News from LabRulezLCMS Library - Week 32, 2024

Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 5th 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 applications, brochures and specifications from Agilent Technologies, Shimadzu, Metrohm, Thermo Fisher Scientific, and Waters Corporation!

1. Shimadzu: How Shimadzu is simplifying your compliance workload for the new IVD regulation!

• Technical Note

What is the New EU In Vitro Diagnostic Regulation (IVDR)?

New In Vitro Diagnostic Medical Device Regulation (IVDR) (EU) 2017/746 was published in EU Official Journal in May 2017 in order to ensure a high level of safety and health whilst supporting innovation. It applies from 26 May 2022 and replaced the previous IVD Directive (98/79/EC). This regulation set rules for in vitro diagnostic devices such as reagent, calibrator, kit, instrument, and software intended by the manufacturer to be used for diagnostic purposes.

IVDR retains some concepts of IVD Directive such as CE marking or essential requirements (under IVDR called “general safety and performance requirements”). As in the past, the manufacturers have to demonstrate that they and their devices meet the requirements through conformity assessment procedure.

Different from IVD Directive, however, IVDR introduced the riskbased classification (Figure 1). Under IVDR devices are classified in four: The devices with highest risks are classified in class D. The class A devices have low risks and include products for general laboratory use, accessories which possess no critical characteristics.

2. Shimadzu: Determination of 30 PFAS in Seafood by Liquid Chromatography Triple Quadrupole Mass Spectrometry (LC-MS/MS)

• Application

User Benefits

• Validated method for 30 PFAS in seafood (shrimp) meeting all criteria of AOAC SMPR 2023.003
• High precision, excellent recovery, low Limit of Quantification (LOQ)
• Simple and rapid extraction using QuEChERS

Introduction: Per-and polyfluorinated substances (PFAS) are a diverse group of man-made chemicals used in numerous products since the 1950s. PFAS can enter the food supply by contact in environmentally contaminated areas, during food processing, or exposure to packaging. Because PFAS have been linked to serious health effects, accurate methodology is needed. In this application news, we describe a single laboratory validation study with a rapid extraction of low concentrations of 30 PFAS in shrimp using the QuEChERS technique followed by analysis using the Shimadzu Nexera Liquid Chromatograph coupled to a Shimadzu LCMS-8060NX triple quadrupole mass spectrometer (Figure 1).

We optimized the chromatography and instrument operating parameters to achieve excellent peak shape, separation, and sensitivity. Sensitivity was improved for 4 PFAS, PFOA, PFHxS, PFNA, and PFOS.

In this study, we spiked samples at three concentrations in triplicate. For greater accuracy, standards were matrixmatched and extracted and spikes were quantified using the isotope dilution technique. Recovery and precision were compared to the requirements of AOAC SMPR 2023.003. In addition, we determined the Limit of Quantitation (LOQ) as the lowest concentration meeting accuracy and precision, ion ratio, retention time, and signal-to-noise ratio criteria of the qualifier ion. All recovery, precision, and LOQ’s met the acceptance criteria of the SMPR. The target analytes, their acronym, chemical abstract number, and experimentally determined LOQ are shown in Table 1.

Conclusion: The Shimadzu LCMS-8060NX Triple Quadrupole Mass Spectrometer coupled with a Shimadzu Nexera Liquid Chromatograph was used in a single laboratory study to measure 30 PFAS compounds in a shrimp matrix and compared to criteria set by AOAC SMPR 2023.003. Chromatography conditions and the mass spectrometer were optimized to achieve excellent separation of all analytes, baseline resolution between linear and branched isomers, and a two-minute separation between PFOS and potentially interfering cholic acids.

Precision and recovery and the experimentally determined LOQ are well within the requirements of the SMPR.

3. Agilent Technologies: Boost Efficiency and Productivity with Instrument Intelligence and Workflow Automation

• Technical Note

Introduction: Ever-increasing pressure for laboratories to minimize operating costs while maximizing output demands innovative approaches to their daily work. Recent developments in artificial intelligence (AI) and automation can help to alleviate this growing burden. The goal is to get technology to contribute more and more to day-to-day operations by reducing errors and ensuring consistent high-quality data generation. For workflows using Agilent LC/TQ and LC/(Q-)TOF instruments, this goal is becoming a reality.

The latest generation of Agilent LC/TQ and LC/(Q-)TOF instruments offers a suite of intelligent automation features to keep the instruments at peak performance. SWARM autotune allows instruments to be tuned for specific applications and desired performance. Auto- and check tunes can be scheduled to run during downtime so that the instruments are always ready to go. Instrument status is actively monitored using Early Maintenance Feedback (EMF) with constant feedback so that users can anticipate and plan for maintenance. Data are actively examined to ensure high quality. And, if data-related issues are suspected, Intelligent Reflex reinjection workflows automatically work to correct the situation. The culmination of these instrument intelligence features reduces errors, rework, and manual intervention—boosting laboratory efficiency and productivity.

Conclusion: Agilent LC/TQ and LC/(Q-)TOF instruments include numerous intelligence and workflow automation features to boost productivity and efficiency of demanding routine analysis applications. These intelligence features:

• Maximize instrument performance and analytical sensitivity (application-based SWARM autotune)
• Minimize downtime (early maintenance feedback and scheduled tuning)
• Maximize sample throughput (intelligent reflex and fast screening workflows)
• Ensure that injected samples do not require repeat work(intelligent reflex carryover detection and above calibration range workflows)
• Improve sample characterization (intelligent reflex targeted MS/MS confirmation and iterative MS/MS workflows)

4. Agilent Technologies: Confirmation of Peptide-Related Impurity Intact Mass Using Agilent 1290 Infinity II Bio 2D-LC and InfinityLab LC/MSD XT

• Application

Abstract: This application note presents the analysis of peptide-related impurities for Semaglutide using an Agilent 1290 Infinity II bio 2D-LC and an Agilent InfinityLab LC/MSD XT system. Peptide-related impurities are challenging to separate from peptide-active pharmaceutical ingredients due to their structural diversity and relatively large molecular weights. To address this challenge, an Agilent AdvanceBio Peptide Plus column, which is stable under high-concentration acidic conditions, was used to achieve good separation of Semaglutide impurities. A solvent system compatible with mass spectrometry (MS) was used with the 1290 Infinity II bio 2D-LC and InfinityLab LC/MSD XT, enabling the acquisition of MS spectra. The deconvolution feature in Agilent OpenLab CDS software, version 2.8, facilitated the easy determination of molecular weights for impurity peaks analyzed by two-dimensional liquid chromatography (2D-LC).

Conclusion: Generally, difficult-to-separate peptide-related impurities can benefit from improved resolution through optimizing the concentration of acidic modifiers and fine-tuning the column performance. However, confirming the molecular weight of impurities can be challenging due to mobile phases not being suitable for MS analysis. In this application note, a stability indicating test for Semaglutide demonstrated efficient transition to LC conditions compatible with MS analysis using the Agilent InfinityLab multiple heart-cutting valve feature of an Agilent 1290 Infinity II bio 2D-LC. An Agilent InfinityLab LC/MSD XT exhibited the sensitivity to confirm MS spectra, even when analyzing impurities at low concentrations in the second dimension, and the deconvolution feature of Agilent OpenLab CDS software, version 2.8, facilitated easy confirmation of accurate molecular weights. Based on these results, it is anticipated that this method will provide foundational data for the management of peptide-related impurities not only for formulation compatibility but also for stability studies.

5. Thermo Fisher Scientific: Revolutionize your oligonucleotide analysis with game-changing DNAPac RP columns

• Other

The semi-preparative format – ideal for nucleic acid purification

Product features

• Designed for ion-pair reversed-phase (IP-RP) LC and LC/MS separations of both single-stranded oligonucleotides and double-stranded nucleic acids
• Unique selectivity of the DNAPac RP chemistry, now available for purification
• Bioinert hardware to eliminate analyte adsorption and avoid the need for column passivation
• Wide operating pH range and high-temperature stability (up to 110 °C)
• High resolution and high throughput

6. Thermo Fisher Scientific: Thermo Scientific DNAPac columns - Ultra-high resolution oligonucleotide separation

• Other

• Thermo Scientific™ DNAPac™ PA200 and DNAPac™ PA200RS Columns offer the highest resolution charge based oligonucleotide separations. These anion-exchange columns provide unsurpassed resolution of full length from n–1, n+1 (and other failure sequences).

• Thermo Scientific™ DNAPac™ RP (U)HPLC Columns are high-throughput, high-resolution columns designed for ion-pair reversed-phase separations. They are compatible with LC-MS, have long-lasting polymeric columns, and can operate up to pH 14. They also have temperature stability up to 110 °C. These columns can easily resolve both small polynucleotides and large dsDNA fragments up to 10,000 bp due to pores varying from 50-2000Å. Now available in semi-preparative format.

• Thermo Scientific™ DNASwift™ SAX-1S Monolith Column provide high loading capacity for easy laboratory scale charge-based purification of oligonucleotide samples.

7. Waters Corporation: VISUALIZATION AND QUANTIFICATION OF DRUG/METABOLITES BY SENSITIVE AND FAST TARGETED DESI IMAGING ON TQ SYSTEM

• Poster / ASMS 2024

INTRODUCTION: Mass spectrometry (MS) coupled to liquid chromatography (LC) has been typically used for drug metabolism and pharmacokinetics (DMPK) studies. However, tissues are homogenised and therefore the molecular localisation is lost.

Desorption electrospray ionization (DESI) mass spectrometry imaging (MSI) has proven to successfully map and independently quantify small molecules without radioactive labelling. Including small pharmaceutical molecules, as well as endogenous metabolites directly from tissue without the need of sample preparation after sectioning.

Tandem quadrupole (TQ) MS are renowned for their sensitivity and specificity in targeted applications using Multiple reaction monitoring (MRM) mode of acquisition.

Here, we used a commercial central nervous system (CNS) active drug named pitolisant that treats narcolepsy. It was dosed at pharmacological level and no observed adverse effect level (NOAEL) to show the distribution of the drug and its main metabolite in different organs such as liver and brain using the sensitive and fast DESI TQ system.

CONCLUSION

• Pitolisant and its metabolites were detected sub 0.1 ng when spotted onto tissue section, demonstrating the high sensitivity and specificity of targeted MRM MS Imaging workflow.
• A highly flexibility workflow is evidenced allowing high speed DESI MSI experiments with no compromise in sensitivity or including more MRM transitions to map biomarkers as well as drug.
• Pitolisant and its oxidized metabolite were easily detected at pharmacological dose, illustrating the different distributions in the liver and brain murine tissue sections, information that wouldn’t be discovered by traditional DMPK studies.
• MSI quantification workflow is demonstrated with the in-house MicroApp MSI Quantify.

8. Waters Corporation: EVALUATION OF DESI MOUNTED ON Q-TOF AND TQ MASS SPECTROMETERS TO IMAGE PFAS COMPOUNDS IN DIFFERENT MATRICES

• Poster / ASMS 2024

INTRODUCTION: Per- and polyfluoroalkyl substances (PFAS) are a well- known group of chemicals that have a variety of commercial and consumer uses. They have been found to be highly persistent and can accumulate in humans, animals and the environment.

Traditionally, analysis of PFAS has focused on determination with LC-MS/MS. Often laborious SPE techniques are required before analysis of solid matrices, such as food, plants or animal tissue to assess exposure routes. Furthermore, samples are homogenized, and therefore the information of where the PFAS accumulate is not available.

Here we evaluate mass spectrometry imaging (MSI) to detect PFAS directly from sample surfaces. Figure 5. Multiple conformations of N-EtFOSAA and corresponding spe- cific drift time ion images of the dilution series. Analyzed using SE- LECT SERIES Cyclic IMS MS in IMS mode . A desorption electrospray ionization (DESI) source combined with two types of Q-Tof mass spectrometers operating in full scan MS (with and without, and a high performance tandem quadrupole mass spectrometer operating in targeted multiple reaction monitoring (MRM) mode were used to evaluate limits of detection (LODs) of several PFAS compounds.

CONCLUSION

• 61% and 55% of the PFAS compounds were detected as low as 10 ng/mL spotted on tissue when analyzed by negative mode DESI in discovery full scan MS acquisition mode.
• The targeted MRM acquisition mode on the DESI XS Xevo TQ Absolute MS is the most sensitive system with 77% of the PFAS compounds detected as low as 10 ng/mL spotted on tissue.
• PFAS compounds have a different ion mobility, with faster arrival time in the gas phase, allowing differential selection and a decrease of interference signal.
• Different conformations of several PFAS were detected by ion mobility with only a single pass in the cyclic device.
• Distinct distributions of PFAS in the seed and root of contaminated PFAS lentils were demonstrated.

9. Metrohm: Mott-Schottky Analysis - Using EIS for semiconductor applications

• Application

Using EIS for semiconductor applications: Semiconductors are intrinsic to modern life, but they stand to play an even greater role in the coming energy transition. Several techniques are employed in the pursuit of improved materials for energy production and storage. One particularly popular method for studying the electronic properties of potential new semiconducting materials is MottSchottky Analysis. The technique can be best understood as an extension of electrochemical impedance spectroscopy (EIS). By performing EIS at a range of different DC bias or offset potentials, the reciprocal of the square capacitance can be derived from the data and then plotted against the DC offset itself, producing a so-called Mott-Schottky Plot. From such a plot, critical parameters such as the carrier concentration, the doping profile, and the flatband potential can be conveniently extracted. This technique is popular across different fields, as it is versatile, non-destructive, and relatively fast. Compared to other options, Mott-Schottky Analysis also requires relatively little specialist equipment. This Application Note presents an example of MottSchottky measurement on a popular semiconducting material using VIONIC powered by INTELLO.

CONCLUSIONS: In this Application Note, an example Mott-Schottky measurement is shown using VIONIC powered by INTELLO. The flatband potential and doping density have been calculated using NOVA and are comparable to values for this type of sample found in the literature.