Zhang Ying-Ying, Jin Lin, Wu Yu-Ying, Li Rui, Wang Kai-Ling, Li Hai-Feng
College of Pharmacy, Dali University Dali 671000, China Institute of Materia Medica, Dali University Dali 671000, China Yunnan Key Laboratory of Screening and Research on Anti-pathogenic Plant Resources from Western Yunnan (Cultivation) Dali 671000, China.
Infectious Disease Hospital of Lijiang Lijiang 674100, China.
Zhongguo Zhong Yao Za Zhi. 2021 Oct;46(19):4936-4944. doi: 10.19540/j.cnki.cjcmm.20210722.101.
In this study, liquid chromatography-mass spectrometry(LC-MS) and high performance liquid chromatography(HPLC) were employed for qualitative and quantitative analysis of the steroidal saponins in rhizomes of Paris polyphylla var. yunnanensis from three different habitats cultured in vitro, in an attempt to explore whether the rhizomes of the medicinal herb cultured in vitro can synthesize the steroidal saponins, including polyphyllinsⅠ, Ⅱ, and Ⅶ, the quality markers specified in Chinese Pharmacopoeia(2020 edition). A total of 20 steroidal saponins were identified in the rhizomes from Changxin, Yunlong(S1), Fengyi, Dali(S2), and Niujie, Eryuan(S3): parisyunnanoside A and parisyunnanoside D or E, proto-polyphyllin Ⅱ, polyphyllins G and H, polyphyllinsⅠ, Ⅱ, Ⅴ, Ⅵ, and Ⅶ, dioscin, gracillin, prosapogenin A, Tg, isomer of Th, saponin Th, reclinatoside, proto-pairs D, pseudoproto-dioscin, and 23-O-glc-(23S,25R)-spirost-5-en-3β,23α,27-triol-3-O-rha-(1→2)-[ara(1→4)]-glc or 27-O-glc-(23S,25R)-spirost-5-en-3β,27α-diol-3-O-rha-(1→2)-[ara(1→4)]-glc. Among them, polyphyllinsⅠ, Ⅱ, and Ⅶ were detected in the rhizomes from S1, with the mass fraction of 0.109 1%, 0.165 2%, and 0.051 03%, respectively(total 0.325 3%). Polyphyllins Ⅱ and Ⅶ were identified in the rhizomes from S2 with the respective mass fraction of 0.192 2% and 0.074 23% and total content of 0.266 5%. Moreover, polyphyllins Ⅱ and Ⅶ were also found in the rhizomes from S3, which had the mass fraction of 0.207 7% and 0.186 9%, separately, with the total content of 0.394 6%. Thus, steroidal saponins, including the quality makers polyphyllins Ⅰ, Ⅱ, and Ⅶ recorded in Chinese Pharmacopoeia(2020 edition) can be synthesized in rhizomes of Paris polyphylla var. yunnanensis cultured in vitro, but their total content fails to meet the standard(0.60% in Chinese Pharmacopoeia). Therefore, in vitro culture of the Paris polyphylla var. yunnanensis is feasible, but the culture conditions need to be further improved.
本研究采用液相色谱 - 质谱联用（LC - MS）和高效液相色谱（HPLC）对三种不同生境下的滇重楼组培苗根茎中的甾体皂苷进行定性和定量分析，以探究组培苗根茎能否合成甾体皂苷，包括《中国药典》（2020年版）规定的质量标志物重楼皂苷Ⅰ、Ⅱ和Ⅶ。在来自长新乡、云龙（S1）、凤仪、大理（S2）和牛街、洱源（S3）的根茎中共鉴定出20种甾体皂苷：滇重楼苷A、滇重楼苷D或E、原重楼皂苷Ⅱ、重楼皂苷G和H、重楼皂苷Ⅰ、Ⅱ、Ⅴ、Ⅵ和Ⅶ、薯蓣皂苷、纤细皂苷、原皂苷元A、Tg、Th的异构体、皂苷Th、偏诺皂苷、原重楼皂苷D、伪原薯蓣皂苷，以及23 - O - 葡萄糖 -（23S，25R）- 螺旋甾 - 5 - 烯 - 3β，23α，27 - 三醇 - 3 - O - 鼠李糖 -（1→2）-[阿拉伯糖（1→4）]- 葡萄糖或27 - O - 葡萄糖 -（23S，25R）- 螺旋甾 - 5 - 烯 - 3β，27α - 二醇 - 3 - O - 鼠李糖 -（1→2）-[阿拉伯糖（1→4）]- 葡萄糖。其中，在S1的根茎中检测到了重楼皂苷Ⅰ、Ⅱ和Ⅶ, 质量分数分别为0.109 1%、0.165 2%和0.051 03%（总计0.325 3%）。在S2的根茎中鉴定出了重楼皂苷Ⅱ和Ⅶ，质量分数分别为0.192 2%和0.074 23%，总含量为0.266 5%。此外，在S3的根茎中也发现了重楼皂苷Ⅱ和Ⅶ，质量分数分别为0.207 7%和0.186 %，总含量为0.396 %。因此，《中国药典》（2020年版）中记录的甾体皂苷，包括质量标志物重楼皂苷Ⅰ、Ⅱ和Ⅶ，能够在滇重楼组培苗根茎中合成，但它们的总含量未达到标准（《中国药典》中为0.60%）。因此，滇重楼组培可行，但培养条件有待进一步优化。
