title: Optimisation of high performance liquid chromatography separation of neuroprotective peptides. Fractional experimental designs combined with artificial neural networks.
abstract: The study of experimental design conjunction with artificial neural networks for optimisation of isocratic ion-pair reverse phase HPLC separation of neuroprotective peptides is reported. Different types of experimental designs (full-factorial, fractional) were studied as suitable input and output data sources for ANN training and examined on mixtures of humanin derivatives. The independent input variables were: composition of mobile phase, including its pH, and column temperature. In case of a simple mixture of two peptides, the retention time of the most retentive component and resolution were used as the dependent variables (outputs). In case of a complex mixture with unknown number of components, number of peaks, sum of resolutions and retention time of ultimate peak were considered as output variables. Fractional factorial experimental design has been proved to produce sufficient input data for ANN approximation and thus further allowed decreasing the number of experiments necessary for optimisation. After the optimal separation conditions were found, fractions with peptides were collected and their analysis using off-line matrix assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF-MS) was performed.
authors.affiliation: Masaryk University, Faculty of Science, Department of Analytical Chemistry, Kotlárská 2, 611 37 Brno, Czech Republic.
authors.forename: Klára
authors.initials: K
authors.lastname: Novotná
authors.forename: Jan
authors.initials: J
authors.lastname: Havlis
authors.forename: Josef
authors.initials: J
authors.lastname: Havel
mesh_terms: D019943:Amino Acid Substitution; D005591:Chemical Fractionation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000302:isolation & purification; D016571:Neural Networks, Computer; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; C427211:humanin