Makoto Ichikawa

Preparative separation of the saponin lancemaside a from Codonopsis lanceolata by centrifugal partition chromatography

INTRODUCTION Lancemaside A is a saponin that inhibits decreases in blood testosterone level and thus prevents or ameliorates symptoms associated with male climacteric disorder. Our initial attempt to preparative isolation of lancemaside A from the saponin fraction of Codonopsis lanceolata roots by a preparative HPLC did not give a clear result. OBJECTIVE To develop a simple and efficient method for the preparative isolation of lancemaside A from the hot water extract of C. lanceolata roots using centrifugal partition chromatography (CPC). METHODOLOGY The saponin fraction obtained from the hot water extract of C. lanceolata roots was used as the sample for preparative-scale separation of lancemasides by CPC using n-hexane:n-butanol:methanol:0.1% aqueous formic acid (3:4:1:6, v/v) as the two-phase solvent system. The upper phase (organic phase) of the two-phase solvent system was used as the mobile phase, and 0.5 g of saponin fraction was applied for separation by CPC. Each fraction that was separated by CPC was analysed by HPLC, and the fractions containing each of the separated compounds were pooled together, and then were purified by simple preparative HPLC. RESULTS The demonstrated separation sequence, hot water extraction, DIAION HP-20 column chromatography, CPC and preparative HPLC, yielded lancemaside A, foetidissimoside A and astersaponin Hb in their pure forms. CONCLUSION The simple and efficient method for the preparative isolation of lancemaside A along with two other saponins, foetidissimoside A and astersaponin Hb, from the saponin fraction of C. lanceolata was established using CPC.

Rapid identification of triterpenoid saponins in the roots of Codonopsis lanceolata by liquid chromatography–mass spectrometry

Liquid chromatography coupled with sequential mass spectrometry (LC–MSn) has been used to identify 3,28-bidesmosidic triterpenoid saponins, lancemaside A (1), foetidissimoside A (2), aster saponin Hb (3), lancemaside E (4), lancemaside B (5), lancemaside F (6), lancemaside G (7), lancemaside C (8), and lancemaside D (9) in the roots of Codonopsis lanceolata. Structural information about both the aglycone and the sugar moiety at the C-3 position of saponins was obtained in the negative-ion mode. On the other hand, positive-ion spectra mainly provide structural information about the sugar chains of saponins, especially the oligosaccharide moiety at the C-28 position. During subsequent fragmentation of the product ions derived from the oligosaccharide moiety at the C-28 position, fragments produced by sequential loss of a monosaccharide unit were observed. Furthermore, the structural features of two unknown saponins in the roots of C. lanceolata were assigned on the basis of the fragmentation patterns of the known saponins. These studies demonstrate that LC–MSn analysis has great potential for the identification and characterization of triterpenoid saponins in plant extracts.

Antioxidants in processed garlic: II. Tetrahydro-β-carboline derivatives identified in aged garlic extract

Abstract The antioxidant effects and the changes in concentrations of tetrahydro-β-carboline derivatives (THβCs) identified in aged garlic extract (AGE) were determined using several in vitro assay systems and liquid chromatography mass spectrometry (LC-MS). Our data indicate that THβCs are also potent antioxidants, which increase during the natural aging process of garlic and, thus, may contribute to the biological activities of processed garlic such as AGE.