The study on the chromatographic fingerprint of Fructus xanthii by microwave assisted extraction coupled with GC–MS
Introduction
Fructus xanthii, the fruit of Xanthium strumarium, commonly known as Cang’erzi in China, is used for the treatment of cramping and numbness of the limbs, ulcer, sinusitis, catarrhs and pruritus [1]. The chemical components of F. xanthii are complex, including volatile components in essential oils and medium extraction components such as amino acid, organic acid and glycosides [2]. In the previous studies on X. strumarium, some compounds such as atractyloside (Atr) [3], carboxyatractyloside (carboxyAtr) [3], thiazinedione [4], sesquiterpene lactones e.g. xanthatin [5] and xanthinin [6] were isolated and identified from the different parts of X. strumarium, such as leaf, stem and fruit. Despite its healing/curing effects, F. xanthii is toxic at high dose and has been shown to cause fatal nephrosis and hypohepatia [2]. The Atr and carboxyAtr are suspected to be the main toxic components of F. xanthii [7], [8]. In clinical application, F. xanthii is always toasted in order to decrease the toxicity [1], [2]. However, the toasting procedure affects the chemical components and efficacy of F. xanthii. Therefore, it is highly desirable to investigate the chemical composition of F. xanthii and the effects of the toasting procedure.
A chromatographic fingerprint is a chromatographic pattern of the extract of some common chemical components of pharmacologically active and/or chemically characteristics in Traditional Chinese Medicines (TCM) sample [9]. In the TCM research, the drugs are complex mixtures, usually containing hundreds of chemical constituents. However, only a few separate markers or pharmacologically active constituents can demonstrate to stand for the complex medicine or their preparations. Therefore, fingerprint technology has been introduced to achieve quality control of herbal medicines in recent years [9], [10], [11], [12], [13], [14].
Soxhlet and normal boiling-water extraction method are the common methods in TCM extraction [1], but these methods are time consuming and require large amounts of the organic solvent. Therefore, some efficient and selective methods such as microwave assisted extraction (MAE), supercritical fluid extraction (SFE) [15] and accelerated solvent extraction (ASE) [16] have been adopted for the component study in recent years. Among these methods, MAE is an effective extraction method for many kinds of herbs especially when the target compounds for extraction are polar. MAE utilizes the energy of microwaves to arouse molecular movement and rotation of liquids with a permanent dipole leading to a very fast heating of the solvent and sample [17]. Some applications of MAE for biologically active compounds have been reported, such as the extraction of taxanes from Taxus biomass [18], tanshinones from the root of Salvia miltiorrhiza [19], glycyrrhizic acid from licorice root [20], quercetin and its glycosides from guava leaves [21], and ginsenosides from ginseng root [22], etc.
Due to the powerful separation efficiency and the sensitive detection, GC–MS has become a popular and useful analytical tool in the research field of herbal medicines [23], [24], [25], [26]. The method of MAE coupled with GC–MS has been used for some desirable components analysis in TCM studies, such as camphor and borneol in Flos Chrysanthemi Indici [27],and paeonol in Cynanchum paniculatum and Paeonia suffruticosa [28]. In this study, we developed a method for investigating the chromatographic fingerprint of F. xanthii by MAE coupled with GC–MS. Chromatographic fingerprint based on this method was established for the characteristic analysis of samples from different producing areas and for the diversity of toasted samples. The fingerprints obtained from this method can provide useful information for the evaluation of the difference in F. xanthii grown in the various areas of China and for further interpretation of the quality changes in toasting process.
Section snippets
Samples
Twenty-five batches of F. xanthii crude samples were purchased from different pharmacies in three producing areas of China (4 batches in Guangxi, 8 batches in Henan and 13 batches in Guangdong). All the samples are in clinical use and have been identified in the pharmacies. Among them, one sample collected from Guangdong was used for the MAE parameters optimization experiment and for the analysis of toasting process. The toasted samples were prepared in an oven by controlling the oven
The optimization of MAE conditions
The ability of microwave-absorption of extracting solvents and the microwave-heat transformation obviously affects the extraction efficiency. The energy transfer between the polar molecules and the solvents, which is coupled with microwave and nonpolar solvents, is rapid and provides an effective mechanism for increasing cell rupture [30]. Because the major components of F. xanthii are polar molecules such as organic acids, alcohols, glucosides, the corresponding derivatives, ethanol, methanol
Conclusion
F. xanthii is a common Traditional Chinese Medicine. Although there are morphological differences and qualitative and quantitative variations in chemical compositions as a result of different origins and toasting methods, the chromatograms of different products were found generally consistent with some common characteristics both in retention times and constitution of components.
Microwave assisted extraction could be considered as an effective technique for rapid and selective extraction for
Acknowledgements
The authors would like to thank the National Natural Science Foundation of China and the National Basic Research Program of China for financially supporting this research under Contract No.20375050 and No. G2000026302, respectively.
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