Agilent 1260 Infinity II 制备型液相色谱

In analytical HPLC good resolution is required over the entire gradient; in preparative HPLC it is only important to achieve good resolution around the target compound, which should be collected in highest purity. With a small set of optimized gradient methods a much better resolution can be achieved than with a single generic method. The generation of a small method set based on retention time windows in the pre-preparative analytical run is described in this Application Note. Using Agilent standards with known elution properties makes this an easy four-step process, which can be accomplished in less than one hour. An application example demonstrating how the resolution of the target compound can be improved is also shown.

Synthesizing novel compounds or isolating natural products can be a laborious and time-consuming process. After analyzing the precious sample on an analytical UHPLC system, the crucial step is to transfer the method to a preparative system with a minimal risk of losing valuable work or collecting impure compounds. This Technical Overview describes a practical way to optimize the scale-up process for reversed-phase chromatography from an analytical UHPLC system to preparative LC systems using a focused gradient to increase the sample load on the preparative column to achieve optimum purity.

This Technical Overview describes a solution to monitor and trigger the collection of preparative samples with the Agilent 1260 Infinity Evaporative Light Scattering Detector (ELSD) using a flow splitter. The Agilent 1260 Infinity ELSD is the detector of choice to monitor low UV-absorbing compounds or compounds with no UV-chromophore. Furthermore, this detection method is able to detect all compounds less volatile than the mobile phase, suitable for the detection of small molecule mixtures. A drug-like sample mixture has been purified on an Agilent 1260 Infinity Preparative-scale Purification System by triggering the fraction collection with the Agilent 1260 Infinity ELSD. 99 % of purity and 91 % recovery were achieved.

The purification of a valuable product or product library after a long and complex synthesis is a very important process. The Agilent Automated Purifi cation Software introduces an Open-Access-based solution for isolating substances, with automated focused gradient to simplify the pre- and post-purification steps. This Technical Overview describes the easy steps to follow, from crude sample analytical scouting submission to pure target product archiving or screening. For example, a purification sample containing a target peak overlapped by an impurity was purifi ed on the Agilent 1260 Infinity Preparative-Scale LC/MS Purifi cation System using the Automated Purifi cation Software. This purifi cation produced a pure target compound with 88 % recovery, saving 40 % of solvent and runtime compared to a standard purifi cation method.

On the Agilent OpenLAB CDS, fraction collection can be performed with a UV detector, or any additional detector, such as an Evaporative Light Scattering Detector (ELSD). In another Technical Overview1 , we described a simple way to split the preparative fl ow on the Agilent preparative LC systems to trigger the collection with an ELSD. This Technical Overview shows how to optimize peak-based fraction collection triggered by an ELSD. We describe the system setup, the delay calibration between detector and fraction collector, and the use of the variable light sensitivity of the Agilent 1260 Infi nity Evaporative Light Scattering Detector. We also discuss the peak-based fraction collection settings required to achieve the best purity and recovery.

Peak-based fraction collection triggered by a UV signal threshold leads to satisfactory results in most cases. Some samples and applications, however, require special attention when fraction collection parameters are set. This Technical Overview describes different strategies on how to achieve best collection results using different settings on an Agilent 1260 Infinity II Preparative LC/MSD System. UV and mass-based fraction collection, fraction pooling, as well as recovery collection are discussed in detail to provide knowledge that can be turned into successful and confident fraction collection.

?This Application Note demonstrates: • Configuration of the Agilent 1200 Series Rapid Resolution LC (RRLC)analytical scale (AS) purification system. • Use of the Agilent 1200 Series RRLC AS purification system with anoptimized method for purification of peptide nucleic acids from solidphase synthesis.

在制备型液相色谱中成功使用大直径色谱柱，需要将高质量填料与合适的色谱柱硬件结合。本文讨论了如何在 Load & Lock 柱上装填 YMC Prep C18 10 μm 填料，以获得稳定的柱床，同时使柱效达到不低于 30000 理论塔板数/米。

近来，水溶性维生素的分析和定量在制药领域受到越来越多的关注。反相梯度（无离子对）分离可替代常规的等度分离，能够分析包含B 族维生素、泛酸、叶酸和维生素C 等物质的8 组分样品。本研究使用 ZORBAX SB-C8 柱进行分析。