title: Development of a fast extraction method and optimization of liquid chromatography-mass spectrometry for the analysis of phenolic compounds in lentil seed coats.
abstract: A systematic set of optimization experiments was conducted to design an efficient extraction and analysis protocol for screening six different sub-classes of phenolic compounds in the seed coat of various lentil (Lens culinaris Medik.) genotypes. Different compounds from anthocyanidins, flavan-3-ols, proanthocyanidins, flavanones, flavones, and flavonols sub-classes were first optimized for use as standards for liquid chromatography mass spectrometry (LC-MS) with UV detection. The effect of maceration duration, reconstitution solvent, and extraction solvent were investigated using lentil genotype CDC Maxim. Chromatographic conditions were optimized by examining column separation efficiencies, organic composition, and solvent gradient. The results showed that a 1h maceration step was sufficient and that non-acidified solvents were more appropriate; a 70:30 acetone: water (v/v) solvent was ultimately selected. Using a Kinetex PFP column, the organic concentration, gradient, and flow rate were optimized to maximize the resolution of phenolic compounds in a short 30-min analysis time. The optimized method was applied to three lentil genotypes with different phenolic compound profiles to provide information of value to breeding programs.
authors.affiliation: Plant Sciences Department, University of Saskatchewan, Saskatoon, SK, Canada S7N 5A8.
authors.forename: Mahla
authors.initials: M
authors.lastname: Mirali
authors.affiliation: ACRD, National Research Council of Canada, Saskatoon, SK, Canada S7N 4K9.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Ambrose
authors.affiliation: ACRD, National Research Council of Canada, Saskatoon, SK, Canada S7N 4K9.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Wood
authors.affiliation: Plant Sciences Department, University of Saskatchewan, Saskatoon, SK, Canada S7N 5A8.
authors.forename: Albert
authors.initials: A
authors.lastname: Vandenberg
authors.affiliation: Plant Sciences Department, University of Saskatchewan, Saskatoon, SK, Canada S7N 5A8; ACRD, National Research Council of Canada, Saskatoon, SK, Canada S7N 4K9. Electronic address: randy.purves@usask.ca.
authors.forename: Randy W
authors.initials: RW
authors.lastname: Purves
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005419:Flavonoids / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D027806:Lens Plant / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D015203:Reproducibility of Results; D012107:Research Design; D012639:Seeds / Q000737:chemistry*
chemical_list: D005419:Flavonoids
keywords: Core–shell column; Extraction; HPLC condition optimization; Lens culinaris; Polyphenols