Gilson Connect
In the pharmaceutical and biotechnology industries, achieving high purity and high yield when developing treatment candidates is of utmost importance. Any impurities present can interfere chemically and physically with a candidate, leading to inaccurate or unpredictable results, and, at worst, they pose a potential safety concern for patients.
Further complicating matters, the processes involved in drug development are often multi-step. Attrition at each purification stage can lead to a low yield and require candidate resynthesis — which is time-consuming, resource-intensive, and costly.
Liquid chromatography (LC) is the standard method used for purifying products in the discovery stages, as it is cost-effective, relatively simple to perform, and reliable. However, obtaining high purity and yield using this approach often comes at the cost of throughput, as purification runs must be lengthened to achieve greater separation between components. Such a trade-off between purity, yield, and throughput presents a significant challenge for researchers and scientists. How can labs limit this trade-off in their workflows?
This article explores how users can enhance fraction collection efficiency and streamline the purification process by using the latest advances. By doing so, users maintain purity and yield while minimizing re-analysis, ultimately saving valuable time and resources.
When trying to maximize the purity and yield of products, labs often must choose between two approaches to collect fractions in liquid chromatography:
Purity can generally be enhanced by improving the resolution between peaks. However, it means that separation takes a longer time, thereby limiting throughput.
Critical at the pilot or process scale, recovery and throughput are usually enhanced using advanced purification modes. This compromises peak resolution, however.
Such considerations can be summarized by the Purity-Recovery-Throughput triangle (Figure 1).
Figure 1. The relationship between purity, recovery or throughput, and throughput in LC workflows (left), and visualizing the trade-off between the different factors (right).
To strike the right balance between yield and throughput, different collection capabilities can be employed: Level (Threshold), Peak, Apex, or Conditional Peak (Figures 2 and 3), each of which can impact the purity of the final compound.
Figure 2. Demonstration of Level, Peak, and Apex collection.
Figure 3. Demonstration of Conditional Peak collection with a peak composed of three different compounds.
Gilson's TRILUTION LC v4 software— which is used by modular Gilson preparative chromatography systems— provides complete control over the fractionation and destination of chromatography fractions, significantly improving the purification process. The advanced peak collection approach described reduces the need to compromise between recovery, purity, and throughput.
TRILUTION LC software splits the peak into three sections: front, middle, and rear (Figure 4). The different parts of the peak can then be directed into different vessel types, giving you control over the purification process. The most concentrated part of the peak can be collected into a large bottle(s) as a single fraction, reducing the number of fractions to dry down or re-analyze.
To improve yield further, the front and rear of the peak can be sliced and collected into multiple tubes, allowing for further analysis and re-purification if the fraction(s) do not meet the purity requirements. It is worth noting that fraction reprocessing will impact throughput, demonstrating the fine balancing between purity, recovery, and throughput.
Overall, the advanced peak collection method reduces re-analysis time, as most of the product is collected into a single bottle, which enables greater throughput. The capabilities of the approach make it ideal for the final stage of purity enhancement.
Figure 4. TRILUTION LC software’s advanced peak collection method splits the peak into front, middle, and rear.
To optimize your purification work and benefit from advanced fraction collection using TRILUTION LC software, consider the following requirements and capabilities for your LC setup:
To ensure that the collection process is both efficient and targeted, consider installing a dual setup. By doing so, one unit can collect liquid into tubes on the liquid handler, while the other collects the pure product into bottles.
Using mass detection allows for targeted compound identification, thereby guaranteeing that compounds of interest will not be lost. Here, fraction collection can be triggered to segregate pure fractions from lower purity ones. This targeted fractions collection mode reduces the number of fractions to dry down or re-analyze.
If you have specific requirements for your synthetic compound purification, you can customize TRILUTION LC software to set the conditions you need. By tailoring the software to your unique purification requirements, you can ensure your process is optimized for maximum efficiency.
TRILUTION LC is a user-friendly software package designed to control high-pressure LC and automated liquid handling instruments, making it an invaluable tool for researchers and scientists across pharma and biotech labs. The software offers a range of features that enable users to take full control of their purification processes:
Traditionally, improving compound purity and yield has come at the cost of throughput, presenting a significant challenge for researchers and scientists across the pharma and biotechnology industries. However, TRILUTION LC software offers an innovative solution, allowing you to control fractionation and the destination of your fractions to maintain purity and yield while minimizing re-analysis time. Equipped with this latest advance, you can ensure maximum productivity, profitability, and confidence in your final product purification process.
