Practical use of an optimization strategy in reversed-phase high-performance liquid chromatography for the separation of a limited subset of components in a reaction mixture

✍ Scribed by Akos Bartha; Hugo A.H. Billiet; Leo De Galan

• Publisher: Elsevier Science
• Year: 1991
• Tongue: English
• Weight: 881 KB
• Volume: 464
• Category: Article
• ISSN: 1873-3778
• DOI: 10.1016/s0021-9673(00)94242-0

✦ Synopsis

A step-by-step development of a high-performance liquid chromatographic method for the optimized separation of a largely unknown sample is presented. A limited subset of two main components is separated from a ten-component reaction mixture of pyrroloquinoline quinone and cyclopropanol, using tetrahydrofuranwater binary eluents buffered to pH 2.5 with triethylamine phosphate on a reversedphase column packed with 5-pm ODS-Hypersil. After the definition of the separation problem, the mobile phase parameters are selected rationally on the basis of a systematic gradient scouting procedure. The optimization search area is defined in accordance to the complexity of the sample mixture (two out of ten components are of interest) by utilizing a statistical approach, while the selectivity optimization is carried out using binary eluents of variable eluotropic strength.