重楼药材和混伪品中5种皂苷的含量及对《中国药典》2015年版重楼含量测定修订的探讨

重楼为百合科植物云南重楼Paris polyphylla var.yunnanensis或七叶一枝花P.polyphylla var.chinensis的干燥根茎。针对现行重楼药材标准存在的问题,前期研究提出"删除重楼皂苷Ⅵ,增补重楼皂苷H,以重楼皂苷Ⅰ,Ⅱ,H和Ⅶ的总量进行限度控制"。为了考察更广范围内重楼样品中重楼皂苷H的含量以及重楼皂苷Ⅰ、重楼皂苷Ⅱ、重楼皂苷H和重楼皂苷Ⅶ的总量,该研究基于2015年版《中国药典》重楼标准定量方法的色谱条件,进行了同步测定重楼皂苷Ⅰ、重楼皂苷Ⅱ、重楼皂苷H和重楼皂苷Ⅶ含量的方法学验证,测定了69批不同来源重楼样品的含量;同时,测定了易混伪药材头顶一颗珠以及重楼属7种近缘植物根茎中5种重楼皂苷的含量。结果表明重楼皂苷Ⅰ、重楼皂苷Ⅱ、重楼皂苷H和重楼皂苷Ⅶ分别在0.006 48~0.828,0.006 52~0.834,0.006 17~0.790,0.006 31~0.808 g·L-1线性关系良好,平均加样回收率在100.2%~101.4%,RSD均小于3.5%。方法学验证均符合含量测定要求。36批不同产地的七叶一枝花样品中重楼皂苷Ⅰ、重楼皂苷Ⅱ、重楼皂苷H和重楼皂苷Ⅶ的总量在0.050 9%~3.99%(不含3批未检出);33批不同产地的云南重楼样品中4种成分总量在0.115%~3.23%。同属近缘植物中,球药隔重楼和长柱重楼中甾体皂苷含量较高,狭叶重楼、金线重楼、黑籽重楼含量较低,毛重楼几乎不含皂苷。饮片易混伪品头顶一颗珠中重楼皂苷Ⅵ在0.526%~1.55%,并且不含重楼皂苷H。建议以干燥品计,含重楼皂苷Ⅰ、重楼皂苷Ⅱ、重楼皂苷H和重楼皂苷Ⅶ的总量不得少于0.80%进行限度控制,以提升商品重楼药材质量,遏制低含量的重楼属近缘种(比如毛重楼、黑籽重楼和狭叶重楼等)以及头顶一颗珠的混伪现象。

Determination of five steroidal saponins in Paridis Rhizoma and its adulterants as well as consideration on its quantitative method described in Chinese Pharmacopoeia (2015 edition)

Paridis Rhizoma is prepared from the dried rhizoma of Paris polyphylla var. yunnanensis or P. polyphylla var. chinensis. For the improvement of the quality standard of Paridis Rhizoma described in Chinese Pharmacopoeia(2015 edition), it is proposed that the quality marker no longer contains polyphyllin Ⅵ, and instead, polyphyllin H is an alternative for the quantitative analysis. To determine polyphyllin Ⅰ, Ⅱ, H and Ⅶ in the Paridis Rhizoma samples collected from the different growing area in China, HPLC method was established using the same chromatographic conditions as those for simultaneous determination of polyphyllin Ⅰ, Ⅱ, Ⅵ and Ⅶ described in Chinese Pharmacopoeia(2015 edition). The methodology validation indicated that there was a good linearity among the ranges of 0.006 48-0.828, 0.006 52-0.834, 0.006 17-0.790, 0.006 31-0.808 g·L-1 for polyphyllin Ⅰ, Ⅱ, H and Ⅶ, respectively. The average recoveries of four components were 100.2%-101.4%, with RSD less than 3.5%. The total amount of polyphyllin Ⅰ, Ⅱ, H and Ⅶ in the analyzed samples of P. polyphylla var. chinensis and P. polyphylla var. yunnanensis ranged from 0.050 9% to 3.99% and from 0.115% to 3.23%, respectively. In the tested samples collected from other Paris plants, there are high content of steroidal saponins in the samples of P. fargesii and P. forrestii, low content in the samples of P. polyphylla var. stenophylla, P. delavayi and P. thibetica, and almost not occurrence in the sample of P. mairei. As a representative adulterant of Paridis Rhizoma processed slices, 7 batches of Trillium samples contained high amount of polyphyllin Ⅵ and did not have polyphyllin H. Based on the present investigation, it is recommended that polyphyllin H together with polyphyllin Ⅰ, Ⅱ and Ⅶ are suitable for the improvement of quality standard of Paridis Rhizoma and the total amount of four components are not less than 0.80%.