Preparative HPLC Systems

What is preparative HPLC?

Preparative high-performance liquid chromatography (prep HPLC) is a technique used to isolate and purify compounds from complex mixtures.
When discussing HPLC, there are three approaches to consider:

• Semi-preparative HPLC
• Preparative HPLC
• Analytical HPLC

The analytical, semi-prep and prep HPLC principles rely on interactions between compounds, the stationary phase and the mobile phase, but there are a couple of key differences specifically between analytical and prep HPLC:

• As a scaled-up version of analytical HPLC, prep HPLC relies on instruments with larger columns, higher flow rates and larger sample injections.
• Unlike analytical HPLC, prep HPLC can purify and collect purified compounds for further use following identification for evaluation, analysis and processing in the shortest possible time.
• Prep HPLC is a highly precise analytical process that can handle a wide variety of sample volumes and high sample concentrations for greater precision in analysis.
• The system setup of prep HPLC often includes automated loading, fraction collection and wide-bore columns.

 

How does prep HPLC work?

HPLC is used to separate and refine high-purity target compounds from a mixed solution after a synthesis reaction or from natural extracts. A SFC system uses carbon dioxide as the mobile phase and shows several advantages over traditional HPLC systems:

• Shorter analysis time
• Almost 70% reduction in mobile phase organic solvent consumption
• No need for evaporation of collected fractions
• Improved isomer separation capabilities

A preparative system (HPLC or SFC) is used to separate and collect high-purity compounds and in some cases, large quantities of compounds required for subsequent evaluation, analysis, and processing in the shortest possible time.

Shimadzu preparative HPLC systems

Shimadzu has been a world leading manufacturer of high-quality preparative HPLC systems that meet the application requirements of a broad range of customers.
Our portfolio includes:

• Nexera Prep (HPLC Preparative System)
• Nexera UC Prep (SFC Preparative System)
• UFPLC (Ultra-Fast Preparative and Purification Liquid Chromatography System) – a fully automated system capable of performing preparation to recovery for LC applications.

We also provide related instruments:

• LH-40 liquid handler
• FRC-40 fraction collector
• Flow Splitters for preparative LC-MS analysis

Applications

Shimadzu’s suite of instruments geared towards prep HPLC – the Nexera UC Prep and Nexera Prep range – offers shorter analysis time, an almost 70% reduction in mobile phase organic solvent consumption, no need for evaporation of collected fractions, and improved isomer separation capabilities. And, for their ability to isolate and purify target compounds from complex mixtures, these instruments are utilized across numerous industries, from research and manufacturing to quality-control environments.
You’ll find these prep HPLC instruments used in the following applications:

• Pharmaceuticals: For active pharmaceutical ingredient (API) purification, impurity removal and drug formulation.
• Research and development: For isolating natural compounds and the purification of synthetic targets.
• Biotechnology and life sciences: For peptide/protein purification and DNA/RNA isolation.
• Food, beverage and cosmetics: For the extraction of flavors and fragrances, as well as quality testing.
• Chemical and industrial applications: For fine chemical purification and pilot-scale production.

 

FAQs

What is prep HPLC used for?

Preparative high-performance liquid chromatography (prep HPLC) is used to isolate and purify compounds from complex mixtures. It is ideal for downstream use in pharmaceuticals, biotech and a variety of broader scientific industry applications for research, quality control and manufacturing.

What is the difference between prep HPLC and analytical HPLC?

The difference between prep HPLC and analytical HPLC lies in the goals of each process. Analytical HPLC focuses on only qualifying and quantifying components in a sample mixture, whereas prep HPLC is able to purify and then collect purified compounds for further evaluation, analysis and processing. Prep HPLC relies on instruments with larger columns, higher flow rates and larger sample injections than those required for analytical HPLC, with system setups prioritising automated loading, fraction collection and wide-bore columns.

What to do before HPLC

Before running HPLC analyses, you should prepare the sample for testing, ensuring you’ve diluted it to a suitable concentration for this technique and confirmed your sample is compatible with the mobile phase. Don’t disregard the importance of sample filtration to remove particles that could clog the column. You need to fill the solvent reservoirs with the mobile phase, set the pump to align with the ideal flow rate and prepare the injector to introduce the sample to the mobile phase stream. Then, set the column to the temperature that ensures optimal separation for your sample and check your flow rate before running the analysis.

What is the difference between flash and prep HPLC?

Flash and prep HPLC are liquid chromatography approaches that achieve purification during the analysis. Flash prioritizes speed and efficiency without compromising on accuracy. While prep HPLC may be a little slower with lower loading capacity, it is highly efficient at providing very high-purity compounds. Further, prep HPLC is ideal for purifying complex mixtures. Flash is frequently used in pharmaceutical research, as is prep HPLC, joining biopharma and chemical industry applications.