Benefits of Ascentis Express 5 um Columns in Analysis of Pharmaceutically Relevant Compounds
Supelco/Sigma-Aldrich
Non-steroidal anti-inflammatory drugs (NSAIDs) are pharmaceutically relevant compounds that inhibit the inflammation process without the use of steroid drugs. The most commonly used NSAIDs are aspirin, ibuprofen, and naproxen.
NSAIDs often cause dangerous side effects. As reported to the US Food and Drug Administration, the majority of the undesired side effects are related to damage to the gastrointestinal (GI) tract and kidneys (1). As a result, there is continuous interest in improving the analysis and characterization of NSAIDs.
In this study a simple UV-based HPLC method was developed using the new Ascentis® Express 5 μm Fused-Core HPLC column. A comparison was then made to a traditional 5 μm fully-porous HPLC column.
Experimental Conditions
An aliquot (2 μL) of NSAIDs mixture was injected on an Ascentis Express 5 μm and a regular porous 5 μm C18, 15 cm × 4.6 mm I.D. column. Figure 1 shows the two chromatograms and details the experimental conditions.
Figure 1: Comparision of NSAIDs analysis on Ascentis Express 5 μm (50537-U) and a regular porous 5 μm, C18, 15 cm à 4.6 mm ID column, mobile phase A: 20-mM pH 2.5 potassium phosphate, B: 50/50 ACN/MeOH; A:B = 48% A:52% B; instrument: Shimadzu Prominence UFLC XR; flow rate: 2.0 mL/min, injection volume: 2 μL, det: 254 nm; temp: 35 °C. Peaks: 1. Acetaminophen, 2. Aspirin, 3. Salicylic acid, 4. Tolmetin, 5.Ketoprofen, 6. Naproxen, 7. Fenoprofen, 8. Diclofenac, 9. lbuprofen.
Conclusion
The Fused-Core column shows same selectivity as a regular 5 μm porous C18 HPLC column, but outperforms the fully-porous column in terms of efficiency, with almost twice the theoretical plates (N = 20,500 vs. N = 11,000). In addition to this, the 5 μm Fused-Core column exhibits greater resolution in about half the run time when compared to the fully porous column, all at about the same backpressure. The Fused-Core particle design provides an easy way to realize higher efficiencies and shorter run times.
Reference
(1) A.F. Spac and V. Dorneanu, "HPLC Analysis of NSAIDs", Encylopedia of Chrom, Edition 3, Chapter 129, 495 (2009).
Ascentis is a registered trademark of Sigma-Aldrich Co. LLC. Fused-Core is a registered trademark of Advanced Materials Technology, Inc.
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SEC-MALS of Antibody Therapeutics—A Robust Method for In-Depth Sample Characterization
June 1st 2022Monoclonal antibodies (mAbs) are effective therapeutics for cancers, auto-immune diseases, viral infections, and other diseases. Recent developments in antibody therapeutics aim to add more specific binding regions (bi- and multi-specificity) to increase their effectiveness and/or to downsize the molecule to the specific binding regions (for example, scFv or Fab fragment) to achieve better penetration of the tissue. As the molecule gets more complex, the possible high and low molecular weight (H/LMW) impurities become more complex, too. In order to accurately analyze the various species, more advanced detection than ultraviolet (UV) is required to characterize a mAb sample.
