An automated multidimensional screening approach for rapid method development in high performance liquid chromatography (original) (raw)
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Journal of Chromatography A, 2011
An analytical HPLC-MS screening methodology has been developed to improve preparative RP-HPLC-MS purifications in medicinal chemistry laboratories. Although several approaches have been previously described to optimize analytical separations, none of them met our needs for the optimization of preparative conditions. Our screening protocol is based on searching among several orthogonal conditions to find the optimum preparative separation. Five different buffer conditions, from low to high pH, two organic solvents, acetonitrile and methanol, and five stationary phases of different polarities and characteristics were used. The orthogonality of the system was demonstrated using both, a standard mixture and mixtures from synthesis. To carry out the screening one of the analytical "open access" HPLC-MS systems was modified to perform the analytical screening while maintaining the open-access functionality for synthesis reaction monitoring. A software tool for automated sample programming and data reporting was also developed.
2015
Many different strategies of high performance liquid chromatographic method development are being used today. This overview describes a strategy for the systematic development of High performance liquid chromatographic (HPLC) methods. It is an analytical tool which is able to separate, detect and quantify the drug, its various impurities and drug related degradants that can form on synthesis or storage. HPLC involves the understanding of chemistry of drug substance and facilitates the development of the analytical method. Many chromatographic parameters were evaluated in order to optimize the method. Appropriate mobile phase, stationary phase, column, column size, temperature, wavelength and gradient must be found that affords suitable compatibility and stability of drug as well as impurities and degradants. In this paper we have discussed the different physical and chemical parameters that govern the HPLC process and operation and suggested method development for the most optimum c...
Journal of Chromatography A, 2009
In this paper a comparison of two column characterisation systems is reported: the method based on the hydrophobic-subtraction model of Dolan and Snyder (HS method) versus the method developed at the Katholieke Universiteit Leuven (KUL method). Comparison was done for seven different pharmaceutical separations (fluoxetine, gemcitabine, erythromycin, tetracycline, tetracaine, amlodipine and bisacodyl), using a set of 59 columns. A ranking was built based on an F value (KUL) or F s value (HS) versus a (virtual) reference column. Both methods showed similar probabilities of ranking patterns. Correlation of the respective test parameters of both approaches was poor. Both methods are not perfect and do not match well, but anyhow yield results which allow, with a relatively high certainty, the selection of similar or dissimilar columns as compared to a reference column. An analyst that uses either of the two methods will end up with a similar probability to choose an adequate column. From a practical point of view, it must be noted that the KUL method is easier to use.
Reviews in Analytical Chemistry
This review presents the essential brief annals, crucial analytics, precise applications and noteworthy implications of design of experiments which enrouted to liquid chromatography (LC) in the midst of utmost focus on high-performance liquid-chromatography (HPLC) and broadened its impressions on allied techniques in pharmaceutical analysis. Being most widely applied statistical methodologies for such purpose, its use was started in 1970 and heightened after Fischer’s precious input in 1981. The persistent use of statistical approaches one after another led to the efficient attention of pharmaceutical analysts. Hence, in order to fine-tune the trail impressed by the cumulative trends, the use of statistical designs in HPLC analysis has been reviewed and efforts were made to recognize its relative impact and corresponding future prospects. Applications of precise methodologies have been reassessed with respect to the need established by recent regulatory perspectives with a fanatical...
Critical Reviews in Analytical Chemistry, 2012
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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.
High‐throughput screening of chromatographic separations: I. Method development and column modeling
Biotechnology and Bioengineering, 2008
Ion-exchange (IEX) chromatography steps are widely applied in protein purification processes because of their high capacity, selectivity, robust operation, and wellunderstood principles. Optimization of IEX steps typically involves resin screening and selection of the pH and counterion concentrations of the load, wash, and elution steps. Time and material constraints associated with operating laboratory columns often preclude evaluating more than 20-50 conditions during early stages of process development. To overcome this limitation, a high-throughput screening (HTS) system employing a robotic liquid handling system and 96-well filterplates was used to evaluate various operating conditions for IEX steps for monoclonal antibody (mAb) purification. A screening study for an adsorptive cation-exchange step evaluated eight different resins. Sodium chloride concentrations defining the operating boundaries of product binding and elution were established at four different pH levels for each resin. Adsorption isotherms were measured for 24 different pH and salt combinations for a single resin. An anion-exchange flowthrough step was then examined, generating data on mAb adsorption for 48 different combinations of pH and counterion concentration for three different resins. The mAb partition coefficients were calculated and used to estimate the characteristic charge of the resinprotein interaction. Host cell protein and residual Protein A impurity levels were also measured, providing information on selectivity within this operating window. The HTS system shows promise for accelerating process development of IEX steps, enabling rapid acquisition of large datasets addressing the performance of the chromatography step under many different operating conditions.