| 摘要: |
| 目的 采用高效液相-飞行时间质谱(HPLC-TOF/MS)联用技术快速分离并鉴别大鼠灌胃给予重楼提取物后尿液中甾体皂苷类成分,以探索重楼皂苷在大鼠体内的代谢途径。方法 选用SD大鼠,按照1.6 g/kg体质量灌胃给予重楼提取物,收集给药后24 h内的尿液。样品分析采用MG-C18柱(3.0 mm×100 mm,3.0 μm),乙腈-0.1%甲酸水为流动相梯度洗脱,在TOF/MS电喷雾离子源下采集正、负离子模式下的数据。利用TOF/MS得到的精确相对分子质量,对照化学成分数据库,对尿液中的甾体皂苷类成分进行鉴别。结果 共鉴别出尿液中20个甾体皂苷原型成分,通过碎片离子以及与对照品相比较鉴别出2对同分异构体: 重楼皂苷Ⅶ和偏诺皂苷元-3-O-α-L-鼠李吡喃糖基(1→4)-α-L-鼠李吡喃糖基(1→3)\[α-L-鼠李吡喃糖基(1→2)\]-β-D-葡萄吡喃糖苷,纤细薯蓣皂苷和偏诺皂苷元-3-O-α-L-鼠李吡喃糖基(1→2)\[α-L-鼠李吡喃糖基(1→4)\]-β-D-葡萄吡喃糖苷。结论 该方法准确、可靠,可成功用于重楼体内成分的鉴别,可为药动学和药效学研究提供参考。 |
| 关键词: 高效液相-飞行时间质谱 重楼 甾体皂苷 尿 |
| DOI：10.3724/SP.J.1008.2013.00421 |
| 投稿时间：2012-11-29修订日期：2013-01-05 |
| 基金项目: |
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| HPLC-TOF/MS in identifying steroidal saponins in rat urine after oral administration of Paris polyphylla extract |
| WANG Ben-wei,ZHANG Jie-jin,LI Yue-yue,ZHANG Hai,ZHAO Liang,Lü Lei,ZHANG Guo-qing* |
(Department of Pharmacy, Eastern Hepatobiliary Hospital, Second Military Medical University, Shanghai 200438, China
*Corresponding author.) |
| Abstract: |
| Objective To use high-performance liquid chromatography-time of flight mass spectrometry (HPLC-TOF/MS) for examining the steroidal saponins in rat urine after oral administration of Paris polyphylla extract, so as to lay a foundation for studying the metabolism of steroidal saponins in vivo. Methods SD rats were administered with an oral dose of 1.6 g Paris polyphylla extracts/kg body weight. The urine samples were collected 24 h after administration by oral gavage. The sample analysis was carried out on a reverse phase MG-C18 column (3.0 mm×100 mm, 3.0 μm) using a gradient mobile phase system of acetonitrile-water containing 0.1% formic acid. TOF/MS was applied for qualitative analysis under positive and negative ion modes. The steroidal saponins in rat urine were identified by using the accurate molecular weight obtained by TOF/MS and formula database. Results A total of 20 steroidal saponins were identified in the rat urine, and two pairs of isomers were deduced through their fragment ions and standards: polyphyllin Ⅶ and pennogenin-3-O-α-L-rhamnopyranosyl(1→4)-α-L-rhamnopyranosyl (1→3)-\[α-L-rhamnopyranosyl(1→2)\]-β-D-glucopyranoside; and gracillin and pennogenin-3-O-α-L- rhamnopyranosyl(1→4)-α-L-rhamnopyranosyl(1→4)-β-D-glucopyranoside.Conclusion The established analysis method is accurate, reliable for identifying steroidal saponins in vivo, which paves a way for further pharmacokinetics and pharmacodynamics study of Paris polyphylla. |
| Key words: high-performance liquid chromatography-time of flight mass spectrometry Paris polyphylla steroid saponin urine |
