This is one from a set of two application notes. In this application note, we determine the purity of collection of a small (0.2%) impurity in a concentrated sample mix by mass-directed fraction collection using supercritical fluid chromatography (SFC) coupled with the Advion expression Compact Mass Spectrometer (CMS).
For the other application note on the recovery and purification of two, three component standard mixtures, click here.
Advion co-authored with a major pharmaceutical partner to create two application notes highlighting the benefits of supercritical fluid chromatography (SFC) coupled to a compact mass spectrometer for mass directed fraction collection. These application notes conclude that mass‐directed fraction collection with a simplified passive splitter on the Preparative JASCO SFC coupled with expression CMS system increases purification selectivity by mass directed fraction collection while maintaining excellent recovery and purity. The MS accepted high concentrations of sample material with peak shapes trailing edges almost indistinguishable from the UV trace. Low, Middle and High isocratic flow rates all showed comparable MS source clearance leading to minimal tailing beyond the UV. Gradients further minimized peak tailing as expected on the UV and this was also seen on the TIC and XIC. Peak purities for all main band purifications were ≥99.9% pure and minor impurity collections showed equivalent success yielding 100% pure fractions. Acceptable recoveries for all collected fractions were 94% or better.
Merck & Co.
Abstract
The evolution of supercritical fluid chromatography (SFC) instrumentation, improved detection capability, and expanded modifier range has led to extending the reach of SFC to the analysis of a broader spectrum of analytes beyond enantioselective separations. However, preparative SFC has yet to see the same technological revitalization, especially in regards to the purification of highly polar analytes. Enhanced fluidity liquid chromatography (EFLC) has been demonstrated as one of the ways to extend the applicable range of SFC instrumentation to highly polar analytes such as proteins, carbohydrates, and nucleotides. Despite recent applications of EFLC for challenging mixtures of hydrophilic metabolites and analogs, its viability in preparative purification, which is of great importance to the pharmaceutical industry, remains unknown. Herein, multiple chromatographic parameters that are critical to achieve feasible EFLC purification methods were investigated, including system pressure as a function of modifier composition (for several MeOH:H2O ratios), effect of diluent injection conditions on peak shape, and optimization of mass load with diluent composition. The usage of 50% acetonitrile or methanol diluents provided the most volumetric loading capacity. In the case of sucrose, leveraging higher analyte solubility in water proved to be more favorable than the volumetric loading capacity of diluents with higher organic content. In fact, an 80 mg injection of sucrose was possible on a 2 cm preparative HILIC column with minimal peak shape degradation. The combined information led to the successful demonstration of EFLC for the preparative separation of sugars using readily available MS-directed SFC instrumentation.
Quant Express, an addition to Advion’s user-friendly Mass Express 4.0 software suite, features quantitation to create a fully integrated software system for expression CMS users.
With decades of mass spectrometry and chemistry expertise, Advion Interchim Scientific has produced a family of compact mass spectrometers designed for the chemist. The affordability, small size and ease of use make them ideal for use directly at the chemist’s bench, giving immediate answers and informed decisions instead of waiting.
The Advion Interchim Scientific expression® compact mass spectrometer (CMS) is a fast and easy analytical tool for the organic chemist. Ideal for fast reaction monitoring, the expression® CMS features a single quadrupole that can adapt to multiple ionization sources in seconds, including both ESI and APCI. The mass spectrometer works in a variety of applications, including food and beverage, pharmaceutical, biomedical, peptides and proteins, drug discovery and more. The expression® CMS offers a variety of novel sampling techniques, including:
- Direct mass analysis of TLC plates in 30 seconds at the push of a button with Plate Express™
- One-touch analysis of solids and liquid samples with the ASAP® probe
- LC/CMS
- Many more options and configurations to suit your needs
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This is one from a set of two application notes. In this application note, we review recovery and purity determination of two, three component standard mixtures by supercritical fluid chromatography (SFC) couped with the Advion expression Compact Mass Spectrometer (CMS).
For the other application note on the fraction collection of a 0.2% impurity, click here.
Advion co-authored with a major pharmaceutical partner to create two application notes highlighting the benefits of supercritical fluid chromatography (SFC) coupled to a compact mass spectrometer for mass directed fraction collection. These application notes conclude that mass‐directed fraction collection with a simplified passive splitter on the Preparative JASCO SFC coupled with expression CMS system increases purification selectivity by mass directed fraction collection while maintaining excellent recovery and purity. The MS accepted high concentrations of sample material with peak shapes trailing edges almost indistinguishable from the UV trace. Low, Middle and High isocratic flow rates all showed comparable MS source clearance leading to minimal tailing beyond the UV. Gradients further minimized peak tailing as expected on the UV and this was also seen on the TIC and XIC. Peak purities for all main band purifications were ≥99.9% pure and minor impurity collections showed equivalent success yielding 100% pure fractions. Acceptable recoveries for all collected fractions were 94% or better.
The expression CMS was developed with the chemist’s objectives and laboratory workflow in mind. Organic, Natural Product and Peptide chemists utilize many techniques to determine the progress, yield and purity of their reactions.
Supercritical fluid chromatography (SFC) is a fast growing technique. Initially used almost exclusively for only thermally labile and chiral compounds it is now becoming accepted as a versatile analytical technique applied across a broad range of compounds. It is faster and higher-throughput than regular HPLC and Preparative LC and becoming popular as a ‘green’ technique, with far lower solvent consumption and lower consumable cost than HPLC. SFC is also compatible with a wide range of detection methods including electrospray (ESI) and atmospheric pressure chemical ionization (APCI) mass spectrometry.
Download this product note to see how SFC combined with compact mass spectrometry (CMS) offers chemists several benefits.
In this study we evaluated the performance of a Preparative SFC‐MS system combining the JASCO Preparative SFC with the Advion expression® CMS (Compact Mass Spectrometer) utilizing a simplified passive splitter for mass‐directed fraction collection. The initial test explored a wide range of flow rates both isocratic and gradient to test performance and reliability of the passive splitter, coinciding timing of MS and UV signal and the clearance of the source with highly concentrated samples.
