News from LabRulezLCMS Library - Week 44, 2024

Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 28th October 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 you applications, technical notes, and brochures by Watrex Praha, Thermo Fisher Scientific, Shimadzu, Waters Corporation, and Agilent Technologies!

1. Watrex Prague: Watrex HPLC COLUMNS

• Brochures and specifications

Ion chromatography - Analysis of cations

• Watrex IonPlus™ CS19 - Styrene-divinylbenzene, 55% cross-linking (7 μm particle i.d.)

Ion chromatography - Analysis of anions

• Watrex IonPlus™ AS19 - Styrene-divinylbenzene, 55% cross-linking (12 μm particle i.d.).
• Watrex IC Anion I - Styrene-divinylbenzene (7 μm particle i.d.)

Reversed phase chromatography

• Watrex DeltaSil™ 100 C18 column is a standard reversed phase for wide field of applications in analysis of drugs, organic compounds, peptides and others. Stationary phase is based on synthetic 5 μm silica-gel with 100 Å pores and 15% carbon content. The column has a good peak symmetry and efficiency. Theoretical plate count value for non-polar compounds, such as naphthalene, reaches up to 90.000 theoretical plates per meter.

Carbohydrate analysis

• Watrex Polymer IEX™ is the Europe’s leading sorbent for ion exclusion analysis of alcohols, carboxylic acids, sugars, and polysacharides.

Gel-permeation chromatography

• Watrex DeltaGel ™ - Mixed-bed GPC columns for SEC chromatography in organic solvents.

2. Agilent Technologies: Agilent Advanced Sample Linking (Infinity III)

• Technical note

Abstract

With the release of the Agilent Infinity III LC series, the Agilent 1260 and 1290 Infinity III Multisampler can optionally be equipped with the Agilent InfinityLab Sample ID Reader. The integration of the LC system into a client-server environment allows the use of additional software, such as Agilent Sample Linking software and Agilent Sample Scheduler for OpenLab, enabling a complete workflow that links samples from any laboratory information management system (LIMS) to the vials and HPLC analysis results. This white paper highlights the benefits of the integrated analytical workflow. In this workflow, barcodes are used to track samples from the sample list to their vial position, and the presence of the correct sample is recognized without manually entering its position. This saves time and improves ease of use, with fewer errors over the complete workflow.

Conclusion

This white paper describes a workflow that begins with an analytical sequence initiated by any LIMS, then links the barcodes of sample containers to the barcodes of target pre-barcoded vials. The samples are analyzed to get the final data results, and a report is transmitted back to LIMS. This closed-loop, error-free, and time-saving workflow is enabled by Agilent Sample Linking software together with the Agilent Sample Scheduler for OpenLab. Samples can be tracked through the complete workflow using barcodes to transfer them accurately from their original containers to a final barcoded vial, managed completely by the Sample Linking software. This saves time, enhances ease of use, and results in fewer errors over the complete workflow.

3. Waters Corporation: Improving USP Monograph Analysis Time Through Scaling by N on an HPLC System Using CORTECS™ Premier Columns

• Application

Abstract

The United States Pharmacopeia General Chapter <621> allows certain changes to monograph methods. Changes in column length and particle size are employed as part of modernization efforts from HPLC to UPLC™ instrumentation. However, some modernization activities can be performed without switching LC systems. For HPLC analyses, reducing particle size from 5 μm to 3.5 μm can improve throughput and run times, however those columns increase backpressure of the system. Additionally, depending on the monograph, modernization may not be possible due to available column configurations, and the guidelines set in General Chapter <621> for scaling by L/dp. Modernizing by plate count (N) is one path forward, especially if the column being modernized to takes advantage of highly efficient particles. This application note scales the USP impurities assay of
zidovudine from the original 4.6 x 250 mm column using fully porous 5 μm particles to a 4.6 x 150 mm column using solid-core particles to improve both throughput and solvent usage.

Benefits

• USP Monograph for zidovudine impurities analyzed on various columns
• Vastly improved separation efficiency using CORTECS Premier 5 μm columns with original USP monograph column configuration
• 40% improvement in solvent usage and up to 60% improvement in analysis run time using CORTECS Premier 5 μm Column

Introduction

Monograph methods have traditionally employed the use of 5 μm particle size columns due to pressure limitations of older HPLC instrumentation. In combination with the larger particle stationary phases, longer column dimensions were needed in order to achieve appropriate column efficiency. This leads to lengthy analyses with high volumes of solvent being consumed. Monograph methods can be modernized however, and the United States Pharmacopeia (USP) outlines two routes for modernization as outlined in USP General Chapter <621>.1 The first is by changing particle size and column length (L/dp) which can be done but may require the use of more modern LC technology like UPLC. The second route to modernization involves scaling a column based on the plate count (N) for the assay. Changes can be made to either the L/dp or N by -25% and up to 50%. Modernizing by N allows a method to be changed to any configuration as long as the plate count for the new conditions falls within the limits outlined in General Chapter <621>.

For some assays, modernizing by N can be possible by using higher efficiency stationary phases, like solid-core particles. Solid-core particle columns, like CORTECS columns, have been shown in the past to boost separation efficiency due to particle morphology.2–4 By taking advantage of the solid-core particles, a shorter column may be used while still achieving acceptable plate counts. Thus, improvements to run time and solvent usage could
be possible without changing particle size, therefore avoiding the potential increases to system pressure accompanied with smaller particle sizes. In this application note, the assay and impurities monographs for zidovudine were scaled from a 250 mm column down to a 150 mm column using the modernize by N approach. Using the new CORTECS Premier 5 μm columns results in a 40% decrease in solvent usage with up to a 60% reduction in analysis time per sample.

Conclusion

Validated monograph methods often employ HPLC systems with larger particle size columns due to pressure limitations of the system. Modernization of these monograph methods can be performed under USP General Chapter <621> using either the length to particle size ratio (L/dp) or Plate Count (N). In either case, the new column configuration must be within -25% to 50% of the original conditions. Once a new column is selected, changes to gradient profile and flow rate are calculated. This application note focused on the assay and impurity test for zidovudine, which calls for a 250 mm column in the original monograph.

By using CORTECS Premier 5 μm Column the assay and impurities test for zidovudine was successfully scaled down to a 150 mm column while maintaining the particle size. This greatly improved sample throughput with a 40% reduction in run time and solvent usage. Further improvements to throughput were realized by increasing flow rate from 1.5 mL/min to 2.25 mL/min. The increased flow rate, while not acceptable under USP General Chapter <621>, is still able to be run due to the lower system pressure generated by the 150 mm column. A 60% reduction in run time is realized with the higher flow rate. CORTECS Premier 5 μm Columns offer highly efficient solid-core particles combined with MaxPeak Premier High Performance Surface Technology hardware.

4. Shimadzu: Analysis of Per- and Polyfluoroalkyl Substances (PFAS) using Triple Quadrupole Mass Spectrometer Part 1 - Fish Fillet

• Application

User Benefits

• The optimized procedure for pretreatment and LC-MS/MS analytical conditions enable accurate quantification of thirty major PFASs targeted by AOAC SMPR from 0.1 μg/kg.
• The method allows the initiation of PFASs analysis in food

Introduction

Per- and Polyfluoroalkyl Substances (PFASs) are a collective name for more than four thousand organofluorine compounds. Perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) are representative compounds of PFAS. They are used in a wide range of applications, such as fire retardants, food packaging materials, and non-stick coatings, due to their water-repellent, oil-repellent, heat-resistant, and chemical-resistant properties. Due to their structural stability, PFAS widely remains in the environment. There are concerns about health risks caused by human ingestion of fish that have ingested seawater, river water, or feed contaminated with PFAS. Therefore, quantitative assessment of PFAS levels in fish should be important.

This application news introduces a quantitative analysis of PFAS in tuna fillets with LC-MS/MS. Thirty principal PFASs targeted by AOAC INTERNATIONAL, a North American organization that standardizes food testing methods and verifies analytical methods, were analyzed using newly created method, which was investigated from the pretreatment procedure then validated by recovery test. By reducing losses during the pretreatment and optimizing the analytical conditions, good recovery rates were obtained for all compounds.

Conclusion

This application news described a quantitative analysis of thirty PFAS targeted by AOAC INTERNATIONAL in tuna fillet. The analysis was performed using a quadrupole mass spectrometer LCMS-8060NX equipped with ultra-high performance liquid chromatograph Nexera X3 UHPLC system. Shim-pack Scepter, which provides good separation and peak shape, was used as the column. A spiked recovery test were conducted, and all compounds showed recovery rate within 80-120% and repeatability below 20% at spiked concentrations of 0.1, 1, 5 μg/kg. In particular, recovery rates of PFOS, PFOA, PFNA, and PFHxS were within 95.5-108.4% at all spiked concentrations. Using optimized pretreatment and analytical methods, accurate quantitation is possible from 0.1 μg/kg.

5. Thermo Fisher Scientific: Highly sensitive method for the determination of 12 nitrosamine impurities in multiple ARBs, the class of sartan drug formulations

• Application

Application benefits

• Single method applicable for highly sensitive quantitation of 12 nitrosamines in three ARB drug substances.
• A robust gradient method with significant chromatographic separation between nitrosamines and ARB drug substances.
• Better retentivity and resolution of nitrosamines and APIs with comparatively shorter runtime, ideal for high throughput screening.

Goal

Development of a sensitive and robust LC-MS/MS method to quantitate the following 12 nitrosamine impurities in three ARB drug substances: N-nitrosodimethylamine (NDMA), N-nitroso-diethylamine (NDEA), N-ethyl-N-nitroso-2-propanamine (NEIPA), N-nitroso-diisopropylamine (NDIPA), N-nitroso-di-n-propylamine (NDPA), N-nitroso-methylphenylamine (NMPA), N-nitroso-di-n-butylamine (NDBA), N-nitroso-N-methyl-4-aminobutyric acid (NMBA), N-nitrosomorpholine (NMOR), N-nitrosopyrrolidine (NPYR), N-nitrosomethylethylamine (NMEA), and N-nitrosopiperidine (NPIP).

Introduction

Angiotensin receptor blockers (also called ARBs or angiotensin II inhibitors) are medicines that dilate blood vessels and are used in the treatment of high blood pressure (hypertension), heart failure, or kidney disease in people with diabetes.¹ These ARBs, also known as sartan medicines, have a specific tetrazole ring-containing structure whose synthesis could potentially lead to the formation of nitrosamine impurities.²

If the content of nitrosamine impurities is found to be greater than the acceptable limit, then regulatory authorities need to be informed, and that may ultimately lead to the drug substance product being recalled. There have been numerous drug recalls associated with nitrosamine impurity levels exceeding the allowable limits in drug substances of various pharmacological categories, including ARBs.³ In 2018 and in 2019, the U.S. Food and Drug Administration (FDA) found traces of NDMA and NDEA impurities in the ARB drug products valsartan, losartan, and irbesartan. Since July 2018, more than two dozen specific ARB products have been recalled owing to the unacceptable presence of potentially carcinogenic nitrosamine impurities.⁴

In 2018, the FDA issued guidance to the industry on how to assess and control impurities. EMA finalized a review under Article 5(3) of Regulation (EC) No 726/2004 in June 2020 to provide guidance to marketing authorization holders on how to avoid the presence of nitrosamine impurities in human medicines.

The CHMP asked marketing authorization holders to review all chemical and biological human medicines for the possible presence of nitrosamines and test products at risk. Regulators across the globe have been vigilant on nitrosamine testing and subsequent modification of guidelines or testing procedures to ensure the best quality products reach the market. Recently, the FDA⁵ and EMEA⁶ updated guidelines with a more comprehensive and categorized approach towards nitrosamine testing.

In this application note, the goal was to develop an LC-MS/MS solution to the increased frequency of demands for higher sensitivity and quantification of an extensive array of nitrosamine impurities in pharmaceutical formulations with a one-method approach for multiple products. From the multiple techniques available today, liquid chromatography-tandem mass spectrometry is one of the best for highly sensitive and reproducible quantification of nitrosamines. It not only benefits sensitivity in sub-nanogram concentrations but also provides a more robust, linear, and reproducible solution for nitrosamine detection. The guidelines followed in this application are from the ICH Q2(R2)⁷, FDA⁵, and EMEA⁶.

As demonstrated by the excellent results observed in this study, the Thermo Scientific™ TSQ Altis™ Plus Triple Quadrupole mass spectrometer connected to the Thermo Scientific™ Vanquish™ Flex UHPLC system makes a suitable configuration for the highly sensitive analysis of multiple nitrosamines in a single method applicable for 3 ARBs—olmesartan, irbesartan, and valsartan. For data processing and reporting, Thermo Scientific™
Chromeleon™ Chromatography Data System (CDS) provides a complete solution, which fulfils all compliance requirements and ensures data integrity and security requirements are met, with a detailed workflow that includes user management along with instrument and data audit trails.