桦褐孔菌子实体和发酵菌丝体中甾类化合物的定量测定

色谱 , 2009, 27(6): 745-749. doi: 10.3321/j.issn:1000-8713.2009.06.002

桦褐孔菌子实体和发酵菌丝体中甾类化合物的定量测定

1.江南大学医药学院制药工程研究室,江苏,无锡,214122;江南大学工业生物技术教育部重点实验室,江苏,无锡,214122

2.江南大学医药学院制药工程研究室,江苏,无锡,214122

3.江南大学工业生物技术教育部重点实验室,江苏,无锡,214122

4.江南大学医药学院制药工程研究室,江苏,无锡,214122;江南大学工业生物技术教育部重点实验室,江苏,无锡,214122

基金项目: the Program for New Century Excellent Talents in the Universities of China(NCET-07-0380) 国家重点基础研究发展规划(973计划)(2007CB707804) National High-Tech Program of China(2007AA021506)

关键词：高效液相色谱 , 甾类化合物 , 子实体 , 菌丝体 , 桦褐孔菌

Quantitative determination of steroids in the fruiting bodies and submerged-cultured mycelia of Inonotus obliquus

Keywords： high performance liquid chromatography (HPLC) , steroids , fruiting bodies , mycelia , Inonotus obliquus

建立了采用高效液相色谱(HPLC)定量测定桦褐孔菌子实体和发酵菌丝体中白桦脂醇、麦角甾醇、胆甾醇、羊毛甾醇、豆甾醇和谷甾醇含量的方法.色谱条件:以C_(18)柱进行分离,流动相为不同浓度梯度的水-甲醇(0-10 min,体积比为10∶90;10-40 min,体积比为3∶97),流速为1.4 mL/min,检测波长为202 nm,整个分析在40 min内完成.结果表明所建立的方法具有很好的重复性和回收率.甾类化合物分析测定的日内相对标准偏差为2.10～2.94%(n=5),在0.4～4.8 μg范围内有很好的线性关系.白桦脂醇、麦角甾醇、胆甾醇、羊毛甾醇、豆甾醇和谷甾醇的回收率分别为100.05% ～100.72%,99.31% ～101.04%,97.52% ～101.63%,96.61% ～100.08%,96.21% ～100.76% 和100.04% ～100.51% .本方法可快速、准确地定量测定桦褐孔菌子实体和发酵菌丝体中的甾类化合物.

This study describes the method of quantitative determination of betulin,ergosterol,cholesterol,lanosterol,stigmasterol and sitosterol in the fruiting bodies and submerged-cultured mycelia of Inonotus obliquus. A high performance liquid chromatographic (HPLC) method was applied to separate these steroids. The procedure was carried out on a reversed-phase C_(18) column,using a stepwise gradient of water-methanol as mobile phase with the following profile: 0-10 min,10% water,90% methanol; 10-40 min,3% water,97% methanol. The flow rate was 1.4 mL/min and the detection wavelength was 202 nm. The analysis was completed within 40 min. The results showed that this method has good reproducibility and satisfactory recoveries for the determination of steroids. The relative standard deviations of the peak areas were less than 2.94% (n=5) for intraday assays. A good linear correlation was obtained in a range of 0.4-4.8 μg. The recoveries of betulin, ergosterol, cholesterol,lanosterol, stigmasterol,and sitosterol were 100.05% -100.72%,99.31% -101.04%,97.52% -101.63%,96.61% -100.08%,96.21% -100.76% and 100.04% -100.51%,respectively. This method can be applied to evaluate real samples,and it is rapid,accurate and suitable for the quantitative determination of steroids in the fruiting bodies and submerged-cultured mycelia of Inonotus obliquus.

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