News from LabRulezLCMS Library - Week 19, 2025

Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 5th May 2025? 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, Metrohm, and Shimadzu, other document by Thermo Fisher Scientific and poster by Waters Corporation / SETAC!

1. Agilent Technologies: A Direct Aqueous Injection Method for Polar Pesticides in Drinking and Nonpotable Water

• Application note
• Full PDF for download

A fast, simple method using the Agilent 6495D LC/TQ

Glyphosate is a broad-spectrum systemic herbicide and, by volume, is one of the most widely used herbicides throughout the world. It is an organophosphorus compound, specifically a phosphonate. There is a great demand for a sensitive method at the low ppb level for food, and even lower levels for environmental water analysis. Reliable sample preparation and analysis are needed to routinely analyze glyphosate and its major metabolite, AMPA. However, glyphosate and its metabolites have high polarity and chelating properties, so they can be challenging to extract from food or water and for analysis. Of concern is the affinity of these compounds to stainless steel and other surfaces, making system-to-system reproducibility difficult. This application note shows the analysis of glyphosate, plus eight other metabolites and polar pesticides. All the pesticides are phosphonates. Fosetyl aluminum, a postharvest fungicide, is also important to include, as it can be mistaken for AMPA. The analysis uses a 1290 Infinity II LC coupled to a 6495D LC/TQ for the analysis of water samples to 10 ppt.

Results and discussion

For each compound, MRM transitions, as well as collision energies and ionization polarity, were optimized using Agilent MassHunter Optimizer software. The two or three most abundant product ions per compound were selected automatically, where available. Depending on the fragmentation behavior of the individual compound, two or three MRM transitions were selected per compound. This selection was done for confident identification and confirmation by the LC/TQ. The most abundant fragments were defined as primary transitions. The chromatographic method was optimized using the Metrohm A Supp 5 column, which resulted in good separation and distribution of polar pesticides analyzed within an 18.2-minute HPLC gradient. The flow rate offered effective desolvation of target ions using the AJS ion source. The selected cycle time ensured that sufficient data points were collected across the chromatographic peaks for reproducible quantitation and confirmation of results.

Conclusion 

This application describes a highly sensitive and reproducible method for the fast and reliable quantitation of polar pesticides in water by direct injection. The dMRM method was created and optimized using Agilent MassHunter software and allows the addition of more MRM transitions if future development is required for additional compounds. An Agilent 1290 Infinity II LC coupled to an Agilent 6495D LC/TQ was used for the analysis. The 18.2-minute LC gradient method using a Metrohm A Supp 5 column offered good chromatographic separation and retention time distribution of all targets. The LC/TQ data acquisition was in dMRM mode with fast polarity switching for the most efficient use of instrument cycle time. The method performance was verified based on requirements for calibration curve linearity, instrument LOD, recovery, and precision. The results demonstrate the capability of the quantitative analytical method for polar pesticides in water by direct injection. The pesticide metabolite data are listed in the Appendix.

2. Metrohm: Anions in lithium-ion battery solvents

Determination of anions in N-methylpyrrolidone (NMP) by ion chromatography (IC)

• Application note
• Full PDF for download

N-Methylpyrrolidone (also known as N-methyl-2- pyrrolidone or NMP) is an organic solvent used to make slurry in battery manufacturing and is a key raw material for the lithium-ion battery (LIB) industry. It serves as an effective solvent for electrode binders, such as polyvinylidene fluoride, which are essential for maintaining electrode stability [1,2]. NMP is completely removed during the manufacturing process and can be recycled efficiently [3]. Global demand for NMP is high and it accounts for a substantial percentage of lithium-ion battery manufacturing costs [4]. NMP impurity analysis is crucial to assess the quality of both newly fabricated and recycled NMP. Ion chromatography (IC) with matrix elimination is a robust and reliable technique to quantify impurities in NMP in the μg/L range. Using this method, battery manufacturers can ensure the proper composition and electrochemical behavior of the electrolyte and evaluate Li-ion battery stability and safety. 

Metrohm's intelligent Preconcentration Technique with Matrix Elimination (MiPCT-ME) quantifies anions in N-methyl pyrrolidone down to the μg/L range without sample treatment or dilution steps.

RESULTS

Anions were separated and eluted from the Metrosep A Supp 7 column in less than 34 minutes under isocratic conditions. Concentrations ranged from 11−76 μg/L. The undiluted NMP sample was measured both unspiked and spiked with 30 μg/L standard ions, reaching a recovery of 90−120 % even for the very low concentrated ions (Table 2). Figure 2 shows the separation of anions in NMP. Baseline separation is achieved for the indicated anions. The chromatogram shows two early eluting peaks which were not identified. Most likely these peaks account for acetate and formate showing the enormous potential for further development and thereby allowing quantification of other relevant anions.

CONCLUSION 

The concentrations of the measured anions in NMP range from 11 to 76 μg/L. Such low analyte concentrations in combination with an interfering matrix can be challenging for chromatography. Metrohm MiPCT-ME is capable of measuring trace anions in a widely used solvent of the lithium battery manufacturing process. This analytical technique can make a major contribution to guarantee the quality, lifetime, and safety of lithium batteries. The method can easily be transferred to other relevant solvents like methanol, ethanol, acetone, and 2‐ propanol.

3. Shimadzu: Robustness Evaluation of PFAS Analysis in Soil Using LCMS -8060RX 

• Application note
• Full PDF for download

User Benefits

• The LCMS RX series, which improved stability and robustness with CoreSpray technology, showed excellent peak area repeatability when measuring PFAS at low concentrations 500 consecutive times in a soil matrix.
• Quality control samples measured for every 20 soil sample also showed good recovery rates of all PFAS remaining between 80 and 120 %.

Perfluoroalkyl and polyfluoroalkyl substances (PFAS) are widely used in various field and indistries. But their tendency to accumulate in the environment due to their highly stable structures, which resist degradation, raises concerns about their effects on humans. Therefore, the Environment Protection Agency (EPA) in the USA and the European Chemicals Agency (ECHA) have recently strengthened regulations on PFAS. Research has shown they accumulate in the bodies of humans from drinking water and in livestock, agricultural, and marine products from water and soil and that they may have a potential adverse effect on health. 1) This has created a need for highly accurate and reliable methods of measuring PFAS in not only water but also in soil and other samples with relatively complex matrixes. 

This article presents a robustness study that added 30 PFAS (Table 1) to a soil matrix and used the LCMS-8060RX to measure these PFAS in the soil matrix 500 consecutive times. The LCMS8060RX produced good results and provided a robust analytical platform for the measurement of low concentrations of PFAS in a soil matrix sample.

Robustness Evaluation with a Soil Matrix 

A soil sample was prepared using soil preparation procedures published by the National Agricultural and Food Research Organization. 2)Once the soil matrix was ready, the standard was added to create a soil sample with a target compound concentration of 0.1 µg/L in solution. The soil matrix content of the resulting soil sample was at least 90 %. This soil sample was then analyzed 500 times in succession. Fig. 4 shows the normalized peak areas of five major compounds (HFPO-DA, PFOA, PFHxS, PFNA, and PFOS) in the spiked soil sample. MRM chromatograms from the first and 500th analysis for these five compoundsare shown in Fig. 5. Good peak shapes were obtained at the start and end of the 500 analyses and peak area repeatability was good for PFAS in the soil matrix sample. Table 4 shows the %RSD and the detection limit in the soil matrix sample (based on the 500 consecutive analyses) for all 30 target PFAS. Peak area repeatability was good with %RSD below 8.5 for all target compounds. Quality control samples were also analyzed (n = 3) every 20 analyses. Recovery from the quality control samples was within 80 to 120 % for all target compounds for the duration of the 500 consecutive analyses(Fig. 6).

Conclusion 

The LCMS-8060RX performed 500 consecutive measurements of 30 PFAS added to a soil matrix sample. The results showed good peak area repeatability, good peak shapes, and good recovery. The LCMS-8060RX, which is now equipped with CoreSpray technology for improved stability and robustness, provides reliable analysis even with samples containing numerous impurities,such assoil matrix samples.

4. Thermo Fisher Scientific: High Force Research streamlines operations, ready for growth with Chromeleon CDS 

• Other document (Case study)
• Full PDF for download

High Force Research, Ltd. (HFR) is an independent contract research organization (CRO)/contract development and manufacturing organization (CDMO) that exists to solve client chemistry challenges across multiple scientific industries. Operations are carried out in the company’s purpose-built manufacturing facility in Bowburn, UK, which includes two ISO Class 8 laboratories in which multi-stage chemical synthesis according to cGMP standards is carried out. Further capacity at NETPark, Sedgefield, UK, allows initial project research and development activities to be undertaken. The company fulfills client needs offering dependable, flexible, and affordable high-quality services and products. Toward this goal, the company continues to invest in new equipment, techniques, and processes. Thermo Scientific™ Chromeleon™ Chromatography Data System (CDS) is one such investment that has dramatically streamlined operations while enhancing the quality of work, ensuring HFR is prepared for continued business growth and competitiveness.

Ease of use influenced the decision 

Chromeleon CDS is designed to simplify tasks, reduce errors, and improve lab efficiency. Before HFR finalized its decision to make the transition, Thermo Fisher Scientific staff provided training so users could experience the benefits firsthand. Jude Humphrey, Senior QC Scientist at High Force Research, said, “The technical team at Thermo Fisher initiated an on-site training course, allowing us to gain user experience early. The simplicity, user-friendly nature of the software and assurance of compliance persuaded us to transition, and it has been of great benefit to the department since. As an apprentice at the time of install, it was great to see HFR investing in such great analytical software.” 

eWorkflow procedures increase efficiency, reduce errors, and make analyses easier 

Chromatography workflows are very similar. Samples are injected, separations are performed, signals are captured, and results are generated. Where they differ is in the method details, such as the instrument conditions, injection sequence requirements, and the way results are calculated. These differences create complexity for operators, reducing their efficiency and increasing the risk of error. 

Chromeleon CDS solves these problems with an eWorkflow procedure, which contains a set of rules to capture all the unique aspects of a chromatography workflow and guides the operator through a minimal number of choices to create the correct sequence. The operator simply selects an instrument, specifies the number of samples and the software runs the sequence, processes the data, and generates the results, ensuring that the lab’s procedural rules and guidelines are followed. 

HFR uses eWorkflow procedures to increase the number of samples that can be measured per day. Scott Little, Laboratory Instrumentation Specialist at High Force Research explained, “We used to input data manually and it would take 20 minutes to start the sequence. Now, sequences are set up and ready for review within a few minutes.” Little expanded on the benefits of an eWorkflow procedure, “We’ve added new members into our QC team recently who have all agreed that the Chromeleon eWorkflow procedures are far simpler to use.”

5. Waters Corporation / SETAC: SETAC: MULTI-RESIDUE METHOD FOR PESTICIDES, PHARMACEUTICALS AND PERSONAL CARE PRODUCTS (PPCPs) IN WATER BY DIRECT INJECTION USING UHPLC-MS/MS

• Poster
• Full PDF for download

In recent years, there has been increasing concern about the presence of pesticides, pharmaceutical and personal care products (PPCPs) in water bodies throughout the world. The effect of these emerging contaminants on human health and their potential impact on the environment is not yet fully understood. As concerns continue to grow, many government agencies around the world are funding studies to assess if PPCPs can cause harmful ecological effects.

Many publications have shown that PPCPs are present at parts-per-trillion (PPT) levels in rivers and streams highlighting the need for methods that are able to detect compounds at these trace levels. In addition to the low-level detection of these compounds, a major analytical challenge for analysis lies in the wide chemical diversity of compound classes and structures, examples of which are shown in Figure 1. Furthermore, the complexity of the water samples requiring analysis can be very diverse.

The recast Drinking Water Directive entered into force in January 2021 and is the EU's main law on drinking water.  It commits EU member states to achieve a common goal of 'good chemical and good ecological status' for all ground and surface waters.  It concerns the access to, and the quality of water intended for human consumption in order to protect human health and ensures an integrated approach to water management, respecting the integrity of entire ecosystems, and by regulating individual pollutants and setting corresponding regulatory standards.

Another layer committing to water quality is a watch list mechanism which was established to improve the information around identifying and monitoring emerging substances of concern. Member states must monitor the listed substances at least once a year for up to four years. The watch list was established in 2015, and updated most recently in 2022, and on these lists are 
increasing amounts of pharmaceuticals as well as personal care products. 

Instrumental conditions:

• UHPLC System: ACQUITY^TM Premier System 
• MS System: Xevo^TM TQ Absolute MS
• Column: ACQUITY Premier HSS T3 Column 1.8 µm, 2.1 x 100

RESULTS AND DISCUSSION

Sample diluent influence on peak shape and sensitivity: for maximum comprehensive performance, a careful balance is required when simultaneously analysing for a variety of compounds with a range of physiochemical properties.

• Highest overall sensitivity was seen when a higher aqueous sample composition was used, and this was essential for compounds such Acephate and Trimethoprim.
• Compounds that are less soluble in water such such as β-Estradiol, Erythromycin and Phenol require an elevated organic content but ratios of 10 % acetonitrile and above in the injected sample were not optimum for the majority, especially when this composition created solubility issues and distortion in chromatography for the earlier eluting compounds.

Desolvation and source temperature investigation: Some compounds were difficult to detect at 10 ng/L. Upon troubleshooting, lower desolvation temperatures were studied and compounds such as MCPB gave a better response at 300°C compared to compounds such as Diazepam which favoured 650°C.  Soft transmission was also investigated, and sensitivity improvements were seen in labile compounds such as Dicamba that benefit from a gentler StepWave environment.

There are many benefits to running multi-residue screening methods including streamlined workflows and more efficient Figure 3. Peak response of Metconazole over a range of matrix asset utilisation.  These methods can also be further customised to accommodate challenging compounds with specific types demonstrated at blank, 10 ng/L and 100 ng/L levels. preferences to parameters such as sample composition and source temperatures that deviate from that of the majority. S/N ratio >100 at 10 ng/L for all matrices.

CONCLUSIONS

• A sensitive multi-method has been developed to determine residues at concentrations as low as 5 ng/L in drinking and surface water without the need for lengthy clean-up or concentration steps.
• 200 compounds can be quantified at ≥100 ng/L and 181 compounds at ≥10 ng/L, this exceeds the EU Drinking Water Directive requirements, and the proposed levels in the Water Framework watch list and
• Environmental Quality Standard Directive for many relevant compounds. 
• This method offers sufficient chromatographic retention, separation and detection for compounds in matrix-matched calibrants.
• Multi-methods are a useful screening tool and can be customised to accommodate for challenging compounds where optimum conditions may deviate from the overall suite.  You can find many out of the box solutions and resources on the Waters website which can be adapted or used as starting points of reference.