Endale A., Kammerer B., Gebre-Mariam T., Schmidt P.C.
Department of Pharmaceutical Technology, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany; Institute of Pharmacology and Toxicology, Division of Clinical Pharmacology, University Hospital of Tübingen, Otfried-Müller-Str. 45, 72076 Tübingen, Germany; Department of Pharmaceutics, School of Pharmacy, Addis Ababa University, P.O. Box 1176, Addis Ababa, Ethiopia
Endale, A., Department of Pharmaceutical Technology, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany; Kammerer, B., Institute of Pharmacology and Toxicology, Division of Clinical Pharmacology, University Hospital of Tübingen, Otfried-Müller-Str. 45, 72076 Tübingen, Germany; Gebre-Mariam, T., Department of Pharmaceutics, School of Pharmacy, Addis Ababa University, P.O. Box 1176, Addis Ababa, Ethiopia; Schmidt, P.C., Department of Pharmaceutical Technology, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany
The total flavonoids and saponins of the seeds of Glinus lotoides in the crude extracts and tablet formulation thereof were quantified by reversed-phase high-performance liquid chromatographic (RP-HPLC) methods with UV detection. The saponins were analyzed after acid hydrolysis in 3 M HCl at 100°C for 1 h. Vicenin-2 and mollugogenol B were isolated and used as reference substances for the quantification of total flavonoids and saponins, respectively. The identity and purity (>97%) of the standards were confirmed by spectroscopic (UV, MS, and NMR) and chromatographic (HPLC) methods. The flavonoids and saponins of the crude extract of the seeds and tablet formulation were separated by RP-HPLC (Nucleosil RP-18 column, 250 mm × 4.6 mm) using linear gradient elution systems of acetonitrile-water-0.1 M H3PO4 for flavonoids and methanol-water for saponins. Satisfactory separation of the compounds was obtained in less than 30 and 25 min, for the flavonoids and saponins, respectively. The methods were validated for linearity, repeatability, limits of detection (LOD) and limits of quantification (LOQ). Repeatability (inter- and intra-day, n = 6 and 9, respectively) showed less than 2% relative standard deviation (RSD). The LOD and LOQ were found to be 0.075 and 0.225 mg/mL, respectively, for vicenin-2 and 0.027 and 0.082 mg/100 mL, respectively, for mollugogenol B. The content of flavonoids and saponins of six single tablets was between 95 and 103% for flavonoids and 94-98% for saponins. The validated HPLC methods were employed to standardize a fingerprint of a laboratory produced purified extract, which could be used as a secondary standard for the routine quality control. Accordingly, the purified extract was found to contain 21.3% flavonoids (vicenin-2, 10%) and 25.4% saponins (glinuside G, 14.2%). © 2005 Elsevier B.V. All rights reserved.
Acetonitrile; Aromatic compounds; Extraction; Hydrolysis; Mass spectrometry; Nuclear magnetic resonance spectroscopy; Purification; Seed; Ultraviolet radiation; Ultraviolet spectroscopy; Crude extracts; Limits of detection (LOD); Limits of quantification (LOQ); Relative standard deviation (RSD); Tablet formulation; High performance liquid chromatography; acetonitrile; flavonoid; Glinus lotoides extract; glinuside G; hydrochloric acid; methanol; mollugogenol B; plant extract; saponin derivative; unclassified drug; vicenin 2; vitexin 2'' o glucoside; water; analytical equipment; article; drug purity; elution; glinus lotoides; high performance liquid chromatography; hydrolysis; mass spectrometry; medicinal plant; nuclear magnetic resonance; plant seed; priority journal; quality control; reproducibility; reversed phase high performance liquid chromatography; separation technique; standard; tablet formulation; temperature; ultraviolet radiation; ultraviolet spectroscopy; validation process; Apigenin; Chromatography, High Pressure Liquid; Flavonoids; Glucosides; Molluginaceae; Reference Standards; Saponins; Seeds; Tablets; Extractives; Flavonoids; Liquid Chromatography; Saponins; Seeds; Ultraviolet Radiation; Glinus lotoides