News from LabRulezLCMS Library - Week 35, 2024

Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 26th August 2024? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!

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This week we bring to you posters from ASMS 2024 and 52nd HPLC by Agilent Technologies, Shimadzu, Thermo Fisher Scientific, and Waters Corporation!

1. Agilent Technologies: Enhancing High Throughput Preparative HPLC: Offline Regeneration of Identical Superficially Porous Particle Preparative Columns Utilizing The 1290 Infinity II Preparative LC System and Agilent InfinityLab Poroshell 120 Preparative LC Columns to extend Column Lifetime, Minimize Sample Carryover and Increase Throughput

• poster / HPLC

Abstact: High throughput sample purification and analysis are important tasks in many industries. With the increasing number of samples synthesized– HPLC purification should not be the bottleneck in discovery and development. The throughput on the Prep system can be increased by optimizing the chromatographic method and by optimizing the instrument hardware. In this poster we show how the throughput can be enhanced even further by using two identical Poroshell Preparative HPLC columns in the system and operating them alternately optimizing the 1290 Infinity II Preparative LC System for Off-Line Column Regeneration. The greatest benefit of the offline column cleanup and equilibration is the increase in sample throughput, but added benefits include extended column lifetime and a reduction in sample carryover.

Conclusions: Faster run times and higher throughput is desirable when doing preparative HPLC. Using higher flow rates, shorter and lower pressure columns as well as optimized equipment can help to achieve these goals. Higher backpressure limits of the Agilent Inf II Prep pumps along with high pressure valves further reduce cycle times by allowing configuration of automated offline column regeneration. This means two columns are alternatively used via a 2-position/14-port valve. Cleaning and equilibration time can be excluded from the cycle time and 50% more runs were performed.

2. Waters Corporation: Scaling Reverse-Phase and Hydrophilic Interaction Liquid Chromatography (HILIC) Methods to HPLC

• poster / HPLC

INTRODUCTION:

Achieving identical chromatographic separation and equivalent results are critical when scaling and migrating methods between different liquid chromatography (LC) instruments.

In this work, two different methods developed on 1.7 μm particle size columns using a low dispersion system were scaled to HPLC. In first example, a reverse-phase method for naphazoline hydrochloride, pheniramine maleate and associated related substances was scaled from 1.7 μm to a 2.5 μm particle column. A second method for the analysis of ribavirin and related compounds, operated under HILIC conditions, was scaled from 1.7 μm to 2.5 and 3.5 μm particle columns and run on different HPLC platforms. The scaled methods generated equivalent chromatographic performance and assay results as the original method.

CONCLUSION:

• Reverse-phase and HILIC methods were successfully scaled from 1.7 µm particle size columns to HPLC.
• The target LC systems including ACQUITY Arc, Arc HPLC and Alliance iS HPLC Systems generated equivalent performance as the original method run on an ACQUITY UPLC H-Class Plus System.
• Dwell and extra-column volumes were accounted for to ensure quality of the chromatographic data for method scaling and migration across different LC platforms.

3. Thermo Fisher Scientific: Performance evaluation of a new reversed-phase column for peptide mapping and monitoring of monoclonal antibodies

• poster / HPLC

Abstract

Purpose: Demonstrating the benefits of the newly introduced Thermo Scientific Hypersil GOLD Peptide C18 column with 1.9 µm particle and 175 Å average pore size, in the dimension of 150 x 2.1 mm (L x ID) for the analysis of tryptic mAb digest samples.

Methods: UHPLC-MS/MS peptide mapping and UHPLC-UV-MS peptide monitoring.

Results: The Hypersil GOLD Peptide column enables to uniquely meet the demands of peptide level analysis where consistent lot-to-lot and column-to-column performance is required.

Conclusions

The two studies of peptide mapping and peptide monitoring demonstrate the benefits of the Hypersil GOLD Peptide column for peptide level biopharma analysis based on a comprehensive dataset obtained from rituximab and adalimumab digest samples.

• SET and SST criteria of retention time and peak area precision are easily met when using the Hypersil GOLD Peptide column.
• The Hypersil GOLD Peptide column provides
  • peaks with good asymmetry values, strong retention time and peak area reproducibility.
  • retains and separates hydrophilic peptides and has very good chromatographic resolution values for deamidated species in the PENNYK region.
  • excellent column lot-to-lot reproducibility with % RSD RT ≤ 0.6, and % RSD peak area ≤ 6.2 for peptides across the entire elution window.
  • consistent column-to-column results for the quantification of glycopeptides with % RSD of relative abundance ≤ 3.0.

4. Shimadzu: Development of a blood-based multiplex quantitative LC-MS method for neurodegenerative diseases patient stratification : towards an innovative diagnostic tool

• poster / ASMS 2024

Introduction and objectives: Current neurodegenerative biomarkers measurements for diagnosis purposes are performed using CSF samples which requires invasive procedures for the patient. Besides, AD and other associated dementias are very complex pathologies that can have intricate symptoms and pathological deleterious effects. Therefore, the developed quantitative multiplex method includes 22 blood neuroinflammatory, brain function-linked, genetic and general state biomarkers for disease state evaluation and monitoring, patient stratification and differential diagnosis purposes.

Conclusions and Perspectives: Further analytical validation would be performed to fully assess method performances (stabilities, intra-laboratory reproducibility). Then, large patient cohorts would be analyzed to evaluate and clinically validate this innovative tool and its capacity to differentiate diseases, severity and co-pathologies. Such tool would bring dementia diagnosis within the landscape of personalized medicine fostering better patient care.