Emerging Analytical Separation Techniques with High Throughput Potential for Pharmaceutical Analysis, Part I: Stationary Phase and Instrumental Developments in LC

Abstract

In recent years, a trend of change has been observed within pharmaceutical industry. As modern drug discovery has reached a remarkable level of complexity and drugs need to be discovered, developed and produced against strict timelines and within cost- and regulatory constraints, industry seeks "lean" solutions to increase productivity. Among them, increasing the sample throughput of the ever-growing number of necessary (routine) analyses has become a popular target to cut precious time. For the last thirty years, High-Performance Liquid Chromatography (HPLC) has been the leading technology when it comes to various analyses in pharmaceutical industry; however, its necessity of serial analyses taking typically 10-45 min has been a sample throughput-limiting barrier. Lately, the fundamentals of HPLC have been exploited to raise new technologies that can speed up analyses to ground breaking limits, without compromising separation efficiency. This paper reviews some promising technologies, i.e. totally porous sub-2microm particles accompanied by pressures up to 1000 bar (Ultra-Performance Liquid Chromatography or UPLC), fused-core particle technology, monolithic supports and High Temperature Liquid Chromatography (HTLC), having the potential to take LC to the next level in pharmaceutical industry. As each analytical method has its own demands, the advances of the above technologies are discussed for different applications in pharmaceutical analysis where high-throughput analysis can be meaningful, i.e. in a drug discovery and development setting, and in quality operations. Both chemical and biological pharmaceuticals are considered. We discuss the perspectives of these technologies and their realizations up to now in high-throughput pharmaceutical analysis.

The role of liquid chromatography–tandem mass spectrometry in the clinical laboratory

“…Chromatographic run-time can be shortened by using ultra-high-performance liquid chromatography (UHPLC) coupled to MS. UHPLC is a variant HPLC technique that allows the system to handle the high backpressure resulting from the stationary phase with sub-2 m particles, offering advantages in chromatographic resolution, speed, and sensitivity over conventional HPLC systems [77]. An alternative approach to alleviate the restrictions on LC-MS/MS throughput, is the use of a multiplexed LC system, in which multiple separate LC-systems operate simultaneously, but in a staggered fashion, allowing analytes to enter the MS only at the time when the peak of interest elutes, thereby increasing throughput up to four-fold when compared to a single LC system [49,78,79].…”

Proteomics and Non-proteomics Approaches to Study Stable and Transient Protein-Protein Interactions

“…However, since the introduction of nano-LC-MS/MS, sensitivity has improved, but the chromatographic time still require improvements. Nano-LC can achieve narrow peak widths and high reproducibility of the separation of complex peptide samples [159][160][161] and a reduction of the time required for analysis will help make LC the pre-fractionation method of choice over gel electrophoresis.…”

The proteomic future: where mass spectrometry should be taking us

“…A progressive shift from ion spray (millilitre flow rates) to capillary (microlitre flow rates) to nanospray (nanolitre flow rates) chromatography has been afoot since the advent of proteomics. Higher-pressure lower-flow chromatography for online nLC (nano-LC)-MS/MS [sometimes referred to as UPLC (ultra-performance liquid chromatography)] improves sensitivity and also reduces chromatography time by reducing peak width, particularly for targeted proteomics [24,25]. Nanoflow LC has the advantage of near-zero dead volume coupled to ultra-narrow peak widths (5 s) and is capable of reproducible separation of complex peptide samples in the 10-100 ng range.…”