•Three batches Farfarae Flos were compared by NMR based metabolomic approach.•Chemical and the corresponding metabolic response comparisons were both conducted.•Chemical differences and metabolites variation in serum were correlated.•Pathway analysis was used to reveal the subtle drug induced metabolic difference.Farfarae Flos (FF) is a commonly used herbal drug which has been used for a long time in the Tradition Chinese Medicines (TCM). Nowadays, the cultivated FF in the Northern China is the main source of FF used in the clinic of TCM. The chemical compositions of the herbal drugs are always influenced by the weather, geographic location, soil conditions, and cultivation patterns. Thus it is difficult to guarantee the homogeneity or uniformity of the herbal drugs. In this study, a metabolomic approach was used to compare three batches of FF collected from different growing regions. The results showed that three batches of FFs differed from each other both in the primary and secondary metabolites, and there also existed in vivo differences among three groups of FFs. The clustering pattern of n-butanol extracts was similar to those of crude water extracts and serum, indicating that the polar compounds, such as phenylpropanoids and flavonoids, play an important role in the water extracts of FF. The results presented here suggested that the metabolomic approach can be used as a valuable method to evaluate the difference of herbal drugs from various origins.Download high-res image (314KB)Download full-size image

Farfarae Flos (FF) is widely used in china to treat pulmonary disorders such as coughs, bronchitis, tuberculosis and asthmatic disorders. In this study, a 1H NMR based metabolomic approach coupled with biochemical assay as well as histopathological inspection had been employed to study the protective effect of petroleum extracts of FF (PEFF) against OVA-induced asthma using a rat model. Multivariate analysis revealed that 19 of the perturbed endogenous metabolites in the lung could be reversed by PEFF, and MetPA analysis revealed that the anti-asthamtic effect of PEFF was probably related with regulation of arginine and proline metabolism, pyruvate metabolism and D-glutamine and D-glutamate metabolism. The regulatory effect on the cytokines (IL-4, IL-5, IL-13, and IL-17) and serum HO-1 suggested that the anti-asthmatic action of PEFF was also probably related with the balancing of the Th1/Th2 cells and the Nrf2/HO-1 pathway. The results of chemical analysis showed that the sesquiterpenes were present as the major components in PEFF, and further studies are needed to validate the bioactive compounds responsible for the anti-asthmatic effect of PEFF.

•DOX-induced toxicity was analyzed by echocardiography and NMR based metabonomics.•DOX caused disturbances of a wide range of metabolic pathways.•The changed metabolites were correlated with the echocardiography parameters.Doxorubicin (DOX) is used as a chemotherapy drug with severe carditoxicity. In this study, an integrated echocardiography along with pathological examination and 1H NMR analysis of multiple biological matrices (urine, serum, heart, and kidney) was employed to systemically assess the toxicity of DOX. Echocardiographic results showed that impaired left ventricular contractility and degenerative pathology lesions in DOX group, which were in consistent with pathology. The endogenous metabolites in the urine, serum, heart and kidney was identified by comparison with the data from the literature and databases. Multivariate analysis, including PCA and OPLS, revealed 8 metabolites in urine, including succinate, 2-ketoglutarate, citrate, hippurate, methylamine, benzoate, allantion, and acetate were the potential changed biomarkers. In serum, perturbed metabolites include elevation of leucine, β-glucose, O-acetyl-glycoprotein, creatine, lysine, glycerin, dimethylglycine, trimethylamine-N-oxide, myo-inositol, and N-acetyl-glycoprotein, together with level decreases of acetone, lipid, lactate, glutamate, phosphocholine, acetoacetate and pyruvate. For heart, DOX exposure caused decline of lipid, lactate, leucine, alanine, glutamate, choline, xanthine, glycerin, carnitine, and fumarate, together with elevation of glutamine, creatine, inosine, taurine and malate. Metabolic changes of kidney were mainly involved in the accumulation of α-glucose, lactate, phosphocholine, betaine, threonine, choline, taurine, glycine, urea, hypoxanthine, glutamate, and nicotinamide, coupled with reduction of asparagine, valine, methionine, tyrosine, lysine, alanine, leucine, ornithine, creatine, lipid, and acetate. In addition, alterations of urinary metabolites exhibited a time-dependent manner. Complementary evidences by multiple matrices revealed disturbed pathways concerning energy metabolism, fatty acids oxidation, amino acids and purine metabolism, choline metabolism, and gut microbiota-related metabolism. In addition, the change of endogenous metabolites in rats urine, serum, heart and kidney were correlated with the echocardiography parameters. This integrative study should help to develop a systematic understanding of cardiomyopathy-related diseases and their metabolic events.

Astragali Radix (AR) is a commonly used herbal drug in traditional chinese medicine and is widely used for the treatment of diabetes, cardiovascular diseases, nephropathy, and neuropathy. The main source of AR in China is the dried root of Astragalus membranaceus var. mongholicus (Bge.) Hsiao, and both cultivated and wild ARs are used clinically. A systematic comparison of cultivated AR (GS-AR) and wild AR (SX-AR) should be performed to ensure the clinical efficacy and safety. In this study, the chemical composition of the two different ARs, which were collected in the Shanxi (wild) and Gansu (cultivated) provinces, were compared by NMR-based metabolic fingerprint coupled with multivariate analysis. The SX-AR- and GS-AR-induced metabolic changes in the endogenous metabolites in mice were also compared. The results showed that SX-AR and GS-AR differed significantly not only in the primary metabolites but also in the secondary metabolites. However, alterations among the endogenous metabolites in the serum, lung, liver, and spleen were relatively small. This study provided a novel and valuable method for the evaluation of the consistency and diversity of herbal drugs, and further studies should be conducted on the difference in polysaccharides as well as the biological effects between the two kinds of AR.

•Chemical compositions of Danggui and European Danggui differed significantly.•Different metabolites were determined by multivariate analysis.•Danggui showed better blood enriching effect than European Danggui.•Compounds other than ferulic acid and Z-ligustilide should be further investigated.Danggui is a widely used herbal drug in traditional Chinese medicine, and adulteration with European Danggui is frequently encountered in the market. We compared the chemical compositions and biological effects of Danggui and European Danggui using proton nuclear magnetic resonance spectroscopy coupled with multivariate analysis. Results showed that Danggui and European Danggui differed in both primary and secondary metabolites. Danggui contained higher levels of alanine, γ-aminobutyrate, adenosine, arginine, sucrose, α-glucose, β-glucose, tryptophan, and cis-Z,Z′-3a.7a′,7a.3a′-dihydroxyligustilide than European Danggui. Meanwhile, European Danggui contained higher contents of valine, proline, fumaric acid, phenylalanine, nicotinamide derivative, Z-butylidenephthalide, coniferyl ferulate, ferulic acid, Z-ligustilide, and Z,Z-6,6′7,3a-diligustilide than Danggui. A blood deficiency model was used to compare the biological effects of the two drugs. Despite its higher levels of Z-ligustilide and ferulic acid, European Danggui showed a weaker blood enriching effect than Danggui. Thus, the bioactive compounds responsible for the blood enriching effect in Danggui and their possible synergistic effects should be further studied.

The roots of Rehmannia glutinosa Libosch, Rehmanniae Radix, also known as Dihuang in China, are an important herbal drug in Traditional Chinese Medicine (TCM). According to the theory of TCM, the raw and processed Rehmanniae Radix are used quite differently. In this study, the chemical change of Rehmanniae Radix induced by processing is investigated by 1H NMR spectroscopy based fingerprinting coupled with multivariate data analysis. The results indicated that big chemical change had occurred after the processing of Rehmanniae Radix. Due to the high concentration of sugars, liquid–liquid partition approach was used to eliminate the interfering effect from sugars, in which more metabolites could be detected, and phenethylalcohol glycosides were concentrated in the EtOAc fractions, while the iridoid glycoside were enriched in the n-BuOH part. In addition, various statistical methods were used to find the differential metabolites between the groups. The results obtained in this study suggest that more than one extraction and statistical method should be used for comprehensive metabolic profiling of medical plants, which contained various metabolites in different concentrations.

Tussilago farfara L. is a common herbal medicine which has been widely used to relieve cough and resolve phlegm. In Traditional Chinese Medicine, the flower buds are used, while, in Europe, the leaves are preferred. In this study, the chemical profiles of its leaves and flowers were systematically characterized using gas chromatographic mass spectrometry (GC–MS) and multivariate analysis. Principle component analysis (PCA) and orthogonal projection to latent structures discriminate analysis (OPLS-DA) of GC–MS data provided a clear separation between those samples. The results obviously showed that the metabolome of leaves, flower buds, and fully open flowers are different, and the corresponding loading S-plots revealed that the differential metabolites between the leaves and flowers, and between the fully open flowers and flower buds, were picked out and identified. In conclusion, GC–MS spectroscopy in combination with multivariate analysis has a great potential for chemical comparison study of the metabolome of herbal drugs.Highlights► Chemical comparison between leaves and flowers of T. farfara were first studied. ► A total of 30 nonpolar compounds were identified. ► GC–MS and multivariate analysis were applied to characterize the metabolic difference.

The roots of Rehmannia glutinosa Libosch, Rehmanniae Radix, also known as Dihuang in China, are an important herbal drug in Traditional Chinese Medicine (TCM). According to the theory of TCM, the raw and processed Rehmanniae Radix are used quite differently. In this study, the chemical change of Rehmanniae Radix induced by processing is investigated by 1H NMR spectroscopy based fingerprinting coupled with multivariate data analysis. The results indicated that big chemical change had occurred after the processing of Rehmanniae Radix. Due to the high concentration of sugars, liquid–liquid partition approach was used to eliminate the interfering effect from sugars, in which more metabolites could be detected, and phenethylalcohol glycosides were concentrated in the EtOAc fractions, while the iridoid glycoside were enriched in the n-BuOH part. In addition, various statistical methods were used to find the differential metabolites between the groups. The results obtained in this study suggest that more than one extraction and statistical method should be used for comprehensive metabolic profiling of medical plants, which contained various metabolites in different concentrations.