News from LabRulezLCMS Library - Week 02, 2026

Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 5th January 2026? 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 application notes by Agilent Technologies, Shimadzu, Thermo Fisher Scientific and Waters Corporation!

1. Agilent Technologies: Automated Optimization and Intelligent Reflex Screening on the Agilent 6495D LC/TQ

• Application note
• Full PDF for download

Forensic toxicology panels evolve, and labs must update methods quickly without sacrificing confidence. Manual tuning of MRM transitions and source parameters is time‑consuming and variable across analysts. Two-tier screening and confirmation often require separate runs and manual intervention. The onboard intelligence suite for Agilent LC/MS systems addresses these pain points. Automated compound and source optimization shorten method setup, and intelligent reflex applies real-time, data-driven decisions to trigger confirmatory runs and reporting without manual steps. 

This application note establishes an orthogonal, automated workflow on the 6495D LC/TQ for forensic toxicology screening. The benefits of MassHunter Optimizer software to develop MRM transitions, MassHunter Source Optimizer software, and intelligent reflex fast screening reinjection logic are demonstrated. 

Experimental 

This workflow was performed on an Agilent 6495D LC/TQ equipped with an Agilent Jet Stream ESI source and paired with an Agilent 1290 Infinity II LC. MassHunter Acquisition and Quantitative Analysis software controlled the system and enabled automated optimization and intelligent reflex workflows. 

Compound optimization for MRM transitions was completed using MassHunter Optimizer software, which automatically selected precursor/product ions and collision energies for each analyte. MassHunter Source Optimizer then refined LC/MS ionization conditions by scanning gas temperature, sheath gas temperature, and flow rates. These optimized parameters were applied to both screening and confirmation methods.

Conclusion 

Automated, intelligent features in Agilent MassHunter software on the Agilent 6495D LC/TQ shorten method development and make two-tier screening routine. MassHunter Optimizer and Source Optimizer deliver consistent transitions and robust source settings. Intelligent reflex converts tier-1 screen results into automatic tier-2 confirmations and a single combined report—with no manual intervention. The result is faster turnaround, less rework, and higher confidence from orthogonal chromatographic confirmation—using one instrument and one workflow.

2. Shimadzu: Analysis of Dietary Fiber in Functional Beverage Using Automated Dietary Fiber Analyzer Combined with Integrated HPLC

• Application note
• Full PDF for download

Dietary fiber is an indigestible carbohydrate present in foods such as grains, legumes, fruits, and vegetables1). A high intake of dietary fiber has been associated with a reduced risk of mortality from heart disease, stroke, type 2 diabetes, and certain cancers including breast, colorectal, and pancreatic cancer2). Adequate dietary fiber intake also helps support healthy weight management and improvesthe intestinal environment 2). Dietary fiber can be classified into two types: IDF and SDF. IDF, such as cellulose, hemicellulose, and lignin, is abundant in grains and contributes to water absorption and intestinal function. SDF, including glucomannan, gum arabic, pectin, fucoidan, and β-glucan, is found in foods such as fruits, vegetables, konjac, seaweed, and mushrooms, and is known for its cholesterol-lowering effects 3) . 

Traditionally, the enzymatic-gravimetric method (Prosky method) has been widely used for dietary fiber measurement. However, this method often shows low recovery rates for SDFS such as resistant dextrin and polydextrose. AOAC 2022.014) analyzes the filtrate from the enzymatic-gravimetric method using HPLC, quantifying fractions of trisaccharides and larger as SDFS. 

This article introduces dietary fiber analysis based on the AOAC 2022.01 method, utilizing an automated dietary fiber analyzer (ANKOM Technology)5) with integrated HPLC.

HPLC Analysis of SDFS

Three commercially available functional soft drinks containing resistant dextrin were processed using an automated dietary fiber analyzer. Diethylene glycol (DEG) was used as the internal standard. As these samples consisted mainly of SDFS, HPLC analysis was conducted specifically for this analyte. When determining TDF content in foods that contain other types of dietary fiber in addition to SDFS, the procedure shown in Fig. 1 can also be applied to measure IDF and high-molecular-weight water-soluble dietary fiber (SDFP) by analyzing protein and ash content. 

The enzymes used for pretreatment and the reagents for confirming HPLC retention times were supplied in the Rapid Integrated Total Dietary Fiber Assay Kit (reference code: KRINTDF) from Megazyme. For HPLC analysis, a Bio-Rad desalting cartridge was used as the ion-exchange guard column, Shim-pack SUR-Na(G) as the guard column, and two Shimpack SUR-Na columns connected in series as the analytical columns. Detection was performed using a differential refractive index detector. Fig. 3 shows the chromatogram of the “LC retention time standard” included in the Megazyme K-RINTDF kit. The SDFS fractionation interval was determined based on maltodextrin elution. Chromatograms of the functional beverages are shown in Figs. 4 to 6. HPLC analytical conditions are summarized in Table 1.

Conclusion 

Shim-pack and Shim-vail are trademarks of Shimadzu Corporation or its affiliated companies in Japan and/or other countries. High-performance liquid chromatography (HPLC) analysis was performed to quantify low-molecular-weight soluble dietary fiber (SDFS) in functional beverages containing resistant dextrin. Sample pretreatment was conducted using a fully automated dietary fiber analyzer (ANKOM Technology), which markedly reduced manual labor and processing time by automating the pretreatment workflow. The adoption of disposable filter bags further minimized processing time, thereby decreasing operator workload and conserving valuable working hours. 

Quantification of SDFS was achieved using integrated HPLC. The Shim-pack SUR-Na column, utilized for this analysis, provides high-resolution separation of target saccharides by combining size-exclusion and sodium-type ligand-exchange chromatographic modes. This configuration is particularly suitable for the analysis, as SDFS is operationally defined as trisaccharides and larger oligosaccharides. 

A comparative assessment between the labeled resistant dextrin content on product packaging and the measured SDFS values in the functional beverages demonstrated strong concordance. Additionally, spike-and-recovery experiments confirmed the favorable accuracy of the analytical method.

3. Thermo Fisher Scientific: Screening and quantitation of perfluoroalkyl and polyfluoroalkyl substances (PFAS) residues in foods using LC-MS/MS

• Poster
• Full PDF for download

Per- and polyfluoroalkyl substances (PFAS) are a group of compounds with variable carbon chains and fluorine. The carboxylic and sulfonic acid functional groups exhibit hydrophilic and hydrophobic properties. PFAS compounds exhibit high thermal stability, low reactivity, and strong carbon-fluorine bonding. They have been widely used in applications such as construction, electronic manufacturing, fire-fighting foams, cookware, packaging, textiles, and pharmaceuticals.1 These chemicals are nonbiodegradable, so they remain permanent, bio-accumulating substances.2

Exposure to these chemicals is a significant risk for human beings due to their effect on the reproductive and immune systems. Human exposure to PFAS can occur by inhalation of dust and airborne volatiles or by ingestion from food packaging. However, the most prevalent route is the consumption of contaminated food or drinking water. PFAS can enter the animal food chain through feed, water, and soil ingestion by foraging farm animals, thereby resulting in the contamination of products such as milk, eggs, and meat. Food can also get contaminated through the presence of PFAS in food packaging or processing equipment.3

The European Food Safety Authority (EFSA) CONTAM panel assessed the risk associated with the consumption of PFAScontaminated food, looking at four substances—PFOA, PFOS, PFNA, and PFHxS. Fish (tissue) was identified as the principal contributor to the exposure, followed by fruit and fruit products, and eggs and egg products.4 The European Union Reference Laboratory (EURL) published the “Guidance Document on Analytical Parameters for the Determination of Per and Polyfluoroalkyl Substances (PFAS) in Food and Feed” to harmonize the analytical performances.5 The Guidance Document principles were then adopted by EU Regulation 2022/1428. Finally, the limits were set in foods by Commission Regulation (EU) 2022/2388 of 7th December 2022, entering into force on 1st January 2023, and amending Regulation 1881/2006 (repealed and substituted by Commission Regulation (EU) 2023/915 of 25 April 2023).6,7 The recommended limit of quantitation (LOQ) for PFOS, PFOA, PFNA, and PFHxS in fish meat and meat is 0.1 µg/ kg for monitoring purposes as per the EC 2022/1431 document.8 

This work reports a modified version of the QuEChERS method. A sensitive, selective, and robust analytical workflow was developed for the determination of PFAS compounds in fish tissue and egg using a TSQ Quantis Plus MS, which met low levels (μg/kg) of detection, thereby fulfilling the LOQ requirement. Finally, the developed method was applied to PFAS monitoring of real food samples according to recommendation 2022/1431.

Experimental

Analysis 

The LC-MS/MS analysis was performed using a Thermo Scientific™ Vanquish™ Flex UHPLC coupled to the TSQ Quantis Plus triple quadrupole mass spectrometer, including a heated electrospray ionization (HESI) ion source. The best ionization was achieved in the negative polarity (HESI-). The chromatographic separation was performed on a Thermo Scientific™ Accucore™ C18 Column (100 × 2.1 mm, 2.6 μm) with a Thermo Scientific™ Hypersil GOLD™ Column as a PFAS delay column (50 × 2.1 mm, 1.9 μm). The LC mobile phase consisted of 2 mM ammonium acetate in water (A) and 2 mM ammonium acetate in methanol (B). The detailed UHPLC and mass spectrometer parameters are reported in Table 1. A short length delay column with similar characteristics was utilized to reduce the isobaric contamination. This delay column was connected between the mobile phase mixer and the injector port. The delay column tends to have good retention and separates the PFASinterfering signals coming from either the chromatographic system or solvents. The chromatographic separation was achieved for the branched and linear isomers. The total PFOS was obtained by the sum of linear and branched PFOS, and the branched PFOS isomers were quantified against the linear PFOS standard in the hypothesis of an equimolar response.5 

Data analysis 

The data acquisition was performed by using the instrument conditions in Table 1 with the selected reaction monitoring (SRM) and processing carried out by using Thermo Scientific™ Chromeleon™ Chromatography Data System (CDS) 7.3.2.

Conclusions 

This work offers a complete workflow for the trace-level detection and quantification of PFAS concentration in fish and egg by using a Thermo Scientific™ LC-HESI-MS/MS (TSQ Quantis Plus MS). A generic approach for sample preparation, the EN-QuEChERS method, is followed by sample extraction and clean-up and then enrichment, achieving the desired sensitivity (0.1 ng/g), which is below the EU requirements, and acceptable performance (recoveries and precision). This approach enables compliance with the EU MRLs and method performance as per the EU requirements. The sample preparation and analytical conditions offer a total workflow without any interference or carryover issues during analysis. Also, this approach helps to keep the LC-MS/MS clean and operate without a breakdown, which can assist commercial labs in further increasing their high throughput.

4. Waters: Analysis of Per- and Polyfluoroalkyl Substances in Groundwater by Direct Injection Using the Benchtop Multi- Reflecting Time-of-Flight Xevo™ MRT Mass Spectrometer

• Application note
• Full PDF for download

Benefits 

• Exceptional mass measurement accuracy: Xevo MRT Mass Spectrometer achieves RMS mass accuracy consistently ≤ 0.5 ppm, effectively reducing
• Broad PFAS Profiling: Identification and quantification of 30 PFAS compounds, including linear and branched isomers, was acheived with high confidence using HRMS and data-independent acquisition (MSE).
• High sensitivity and wide dynamic range: Low limits of detection of ≤ 5 ng/L for 24 out of 30 PFAS compounds and a dynamic range spanning four orders of magnitude (1 ng/L to 10,000 ng/L) were achieved.
• Absolute quantitation using direct injection of water samples: Similar to TQ systems, the Xevo MRT Mass Spectrometer provides sufficient sensitivity to support absolute quantification of PFAS via direct injection of a 10 µL sample.
• Integrated screening and quantification platform: The waters_connect™ Software Platform, including the UNIFI™ Application, supports non-targeted screening and absolute quantification within a unified platform, enhancing productivity, and data traceability.

Sample preparation methods such as solid phase extraction (SPE) are commonly recommended prior to analysis to concentrate the analytes and facilitate sample cleanup. However, SPE can introduce bias by favoring compounds with strong adsorption affinity to the cartridges. Additionally, many PFAS-related compounds are either proprietary or result from biotransformation processes and may therefore be absent in existing databases. 2,3,4 Comprehensive identification of these compounds without reference standards would require complementary techniques like nuclear magnetic resonance (NMR)5 , which poses challenges due to its need for large sample volumes and high analyte concentrations. In contrast, liquid chromatography coupled with HRMS (LC-HRMS) direct injection methods offer an unbiased approach to sample interrogation, enabling putative identification through accurate mass measurements, isotopic patterns, fragment ion data, and mass defect filtering.6,7 HRMS, particularly when coupled with NTS workflows, offers a powerful solution for comprehensive PFAS analysis. The Xevo MRT Mass Spectrometer employs multi-reflecting time-of-flight technology to deliver high mass accuracy and sensitivity, thereby enabling the direct injection analysis of environmental samples, eliminating the need for SPE sample preparation. 

This study evaluates the performance of the Xevo MRT Mass Spectrometer for non-target screening and quantification of PFAS compounds in groundwater from the Channel Islands, demonstrating the instrument’s sensitivity, mass accuracy, and suitability for both quantitative and NTS workflows. 5 PFAS compounds were confidently identified and quantified, including perfluorobutane sulfonate (PFBS), perfluoroheptanesulfonic acid (PFHpS), perfluoropentanesulfonic acid (PFPeS), and both linear and branched isomers of perfluorooctanesulfonic acid (PFOS) and perfluorohexane sulfonate (PFHxS). Detected concentrations ranged from 13 ng/L to 1,000 ng/L. All 5 compounds are among the EU’s PFAS watch list. The amount of total PFAS quantified in the sample exceeds the European Commission proposed Environmental Quality Standards (ECEQS) and the UK guidance thresholds for groundwater.8,9 These results highlight the value of a sensitive HRMS platform like Xevo MRT Mass Spectrometer in delivering both non-target screening and quantitative analysis, supporting the transition from legacy TQ targeted methods to a more holistic view of the PFAS content.

Experimental

• LC System: ACQUITY™ Premier LC System modified with PFAS Kit (p/n: 205000588, 205000589)
• Analytical column: ACQUITY Premier BEH C18 Column, 1.7 µm, 2.1 × 100 mm, 90 Å (p/n: 186009453)
• MS system: Xevo MRT Mass Spectrometer System

Software Tools 

Data acquisition and processing were performed using the waters_connect Software Platform with the UNIFI Application, supporting both targeted quantification and non-targeted screening workflows.

Conclusion 

This study demonstrates the robust analytical performance of the Xevo MRT Mass Spectrometer for the direct quantification of PFAS in groundwater. The platform consistently delivered high mass accuracy and exceptional sensitivity, with LOD below 5 ng/L for the majority of the 30 PFAS compounds evaluated. Precision was maintained across a dynamic range spanning 4 orders of magnitude, enabling confident quantification from trace to elevated concentrations. To enhance the sensitivity of the method, the injection volume can be increased from 10 µL to 50 µL.

This approach proved effective in detecting and quantifying PFAS in real-world groundwater samples from the Channel Islands, where concentrations of PFOS and PFHxS exceeded regulatory thresholds set by EU Directives. 8,9 The integration of the waters_connect Software Platform and UNIFI Application facilitated both targeted quantification and non-targeted screening within a unified data environment, enhancing throughput, traceability, and confidence in compound identification. The Xevo MRT System is well-suited for regulatory compliance workflows and exploratory screening, offering a scalable solution for laboratories addressing the growing demand for PFAS analysis in complex environmental matrices.