Dan Mo, Xie Guoxiang, Gao Xianfu, Long Xiangbao, Su Mingming, Zhao Aihua, Zhao Tie, Zhou Mingmei, Qiu Yunping, Jia Wei
School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
Phytochem Anal. 2009 Jan-Feb;20(1):68-76. doi: 10.1002/pca.1099.
Saponins are bioactive compounds employed in the prevention and treatment of cardiovascular and cerebrovascular diseases. The adventitious roots of Panax notoginseng may offer an alternative source of saponins. Identification and determination of saponins in the crude extract is challenging owing to their similar structures and the lack of standards.
To develop a rapid, sensitive and accurate method based on solid-phase extraction followed by ultra-performance liquid chromatography-electrospray ionisation mass spectrometry (UPLC-ESI-MS) for the identification and quantification of saponins in P. notoginseng.
Following extraction using Waters Oasis(TM) HLB cartridges, the analytes were subjected to a UPLC system with a Waters Acquity BEH C(18) chromatographic column and a binary mobile phase system consisting of 0.05% formic acid in water and acetonitrile under gradient elution conditions, with final detection by ESI-MS in the positive ion mode.
The UPLC-ESI-MS method gave limits of detection and quantification within the range 0.015-0.382 and 0.052-1.124 microg/mL, respectively, for 15 studied saponins. The instrumentation/injection precision (RSD) was 4.5% for a low concentration and 3.2% for an intermediate concentration sample. The intra- and inter-day repeatability was less than 2.65% (RSD). The method described was validated using spiked samples with different amounts of saponin standards.
This UPLC-ESI-MS assay provides a suitable quality control method for the tentative identification and determination of major biological active constituents in adventitious and native roots of P. notoginseng.
皂苷是用于预防和治疗心脑血管疾病的生物活性化合物。三七的不定根可能是皂苷的另一种来源。由于皂苷结构相似且缺乏标准品,因此对粗提物中皂苷的鉴定和测定具有挑战性。
建立一种基于固相萃取结合超高效液相色谱-电喷雾电离质谱(UPLC-ESI-MS)的快速、灵敏且准确的方法,用于三七中皂苷的鉴定和定量分析。
使用Waters Oasis™ HLB柱进行萃取后,将分析物注入配备Waters Acquity BEH C18色谱柱的UPLC系统中,采用由0.05%甲酸水溶液和乙腈组成的二元流动相体系,在梯度洗脱条件下进行分离,最后采用ESI-MS在正离子模式下进行检测。
对于15种研究的皂苷,UPLC-ESI-MS方法的检测限和定量限分别在0.015 - 0.382 μg/mL和0.052 - 1.124 μg/mL范围内。低浓度样品的仪器/进样精密度(RSD)为4.5%,中等浓度样品为3.2%。日内和日间重复性均小于2.65%(RSD)。该方法通过向样品中添加不同量的皂苷标准品进行了验证。
这种UPLC-ESI-MS分析方法为三七不定根和原生根中主要生物活性成分的初步鉴定和测定提供了一种合适 的质量控制方法。