{"id":3585,"date":"2019-04-03T00:00:00","date_gmt":"2019-04-03T00:00:00","guid":{"rendered":"https:\/\/www.thermofisher.com\/blog\/analyteguru\/hplc-or-uhplc\/"},"modified":"2026-06-11T20:45:36","modified_gmt":"2026-06-11T20:45:36","slug":"hplc-or-uhplc","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/analyteguru\/hplc-or-uhplc\/","title":{"rendered":"HPLC or UHPLC?"},"content":{"rendered":"
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This almost sounds like a line from Shakespeare, but HPLC or high-performance liquid chromatography<\/a> and ultra-high-performance liquid chromatography (UHPLC<\/a>) came a little after the \u2018Bard of Avon.\u2019 Both HPLC and UHPLC are common techniques in the analytical laboratory, but recent years have seen a shift from HPLC to UHPLC. Why is this? What are the differences between the two and how do you transfer a method from HPLC to UHPLC? These are topics I will briefly discuss in this article.<\/p>\n

\"\" <\/span>Differences between HPLC and UHPLC<\/strong><\/h3>\n

These are the top 5 differences between HPLC and UHPLC:<\/p>\n

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  1. Particle sizes \u2013 In HPLC particle sizes of the stationary phase are typically in the order of 3-5 \u00b5m, whilst UHPLC is characterized by particles of 2 \u00b5m or less.<\/li>\n
  2. Column dimensions \u2013 As with particle sizes there is a corresponding reduction in column dimensions with UHPLC. A typical HPLC column has an internal diameter of 4.6 mm and a length of 250 mm, whilst a UHPLC column has internal diameters of 2.1 mm or less and is much shorter, 100 mm for example.<\/li>\n
  3. Flow rates \u2013 UHPLC runs at much lower flow rates than HPLC, for example 0.2 \u2013 0.7 ml\/min against 1-2 ml\/min respectively.<\/li>\n
  4. Backpressure \u2013 With the smaller particles and reduced column diameter then this manifests itself in to higher backpressures in UHPLC compared to HPLC. HPLC instruments typically operate at maximum pressures of 400-600 bar, whilst UHPLC instruments can operate at up to 1500 bar in the case of the Thermo Scientific\u2122 Vanquish\u2122 Horizon UHPLC System<\/a>.<\/li>\n
  5. Detection parameters \u2013 Narrow peaks are produced with UHPLC, requiring a detector that can keep pace and provide the required number of data points per peak for detection. Most modern detectors<\/a>, though, are capable of detection speeds of up to 250 Hz, which is sufficient for both HPLC and UHPLC.<\/li>\n<\/ol>\n

    These are the common differences between HPLC and UHPLC, but they can also be broadly separated by application area. HPLC is commonly used in routine environments, whilst UHPLC is more common in research and development, but this is not exclusive.<\/p>\n

    Why use UHPLC? (And why not?)<\/strong><\/h3>\n

    As alluded to earlier in the article, there has been a gradual shift to UHPLC from HPLC over the last decade. So why is this? There are a number of reasons:<\/p>\n