High-throughput screening , HPLC

The use of direct UV spectrophotometry to measure sample concentrations in pharmaceutical research is uncommon, presumably because of the prevalence and attractiveness of HPLC and LC/MS methods. Consequently, most researchers are unfamiliar with how useful direct UV can be. The UV method is much faster than the other methods, and this is very important in high-throughput screening. 

In reversed-phase HPLC, column temperature is a strong determinant of retention time and also affects column selectivity. A column oven is therefore required for most automated pharmaceutical assays to improve retention time precision, typically at temperatures of 30-50°C. Temperatures >60°C are atypical due to concerns about thermal degradation of the analytes and column lifetimes. Exceptions are found in high-throughput screening where higher temperatures are used to increase flow and efficiency. Ambient or snb-ambient operation is sometimes found in chiral separations to enhance selectivity. Column ovens... 

HPLC applications assays, impurity evaluation, dissolution testing, cleaning validation, high-throughput screening, and chiral separations (Chapters 13-18). 

The coupling of HPLC with NMR represents a powerful method for the high-throughput screening of peptides in mixtures run in stop-flow and continuous-flow modes. It is possible to obtain routine high-quality HPLC/NMR ID NMR data with as little as 5 pg of compound in a chromatogram peak. However, the HPLC/NMR technique cannot be favorably compared to mass spectrometry techniques (HPLC/MS) in terms of sensitivity and speed of analysis. To date, the majority of reports of the use of HPLC/NMR have been for drug metabolites.1 ... 

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