Studies on content determination and extraction method of iridoid glycosides in Morinda of ficinalis How
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摘要: 目的 建立测定巴戟天环烯醚萜苷类成分含量的方法，并研究其最佳提取方法。 方法 以巴戟天环烯醚萜苷类成分水晶兰苷、去乙酰基车叶草苷酸、车叶草苷酸和车叶草苷为对照品，采用HPLC法进行含量测定。色谱柱为艾杰尔Venusil MP Cl8柱（250 mm ×4.6 mm， 5 μm），流动相为乙腈（A）-0.2%磷酸+0.01 mol/L磷酸氢二钠缓冲盐（B）梯度洗脱（0~12 min，1%~2% A；12~30 min，2%~25% A），检测波长为235 nm，流速为1.0 ml/min，柱温为25 ℃，进样量为20 μl。采用单因素考察和正交试验优化巴戟天环烯醚萜苷类成分的提取方法。 结果 水晶兰苷、去乙酰车叶草苷酸、车叶草苷酸和车叶草苷在0.375~12、0.13~4.16、0.016~0.516和0.012~0.384 μg范围内，线性关系良好；重复性、精密度、加样回收率和稳定性试验结果均符合要求。优化的巴戟天环烯醚萜苷类成分的最佳提取方法为：巴戟天药材用16倍10%乙醇浸泡9 h，渗漉提取，流速为0.8 BV/h。 结论 此法可准确、灵敏地测定巴戟天环烯醚萜苷类成分的含量，并可使该类成分得到充分提取。Abstract: Objective To develop a method for determination of iridoid glycosides in Morinda of ficinalis How. and optimize the extraction methods for iridoid glycosides in Morinda of ficinalis How. Methods The iridoid glycosides, including monotropein, deacetyl asperulosidic acid,asperulosidic acid and asperuloside as standards, HPLC method was developed to determine the content of iridoid glycosides in Morinda of ficinalis How. The separation was performed on Venusil MP C18 (250 mm×4.6 mm, 5 μm) column. The mobile phase was acetonitrile (A)-0.2% phosphoric acid and 0.01 disodium hydrogen phosphate buffer salt (B) with gradient elution (0-12 min, 1%-2% A; 12-30 min, 2%-25% A). The detection wavelength was 235 nm. The flow rate was set at 1.0 ml/min and the column temperature at 25 ℃. The injection volume was 20 μl. Single factor analysis and orthogonal test were used to optimize extraction method of iridoid glycosides in Morinda of ficinalis How. Results Monotropein, deacetyl asperulosidic acid, asperulosidic acid and asperuloside showed good linearity (r>0.999 5) in the ranges of 0.375-12 μg, 0.13-4.16 μg, 0.016-0.516 μg and 0.012-0.384 μg, respectively. This validated method has good repeatability, precision, recovery and stability. It was conformed to meet the requirements and regulation. The optimal extraction method included soaking the raw materials with 16 times of 10% ethanol for 9 h, and then extraction by percolation with the flow rate of 0.8 BV/h. Conclusion The HPLC method sensitively and precisely determined the content of iridoid glycosides in Morinda of ficinalis How. The optimized extraction method extracted these constituents effectively.
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Key words:
- Morinda of ficinalis How. /
- iridoid glycoside /
- content determination /
- HPLC /
- orthogonal test /
- percolate /
- extraction method
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