Yamasaki Katsuhiro, Kawaguchi Masami, Tagami Takaomi, Sawabe Yoshiyuki, Takatori Satoshi
Osaka Prefectural Institute of Public Health, 1-3-69 Nakamichi, Higashinari-ku, Osaka, Japan.
J Nat Med. 2010 Apr;64(2):126-32. doi: 10.1007/s11418-009-0377-x. Epub 2009 Dec 17.
The sennoside A (SA) and sennoside B (SB) contents of various samples of crude drugs were determined using solid-phase extraction (SPE) and HPLC. The samples examined were crude drugs (senna leaf, senna pods, and rhubarb), conventional crude drug products, and Kampo formulations. The sample solution was purified using an Oasis MAX cartridge, which has strong anion-exchange and reversed-phase properties. The samples containing SA and SB were dissolved in a solution of methanol-0.2% sodium bicarbonate (7:3, v/v) and applied to the Oasis MAX cartridge. The cartridge was washed with a solution of methanol containing 1% acetic acid. SA and SB were eluted with methanol-water-formic acid (70:30:2, v/v), and the eluate was used as the sample solution for HPLC analysis. SA and SB were analyzed using a conventional octadecylsilyl (ODS) column at a detection wavelength of 380 nm; water-acetonitrile-phosphoric acid (800:200:1, v/v) was used as the mobile phase. The SA and SB components in most samples were completely separated from other interfering constituents within 10 min. In particular, several interfering peaks adjacent to the SB peak were eliminated by SPE using the Oasis MAX cartridge. On subjecting the Kampo extracts to an additional recovery experiment, high recovery rates of SA and SB were obtained. The method employed in this study proved to be a simple and rapid method for the quantification of SA and SB.
采用固相萃取(SPE)和高效液相色谱法(HPLC)测定了各种生药样品中的番泻苷A(SA)和番泻苷B(SB)含量。所检测的样品为生药(番泻叶、番泻荚和大黄)、传统生药产品和汉方制剂。样品溶液用具有强阴离子交换和反相性质的Oasis MAX柱进行纯化。将含有SA和SB的样品溶解于甲醇 - 0.2%碳酸氢钠溶液(7:3,v/v)中,然后加到Oasis MAX柱上。该柱先用含1%乙酸的甲醇溶液洗涤。SA和SB用甲醇 - 水 - 甲酸(70:30:2,v/v)洗脱,洗脱液用作HPLC分析的样品溶液。SA和SB采用常规十八烷基硅烷(ODS)柱在检测波长380 nm下进行分析;水 - 乙腈 - 磷酸(800:200:1,v/v)用作流动相。大多数样品中的SA和SB成分在10分钟内与其他干扰成分完全分离。特别是,通过使用Oasis MAX柱的SPE消除了与SB峰相邻的几个干扰峰。对汉方提取物进行额外的回收率实验时,获得了较高的SA和SB回收率。本研究中采用的方法被证明是一种简单快速的SA和SB定量方法。