今日药学

2018, v.28(06) 387-390

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在线提取HPLC法测定两种虫草中腺苷和虫草素的含量
Determination of Adenosine and Cordycepin in Two Cordyceps Genus Herbs by Online Extraction HPLC

钱正明;李春红;李文庆;宋月林;周妙霞;冯昆;李文佳;
QIAN Zhengming;LI Chunhong;LI Wenqing;SONG Yuelin;ZHOU Miaoxia;FENG Kun;LI Wenjia;Key Laboratory of State Administration of Traditional Chinese Medicine,Sunshine Lake Pharma Co.,Ltd;Modern Research Center for Traditional Chinese Medicine,School of Chinese Materia Medica,Beijing University of Chinese Medicine;Zhuhai Campus,Zunyi Medical College;

摘要(Abstract):

目的建立测定两种虫草样品中腺苷和虫草素含量的在线提取高效液相色谱定量分析方法。方法样品提取条件为:样品粉末和硅藻土均匀混合后置于空预柱芯中,并装入预柱套,组成样品提取池;将样品提取池置于70℃柱温箱中,以水为提取溶剂,提取溶剂以流速1.5 m L·min~(-1)依次通过提取池、保护柱ZORBAX SB-Aq(4.6 mm×12.5 mm,5μm)和分析柱ZORBAX SB-Aq(4.6 mm×150 mm,5μm)产生压力进行在线加压溶剂提取及色谱分析。色谱条件:ZORBAX SB-Aq(4.6 mm×150 mm,5μm)色谱柱,柱温20℃,流动相为水-甲醇梯度洗脱,流速1.5 m L·min~(-1),检测波长260 nm。结果腺苷及虫草素质量浓度分别在1.01~50.32μg·m L~(-1)(r=1.000)和3.98~198.94μg·m L~(-1)(r=1.000)范围内线性关系良好,平均加样回收率分别为103.5%及97.8%,RSD均小于5%;3批冬虫夏草样品的腺苷含量范围为0.24~0.30 mg·g~(-1);3批蛹虫草样品的腺苷含量范围为0.18~0.37 mg·g~(-1);虫草素仅在3批蛹虫草样品中被检测到,其含量为0.74~1.19 mg·g~(-1)。结论该方法准确可靠,可用于冬虫夏草和蛹虫草的质量评价。
OBJECTIVE To develop an online extraction HPLC method for simultaneous determination of adenosine and cordycepin in two Cordyceps herbs. METHODS The powder of sample was mixed with silica gel and packed into a hollow guard column,which then put into a guard column cartridge. To get high pressure and high temperature extract condition,a ZORBAX SB-Aq( 4.6 mm × 12.5 mm,5 μm) column and a ZORBAX SB-Aq( 4.6 mm ×150 mm,5 μm) column were connected to the end of the guard column cartridge,water was used as extraction solvent with flow rate 1. 5 m L·min~(-1),and extraction was operated in column oven( 70 ℃). The separation was performed on a ZORBAX SB-Aq( 4.6 mm×150 mm,5 μm) column with gradient elution by water and methanol,column temperature was20 ℃,flow rate was 1. 5 m L·min~(-1) and detection wavelength at 260 nm. RESULTS The standard curves revealed a good linear relationship in the ranges of 1.01-50.32 μg·m L~(-1)( r= 1.000) for adenosine,3.98~(-1)98.94 μg·m L~(-1)( r= 1.000) for cordycepin,respectively.The average recoveries of the two constituents were 103.5%,97.8%,respectively with RSDs< 5%. The contents of adenosine were 0.24-0.30 mg·g~(-1) for 3 batches of Cordyceps sinensis and 0.18-0.37 mg·g~(-1) for 3 batches of Cordyceps militaris; the contents of cordycepin were 0.74~(-1).19 mg·g~(-1) for 3 batches of Cordyceps militaris. CONCLUSION The method is sensitivity and accuracy,which is suitable for quality evaluation of Cordyceps sinensis( C. sinensis) and Cordyceps militaris( C. militaris).

关键词(KeyWords): 冬虫夏草;蛹虫草;腺苷;虫草素;在线提取;高效液相色谱
Cordyceps sinensis;Cordyceps militaris;adenosine;cordycepin;online extraction;HPLC

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基金项目(Foundation): 国家工信部中药材提升和保障领域项目

作者(Author): 钱正明;李春红;李文庆;宋月林;周妙霞;冯昆;李文佳;
QIAN Zhengming;LI Chunhong;LI Wenqing;SONG Yuelin;ZHOU Miaoxia;FENG Kun;LI Wenjia;Key Laboratory of State Administration of Traditional Chinese Medicine,Sunshine Lake Pharma Co.,Ltd;Modern Research Center for Traditional Chinese Medicine,School of Chinese Materia Medica,Beijing University of Chinese Medicine;Zhuhai Campus,Zunyi Medical College;

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