title: Development and Validation of a New HPLC Method for the Determination of Biphenyl and Dibenzofuran Phytoalexins in Rosaceae.
abstract: A simple, precise, rapid and accurate isocratic reversed-phase high-performance liquid chromatographic method was developed and validated for the simultaneous determination of biphenyl (aucuparin and noraucuparin) and dibenzofuran (eriobofuran) phytoalexin from elicitor treated cell culture of Sorbus aucuparia (mountain ash). These phytoalexins play crucial role in combating scab disease in many commercially important rosaceous plants, such as apple, pear and mountain ash. The isocratic separation was performed in a Luna C18 reversed-phase column (250 × 4.6 mm, 5 μm particle size) using a mobile phase of 1 mM trifluoroacetic acid (TFA) in water with methanol [40:60 (v/v)]. Quantization of phytoalexin was carried out on Shimadzu-HPLC system using a Photo Diode Array (PDA) detector at 254 nm by comparing the peak area. Peak purity and identity were confirmed by UV spectroscopy and ESI-MS-MS in the negative ion mode. The different analytical performance parameters such as linearity, accuracy, precision, limit of detection and limit of quantification were determined according to the International Conference on Harmonization guidelines. Linearity was observed in the concentration range of 3-400 µg/mL with excellent correlation coefficient (R(2) ≥ 0.995). This newly developed method is rapid, easy, cost-effective and can be used for monitoring scab-resistance potential of rosaceous plants.
authors.affiliation: Plant Molecular Biology Group, Biotechnology Department, Indian Institute of Technology Roorkee, Roorkee 247 667, India.
authors.forename: Deepa
authors.initials: D
authors.lastname: Teotia
authors.affiliation: Plant Molecular Biology Group, Biotechnology Department, Indian Institute of Technology Roorkee, Roorkee 247 667, India.
authors.forename: Shashank Sagar
authors.initials: SS
authors.lastname: Saini
authors.affiliation: Institut für Pharmazeutische Biologie, Technische Universität Braunschweig, Mendelssohnstrasse 1, D-38106, Braunschweig, Germany.
authors.forename: Mariam
authors.initials: M
authors.lastname: Gaid
authors.affiliation: Institut für Pharmazeutische Biologie, Technische Universität Braunschweig, Mendelssohnstrasse 1, D-38106, Braunschweig, Germany.
authors.forename: Till
authors.initials: T
authors.lastname: Beuerle
authors.affiliation: Institut für Pharmazeutische Biologie, Technische Universität Braunschweig, Mendelssohnstrasse 1, D-38106, Braunschweig, Germany.
authors.forename: Ludger
authors.initials: L
authors.lastname: Beerhues
authors.affiliation: Plant Molecular Biology Group, Biotechnology Department, Indian Institute of Technology Roorkee, Roorkee 247 667, India debsrfbt@iitr.ac.in dsircar.iitkgp@gmail.com.
authors.forename: Debabrata
authors.initials: D
authors.lastname: Sircar
mesh_terms: D001713:Biphenyl Compounds / Q000032:analysis*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D000072318:Dibenzofurans / Q000032:analysis / Q000737:chemistry; D057230:Limit of Detection; D015203:Reproducibility of Results; D027824:Rosaceae / Q000737:chemistry*; D012717:Sesquiterpenes / Q000032:analysis*; D000094162:Phytoalexins
chemical_list: D001713:Biphenyl Compounds; D000072318:Dibenzofurans; D012717:Sesquiterpenes; D000094162:Phytoalexins; C010574:biphenyl