News from LabRulezLCMS Library - Week 40, 2024

Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 30th September 2024? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!

👉 SEARCH THE LARGEST REPOSITORY OF DOCUMENTS ABOUT LCMS AND RELATED TECHNIQUES

👉 Need info about different analytical techniques? Peek into LabRulezGCMS or LabRulezICPMS libraries.

This week we bring to you applications by Shimadzu, Thermo Fisher Scientific, Waters Corporation, and Metrohm and a tech nical note by Agilent Technologies!

1. Agilent Technologies: Purity, Assay, and Impurity Profiling of Single-Stranded Oligonucleotides Using Agilent Oligo Analysis Accelerator for OpenLab CDS

• Technical note

Introduction

Challenges with therapeutic synthetic oligonucleotides include determining the sample purity, label claim or assay, and quantitation of impurities. Traditional HPLC methods that rely on 260 nm UV detection may not have the necessary sensitivity or specificity for quality control (QC) testing. Using a more selective detector such as a mass spectrometer is a viable approach for QC testing needs. Although LC/MS methods are a mainstay for characterization of oligos, many high-resolution accurate mass detectors may not have the robustness needed in a routine testing environment such as QC. Additionally, a high level of technical expertise is often required for MS operation, data analysis, and data interpretation.

A more practical approach uses a single quadrupole detector that is both sufficiently selective and sensitive enough for purity, assay, and impurity profiling of oligos. Given its relatively high mass range and sensitivity in full scan mode, the Agilent InfinityLab LC/MSD XT can meet the demands for LC/MS QC.

As demonstrated by Rentel et al.,1 the LC/MSD XT can be used and validated for the QC lot release of oligos. However, one challenge that remains is the complex, multistep data analysis which requires the extraction of several extracted ion chromatograms and manual integrations. Performing these operations in a chromatography data system (CDS) while meeting compliance requirements can be time consuming. Manual transcriptions by the analyst may lead to a high margin of error, further delaying decision making.

This technical overview will address the Oligo Analysis Accelerator (OAA) for Agilent OpenLab CDS. This software add-on acts as the data analysis front end to OpenLab CDS, leveraging built-in technical controls that help meet data integrity and compliance requirements. By performing overlays, integrations, and calculations in a fit-for-purpose user interface, data analysis is significantly streamlined. This not only saves time-on-task but also improves data quality.

Summary

The final step in the workflow is to review the results. This includes the SST results shown in previous steps, as well as purity and identity results. Assay results may vary depending on which formulation was selected for the sample.

It is important to note that the summary itself is not the report. An OpenLab CDS report template must still be used to report the results for review. Further, any electronic signatures and approvals occur within OpenLab CDS, not in OAA. Again, this is because OAA leverages the already existing data compliance engine within the OpenLab CDS platform.

Future considerations

LC/MS data analytical workflows have been confined to data systems either used to interpret spectra or chromatograms. However, the need for more user-friendly, fit-for-purpose data analysis is becoming increasingly important for the development of complex therapeutics such as oligonucleotides. 

Agilent Oligo Analysis Accelerator for OpenLab CDS is a paradigm shift in the way software is being developed.

2. Waters Corporation: Repeatability and Reproducibility of the Oasis™ WAX/GCB for PFAS Analysis Cartridges in Aqueous Samples for EPA Method 1633

• Application

Abstract

In January 2024, EPA Method 1633 was finalized. EPA Method 1633 is the first method to incorporate the determination of 40 PFAS compounds across many challenging environmental sample matrices outside of drinking water, including non-potable waters (ground water, surface water, and wastewater) as well as soil, biosolids, and tissue by LC-MS/MS analysis. EPA Method 1633 requires use of both weak anion exchange (WAX) solid phase extraction with graphitized carbon black (GCB) cleanup and is a performance-based method allowing for modifications as long as acceptance criteria for recoveries and %RSDs are met.

This application note highlights the sample extraction for non-potable aqueous samples in EPA Method 1633 using a dual-phase, or bilayer cartridge: Oasis WAX/GCB for PFAS Analysis meeting the method acceptance criteria and displays reproducibility of the cartridges.

Benefits

• Oasis WAX/GCB for PFAS Analysis, a dual-phase cartridge is reproducible and repeatable for EPA Method 1633 aqueous samples in both inter- and intra-batch assays
• Reduction of manual steps, overall sample preparation time by use of the dual-phase SPE cartridge
• Acceptance criteria for recoveries and %RSDs are met for EPA Method 1633 for aqueous samples

Conclusion

This study demonstrates the reproducibility of the dual-phase Oasis WAX/GCB for PFAS Analysis cartridges for determination of 40 PFAS and standards using the ACQUITY UPLC I-Class System and Xevo TQ-XS Mass Spectrometer. The cartridges are suitable for PFAS analysis in accordance with EPA 1633 guidelines for recovery and %RSD. The WAX/GCB cartridges show excellent repeatability across multiple product lots and within multiple replicates of each lot across non-potable water samples. The data demonstrates Oasis WAX/GCB for PFAS Analysis cartridges are ideally suited for PFAS analysis from complex matrices, such as non-potable waters like those described in EPA Method 1633. Out of the box performance is expected lot to lot and within a lot for SPE when using Oasis WAX/GCB for PFAS Analysis.

3. Thermo Fisher Scientific: Dispersive liquid-liquid micro-extraction for the automated sample preparation of PFAS in drinking water

• Application

Application benefits

• Improved sensitivity: DLLME provides high enrichment factors, which can improve the detection sensitivity of PFAS in drinking water samples, down to challenging regulatory detection and reporting limits.
• Cost-effective: DLLME uses a small volume of solvent, which not only reduces the cost but also makes the method more environmentally friendly.
• Versatility: DLLME can be employed for the analysis of a wide range of PFAS classes, followed by either high performance liquid chromatography or gas chromatography as a separation technique, coupled to tandem mass spectrometry or high-resolution accurate mass as detection method.

Goal

To demonstrate a comprehensive method for the analysis of per- and polyfluoroalkyl substances (PFAS) by harnessing the potential of automated sample preparation on the Thermo Scientific™ TriPlus™ RSH SMART liquid handling station, with dispersive liquid-liquid micro extraction (DLLME) as a simple, cost-effective, and versatile extraction and pre-concentration technique. This was performed on LC-MS, but the same workflow can be also applied to GC-MS.

Keywords: DLLME, PFAS, sample preparation, automation, liquid handling, drinking water

Conclusions

The DLLME method presented in this work is a promising technique for the extraction and pre-concentration of PFAS from drinking water samples. Thanks to the described careful optimization and validation of the method, it provides an automated, fast, efficient, and green alternative to traditional and manual extraction methods. As compared to SPE procedures, the only manual steps consist in adding internal standards and an acidic solution to the sample, and then the TriPlus RSH SMART instrument will perform the extraction automatically, while for SPE procedures, the rest of the steps are mostly manual. This workflow allows laboratories to overcome the challenges associated with this analysis, providing high enrichment factors for all the different compound classes studied, as well as good reproducibility and robustness of the extraction process. In summary, the presented automated DLLME sample preparation has the following advantages:

• Reduced sample volume (15 mL) for easy sample handling, transportation, and storage.
• Cost reduced by low solvent usage and no need for filters or SPE cartridges.
• Automation enables reproducible and accurate results, with low inner- and cross-contamination.
• Short runtime and automated process managed by Chromeleon software saves time and allows staff to perform more profitable activities.
• The short runtime of the TriPlus RSH SMART instrument opens the possibility to feed one or more analytical instruments, including HPLC-MS or GC-MS technologies.
• The extraction protocol without using filters and based on the use of low-density solvent for extraction can open the way for the extraction of other matrices such as wastewater.

4. Shimadzu: Efficient Data Analysis with Peakintelligence - Application for Analyzing Per- and Polyfluoroalkyl Substances (PFAS)

• Application

User Benefits

• Peakintelligence peak integration software substantially reduces the time and effort required for data analysis in PFAS analysis.
• Since it does not require parameter settings, Peakintelligence allows even inexperienced users to perform peak integration and achieve results on par with experienced workers.

Introduction

Per- and polyfluoroalkyl substances (PFAS) are lyophobic, heat resistant, and chemically resistant compounds used extensively as coatings, surface treatment agents, emulsifiers, and fire-extinguishing agents. Concerns about PFAS and their persistence in the environment, accumulation in living organisms, toxicity to living organisms, and ability to travel long distances have recently led to fact-finding studies on PFAS and the introduction of regulations. As of October 2023, only three PFAS are subject to regulatory restrictions in Japan (PFOS, PFOA, and PFHxS), but more are subject to restrictions in the USA and Europe. Throughout the world, an increasing number of PFAS are expected to require analysis. These changes will create more analytical data that will have to be processed, creating a need for more efficient methods of data analysis.

Peakintelligence is a peak integration software equipped with peak-finding algorithms that have been developed with the help of artificial intelligence (AI). This Application News describes a case example in which it was used to process chromatogram data of PFAS analysis and reduce the effort required and increase the efficiency of the analysis.

Conclusion

Using Peakintelligence to process peaks in PFAS analysis substantially reduced the number of false detections and falsely identified peaks. It also significantly reduced the number of peaks that required manual processing, and it did not need pre-analysis adjustment of parameters. These improvements enable substantial reductions in the time and effort required to perform data analysis. Avoiding parameter settings by the user also eliminates variability in the results between users, which helps eliminate dependency on specific personnel. As the number of PFAS requiring analysis increases throughout the world, a more efficient method of analyzing peaks will be needed; Peakintelligence is an effective solution for this need.

5. Metrohm: Iron speciation in LiFePO₄ batteries

• Application

Simultaneous determination of Fe(II) and Fe(III) in lithium iron phosphate with the Multi-Mode Electrode pro

Lithium iron phosphate (also known as LiFePO₄ or lfp) batteries last for over 2000 charges and are safer because of their lower risk of overheating. LiFePO4 batteries have a slightly lower energy density compared to lithium-ion batteries. However, high discharge rates of lfp batteries make them ideal for electric vehicles, renewable energy storage, and backup power systems. Lithium iron(II) phosphate is used as a cathode material in lithium iron phosphate batteries. Characterization of lfp and monitoring the oxidation state of iron in lfp batteries is relevant to battery performance in terms of durability, capacity, and safety. Additionally, analyzing chemical composition can be useful for battery research and can aid eco-friendly recycling practices. This is essential for driving battery technology forward and promoting clean energy solutions.

Polarographic speciation of Fe(II) and Fe(III) can be used to evaluate the purity of LiFePO4 and its usability as a cathode material in lithium iron phosphate batteries. It can further be used to study the concentrations of Fe(II) and Fe(III) in the cathode material after several charging and discharging cycles to evaluate the aging behavior.