We, 16.9.2026 | Original article from: LabRulezLCMS Library
This week we bring you application notes by Agilent Technologies and Waters Corporation and posters by Shimadzu / AOAC and Thermo Fisher Scientific / Metabolomics!News from LabRulezLCMS Library - Week 38, 2026
LabRulez / AI: News from LabRulezLCMS Library - Week 38, 2026
Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 14th September 2026? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!
👉 SEARCH THE LARGEST REPOSITORY OF DOCUMENTS ABOUT LCMS AND RELATED TECHNIQUES
👉 Need info about different analytical techniques? Peek into LabRulezGCMS or LabRulezICPMS libraries.
This week we bring you application notes by Agilent Technologies and Waters Corporation and posters by Shimadzu / AOAC and Thermo Fisher Scientific / Metabolomics!
1. Agilent Technologies: Improved Analysis of Intact Proteins and Peptides by Reversed Phase HPLC Using the Altura Ultra Inert PLRP-S 1000 Å Column
- Application note
- Full PDF for download
Intact protein analysis by reversed-phase HPLC (RP-LC) plays a vital role in quality control for determining the purity of biotherapeutic proteins and improving the identification and quantification of critical quality attributes (CQAs). When coupled to mass spectrometric detection, it allows several things. First, an accurate molecular weight to be determined; second, the identification of variants containing post‑translational modifications (PTMs) and proteoforms; and third, the antibody-to-drug ratio in the case of antibody drug conjugates (ADCs).1
While many RP columns have a silica-based stationary phase to enable broader functionalization, residual acidic silanol groups can interact with basic residues in peptides, proteins, and other biomolecules, resulting in significant peak tailing and loss of chromatographic performance. Polystyrene/divinyl benzene (PS/DVB)-based stationary phases do not contain unintentional adsorption sites. Additionally, larger pore sizes have demonstrated superior chromatographic efficiency through less diffusion restriction into the pore volume and reduction of size exclusion mechanisms, degrading the RP-based selectivity.2
Wide-pore columns like the PLRP-S 1000Å provide an exceptional performance with biomolecules, particularly when combined with MS-compatible mobile phase conditions such as formic acid.
Even so, some undesirable interactions may be observed when using stainless-steel column hardware. Molecules that possess multiple negative charges (including oligonucleotides, phosphorylated peptides and proteins, and proteins with low isoelectric point in general) may interact with exposed metal ions in stainless-steel column hardware. This can result in the need to "deactivate" new columns by making multiple injections of sample until reproducible results are obtained.
This application note demonstrates how the new Altura Ultra Inert PLRP-S 1000 Å column serves as an effective high-performance workhorse in the analysis of large, intact biomolecules. Developed to effectively block nonspecific adsorption during separations, Ultra Inert technology reduced passivation time 15-fold and delivered ~ 10% greater recovery for mAbs and ADCs and up to 100% for smaller biomolecules like GLP-1s and small globular proteins.
Experimental
Instrumentation
- Agilent 1290 Multicolumn thermostat (G7116B)
- Agilent 1290 Bio multisampler (G7137A)
- Agilent 1290 Bio high-speed pump (G7132A)
- Agilent 6530 Q-TOF MS system
Software and data processing
- Agilent MassHunter Workstation, version 10.1
- Agilent MassHunter Qualitative Analysis software, version B.07
- Agilent DAR calculator, version B01.01
Conclusion
Intact mass analysis is a critical methodology to determine the purity of biotherapeutics, but due to the chemical complexity of these analytes, nonspecific adsorption from both silica stationary phase and metal column housing can be highly deleterious to separation performance. The Agilent Altura PLRP-S 1000 Å provides a highly robust solution combining the chemical inertness of polystyrene/divinyl benzene stationary phase with the ultra‑inert Altura hardware to dramatically reduce deleterious peak tailing and increase recovery for biotherapeutics. Herein, we see that analyte recovery for both mAbs, ADCs, GLP-1s, and globular proteins are improved versus stainless-steel, while the Altura PLRP-S column is capable of monitoring both quality critical PTMs such as loss of drug payload as well as accurate quantification of ADC DAR.
2. Shimadzu / AOAC: Ambient Ionization Mass Spectrometric Detection of Aroma Released During Gummy Chewing
- Poster
- Full PDF for download
In recent years, there has been growing interest in evaluating flavor using analytical instrumentation, and retronasal aroma during eating is important. In this study, we proposed an analytical method for measuring retronasal aroma during eating using a single quadrupole mass spectrometer (LCMS-2050) equipped with an ambient ion source (Dark Current Discharge Ionization : DCDI) without pretreatment.
Conclusion
We developed a simple method for evaluating retronasal aroma using a single quadrupole mass spectrometer with an ambient ionization source. Ambient ionization mass spectrometry (DCDI LCMS-2050) can rapidly measure retronasal aroma during eating. Time-resolved visualization reveals texture-dependent differences in aroma release dynamics among grape-flavored gummies. It is also expected to lead to the visualization of the marriage between food and alcohol. This approach may facilitate objective evaluation of flavor release during consumption and support product formulation and sensory science.
3. Thermo Fisher Scientific / Metabolomics: Orbitrap Tribrid Real-Time similarity searching of mzCloud library for guided unknown characterization of tea metabolites
- Poster
- Full PDF for download
In metabolomics LC-MS data is processed to generate a list of detected compounds with a level of confidence in the identity of each compound based on reference data. This reference data may be spectral data and/or retention time information stored in a library or in silico information generated based on fragmentation rules. These annotated compounds are then used to evaluate biological changes between normal and disease states. One of the major hurdles between collected data and coming to reliable conclusions about the biology is the limited number of standards available to build these libraries. It is common to have hundreds to thousands of compounds detected in a data set that have no library match because there is no commercial standard for that unknown. Instead, there is just an elemental formula and matches from a structure database. These structures can be compared using in silico fragmentation techniques to determine which isomers plausibly match the data. MSn data can help reduce the list of possible matches, but without guidance, ddMS3 may not consistently collect data on the most relevant MS2 fragments to accomplish this.
In this work we use Real-Time Library Search (RTLS) in a ddMS3 method to help guide the instrument in real time to collect the most relevant data for confirming structural matches.
Materials and methods
Test method(s)
1 μL aliquots were analyzed using a Thermo Scientific Hypersil GOLD column on a Thermo Scientific Vanquish Horizon UHPLC coupled with a Thermo Scientific Orbitrap IQ-X Tribrid mass spectrometer and Thermo Scientific Orbitrap Tribrid Apex Small Molecule mass spectrometer. RTLS was run in similarity mode using the offline version of Thermo Scientific mzCloud library.
Data analysis
Data was processed using Thermo Scientific Freestyle and Compound Discoverer software.
Conclusions
Tea extracts were analyzed using RTLS to find the most similar compound in the offline version of the mzCloud library and trigger MS3 on the highest m/z matching and non-matching fragments. This provided additional information to confirm similarity and provide insight on shared sub-structures to the library compound as well as additional information on the portions of the structure that were unique to the experimental compound.
4. Waters Corporation: The Quantification of Glucagon-like Peptide Receptor Agonists (GLP-1 RAs) in Plasma Extracts Using Capillary UHPLC-MS/MS
- Application note
- Full PDF for download
Benefits
- 10-fold improvement in sensitivity
- Robust methodology for >500 injections
- Reduced solvent usage
- No need for complex nanospray MS source
Obesity represents, perhaps, the largest healthcare challenge, affecting approximately 1.3 billion people globally, contributing to a diverse range of chronic diseases e.g., metabolic, cardiovascular, muscular skeletal, and multiple cancers.1 The rapid growth of obesity and associated global healthcare costs means there is a critical need for safe effective treatments.2 GLP-1 RA initially developed for type 2 diabetes have shown clinical efficacy as weight loss medicines.2 These drugs mimic the action of gut hormones which regulate energy metabolism and appetite by interacting with the hypothalamus, brain stem etc., resulting in weight loss. The success of these GLP-1 RAs is associated with their on-target efficacy and long duration of action,3 e.g., half-life of semaglutide t½ = 160–180 hours3 cf. 1–2 minutes for human GLP-1.4 The long half-life of these GLP-1 RAs results in low, sub ng/mL plasma levels 48–72 hours after dosing, which requires a high sensitivity bioanalytical assay to accurately quantify the plasma concentrations5 and define the PK elimination phase.
Tandem quadrupole electrospray ionization (ESI) mass spectrometry, operated in multiple reaction monitoring (MRM) mode, coupled with 2.1 mm ID ultra-high performance liquid chromatography (UHPLC) is the dominant technology platform for quantitative bioanalysis. As next generation GIP/GLP-1 RAs promise to be more potent, resulting in lower dosing levels and plasma levels, there is a need for more sensitive analytical methodologies to support drug metabolism and pharmacokinetics (DMPK) studies. Capillary/nano LC-MS/MS have been extensively employed in proteomics to increase analytical sensitivity and enable small volume analysis.5 This sensitivity increase is derived from a combination of the superior ionization efficiency at low flow rates and analyte concentration due to the smaller column geometry.6 This application note illustrates the use of capillary (300 µm) UHPLC-MS/MS (MRM) to affect 10-fold improvement in GLP-1-RAs peak response in human plasma extracts, and detection limits as low as 20 pg/mL for tirzepatide.
Experimental
Capillary chromatography was performed on an ACQUITY™ M-class Chromatography System connected to a Xevo™ TQ-XS Mass Spectrometer (Waters Corporation, Wilmslow, UK) equipped with a microflow probe. Capillary UHPLC separations were performed on an ACQUITY Premier Peptide CSH C18, 300 Å, 1.7 µm, 300 µm x 50 mm Chromatography Column.
MS analysis was performed at unit mass resolution using positive ESI at a capillary voltage of 3.0 kV, with a cone voltage of 32 V. The cone gas flow, desolvation gas flow, and desolvation gas temperature were 50 L/h, 300 L/h and 600 °C, respectively. The data were analyzed using waters_connect™ Software, and MRM transitions used to monitored for the MS/MS detection of liraglutide, exenatide, semaglutide and tirzepatide were as follows: liraglutide 938.7 → 1128.4, exenatide 838 → 396, semaglutide 1029.2 → 1238.1, and tirzepatide 1204.2 → 396.2. Data analysis was performed using MS Quan Software.
Conclusion
Bioanalysis plays a critical role in the drug discovery and development processes providing data to support DMPK, safety assessment, pharmacokinetics/pharmacodynamics (PK/PD), and clinical development studies. The development of new, more potent medicines, innovating delivery platforms, micro sampling, and new approach methodologies (e.g., organ on a chip) all require greater levels of analytical sensitivity. Capillary scale UHPLCMS/MS offers the potential to improve bioanalytical detection limits up to 10-fold, without the need for long analysis times and complex operation associated with nano spray LC-MS/MS. Here, the application of capillary (300 µm ID) scale chromatography for the detection of GLP-1 RAs in human plasma extracts at levels as low as 20 pg/mL with robustness over 500 injections was demonstrated.
