Folin-Ciocâlteu, RP-HPLC (reverse phase-high performance liquid chromatography), and LC-MS (liquid chromatography-mass spectrometry) provide complementary information for describing cider (Malus spp.) apple juice

Mineral oil hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons (MOAH) have moved from niche laboratory concerns to central topics in food safety, regulatory policy, and analytical science.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_MOSH_MOAH_Analysis_Technology_Regulation_and_the_Road_Ahead_2_f79d1ed4ee.jpg" alt="Thermo Fisher Scientific: MOSH/MOAH Analysis - Technology, Regulation, and the Road Ahead" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_MOSH_MOAH_Analysis_Technology_Regulation_and_the_Road_Ahead_2_f79d1ed4ee.jpg 245w," sizes="100vw" width="1200" height="600">These compounds—originating from sources such as environmental pollution, lubricants, processing aids, and migration from packaging materials—can enter the food chain and accumulate in both raw ingredients and finished products.<ul><li>MOSH are saturated hydrocarbons, which can accumulate in human tissues but are generally considered of lower toxicological concern.</li><li>MOAH contain aromatic rings, including polycyclic structures; certain high-ring MOAH are suspected to have genotoxic and carcinogenic properties, making them a priority for regulatory control.</li></ul>Growing public awareness, coupled with scientific research and legislative pressure, is pushing the food industry toward more robust monitoring and testing.<h4>The evolving regulatory landscape</h4>For years, MOSH/MOAH testing lacked consistent rules across jurisdictions. Different countries and organizations used their own limits and methods, creating confusion for global supply chains. Over the last decade, several important steps have brought the field closer to harmonization:<h5>2017 – EN 16995 (CEN)</h5><ul><li>The first standardized method for quantifying MOSH and MOAH in foodstuffs using LC-GC-FID technology.</li></ul><h5>2023 – JRC Guidelines v2</h5><ul><li>Expanded scope to include food contact materials, with refinements in sample preparation and measurement.</li></ul><h5>2024 – ISO 20122:2024</h5><ul><li>A dedicated standard for fats and oils, integrating clean-up procedures alongside LC-GC-FID to improve accuracy.</li></ul><h5>Ongoing – EURL/ISO initiatives</h5><ul><li>Collaborative efforts including interlaboratory studies, method refinement, and the potential adoption of GC×GC–MS for enhanced selectivity and molecular insight.</li></ul>A major development still in progress is the EU’s draft regulation for maximum residue limits for MOAH. Expected after 2024, it could introduce strict, enforceable thresholds across the single market, forcing a step-change in testing capacity and method validation.<h4>Analytical challenges in MOSH/MOAH testing</h4>Although standardized methods exist, MOSH/MOAH analysis is not a straightforward “plug-and-play” process. Laboratories face several persistent challenges:<h5>Complex sample matrices</h5><ul><li>Edible oils, processed foods, and packaging extracts contain a mix of natural compounds such as carotenoids, terpenes, and squalene. These can overlap chromatographically with MOSH/MOAH peaks, making quantification more difficult.</li></ul><h5>False positives and overestimation</h5><ul><li>Non-MOSH/MOAH compounds can mimic the target analytes, especially in less selective detection modes, leading to inflated results.</li></ul><h5>Low detection limits</h5><ul><li>Many regulations demand quantification in the low mg/kg range. Achieving this requires both clean sample extracts and highly sensitive analytical instrumentation.</li></ul><h5>Reproducibility across labs</h5><ul><li>Differences in sample handling, clean-up techniques, and instrument tuning can lead to significant interlaboratory variation, complicating compliance and enforcement.</li></ul>These challenges mean that achieving accuracy, repeatability, and regulatory compliance requires optimized workflows that combine precision sample preparation with advanced chromatographic techniques.<h4>A modern workflow: from sample to result</h4>State-of-the-art MOSH/MOAH workflows are typically built around an LC-GC-FID platform supported by automated sample preparation. This integration reduces human error, improves throughput, and delivers reproducible results.<h5>Step 1: saponification</h5>Fats can overwhelm LC columns, limiting injection volumes and obscuring MOSH/MOAH peaks. Saponification removes these fats, enabling:<ul><li>Larger injection volumes.</li><li>Clearer chromatographic baselines.</li><li>Lower quantitation limits without compromising peak integrity.</li></ul><h5>Step 2: epoxidation</h5>For MOAH analysis, carotenoids, terpenes, and other naturally occurring compounds must be removed. Epoxidation—now most commonly performed using PFA reagents—cleans up these interferences while maintaining the integrity of aromatic hydrocarbons.<h5>Step 3: evaporation and concentration</h5>By concentrating the hexane extract after saponification, laboratories can:<ul><li>Achieve lower detection limits.</li><li>Improve recovery from challenging matrices such as palm stearates.</li></ul><h5>Step 4: alu oxide clean-up</h5>Adding an alumina oxide clean-up step can substantially improve MOSH quantification in the presence of n-alkanes. This step is matrix-dependent and should be avoided if n-alkanes are absent to prevent bias.<h5>Step 5: fraction collection for advanced analysis</h5>When further identification is required—such as distinguishing between mineral oil hydrocarbons and plasticizers (POSH)—fraction collection followed by GC×GC-MS analysis can provide a deeper chemical fingerprint.<h4>Beyond “how much”: the role of GC×GC-MS</h4>Conventional LC-GC-FID answers the fundamental question: How much MOSH or MOAH is present in the sample?But it cannot always explain the source or the chemical nature of the detected hydrocarbons.This is where GC×GC-MS adds value:<ul><li>Source tracing – Identifies whether contamination originates from packaging, processing equipment, or environmental exposure.</li><li>Structural characterization – Distinguishes low-ring MOAH (less concerning) from high-ring MOAH (more concerning toxicologically).</li><li>False positive elimination – Confirms the identity of suspect peaks, reducing the risk of misreporting.</li></ul>By combining FID for precise quantification with MS for molecular identification, GC×GC-MS enables both regulatory compliance and investigative insight.<h4>The role of chromeleon software in MOSH/MOAH analysis</h4>Modern analytical systems rely heavily on software to ensure speed, traceability, and accuracy. Chromeleon software offers:<ul><li>Unified control of LC, GC, and sample preparation hardware from a single interface.</li><li>Automated workflow setup – In just a few clicks, vial positions, injection types, and processing methods are assigned.</li><li>Integrated quality checks – Internal standard verification and retention time monitoring are built-in.</li><li>Customizable, regulatory-ready reports – Exportable in PDF, CSV, XML, and other formats without external spreadsheets.</li></ul>The result is streamlined operation, reduced analyst intervention, and consistent compliance with good laboratory practice (GLP) requirements.<h4>Looking ahead</h4>With upcoming EU regulations likely to set strict MOAH limits, and with continued scientific scrutiny of MOSH accumulation, the demand for high-quality testing will only increase.Key trends shaping the future of MOSH/MOAH analysis include:<ul><li>Wider adoption of GC×GC-MS for confirmation and source identification.</li><li>Greater automation in sample preparation to increase throughput and reproducibility.</li><li>Regulatory harmonization, which will help reduce discrepancies between international markets.</li><li>Lower detection thresholds, requiring even more sensitive and selective methods.</li></ul>For laboratories, the challenge is twofold: staying ahead of compliance requirements while building capacity for high-volume, high-accuracy testing. Those that invest in robust, automated, and harmonized workflows now will be better positioned to meet both current and future demands.<blockquote>Acknowledgement: Jorn Hofsteenge, SampleQ, Breda, The Netherlands</blockquote>

MOSH/MOAH Analysis: Technology, Regulation, and the Road Ahead

In the realm of pharmaceutical manufacturing, the detection and analysis of perfluoroalkyl substances (PFAS) have become a growing source of concern due to their persistence and potential health risks. The latest application note from SGS presents a groundbreaking approach to PFAS screening and quantification, leveraging the capabilities of the Thermo Scientific Vanquish UHPLC and Orbitrap Exploris 120 high-resolution mass spectrometer (HRAM-MS).This application note demonstrates the sensitive detection and identification of both known and unknown PFAS compounds in extracts from common pharmaceutical manufacturing and packaging materials. By employing a combined targeted and non-targeted approach, the study highlights how the integration of the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=40&amp;manufacturerItems=3">Vanquish UHPLC </a>and <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/403">Orbitrap Exploris 120 HRAM-MS</a> can revolutionize PFAS analysis.<h5>The key benefits of this method include:</h5><ul><li>Combined Targeted and Non-Targeted Screening: One injection allows for both the quantitation of specific PFAS compounds and the screening for additional, previously unidentified PFAS contaminants.</li><li>Enhanced Analytical Confidence: The use of a PFAS analysis kit and delay column minimizes background PFAS interference, thereby increasing the reliability of the results.</li><li>Regulatory Compliance: Utilizing <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/76">Chromeleon CDS </a>ensures that the data acquisition and quantitative analysis comply with 21 CFR Part 11, a crucial requirement for pharmaceutical applications.</li><li>High Sensitivity and Specificity: The method provides unequivocal identification and quantification of PFAS compounds down to sub-ppb levels.</li></ul>The study underscores the efficacy of the Orbitrap Exploris 120 HRAM-MS in providing a comprehensive solution for PFAS analysis. By integrating both targeted and non-targeted approaches, this method not only enhances the detection sensitivity and specificity but also expands the scope of detectable PFAS contaminants. This dual approach is a significant advancement in extractables screening, offering pharmaceutical manufacturers a robust tool for ensuring the safety and compliance of their products.<blockquote><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_003388_pb_pfas_orbitrap_exploris_120_extractables_an003388_na_en_dd0e422ebd.pdf">Download our joint application note</a> to learn about Comprehensive PFAS screening in pharmaceutical packaging and medical devices by LC-HRAM-MS</blockquote>This collaboration between SGS and Thermo Fisher Scientific illustrates a leap forward in PFAS screening technology, promising higher quality results, increased confidence, and regulatory compliance in the pharmaceutical manufacturing process.<blockquote>Jon BardsleyJon Bardsley is a Market Development Manager for Pharma and Biopharma with Thermo Fisher Scientific.</blockquote>

Advancing PFAS Detection in Pharmaceutical Manufacturing with LC-HRAM-MS

Coupling Liquid Chromatography to Orbitrap Isotope Ratio Mass Spectrometry: Overcoming Isotope Effects of Chromatography and Amount-Dependency by Peak Homogenization

<h3>Introduction</h3>MOSH and MOAH analysis remains one of the most challenging and closely scrutinized areas in food and packaging safety testing. With evolving regulations, updated standards such as ISO 20122:2024, and increasing expectations for data quality and throughput, laboratories are under constant pressure to deliver accurate, reproducible, and compliant results.To help laboratories navigate this complexity, Thermo Fisher Scientific and SampleQ hosted an <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/webinar/5817">Ask the Experts</a> session focused entirely on MOSH/MOAH analysis using LC-GC-FID and GC×GC-FID/MS workflows. Analytical specialists addressed real-world questions from scientists and lab managers—covering sample preparation, quality control, internal standards, data processing, and system selection.This blog brings together the most important expert insights and practical guidance from that session, providing a valuable reference for anyone responsible for MOSH/MOAH testing.<h3>Why MOSH/MOAH analysis is so challenging</h3>Mineral Oil Saturated Hydrocarbons (MOSH) and Mineral Oil Aromatic Hydrocarbons (MOAH) are complex mixtures rather than single compounds. Their broad carbon number distribution, co-elution with natural matrix components, and sensitivity to sample preparation steps make analysis technically demanding.At the same time, regulatory scrutiny continues to increase, particularly for MOAH due to potential carcinogenicity. Laboratories must therefore balance:<ul><li>Regulatory compliance</li><li>Robust quantification</li><li>Reliable identification of markers</li><li>High sample throughput</li></ul>The Ask the Experts session focused on how modern, integrated workflows can help labs meet all these demands.<h3>Are MOSH/MOAH sample preparation steps matrix dependent?</h3>One of the most common questions from laboratories is whether MOSH/MOAH sample preparation can be standardized across all matrices.The expert answer: it depends on the matrix—and validation is essential.For fats and oils, certain steps are always required:<ul><li>Saponification</li><li>Epoxidation</li></ul>However, alumina (Alox) clean-up is matrix dependent. Practical comparisons show that:<ul><li>Some matrices benefit from Alox clean-up</li><li>Others may suffer from analyte loss if Alox is applied unnecessarily</li></ul>For non-fat matrices, sample preparation must be carefully selected, and method validation is critical to ensure accuracy and recovery.Key takeaway: There is no universal sample prep recipe for MOSH/MOAH. Laboratories must adapt workflows to their matrices while staying aligned with ISO 20122 guidance.<h3>Managing multiple sample prep options in a busy lab</h3>Many labs analyze a wide variety of matrices daily. Running different sample prep workflows manually can be time-consuming and error-prone.Experts highlighted how software-driven workflows help solve this challenge:<ul><li>Multiple sample prep methods can run in the same sequence</li><li>Optional steps (such as Alox clean-up) can be toggled per sample</li><li>Overlapping methods maximize instrument utilization</li><li>Analysts only need to weigh samples and load reagents—automation handles the rest</li></ul>E-workflows simplify method selection and ensure the correct prep is applied every time, reducing training requirements and minimizing human error.For lab managers, this translates into:<ul><li>Higher throughput</li><li>More consistent results</li><li>Less hands-on time for skilled analysts</li></ul><h3>Which internal standard is best for MOAH quantification?</h3>Internal standard selection is critical for reliable MOAH quantification. The experts addressed this directly, referencing extensive comparative studies.Three candidates were evaluated:<ul><li>1-Methylnaphthalene (1-MN)</li><li>2-Methylnaphthalene (2-MN)</li><li>Tert-butylbenzene (TBB)</li></ul>Based on repeatability, reproducibility, recovery, and accuracy, TBB emerged as the clear winner and is now specified in ISO 20122.<h3>Why TBB?</h3><ul><li>More stable behavior across sample prep steps</li><li>Better overall performance in routine analysis</li><li>Improved consistency between laboratories</li></ul>For labs updating or validating MOSH/MOAH methods, aligning internal standards with ISO recommendations is strongly advised.<h3>How should ISO criteria for internal standard recovery be interpreted?</h3>A frequent point of confusion is how to interpret internal standard recovery limits defined in ISO methods.The experts clarified an important distinction:<ul><li>Recovery criteria apply to direct injections of internal standards</li><li>All components should fall within 90–110% of the quantification standard</li><li>Perylene is allowed a slightly broader range (80–100%)</li></ul>In real samples, internal standard recoveries can vary due to legitimate chemical effects:<ul><li>Epoxidation may reduce perylene</li><li>Alox clean-up can reduce cholestane</li><li>Saponification may affect methylnaphthalenes</li><li>Some hydrocarbons may be naturally present in the matrix</li></ul>Bottom line: Deviations in sample recoveries do not automatically indicate method failure. Understanding the chemistry behind each step is key to correct interpretation.<h3>Do you still need LC-GC-FID if you have GC×GC-MS?</h3>As GC×GC-MS becomes more accessible, many labs ask whether it can replace LC-GC-FID entirely.According to current EURL guidance:<ul><li>GC×GC is primarily intended for identification, not quantification</li><li>Quantification using GC×GC-FID is technically possible but not yet recommended</li><li>LC-GC-FID remains the reference technique for MOSH/MOAH quantification</li></ul>GC×GC-MS plays a complementary role by:<ul><li>Confirming the presence of aromatic markers and MOSH markers like hopanes, steranes and particular naphthenes</li><li>Differentiating mineral oil components from biogenic interferences</li><li>Supporting risk assessment and regulatory discussions</li></ul>For now, the most robust approach is a combined workflow:<ul><li>LC-GC-FID for quantification</li><li>GC×GC-FID/MS for confirmation and identification</li></ul><h3>Using specific ion masses for marker identification in GC×GC</h3>GC×GC-MS offers powerful identification capabilities when used correctly.Experts highlighted that:<ul><li>The EURL provides recommended ion masses for key MOAH subgroups</li><li>Selective ion chromatograms (SICs) can be generated for markers such as:<ul><li>Naphthalenes</li><li>Hopanes</li><li>Steranes</li><li>DIPNs</li></ul></li></ul>Using multiple SIC views alongside total ion chromatograms (TICs) allows:<ul><li>Faster confirmation of markers</li><li>Clear differentiation from interferences</li><li>More confident reporting of MOAH presence</li></ul>This approach significantly improves interpretability compared to relying on TICs alone.<h3>How is GC×GC-MS data processed efficiently?</h3>One concern with GC×GC-MS is the perceived complexity of data handling.The experts demonstrated a streamlined workflow:<ul><li>Instrument control and acquisition remain in <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/products/76">Chromeleon S/W</a></li><li>Raw data are automatically exported for GC×GC processing</li><li>Templates are applied to identify known marker regions</li><li>Results are organized by vial, run, and detector channel</li></ul>This structured approach ensures:<ul><li>Consistent marker identification</li><li>Reduced manual data handling</li><li>Faster reporting despite large data volumes</li></ul>For labs new to GC×GC, standardized templates and automated processing significantly reduce the learning curve.<h3>Comparing FID and MS chromatograms: why it matters</h3>Accurate comparison between FID and MS data is critical when combining quantification and identification.Thanks to synchronized acquisition:<ul><li>FID and MS chromatograms are recorded simultaneously</li><li>First-dimension retention times are identical</li><li>Second-dimension alignment is highly accurate</li></ul>This enables:<ul><li>Direct correlation of quantified FID regions with identified MS markers</li><li>Greater confidence in results</li><li>Stronger defensibility during audits and regulatory review</li></ul><h3>Why use a single quadrupole MS instead of TOF-MS?</h3>Time-of-flight MS is often associated with GC×GC due to its high scan speed. However, experts explained why <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/products?category=19&amp;manufacturerItems=3">single quadrupole MS </a>can be a practical alternative.By optimizing scan speed:<ul><li>Single quadrupole MS can capture sufficient data points</li><li>Complexity and cost are reduced</li><li>Compliance with EURL guidance is maintained</li></ul>For many routine MOSH/MOAH applications, this offers an excellent balance between performance and operational efficiency.<h3>Is offline sample preparation better for GC×GC?</h3>There is no universal answer—workflow design depends on laboratory needs.Integrated systems offer:<ul><li>Minimal user intervention</li><li>Reduced risk of contamination</li><li>Streamlined operation</li></ul>Separate systems allow:<ul><li>Parallel operation</li><li>Higher overall throughput in some labs</li></ul>The best choice depends on sample volume, staffing, and analytical priorities.<h3>Final thoughts: Building a future-proof MOSH/MOAH workflow</h3>MOSH/MOAH analysis is evolving rapidly, but some principles remain constant:<ul><li>Robust quantification still relies on LC-GC-FID</li><li>GC×GC-FID/MS adds critical confirmation power</li><li>Automation and software workflows reduce complexity</li><li>Alignment with ISO and EURL guidance ensures defensibility</li></ul>By combining expert knowledge with integrated technology, laboratories can confidently meet today’s regulatory requirements—while preparing for what comes next.If you’re looking to optimize or future-proof your MOSH/MOAH analysis, the insights from this Ask the Experts session provide a strong foundation for success.<ul><li>Watch the free webinar on demand: <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/webinar/5817">Talk to an expert about our barrier-free analysis of MOSH/MOAH</a></li><li>Visit us on&nbsp;<a target="_blank" rel="noopener noreferrer" href="https://www.linkedin.com/showcase/72003132/admin/dashboard/">LinkedIn:</a>&nbsp;#automation #MOSH/MOAH</li></ul><blockquote><h4>Petra Gerhards</h4>Petra Gerhards, Dipl-Ing, is Regional Marketing Manager of GC and GC-MS for EMEA at Thermo Fisher Scientific. She has more than 29 years of experience in the fields of GC-MS, SPE and LC-MS. Since joining the regional team she has contributed to workflow solutions combining vials and closures with SPE solutions, GC-MS and LC-MS. She works with KOL's on data for regional specific marketing campaigns, organizes in-house seminars and works on customer specific solutions. Her main expertise is in the field of doping and drugs-of-abuse analysis.</blockquote>

Ask the Experts: Practical Answers to Today’s Biggest MOSH/MOAH Analysis Questions

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