High performance liquid chromatography of natural products. II. Direct biological correlation of components in the fermentation broth.

Connecting Phenotype and Chemotype: High-Content Discovery Strategies for Natural Products Research

“…Thin-layer chromatography (TLC) emerged as the first method for parallelized characterization of natural product extracts and is still widely used as a rapid, low-resolution method for profiling chemical constitution of natural product extracts; however, high-performance liquid chromatography (HPLC) and hyphenated techniques have all but completely replaced TLC for most natural products discovery applications, because of their increased resolution and greater information content. − The use of HPLC retention time in combination with ultraviolet and visible absorbance spectra allows the profiling and comparison of extracts within any screening library and has been used widely by industry and academia. In an early example Miller et al used stream splitting and automated fraction collection in a compound-by-compound bioactivity and dereplication process for the discovery of clavulanic acid, , paving the way for adoption of this approach by many other research groups. The primary disadvantages of these techniques are that HPLC protocols are time-consuming, analysis and dereplication are performed on a compound-by-compound basis, and saving fractions is not practical for very large libraries …”

“…10−12 The use of HPLC retention time in combination with ultraviolet and visible absorbance spectra allows the profiling and comparison of extracts within any screening library and has been used widely by industry and academia. In an early example Miller et al used stream splitting and automated fraction collection in a compound-by-compound bioactivity and dereplication process for the discovery of clavulanic acid, 13,14 paving the way for adoption of this approach by many other research groups. The primary disadvantages of these techniques are that HPLC protocols are time-consuming, analysis and dereplication are performed on a compound-by-compound basis, and saving fractions is not practical for very large libraries.…”

Beta-Lactam Antibiotics

“…A high performance liquid chromatography procedure was developed for separation of ß-lactam antibiotics directly from fermentation broths of C. acremonium, Pénicillium chrysogenum and Streptomyces clavuligerus (88). A high performance liquid chromatography procedure was developed for separation of ß-lactam antibiotics directly from fermentation broths of C. acremonium, Pénicillium chrysogenum and Streptomyces clavuligerus (88).…”