Rapid high performance liquid chromatography method development with high prediction accuracy, using 5cm long narrow bore columns packed with sub-2μm particles and Design Space computer modeling (original) (raw)
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Journal of Separation Science, 2007
Evaluation and comparison of very high pressure liquid chromatography systems for the separation and validation of pharmaceutical compounds Chromatography using sub-2 lm particles is becoming increasingly popular due to the potential for increased speed, resolution, sensitivity, and peak capacity. To meet the demand, various vendors have re-engineered traditional LC systems to operate at pressures of up to 15 000 psi to accommodate the elevated backpressures associated with using sub-2 lm particles. This report investigates and compares the performance of three very high pressure LC (VHPLC) systems: Waters Acquity, Agilent 1200 SL, and Thermo Accela. Specifications for the pump, autosampler, column compartment, detector, and software for each instrument are presented. To assess the chromatographic performance of the three instruments, method development and validation were performed for three pharmaceutical compounds and the results are compared and discussed. The material presented herein serves to highlight the different features of the VHPLC instruments, and assess their suitability for the analysis of pharmaceutical compounds.
Aspects of the “Design Space” in high pressure liquid chromatography method development
Journal of Chromatography A, 2010
The present paper describes a multifactorial optimization of 4 critical HPLC method parameters, i.e. gradient time (t G), temperature (T), pH and ternary composition (B 1 :B 2) based on 36 experiments. The effect of these experimental variables on critical resolution and selectivity was carried out in such a way as to systematically vary all four factors simultaneously. The basic element is a gradient time-temperature (t G-T) plane, which is repeated at three different pH's of the eluent A and at three different ternary compositions of eluent B between methanol and acetonitrile. The so-defined volume enables the investigation of the critical resolution for a part of the Design Space of a given sample. Further improvement of the analysis time, with conservation of the previously optimized selectivity, was possible by reducing the gradient time and increasing the flow rate. Multidimensional robust regions were successfully defined and graphically depicted.
Principles and Applications of Ultra-High-Performance Liquid Chromatography
IntechOpen eBooks, 2023
The science of separation had advanced significantly with the development of ultra-high-performance liquid chromatography (UHPLC), a brand-new type of liquid chromatography. The need for the evolution of HPLC into UHPLC has been driven by the continuously evolving of packing material modifications that affect the separation of mixtures. The separation process of analytes is completed in a substantially decreased amount of time due to the lower particle sizes, which increases surface area of interaction allowing reduction of column length to one-third; thus, shorter columns are employed in UHPLC, which consequently causes the flow rate to be three times higher and subsequently reducing analysis time. Although UHPLC shares the same fundamental idea and instrument layout as HPLC, it differs from HPLC in that it produces narrow peaks and has high spectral quality, allowing for simple compound identification in a variety of analytical applications such as impurity profiling, product formulation, and improved analytical technique and method development. However, high back pressure in UHPLC might lead to decreased column life, and the instrument's higher price compared to HPLC are the disadvantages.
UPLC: a preeminent technique in pharmaceutical analysis
Acta poloniae pharmaceutica
The pharmaceutical companies today are driven to create novel and more efficient tools to discover, develop, deliver and monitor the drugs. In this contest the development of rapid chromatographic method is crucial for the analytical laboratories. In precedent decade, substantial technological advances have been done in enhancing particle chemistry performance, improving detector design and in optimizing the system, data processors and various controls of chromatographic techniques. When all was blended together, it resulted in the outstanding performance via ultra-high performance liquid chromatography (UPLC), which holds back the principle of HPLC technique. UPLC shows a dramatic enhancement in speed, resolution as well as the sensitivity of analysis by using particle size less than 2 pm and the system is operational at higher pressure, while the mobile phase could be able to run at greater linear velocities as compared to HPLC. This technique is considered as a new focal point in...
International Journal of Advances in Pharmacy and Nanotechnology, 2011
Ultra performance liquid chromatography (UPLC) system involves significant technological advances in particle size performance, system optimization, data processing, detector design and control. When all brought together, the specific achievements in each area have created a step-function progress in chromatographic performance. This new technique of analytical separation science uses the principles and practicality of HPLC with increasing the attributes of speed, sensitivity and resolution. Now a day's pharmaceutical industries are in search of new ways to reduce cost and time for analysis of drugs. Analytical laboratories are not exception in this trend. Ultra high performance liquid chromatography (UPLC) with better resolution, assay sensitivity and high sample throughput allows a greater number of analysis to be performed in a shorter period of time and it also impart cost effective advantage over HPLC analysis. So that conventional assay was transferred and optimized for UPLC system. This review introduces the theory of UPLC, and involves some of the most advanced work in the field.
Experimental design in HPLC separation of pharmaceuticals
Arhiv za farmaciju
Design of Experiments (DoE) is an indispensable tool in contemporary drug analysis as it simultaneously balances a number of chromatographic parameters to ensure optimal separation in High Pressure Liquid Chromatography (HPLC). This manuscript briefly outlines the theoretical background of the DOE and provides step-by-step instruction for its implementation in HPLC pharmaceutical practice. It particularly discusses the classification of various design types and their possibilities to rationalize the different stages of HPLC method development workflow, such as the selection of the most influential factors, factors optimization and assessment of the method robustness. Additionally, the application of the DOE-based Analytical Quality by Design (AQbD) concept in the LC method development has been summarized. Recent achievements in the use of DOE in the development of stability-indicating LC and hyphenated LC-MS methods have also been briefly reported. Performing of Quantitative structu...
ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY: A REVOLUTIONIZED LC TECHNIQUE
High Performance Liquid Chromatography (HPLC) is a major technique for qualitative and quantitative drug analysis. More than 90% of drugs prescribed in official pharmacopoeias are being analyzed HPLC. HPLC analyzes the drug content in a sample with high degree of accuracy and precision. Due to the stringent regulatory requirements the number of samples for drug content analysis has been increased significantly. Therefore, pharmaceutical industries need a fast, accurate and affordable method for drug content analysis. Here, Ultra Performance Liquid Chromatography (U-PLC) offers an advancement of HPLC which is based on the principal of use of stationary phase consisting of particles less than 2μm. By using smaller particles; speed and peak capacity can be extended to new limits and the sample can be analyzes in a shorter period of time. It provides good resolution even for congeneric compounds. The present review discusses the various aspects of UPLC in pharmaceutical analysis. Keywords: Ultra Performance Liquid Chromatography (UPLC), HPLC, ACQUITY UPLC.
European Journal of Pharmaceutics and Biopharmaceutics, 2007
Liquid chromatography (LC) is currently considered as the gold standard in pharmaceutical analysis. Today, there is an increasing need for fast and ultra-fast methods with good efficiency and resolution for achieving separations in a few minutes or even seconds. A previous article (i.e. method transfer for fast LC in pharmaceutical analysis. Part I: isocratic separation) described a simple methodology for performing a successful method transfer from conventional LC to fast and ultra-fast LC in isocratic mode. However, for performing complex separations, the gradient mode is often preferred. Thus, this article reports transfer rules for chromatographic separations in gradient mode. The methodology was applied for the impurity profiling of pharmaceutical compounds, following two strategies.
Principle, Instrumentation, and Applications of UPLC: A Novel Technique of Liquid Chromatography
Open Chemistry Journal, 2016
The key focus of the pharmaceutical or chemical industries is to reduce the cost involved in the development of new drugs and to improve the selectivity, sensitivity, and resolution for their detection. The purpose can now be solved by the separation method called UPLC which is the modified HPLC method comprising high pressure and small sized particles (less than 2 µm) used in the column, so the length of the column decreases leading to time saving and reduction in the consumption of solvent. The underlying principle of UPLC is based on van Deemter statement which describes the connection between linear velocity with plate height. UPLC contributes to the improvement of the three areas: speed, resolution, and sensitivity. This is a new advanced category of the HPLC which has the same basic principle and methodology with improved chromatographic performance. This review is an effort to compile the principle, instrumentation, and applications of UPLC.
Journal of chromatography. A, 2017
Large pressure gradients are generated in ultra-high-pressure liquid chromatography (UHPLC) using sub-2μm particles causing significant temperature gradients over the column due to viscous heating. These pressure and temperature gradients affect retention and ultimately result in important selectivity shifts. In this study, we developed an approach for predicting the retention time shifts due to these gradients. The approach is presented as a step-by-step procedure and it is based on empirical linear relationships describing how retention varies as a function of temperature and pressure and how the average column temperature increases with the flow rate. It requires only four experiments on standard equipment, is based on straightforward calculations, and is therefore easy to use in method development. The approach was rigorously validated against experimental data obtained with a quality control method for the active pharmaceutical ingredient omeprazole. The accuracy of retention t...