附子水溶性生物碱提取纯化工艺研究

目的研究附子水溶性生物碱的提取纯化工艺。方法采用UPLC-MS/MS技术,以14种生物碱(多根乌头碱、宋果灵、附子灵、尼奥灵、塔拉萨敏、新乌头原碱、乌头原碱、次乌头原碱、苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、新乌头碱、乌头碱、次乌头碱)的总量为考察指标,采用单因素实验和正交试验方法筛选最佳提取工艺;以8种水溶性生物碱(乌头原碱、新乌头原碱、次乌头原碱、附子灵、尼奥灵、塔拉萨敏、宋果灵、多根乌头碱)的量为考察指标,对5种大孔树脂吸附和解吸附性能进行考察,优选最佳纯化工艺。结果最佳提取纯化工艺为蒸附片加10倍量pH值3.5的酸水,煎煮3次(维持pH值约3.5),每次2 h,合并水煎液,用20%NaOH溶液调pH值10.0,加热回流水解2 h后,放冷,调pH值11.0,上大孔树脂HPD 300,上样量为生药2.5 g/mL树脂,6 BV水洗除杂,4 BV 80%乙醇洗脱,收集乙醇洗脱液,减压浓缩干燥,即得附子水溶性生物碱提取物。该提取物收率为1.69%,其中新乌头原碱、乌头原碱、次乌头原碱、附子灵、尼奥灵、塔拉萨敏、多根乌头碱、宋果灵的总量高达约15%。结论优选得到的提取纯化工艺稳定可行,为附子水溶性生物碱的研究应用提供参考。

Study on process of extraction and purification of water-soluble alkaloids from Aconiti Lateralis Radix Praeparata

Objective To research the process of extraction and purification of water-soluble alkaloids of Aconiti Lateralis Radix Praeparata (ALRP). Methods Orthogonal design L₉(3⁴) and single factor experiments were employed to optimize the extraction conditions using UPLC-MS/MS. The total extraction yield of 14 alkaloids (karacoline, songorine, fuziline, neoline, talatisamine, mesaconine, aconine, hypaconine, benzoylmeaconine, benzoylaconine, benzoylhypaconine, mesaconitine, aconitine, and hypaconitine) was used as an index. The absorption-desorption characteristics of five macroporous resins were evaluated to optimize purification conditions using contents of eight water-soluble alkaloids (aconine, mesaconine, hypaconine, fuziline, neoline, talatisamine, songorine, and karacoline) as indexes. Results The optimized extraction and purification conditions were as follows:steamed tablets were decocted thrice with 10-fold pH 3.5 acidic water for 2 h each time (maintaining pH 3.5). The alkalinity of water decoction was adjusted to pH 10.0 with 20% NaOH and then heated for 2 h. The supernatant was adjusted to pH 11.0 by 20% NaOH and then was subjected to macroporous resin HPD300 whose relative adsorption amount was 2.5 g dried medicinal herb/mL resin, and then the resin was eluted with 6 BV of water and 4 BV of 80% ethanol respectively. The 80% ethanol fraction was evaporated under vacuum to give the water-soluble alkaloids extract. The extract yield was 1.69%. The total contents of mesaconine, aconine, hypaconine, fuziline, neoline, talatisamine, karacoline and songorine were above 15%. Conclusion The optimized extraction and purification process is stable and feasible, and this present study provides the reference for the research and application of water-soluble alkaloids in ALRP.