Molecular characterization of 41 known substances and 6 compounds never detected before in Abelmoschus manihot flowers was performed by Two-Dimensional Microscale Carbon fiber fractionation combined with quadrupole time of flight high-resolution mass spectrometry (2DμCFs-QTOF-HRMS) [1]. The comprehensive analysis was carried out in a single run of only 5 minutes and with a solvent consumption of 1.5 mL. Improved detection sensitivity compared with direct flow-injection MS analyses was achieved by the process of separations into fractions of different polarity (weak, medium, and high) before entering the mass spectrometer. The METLIN database was used to match the accurate experimental MS and MS/MS data. The main components of the flower extracts belong to flavonoids (quercetin, myricetin, quercetin 7-o-glucoside, hyperin, isoquercetin, quercetin 3′-o-glucoside, etc.) amino acids (γ-aminobutyric acid, serine, proline, valine, etc.), nucleosides (adenine, cytidine, 2′- deoxyadenosine, etc.), organic acids (5-(hydroxymethyl)-2-furancarboxylic acid, chlorogenic acid, etc.) and fatty acids (linoleic acid, oleic acid, etc.). The presence of 6 molecules (2-pyrrolidone-5-carboxylic acid, pipecolic acid, trigonelline, glucosamine, 1-aminocyclohexane-carboxylic acid, and 1,2-dioleoyl-sn-glycero-3-phosphocholine) not previously detected in A. manihot flower extracts was observed and confirmed by a deeper investigation based on molecular polarity and mass spectrum fragments. From the dried flowers of Abelmoschus manihot, Huangkui capsule (HKC), a Chinese patent medicine, is prepared and used for the treatment of chronic kidney diseases, including diabetic nephropathy, chronic glomerulonephritis, and membranous nephropathy [2,3]. By 2DμCFs-QTOF-HRMS, a rapid, high-throughput, and comprehensive screening of HKC metabolites in rat plasma was carried out, achieving high sensitivity and metabolite coverage. By excluding the m/z values of blank plasma and HKC extracts, information on HKC plasma metabolites was derived. Most of the metabolites come from the parent hyperoside, which is the main component in HKC. Finally, a comparison was performed between different parts of the Abelmoschus manihot plant. Principal Component Analysis (PCA) was performed using all the results obtained for flowers, buds, and stems, providing valuable insights into the active ingredients of the plant and creating a bridge between phytochemistry and clinical applications.
Bridging the Gap Between HPLC and Direct MS: A Novel 2DμCFs System for High-Throughput and Solvent-Saving Complex Samples Fractionation.
Nardiello, Donatella;Li, Donghao;Quinto, Maurizio
2025-01-01
Abstract
Molecular characterization of 41 known substances and 6 compounds never detected before in Abelmoschus manihot flowers was performed by Two-Dimensional Microscale Carbon fiber fractionation combined with quadrupole time of flight high-resolution mass spectrometry (2DμCFs-QTOF-HRMS) [1]. The comprehensive analysis was carried out in a single run of only 5 minutes and with a solvent consumption of 1.5 mL. Improved detection sensitivity compared with direct flow-injection MS analyses was achieved by the process of separations into fractions of different polarity (weak, medium, and high) before entering the mass spectrometer. The METLIN database was used to match the accurate experimental MS and MS/MS data. The main components of the flower extracts belong to flavonoids (quercetin, myricetin, quercetin 7-o-glucoside, hyperin, isoquercetin, quercetin 3′-o-glucoside, etc.) amino acids (γ-aminobutyric acid, serine, proline, valine, etc.), nucleosides (adenine, cytidine, 2′- deoxyadenosine, etc.), organic acids (5-(hydroxymethyl)-2-furancarboxylic acid, chlorogenic acid, etc.) and fatty acids (linoleic acid, oleic acid, etc.). The presence of 6 molecules (2-pyrrolidone-5-carboxylic acid, pipecolic acid, trigonelline, glucosamine, 1-aminocyclohexane-carboxylic acid, and 1,2-dioleoyl-sn-glycero-3-phosphocholine) not previously detected in A. manihot flower extracts was observed and confirmed by a deeper investigation based on molecular polarity and mass spectrum fragments. From the dried flowers of Abelmoschus manihot, Huangkui capsule (HKC), a Chinese patent medicine, is prepared and used for the treatment of chronic kidney diseases, including diabetic nephropathy, chronic glomerulonephritis, and membranous nephropathy [2,3]. By 2DμCFs-QTOF-HRMS, a rapid, high-throughput, and comprehensive screening of HKC metabolites in rat plasma was carried out, achieving high sensitivity and metabolite coverage. By excluding the m/z values of blank plasma and HKC extracts, information on HKC plasma metabolites was derived. Most of the metabolites come from the parent hyperoside, which is the main component in HKC. Finally, a comparison was performed between different parts of the Abelmoschus manihot plant. Principal Component Analysis (PCA) was performed using all the results obtained for flowers, buds, and stems, providing valuable insights into the active ingredients of the plant and creating a bridge between phytochemistry and clinical applications.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
