Sepbox

2D HPLC/SPE System for Natural Product Isolation

Nature has a practically infinite variety of useful compounds.....

Throughout history, valuable therapeutic ingredients have always been derived from plants. One well-known example is the willow tree, whose bark was used by the famous Greek physician Hippocrates to treat inflammatory diseases around 400 B.C. The effective natural ingredient extracted from this bark has been developed by scientists to make acetylsalicylic acid, which forms the basis of many current pain relievers. Today, we use a substance extracted from a nut tree in the Philippines to treat epilepsy. And Curare, a neurotoxic substance derived from lianas vines in the Amazon jungle, is used to relax muscles and treat tetanus.

Natural compounds still play an increasingly important role in many areas of research and development. Their structural diversity is enhanced by the synthetic potential of combinatorial chemistry. There are many new classes of substances waiting to be discovered, and the diversity of these compounds and their chiral properties means that some of these are certain to be highly marketable. Pharmaceutical research is only just beginning to exploit this diversity.

... unfortunately, they are never found in their pure state

Access to nature’s inexhaustible supply of compounds has increased over recent years. However, unfortunately there isn’t the same limitless supply of ways to process them automatically. Fractionation and separation of samples obtained from nature remain time-consuming, tedious and extremely expensive, even though the assays for testing these samples have become faster and more cost-effective thanks to advanced high-throughput screening processes. Today, high performance Liquid Chromatography (HPLC) is the standard technology used for separating compounds from natural resources.

Isolate Conventionally

That’s why we need state-of-the art solutions...

Processing natural product extracts represents a major bottleneck that is constricting the pace of drug discovery. To ensure that sample preparation is both reproducible and reliable, it would therefore be of immense benefit to have an automated process in place for natural compound isolation and purification; the economic potential of this is tremendous.

...like Sepbox systems for natural product isolation

Sepiatec’s unique Sepbox systems allow natural product extracts to be processed automatically, and significantly accelerate the purification process. Results which usually take weeks or months with conventional methods can be obtained in just a few days – or even one day – with a Sepbox system.

The Sepbox concept is based on a patented combination of high performance Liquid Chromatography (HPLC) and solid phase extraction (SPE) – reliable and proven techniques that provide a universal platform suitable for processing large sample numbers. Up to 600 fractions with a very high yield of pure compounds can be efficiently obtained from subsequent high throughput screening. Using two-dimensional separation, the recovery rate for both polar and non-polar substances is usually above 90 per cent. Using an automated and highly reproducible process, one extract can be completely separated per day. The pure individual components are soluted in suitable solvents and can be collected in microtiter plates or vials.

isolate sepbox

Extensive lab facilities are no longer essential to separate a large number of novel compounds. With a Sepbox system, up to 600 fractions can be produced per day, many of them containing pure compounds, without investing large amounts in human resources or research facilities. This saves time and money and significantly accelerates the early stages of the product development process.

The automated Sepbox 2d-250, 2D-2000 and 2d-5000 systems allow extracts from plants, marine organisms, microbiological or animal materials to be processed in quantities of up to 250, 2,000 or 5,000 milligrams respectively. If required, highly polar substances can be separated using the polar setup. The systems are equipped with two detectors: UV detection (ultraviolet) and optional ELSD detection (evaporative light scattering detection), which can detect almost all substances.

Advantages

  • Isolation of compounds with a speed and accuracy unachievable with conventional chromatography methods
  • On average 20 to 30 per cent pure fractions (ELSD) per run Isolation of large quantities possible
  • Reproducible separations
  • Cost advantages due to automation

Purity of Sepbox fractions 

On average after one Sepbox run, up to 20 per cent of all fractions from plant extracts are 90 to 99 per cent pure (ELSD*), and from extracts made of marine organisms, up to 30 per cent are 90 to 99 per cent pure (ELSD). As well as these highly purified compounds, the user will usually find a large number of fractions with compounds whose purity levels are between 50 and 90 per cent in ELSD detection. Results vary depending on the extract and the examined organism.

* The purity levels quoted here were calculated using the 100 per cent method with ELSD detection, as UV detection does not provide reliable purity levels due to different absorption coefficients.

Sepbox 2000

The Sepbox 2D-250 benchtop model is designed for the automated two-dimensional separation of up to 250 milligrams of extracts. This system offers the choice between liquid and adsorbed (solid) sample injection. Up to 576 fractions can be collected in microtiter plates in just 24 hours. Depending on the extract, up to 30 per cent of all fractions usually contain pure compounds. The Sepbox 250 is suitable for medium to non-polar compounds (polar setup optional).

With the Sepbox 2D-2000 benchtop system up to 2,000 milligram of extract can be processed. If required, highly polar substances can be separated using the Polar Setup. The Sepbox 2D-2000 is equipped with two detectors: UV detection (ultraviolet) and optional ELSD detection (evaporative light scattering detection), which can detect almost all substances. Due to the automated and highly reproducible process, one extract can be completely separated per day using two-dimensional separation. The recovery rate for both polar and non-polar substances is usually above 90 per cent. Up to 576 fractions with a very high yield of pure compounds can be collected in vials for subsequent High Throughput Screening.

The Sepbox 2D-5000 is designed for a large workload of up to 5,000 milligrams of extract made from plants, marine organisms, microbiological or animal material. The automated 2D chromatography provides 576 fractions in 24 hours. The fraction collector is equipped with 12 removable racks, each containing 48 vials with a maximum capacity of 60 milliliters. The Sepbox 2D-5000 is also equipped with software for method development, process control, data acquisition and data analysis. The Sepbox 2D-5000 provides many individual chemical entities in quantities which can be easily used to elucidate structures, screen different targets and build a diverse library.

Sepbox models
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