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| Determination of Ten Ginsenosides in Roots， Leaves and Flowers Buds of Panaxginseng by Solid Phase Extraction/Ultra Performance Liquid Chromatography－Triple Quadrupole Mass Spectrometry |
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| DOI： |
| KeyWord:ginsenosides ultra performance liquid chromatography－triple quadrupole mass spectrometry solid phase extraction ginseng leaves ginseng flowers |
| Author | Institution |
| CHEN Shu-dong， HU Wen-jun， KONG Xiang-ci，FENG Rui， LIANG Tu-jin， CUI Yuan-yuan，ZHONG Man-xia |
1. Guangdong Institute of Traditional Chinese Medicine， Guangzhou ， China;2. Guangdong Medical University， Dongguan ， China |
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| Abstract: |
| A new method based on solid phase extraction/ultra performance liquid chromatography－triple quadrupole mass spectrometry was developed for the determination of ten ginsenosides in roots， leaves and flowers buds of panax ginseng. Firstly， the fragmentation process of the target compounds was determined by analyzing the fragments produced from the ionized lysis of ginsenosides under the negative ion mode of electrospray ion source（ESI－）. Then， the conditions of mass spectrometry and chromatography for the effective separation of 10 ginsenosides were optimized. Meanwhile， an Alumina－N/XAD-2 SPE cartridge composite solid phase extraction column was chosed as the pre-treatment purification column in this study， and XAD－2 macroporous adsorption resin and neutral alumina were also selected as the purification column materials. The principle of this research was based on the molecular exclusion effect of XAD－2 macroporous adsorption resin， which was liable to elute carbohydrates， proteins and organic acids in the sample， while the pigments and flavonoids in the sample were adsorbed by the neutral alumina. Therefore， the ginsenosides were able to further enriched and purified. Finally， the ginsenosides were separated on a Hypersil Gold C18 chromatographic column（100 mm × 2.1 mm， 1.9 μm） using 5 mmol/L ammonium acetate solution containing 0.1% formic acid and acetonitrile as the mobile phases. Furthermore， the ginsenosides were detected with electrospray ionization in negative ion mode under multiple reaction monitoring（MRM） mode. Results showed that there were good linear relationships for ten ginsenosides in the concentration range of 5－2 500 ng/mL， with their correlation coefficients（r） not less than 0.998 0. The limits of detection and limits of quantitation were in the ranges of 0.25－2.5 mg/kg and 0.75－7.5 mg/kg， respectively. For a typical sample of ginseng leaves， the average recoveries at different spiked levels ranged from 87.3% to 110%， with relative standard deviations（RSD） of 1.4%－9.3%. However， the results of different batches of ginseng， ginseng leaves and ginseng flowers showed that the ginsenoside components in leaves and flowers were similar to those in root， while the total contents of some ginsenoside monomers and ten main ginsenosides were more than those in the ginseng root. Overall， the results demonstrated that the proposed method was sensitive and accurate， and was suitable for the ginsenosides quantification in ginseng plants. The method could be applied to the content determination and quality control of ginseng， ginseng flowers and ginseng leaves， and provides a certain basis for the rational development and utilization of ginseng resources. |
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