摘要
目的研究和探讨钩藤药材中钩藤碱与异钩藤碱的最佳提取与含量测定方法。方法采用Phenomenex C18柱(4.6mm×150mm,5μm),以甲醇-0.01mmol·L-1三乙胺水溶液(70∶30,冰醋酸调pH7.0)为流动相;流速:0.5mL·min-1;柱温:30℃;进样器温度:5℃;检测波长为245nm,测定和考察钩藤不同时间的水煎液和不同条件下甲醇冷浸超声提取液中主要有效成分钩藤碱和异钩藤碱含量的变化。结果钩藤碱回归方程Y=76.7170X-0.0727(r=0.9998),在2.5～80.0μg·mL-1范围内线性关系良好,平均加样回收率99.84%～116.91%,RSD小于8.8%;异钩藤碱回归方程Y=87.4729X-0.3666(r=0.9997),在2.0～80.0μg·mL-1范围内线性关系良好,平均加样回收率87.08～104.97%,RSD小于7.3%;甲醇冷浸超声提取液中钩藤碱与异钩藤碱的提取率大约是水煎液中的十倍。在甲醇提取工艺中,影响提取率的因素从大到小依次为超声时间、甲醇用量、冷浸时间,经正交实验优选的最佳提取工艺为用20倍质量的甲醇,冷浸24h,超声60min。结论钩藤药材中有效成分钩藤碱与异钩藤碱的提取,采用甲醇冷浸超声法优于水煎法。该含量测定方法快速、简便、准确,可为评价钩藤药材的质量提供依据。
Objective To explore the optimal method of extraction and determination of rhynchophylline and isorhynchophylline in Ramulus Uncariae cum Uncis. Methods A RP - HPLC was performed on a Phenomenex C18 column （4. 6 mm × 150 ram, 5 pore） at 30 ℃. The mobile phase consisted of methanol - 0. 01 mmol/L triethylamine solution （70： 30, pH 7.0 adjusted by glacial acetic acid） at the flow rate of 0.5 mL/min. The automatic sample injector was set at 5℃ and the ultraviolet detector was operated at 245 nm. And then, the effect of different extraction condition on the contents of rhynehophylline and isorhynchophylline in Ramulus Uneariae cure Uncis was investigated. Results A good linearity of rhynchophylline was in the range of 2.5 p,g/mL to 80.0 μg/mL （Y=76.7170X-0.0727, r=0.999 8）, the average recoveries were from 99.84 % to 116.91% , and the RSD （ n = 6） were less than 8.8 %. A good linearity of isorhynehophylline was in the range of 2.0 μg/mL to 80.0 pug/mL （ Y= 87. 4729X - 0. 3666, r = 0. 999 7）, the average recoveries were from 87.08 % to 104. 97 % , and the RSD （ n = 6） were less than 7.3 %. The contents of rhynchophylline and isorhynehophylline in the methanol - extracted solution were approximately ten times as much as those in water - decocted solution. The main factors which had effect on the extraction of rhynchophylline and isorhynehophylline in methanol solution were supersonic time, methanol amount and colddousing time, in a decreasing sequence. The best condition selected by orthogonal experiment wasb as follows： extracting the medicinal material with 20 - fold volumes of methanol, cold maceration for 24h and supersonic extraction for 1 h. Conclusion The extraction percentage of rhynchophylline and isorhynchophylline in the extraction condition of cold maceration with methanol and supersonic extraction is superior to that in the water - decocting condition. The method is simple, fast and accurate, and it can be used for the quality control of Ramulus Uncariae cure Uncis.
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2009年第4期338-341,共4页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
广东省高等学校中药有效性与安全性重点实验室开放基金(kf07001)
关键词
钩藤
钩藤碱
异钩藤碱
提取方法
含量测定
反相高效液相色谱法
Ramulus Uncariae cure Uncis
Rhynchophylline
Isorhynchophylline
Extraction method
Determination
Reversed phase high performance liquid chromatography （RP - HPLC）
