistent, high-quality chromatographic performance.
Related:
HPLC-grade water is ultrapure water specifically purified for use in high-performance liquid chromatography. It has very low levels of total organic carbon (TOC), ions, particulates, and UV-absorbing impurities to ensure minimal interference with chromatographic analysis
Generally, no. Deionised or distilled water may still contain organic impurities, particulates, or dissolved gases that can affect HPLC performance. Only use ultrapure, Type I water that meets HPLC-grade specifications
Store HPLC water in clean, chemically inert containers (glass or high-purity plastic) in a cool, dark environment. Use within a few days to avoid contamination, and avoid frequent opening of large storage containers.
Impurities in water—like organic molecules, ions, or particulates—can cause baseline noise, ghost peaks, and retention time shifts. These issues compromise sensitivity and reproducibility in chromatographic runs.
Yes. LC-MS often requires even higher purity water, typically referred to as LC-MS grade, which minimises background noise and ion suppression due to trace contaminants.
Yes, if you have an appropriate ultrapure water system (e.g., Milli-Q, PURELAB). Ensure the system is properly maintained and routinely tested for TOC, resistivity, and bacterial contamination.
Pure HPLC-grade water typically has a resistivity of ≥18.2 MΩ·cm at 25°C, indicating a very low concentration of ionic impurities.
No. Mobile phases can degrade or become contaminated over time. Always prepare fresh mobile phase solutions, especially when water is involved, to ensure consistent and reliable chromatographic performance.
Signs of water-related issues include:
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