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


METHODS
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.


CONCLUSIONS
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.
Shirota
Osamu
O
Laboratory of Pharmacognosy and Natural Products Chemistry, Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University, Kagawa 769-2193, Japan. shirota@kph.bunri-u.ac.jp
Nagamatsu
Kumi
K
Sekita
Setsuko
S
Komoto
Noriko
N
Kuroyanagi
Masanori
M
Ichikawa
Makoto
M
Ohta
Sanae
S
Ushijima
Mitsuyasu
M
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D029745:Codonopsis / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D010936:Plant Extracts / Q000737:chemistry; D012503:Saponins / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet
D010936:Plant Extracts; D012503:Saponins; C533150:lancemaside A