超高效液相色谱-飞行时间质谱快速分析人参根中的皂甙化合物

建立了超高效液相色谱-飞行时间质谱快速分析人参根部提取物中的皂甙类化合物的方法。色谱柱为HSS T3超高效液相色谱柱(100 mm×2.1 mm, 1.8 μm);以15 mmol/L甲酸铵水溶液-乙腈为流动相,采用二元梯度洗脱的方式对人参主根的皂甙提取物进行分离。基于待测目标物的多级质谱碎片离子、精确质量等信息,结合9种人参皂甙标准化合物的多级质谱碎片离子质谱图,共鉴定出人参主根提取物中27种皂甙类化合物。在确定的条件下,以9种人参皂甙标样为研究对象,进行了全面的方法学考察,发现它们的线性范围分别为0.33～9.00 mg/L (Rg1), 0.11～9.00 mg/L (Re), 0.02～2.00 mg/L (Rf), 0.07～6.00 mg/L (Rg2), 0.04～3.00 mg/L (Rb1, Rb3), 0.22～6.00 mg/L (Rc), 0.04～9.00 mg/L (Rb2, Rd);在中等加标浓度时,经内标物峰面积校正的9种皂甙标准化合物的峰面积的相对标准偏差(RSD)不高于11.3%;低、中、高3个质量浓度加标水平的回收率范围分别为90%～100%、98%～104%及96%～103%;最低检出限为3.5～18.5 μg/L。该方法具有高分辨、快捷、简便、可靠等特点,并成功地应用于分析同一产地、不同生长时间的人参干燥主根中皂甙的差异。可以预计此方法可进一步应用于各种人参原料和制品中皂甙的快速测定。

An ultra performance liquid chromatography-time-of-flight-mass spectrometric method for fast analysis of ginsenosides in Panax ginseng root

A method for fast analysis of ginsenosides in Panax ginseng roots was developed using ultra performance liquid chromatography-time-of-flight-mass spectrometry (UPLC-TOF-MS). The column used was HSS T3 (100 mm x 2.1 mm, 1.8 microm). The mobile phase consisted of 15 mmol/L ammonium formate and acetonitrile, eluted with the gradient program. The separations of 9 ginsenoside standards and ginseng root extracts were achieved. Based on the MS/ MS fragments and accurate masses of the target compounds and with combination of the MS/ MS fragments of the 9 ginsenoside standards, 27 ginsenosides were identified from the extracts of the ginseng roots. The validation of the analytical method was thoroughly investigated with 9 ginsenoside standards. It was found that 9 ginsenosides had a better linearity in 0.04 - 9.00 mg/L. The recoveries at the three spiked levels (low, medium and high) were 90% - 100%, 98% - 104% and 96% - 103%, respectively. The relative standard deviations (RSDs) of the peak area ratio of 9 ginsenoside standards to internal standard at the medium spiked level were not more than 11.3%, which were satisfactory for profiling analysis of herb extracts. This method is characterized by its high resolution, rapidness, simplicity and reliability, and has been successfully applied to the evaluation of the differentiation between 2- and 6-year-old ginseng roots. It can be expected that this method is also useful for the fast determination of the ginsenosides in other ginseng related samples.