Application Notes: General

HPLC method development of structurally similar components is an arduous task to undertake. Understanding the principles of liquid chromatography and applying them in a structured, streamlined approach not only speeds up the method development process but also provides traceability for the activity in an easy-to-follow format. Having a structured protocol for method development also allows novice users to develop methods independently, without the oversight of expert chromatographers. This app note shows fast method development using a structured protocol called the systematic screening protocol. The protocol used to develop the method relies on MaxPeak™ High-Performance Surfaces (HPS) technology, featured in MaxPeak Premier Columns, to mitigate any non-specific adsorption (NSA) seen between the column and the analytes. Employing this column technology, along with the systematic screening protocol, a method was developed which provides good peak shape and complete separation of all eight dyes.

High efficiency columns increase resolution by reducing peak widths. This enables easier peak integration and identification, as the peaks of interest are better separated from each other and from potential background or excipient peaks. There are several ways for an analyst to improve separation efficiency; one being to use columns packed with smaller particle size stationary phases. Another way is to use longer columns. However, a drawback in the use of both options is that they can be limited by the system operating pressure. Another path forward is the use of solid-core or superficially porous particles (SPP), which have been proven to improve efficiency without sacrificing operating pressure. This app note shows incremental steps on how to improve separation efficiency for a mixture of three analytes.

HPLC method development is not without its challenges. What happens when your favorite column struggles to achieve your peak shape, sensitivity, or reproducibility goals? Whether you’re developing or transferring methods, quickly achieve consistent results with MaxPeak Premier Columns. Available from sub-2 µm to 5 µm HPLC particle sizes, fully porous and solid-core, you can choose the column configuration that meets your needs and eliminate doubt from your chromatographic separations. Find MaxPeak Premier app notes, videos and more at waters.com/tothemax.

Due to their ubiquitous occurrence in aquatic environments, measuring ultrashort-chain per- and polyfluoroalkyl substances (PFAS) in various source waters to monitor their presence and the potential for human exposure has become very important. However, with carbon chain lengths of less than C4, these small, highly polar compounds are difficult to analyze using standard PFAS tests that are based on reversed-phased liquid chromatography (RPLC). In this study, an accurate, reliable analytical LC-MS/MS method for PFAS in water was developed to specifically quantify C1 to C4 PFAS in both potable and non-potable sources. A direct injection workflow was implemented to simplify the testing process and to avoid potential contamination originating from poor sample preparation procedures.

A Survey of Existing PFAS Testing Methods and Guidelines from Around the World

Helium is running out, the worldwide demand is exceeding current production levels. Learn how Hydrogen could be your perfect replacement for Helium.

HPLC is a versatile and widely used method for analyzing various compounds, particularly in the pharmaceutical industry. It's preferred by generic pharmaceutical companies and quality control groups due to its accessibility compared to newer technologies like Ultra-Performance Liquid Chromatography (UPLC™). In recent years, there has been a growing emphasis on making analytical chemistry techniques, such as HPLC, more sustainable. These "green" initiatives aim to reduce the use of toxic reagents, minimize waste, and lower energy consumption, all while maintaining high scientific standards. This app note highlights Development of a new, greener HPLC method for rivaroxaban and impurities, aligned with the USP monograph and using the Analytical Method Greenness Score (AMGS).

High Performance Liquid Chromatography (HPLC) has been a staple in analytical laboratories for several years. The utility of this technique is such that it can be used to analyze a wide variety of analytes. HPLC allows a lower cost alternative compared to UHPLC instruments, which boast higher performance for the higher capital investment. HPLC systems are still used regularly in the pharmaceutical industry for QA and QC type work such as batch release testing and method development.

When scaling an established analytical method across columns packed with different particle sizes and different column configurations (internal diameters and lengths), the amount of time that is required outside of the lab to produce the equivalent method conditions is considerable. When dealing with a gradient method, the calculations required include determining the new flow rate, gradient times, and injection volume. An analyst can perform these calculations manually using the appropriate equations or tools like the Waters Columns Calculator. This app note examines two scaling workflows by first performing a theoretical scale-down experiment manually using the appropriate method scaling equations, and then repeating the experiment using the scaled-down conditions generated by the Waters Columns Calculator. Strong agreement between manual calculations and the results using the Waters Columns Calculator validate the Waters Column Calculator for its use in scaling methods effectively, with a significant improvement in time savings and a reduction in potential calculation errors and/or uncertainties.

To address the Helium shortage GC laboratories have begun looking at possible replacement gases and new approaches to help obtain the carrier gases needed to continue their work. One of the most promising options is using the use of gas generators to provide a steady supply of hydrogen gas as a replacement for helium. Although some concerns—and some myths—persist about using hydrogen, it offers multiple benefits and advantages over helium, notably particularly when a gas generator replaces high-pressure gas cylinders.

A Methods-Based Reference to Lab Supplies for PFAS Testing

Hydrogen is the perfect molecule for gas chromatography, especially for the replacement of the expensive Helium.

Discover the GC 2400™ Platform for analytical precision and robustness for fuel alcohol analysis over a wide range of sample concentrations.

Master Gas Chromatography Troubleshooting - Streamline Your Analysis with Expert Guidance.

Discover how the PerkinElmer GC 2400™ Platform provides fast analysis and high reproducibility.

Discover how the PerkinElmer GC 2400™ Platform helps with the residual solvent in plastic with accuracy and sensitivity, providing reliable and reproducible runs for high throughput laboratories.

Discover how the PerkinElmer GC 2400™ Platform helps improve resolution requirements.

Discover an optimized GC/MS approach for SVOC in complex matrices.

Read how the integrated HS-GC workflow allows a continuous optimization of the pneumatic values for ultimate precision in GC retention time.

Inside the Laboratory is a joint series with LCGC International and Spectroscopy, profiling analytical scientists and their research groups at universities all over the world. This series spotlights the current chromatographic and spectroscopic research their groups are conducting, and the importance of their research in analytical chemistry and specific industries. In this “Inside the Laboratory –– A Look at Environmental Analysis, PFAS, and Contaminants of Emerging Concern” special edition PDF, we profile Dr. Lee Blaney and his laboratory at the University of Maryland Baltimore County (UMBC), in Baltimore, Maryland. This special edition PDF highlights the research Dr. Blaney is spearheading in his laboratory, which include studying contaminants of emerging concern and developing advanced analytical methods that can better improve detection and quantification of CECs in the environment, as well as producing new treatment technologies for diverse contaminants in water and wastewater.

This note summarizes the optimal methods for the HPLC separation of semaglutide and liraglutide under acidic and alkaline conditions.

This application note discusses how to streamline method transfer outcomes using ACD/Labs' method development software, Method Selection Suite.

Discover the supremacy of the bioinert coated hardware of YMC Accura BioPro IEX QF columns for reproducible analysis of oligonucleotides in ion-exchange chromatography.

This application note demonstrates the use of SCION's 8500GC system for the analysis of key greenhouse gases—carbon dioxide, methane, and nitrous oxide—in a single atmospheric air matrix. Highlighting the system's excellent sensitivity and repeatability, this method is crucial for understanding emission sources and combating climate change.

This application note outlines a method for detecting low levels of 1,4-Dioxane, a likely carcinogenic compound, in drinking water using SCION GC-MS technology. It details the procedure for analyzing water samples spiked with 1,4-Dioxane, achieving excellent sensitivity and low detection limits, demonstrating the method's effectiveness for environmental monitoring.

This application note from SCION Instruments delves into the meticulous screening of impurities in plant protection products using GC-FID, with GC-MS for confirmation. It emphasizes the necessity of identifying and quantifying impurities to comply with regulatory standards in industries such as pharmaceuticals, food, and agriculture. The study specifically examines eugenol, showcasing how to determine significant impurities for regulatory submission. The approach integrates GC-MS and analytical standards, ensuring precise impurity identification and quantification critical for product approval.

We show the development of HPLC-ELSD methods for high-throughput and sensitive analysis of the lipids in lipid nanoparticles (LNPs), used to administer mRNA vaccines.

Recycling chromatography is a powerful technique that allows the separation of two closely eluting peaks by increasing the column bed length.