Taking Advantage of Sub-2 μm Core–Shell Technology for Ultra-Fast and Ultra-Efficient Urinary Excretion Profiling
The Application Notebook
Phenomenex
Throughout the drug development process, potential new drug candidates (or new chemical entities; NCEs) and their metabolites must be subjected to rigorous and extensive pharmacokinetic evaluations to determine their rates accumulation, metabolism, and excretion from the body. With regards specifically to the excretion, urinary excretion is typically the predominant route for the elimination of drugs and their metabolites, and urinary excretion profiling is an integral portion of the pharmacokinetic characterization of NCEs.
Liquid chromatography tandem mass spectroscopy (LC–MS-MS) allows scientists to rapidly and accurately quantify specific drugs and their metabolites at extremely low levels from various biological matrices, such as urine. On the liquid chromatography side, the ultra-high efficiency delivered by sub-2 μm UHPLC core–shell media provides analysts with the ability to run their samples in extremely short periods of time while maintaining excellent resolution from sample interferences. In this application note, we present an example of the ability of the Kinetex® 1.7 μm core–shell particle to deliver significantly improved performance over larger core–shell particles and conventional fully-porous media.
Experimental Conditions
LC–MS-MS conditions
Column: Kinetex 1.7 μm C18 30 × 2.1 mm Fused-core 2.7 μm C18 50 × 2.1 mm Fully-porous 3.5 μm C18 50 × 2.1 mm
Mobile phase: A: 10 mM Ammonium formate B: Acetonitrile
Gradient: (95:5) A/B to (0:100) A/B in 2 min, then re-equilibrate at (95:5) A/B for 1 min
Flow rate: 700 μL/min
Temperature: Ambient
Detection: MS using API 4000 detector
HPLC system: Agilent 1200 SL
Concentration: 100 ng/mL for active drug and 50 ng/mL for metabolite.
Analytes: Lorazepam-glucuronide spiked into urine at a concentration of 50 ng/mL.
Figure 1: XIC for oxazepam glucuronide (MRM 463.1 -> 287.0) in urine.
Results
In many instances, suitable MRM transitions for an analyte of interest are hidden because of significant matrix interference. Oxazepam-glucuronide did not suffer from urinary interference (Figure 1), however the 497.2 -> 320.9 MRM transition for the glucuronide metabolite of lorazepam does show significant isobaric interference (Figure 2). In cases such as this, the ultra-high efficiency of the core–shell Kinetex 1.7 μm particle can provide significantly greater peak capacity than standard fully-porous media (3.5 μm in this case) and a larger fused-core particle (2.7 μm fused-core).
Figure 2: Comparison of the performance of the Kinetex 1.7 μm C18 (30 à 2.1 mm) column vs. a fused-core 2.7 μm column (50 à 2.1 mm) and a fully-porous 3.5 μm C18 column (50 à 2.1 mm) for the glucuronide metabolite of lorazepam (MRM 497.2 -> 320.9).
Phenomenex Inc.
411 Madrid Avenue, Torrance, California 90501, USA
tel. +1 (310) 212 0555 +1 (310) 328 7768
Website: www.phenomenex.com
2024 EAS Awardees Showcase Innovative Research in Analytical Science
November 20th 2024Scientists from the Massachusetts Institute of Technology, the University of Washington, and other leading institutions took the stage at the Eastern Analytical Symposium to accept awards and share insights into their research.
Inside the Laboratory: The Richardson Group at the University of South Carolina
November 20th 2024In this edition of “Inside the Laboratory,” Susan Richardson of the University of South Carolina discusses her laboratory’s work with using electron ionization and chemical ionization with gas chromatography–mass spectrometry (GC–MS) to detect DBPs in complex environmental matrices, and how her work advances environmental analysis.
AI and GenAI Applications to Help Optimize Purification and Yield of Antibodies From Plasma
October 31st 2024Deriving antibodies from plasma products involves several steps, typically starting from the collection of plasma and ending with the purification of the desired antibodies. These are: plasma collection; plasma pooling; fractionation; antibody purification; concentration and formulation; quality control; and packaging and storage. This process results in a purified antibody product that can be used for therapeutic purposes, diagnostic tests, or research. Each step is critical to ensure the safety, efficacy, and quality of the final product. Applications of AI/GenAI in many of these steps can significantly help in the optimization of purification and yield of the desired antibodies. Some specific use-cases are: selecting and optimizing plasma units for optimized plasma pooling; GenAI solution for enterprise search on internal knowledge portal; analysing and optimizing production batch profitability, inventory, yields; monitoring production batch key performance indicators for outlier identification; monitoring production equipment to predict maintenance events; and reducing quality control laboratory testing turnaround time.
Infographic: Be confidently audit ready, at any time and reduce failures in pharma QC testing
November 20th 2024Discover how you can simplify the audit preparation process with data integrity dashboards that provide transparency to key actions, and seamlessly track long-term trends and patterns, helping to prevent system suitability failures before they occur with waters_connect Data Intelligence software.
Critical Role of Oligonucleotides in Drug Development Highlighted at EAS Session
November 19th 2024A Monday session at the Eastern Analytical Symposium, sponsored by the Chinese American Chromatography Association, explored key challenges and solutions for achieving more sensitive oligonucleotide analysis.
