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中国药学(英文版) ›› 2015, Vol. 24 ›› Issue (6): 376-382.DOI: 10.5246/jcps.2015.06.048

• 【研究论文】 • 上一篇    下一篇

高效液相色谱法测定小叶莲中8种有效成分的含量

王蔼华1,2, 孔越3, 尚明英1*, 尤荣云4, 刘广学1, 徐风1, 蔡少青1   

  1. 1. 北京大学医学部 药学院 天然药物学系, 北京 100191
    2. 北京赛升药业股份有限公司, 北京 100176
    3. 国家知识产权局 专利审查协作中心, 北京 100080
    4. 青海民族大学, 青海 西宁 810007
  • 收稿日期:2015-03-07 修回日期:2015-04-01 出版日期:2015-06-26 发布日期:2015-04-05
  • 通讯作者: Tel.: 86-10-82802534
  • 基金资助:
    National Key Technology R&D Program “New Drug Innovation” of China (Grant No. 2009ZX09308-004, 2013ZX09103002-006).

Simultaneous quantification of eight active compounds in Sinopodophyllum hexandrum by HPLC-DAD

Aihua Wang1,2, Yue Kong3, Mingying Shang*, Rongyun You4, Guangxue Liu1, Feng Xu1, Shaoqing Cai1   

  1. 1. Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
    2. Beijing Saisheng Pharmaceutical Co., Ltd., Beijing 100176, China
    3. Patent Examination Cooperation Center of the Patent Office, SIPO, Beijing 100080, China
    4. Qinghai University for Nationalities, Xining, Qinghai 810007, China
  • Received:2015-03-07 Revised:2015-04-01 Online:2015-06-26 Published:2015-04-05
  • Contact: Tel.: 86-10-82802534
  • Supported by:
    National Key Technology R&D Program “New Drug Innovation” of China (Grant No. 2009ZX09308-004, 2013ZX09103002-006).

摘要:

运用高效液相色谱法, 对藏药小叶莲中3种木脂素(鬼臼毒素、去氧鬼臼毒素和4'-去甲去氧鬼臼毒素)5种黄酮类成分(8-异戊烯基山柰酚、槲皮素、山柰酚、8,2'-二异戊烯基槲皮素3-甲醚、8-异戊烯基槲皮素)进行含量测定, 采用Agilent Zorbax SB-C18色谱柱, 乙腈–0.05%的磷酸水溶液作为流动相进行梯度洗脱; 流速1 mL/min; 柱温35 °C; 检测波长为290 nm370 nm。该方法呈现良好的稳定性: 日内、日间精密度的RSD值分别小于3%4%。各对照品的线性及相关性良好(R2≥0.9992), 加样回收率测定值在98.29%–101.60%之间。采用上述方法测定了12份小叶莲(桃儿七果实)5份桃儿七根及根茎中8种有效成分的含量。结果表明: 8种被测成分在所测样品中的含量差异明显(总含量值从16.90 mg/g55.68 mg/g间不等), 同时发现异戊烯基黄酮类成分是小叶莲中的代表性成分, 可用于小叶莲的质量控制和鉴别研究。

关键词: 小叶莲, 高效液相色谱, 黄酮, 木脂素, 质量控制

Abstract:

Sinopodophylli Fructus is the commonly used traditional Tibetan medicinal herb. In the present study, we established a reversed-phase high performance liquid chromatography method to simultaneously determine three lignans and five flavonoid constituents, namely podophyllotoxin, desoxypodophyllotoxin, 4′-demethyldesoxypodophyllotoxin, 8-prenylkaemferol, quercetin, kaempferol, 8,2'-diprenylquercetin 3-methylether and 8-prenylquercetin, in Sinopodophylli Fructus. The chromatographic separationwas achieved on a C18 analytical column with a gradient mobile phase consisting of acetonitrile and 0.05% phosphoric acid at a flowrate of 1.0 mL/min. UV detection was set at 290 nm and 370 nm, and the column oven was set at 35 °C. This method provided a good reproducibility, and its overall intra- and inter-day precision was less than 3% and 4%, respectively. The recovery of the method was 98.29%101.60%, and a good linearity (R2≥0.9992) was obtained for all the analytes over a relatively wide range of concentration. A total of 17 samples of S. hexandrum (12 fruits, 5 roots and rhizomes) were collected from different areas and then successfully quantified. The results indicated that the contents of eight compounds significantly varied (the sum content rangedfrom 16.90 to 55.68 mg/g), and prenylated flavonoids could be used as marker constituents in the identification and quality control of Sinopodophylli Fructus.

Key words: Sinopodophyllum hexandrum, HPLC, Flavonoids, Lignans, Quality control

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