A preparative, high-speed, counter-current chromatographic (HSCCC) method for the isolation and purification of bufadienolides from Chansu was successfully developed by using stepwise elution with a two-phase solvent system composed of n-hexane: chloroform: methanol: water (4:1:2.5:5 and 4:1:4:5, v/v). A total of 7.5 mg of cinobufotalin (1), 8.0 mg of bufalin (2), 14.0 mg of cinobufagin (3) and 9.5 mg of resibufogenin (4) were obtained in a one-step separation from 80 mg of the crude extract with purities of 93.2%, 98.7%, 99.2%, and 99.4%, respectively. The chemical structures were determined from 1H NMR and 13C NMR spectroscopic data.
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NingJ, WuTH, TianY, WangCY, TianG, ZhangBJ, LiuKX, MaXC. (2010) Identification of cinobufagin metabolites in the bile of rats. Xenobiotica, 40, 48–54.
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SuZ, WuH, YangY, AisaH, SlukhanU, AripovaS. (2008) Preparative isolation of guaipyridine sesquiterpene alkaloid from Artemisia rupestris L. flowers using high- speed counter-current chromatography. Journal of Separation Science, 31, 2161–2166.
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YeM, GuoH, GuoHZ, HanJ, GuoDA. (2006) Simultaneous determination of cytotxic bufadienolides in the Chinese medicine Chansu by high-performance liquid chromatography coupled with photodiode array and mass spectrometry detection. Journal of Chromatography B, 838, 86–95.
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MaXC, ZhangBJ, DengS, WangCY, ShuXH, HuangSS. (2009) Chemical constituents of bufadienolides from traditional Chinese medicine Veneum bufonis. Progress in Modern Biomedicine, 18, 3519–3522.
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QiaoL, DuanWJ, ChenH, LiuT, PeiYH. (2007) Isolation and identification of bufadienolides from traditional Chinese drug Venenum bufonis. Journal of Shenyang Pharmaceutical University, 24, 611–614.
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ShiSY, ZhangY, ZhaoY, HuangK. (2008) Preparative isolation and purification of three flavonoid glycosides from Taraxacum mongolicum by high-speed counter-current chromatography. Journal of Separation Science, 31, 683–688.
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QiuF, JiangL, YaoS, MaL, KongLY. (2009) Preparative isolation and purification of anthocyanins from purple sweet potato by high-speed counter-current chromatography. Journal of Separation Science, 32, 2146–2151.
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HuangTH, ShenYJ, ShenPN. (2005) Preparative separation and purification of schisandrin and schisantherin from Schisandra chinense Bail by high-speed counter-current chromatography. Journal of Liquid Chromatography & Related Technologies, 28, 2383–2390.
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MaCJ, LiG, ZhengJ, ZhengQS, FanX, WangZH. (2009) An efficient combination of supercritical fluid extraction and high-speed counter-current chromatography to extract and purify homoisoflavonoides from Ophiopogon japonicus Ker-Gawler. Journal of Separation Science, 32, 1949–1956.
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YoungWH, YeongSK. (2009) Preparative isolation of six major saponins from Platycodi radix by high-speed counter current chromatography. Phytochemical Analysis, 20, 207–213.
19.
MaXC, ZhengJ, GuoDA. (2007) Highly selective isomerization and dehydrogenation of three major bufadienolides at 3-OH by Fusarium solani. Enzyme Microbial Technology, 40, 1585–1591.
