Rapid extract dereplication using HPLC-SPE-NMR: analysis of isoflavonoids from Smirnowia iranica.
A novel hyphenated technique, HPLC-SPE-NMR, was used for accelerated identification of isoflavonoids from the roots of Smirnowia iranica. The extract constituents eluted from a HPLC column were automatically trapped on solid-phase extraction (SPE) cartridges, and NMR spectra were acquired with concentrated solutions after solvent change. The structures of 10 new isoflavonoids (1, 4, 5, 7-10, 12, 13, 16) and of seven previously described constituents (2, 3, 6, 11, 14, 15, 17) were elucidated from NMR spectra acquired in the HPLC-SPE-NMR mode. Multiple peak trapping on the same SPE cartridge increased analyte amounts and provided access to 2D NMR data. It was demonstrated that linear accumulation of material is possible in up to seven repeated trapping steps. The use of HPLC-SPE-NMR speeded up dereplication of the S. iranica extract considerably by providing detailed information about the constituents of a complex, essentially crude extract prior to their preparative-scale isolation or extract pre-fractionation, and the information obtained could be used to direct preparative isolation work. In connection with structure elucidation of isoflavonoids containing O-methylated 1,2,3-benzenetriol moieties as the B-ring, O-methylation-induced changes of chemical shifts of aromatic hydrogens were found to depend on the conformation of the resulting methoxy group, i.e., on the number of its ortho substituents. The recognized regularities will be useful in structure determination of partially O-methylated polyphenols based on 1D (1)H NMR spectra obtainable from HPLC-SPE-NMR experiments, diminishing dependence on 2D NMR data and (13)C NMR chemical shifts.
Lambert
Maja
M
Department of Medicinal Chemistry and Department of Analytical Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
Staerk
Dan
D
Hansen
Steen Honoré
SH
Sairafianpour
Majid
M
Jaroszewski
Jerzy W
JW
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D007887:Fabaceae / Q000737:chemistry*; D007492:Iran; D007529:Isoflavones / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D018517:Plant Roots / Q000737:chemistry; D010946:Plants, Medicinal / Q000737:chemistry*
D007529:Isoflavones