Zifeng Pi

Studies on the aconitine-type alkaloids in the roots of Aconitum Carmichaeli Debx. by HPLC/ESIMS/MSn

Studies of aconitine-type alkaloids in the Chinese herb Aconitum Carmichaeli were performed by HPLC/ESIMS/MS(n) and FTICR/ESIMS in positive ion mode. The characteristic fragmentation pathways in the MS(n) spectra were summarized based on previously published research literature and further study. According to the fragmentation pathways of mass spectrometry, results from the analysis of standard compounds and reports from literature, 111 compounds were identified or deduced in a total of 117 found compounds in A. Carmichaeli. In the 11 monoester-diterpenoid alkaloids (MDA), 10 diesterditerpenoid alkaloids (DDA) and 81 lipo-alkaloids, the novel alkaloids including 1 MDA, 2 DDA and 48 lipo-alkaloids were detected. In addition, 1 DDA, 7 lipo-alkaloids and 2 alkaloids with small molecular weights that possess C19-norditerpenoid skeleton were reported in A. Carmichaeli for the first time.

Chemical profiling of Wu-tou decoction by UPLC-Q-TOF-MS.

Wu-tou decoction (WTD), a traditional Chinese medicine (TCM) formula, is composed of Aconiti Radix Cocta, Ephedrae Herba, Paeoniae Radix Alba, Astragali Radix and Glycyrrhiza Radix Preparata, and it has been used for more than a thousand years to treat rheumatic arthritis, rheumatoid arthritis and pain of joints, while the active constitutions of WTD are unclear. In this research, an ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) method in both positive and negative ion mode was established to investigate the major constitutions in WTD. A Waters ACQUITY UPLC BEH C18 column was used to separate the aqueous extract of WTD. Acetonitrile and 0.1% aqueous formic acid (v/v) were used as the mobile phase. 74 components including alkaloids, monoterpene glycosides, triterpene saponins, flavones and flavone glycosides were identified or tentatively characterized in WTD based on the accurate mass within 15 ppm error and tandem MS behavior. All the constitutions were also detected in the corresponding individual herbs. These results will provide a basis for further study in vivo of WTD and the information of potential new drug structure for treating rheumatic arthritis and rheumatoid arthritis.

Screening and Determination for Potential α‐Glucosidase Inhibitors from Leaves of Acanthopanax senticosus Harms by Using UF‐LC/MS and ESI‐MSn

INTRODUCTION Acanthopanax senticosus Harms is a typical Chinese herb with flavonoids existing in all parts of the plant but with the largest content in leaves. However, leaves have been neglected in past research. To investigate the potential use of leaves of A. senticosus Harms for discovering lead compounds to treat type 2 diabetes, the herb leaves were selected for screening the potential of α-glucosidase inhibitors. OBJECTIVE To screen for candidates of α-glucosidase inhibitors from leaves of A. senticosus Harms and evaluate the structure-activity relationship of the α-glucosidase inhibitors. METHODOLOGY Ultrafiltration liquid chromatography/mass spectrometry (UF-LC/MS) assay was developed for screening candidates of α-glucosidase inhibitors from leaves of A. senticosus Harms. The interesting compounds were identified by using reversed-phase high performance liquid chromatography with diode array detector and electrospray ionisation multiple-stage tandem mass spectrometry (RP-HPLC-DAD-ESI-MS(n) ), and confirmed by using electrospray ionisation Fourier transform ion cyclotron resonance multiple-stage tandem mass spectrometry (ESI-FT-ICR-MS(n)). Furthermore, the α-glucosidase inhibitory activity of the compounds detected was estimated using in vitro assays. RESULTS Eight compounds that might bind to α-glucosidase were screened and seven of them were identified successfully. The α-glucosidase inhibitory activity of the related compounds in leaves of A. senticosus Harms was determined. CONCLUSION The results obtained provided new information for the discovery of potential α-glucosidase inhibitors and the potential anti-diabetic application of the leaves of A. senticosus Harms.

Metabonomic study of Wu-tou decoction in adjuvant-induced arthritis rat using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry

A urinary metabonomics method based on the ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) had been established to investigate the holistic efficacy of Wu-tou decoction (WTD), a traditional Chinese medicine (TCM) formula used to treat rheumatoid arthritis (RA), in adjuvant-induced arthritis (AIA) rat model. Multivariate statistical approaches, such as principal component analysis (PCA) and orthogonal projection to latent structures squares-discriminant analysis (OPLS-DA) were used to distinguish healthy control group, AIA model group and WTD treated group and find potential biomarkers. There was a clear separation among the three groups in PCA model. Sixteen potential biomarkers had been identified using OPLS-DA, and 11 of them was considered to be in response to therapeutic effects of WTD involved in tryptophan metabolism, phenylalanine metabolism, tricarboxylic acid (TCA) cycle, bile acid biosynthesis, steroid hormone biosynthesis and valine metabolism. In this study, WTD also showed good anti-inflammatory and antioxidant activities in vivo, and it could suppress histopathological changes of AIA rats. There might be a correlation between these results and the regulation of the disturbed metabolites in urine. This study demonstrates that metabonomics is a powerful methodology to gain insight in the mechanism of TCM formula in therapy.

HPLC/ESI-MSn and ESI-MS studies on the Aconitum alkaloids in three Chinese medicinal herbs

Aconitum lipo-alkaloids (LDAs) peaks in the previous HPLC studies have shown overlapping behavior, making it impossible to identify the total retention behavior of LDAs. The structural diversity of LDAs in Radix Aconiti, Radix Aconiti Kusnezoffii and Radix Aconiti Lateralis Preparata makes it difficult to synthesize and quantify each of them. Therefore, this study has been devoted to develop a HPLC/ESI-MS(n) method with good resolution of Aconitum alkaloids especially LDAs and then set up a fast ESI-MS SIM method to quantify the total contents of specific skeleton LDAs in the three herbs via in-source CID and the capability of mass separation. As a result, the retention behavior of LDAs was summarized based on an optimized chromatographic separation; 32 alkaloids from the three herbs of Aconitum genus were separated and identified according to their fragmentation pathways by using HPLC/ESI-MS(n). In addition, a fast quantitative analysis ESI-MS SIM method was successfully applied to the quantification of the total contents of specific skeleton LDAs in the three herbs.

Differentiation of various kinds of Fructus schisandrae by surface desorption atmospheric pressure chemical ionization mass spectrometry combined with Principal Component Analysis

Various kinds of Fructus schisandrae were studied by surface desorption atmospheric pressure chemical ionization mass spectrometry (DAPCI-MS) without any sample pretreatment. The volatile components in F. schisandrae were detected in the ambient environment and the analytical time for each sample was only 30s. F. schisandrae are produced mainly in 5 different geographical regions (Elunchun, Mudanjiang, Tonghua, Tieling and Shangluo), and they could be successfully differentiated according to their chemical markers by Principal Component Analysis (PCA). A total of 8 components which gave more contribution for PCA analysis were unambiguously identified by comparison of the MS(2) data of chemical markers to the data of reference compounds as reported in the literature. Similarly, wild grown and cultivatable species of F. schisandrae were well separated by the above-mentioned method. In addition, raw and processed cultivatable F. schisandrae (steamed by water, alcohol, vinegar, or honey, and fried by honey) were found to be clustered at different location, respectively. Furthermore, the clustered degree of differently processed products was correlated with their clinical effects. Our results demonstrated that DAPCI-MS in combination with PCA was a feasible technique for high-throughput differentiation of various kinds of F. schisandrae. It is also possible that DAPCI-MS could become a powerful technology in the studies of traditional Chinese medicine studies and in situ analysis of Chinese herbs.

Effect of ginseng polysaccharide on the urinary excretion of type 2 diabetic rats studied by liquid chromatography-mass spectrometry.

Ginseng polysaccharide is known to have anti-hyperglycemic and anti-hyperlipidemic effects in vivo and its precise mechanism of action is not clear. A urinary metabolomics method based on rapid-resolution liquid chromatography/mass spectrometry (RRLC/MS) was developed to investigate the effect of water-soluble ginseng polysaccharide (WGP) on type 2 diabetes in rats. Principal component analysis (PCA) was carried out for pattern recognition and a clear separation between type 2 diabetic rats and those treated with WGP was achieved. Eight potential biomarkers were found and identified. Significantly increased inosine, serotonin, phenylpropionylglycine and dodecanedioic acid showed the effect of WGP on purine metabolism, tryptophan metabolism, fatty acid metabolism and energy metabolism. 1-Methyladenine, 4-deoxyerythronic acid, 5-hydroxyhexanoic acid and tetrahydrocortisol were significantly decreased which indicated that WGP can regulate DNA metabolism, organic acids metabolism and steroid hormone metabolism. This work is helpful in the effect mechanism study of ginseng polysaccharide.

Effect of 20(S)-ginsenoside Rg3 on streptozotocin-induced experimental type 2 diabetic rats: a urinary metabonomics study by rapid-resolution liquid chromatography/mass spectrometry.

RATIONALE 20(S)-ginsenoside Rg3 is an active component of Panax ginseng. It is known that 20(S)-ginsenoside Rg3 has a protective effect against hyperglycemia, obesity and diabetes in vivo, but the precise mechanisms of these actions have not yet been entirely elucidated. METHODS A urinary metabonomics method based on rapid-resolution liquid chromatography/mass spectrometry (RRLC/MS) was developed to investigate the effect of 20(S)-ginsenoside Rg3 on type 2 diabetic rats. RESULTS With multivariate statistical analysis, a clear separation between type 2 diabetic rats and those treated with 20(S)-ginsenoside Rg3 was achieved. Six potential biomarkers were found and identified. CONCLUSIONS This work shows that the mechanism of the effect of 20(S)-ginsenoside Rg3 on type 2 diabetes may be involved with the regulation of nucleic acid metabolism, energy metabolism and gut flora metabolism.

Application of online microdialysis coupled with liquid chromatography-tandem mass spectrometry method in assessing neuroprotective effect of Rhizoma coptidis on diabetic rats

Monitoring the in vivo dynamics of neurochemicals is very important for studies on learning and memory impairment. However, drawbacks in conventional sampling and off-line analysis methods require the employment of a more satisfying system. We present an online microdialysis coupled with liquid chromatography-tandem mass spectrometry method (LC-MS/MS) to determine eight neurochemicals simultaneously, including glutamate, aspartic acid, γ-aminobutyric acid, serine, taurine and acetylcholine, as well as dopamine and serotonin. A self-assembled automated injector with a high-pressure-resisting six-port valve was utilized to control the switch of the two states between loading and injection, which potentially simplified the sample preparation procedures and avoided the restrictions offered by small sample volume. Good linearity (R2 > 0.997) was observed in each analyte dynamic range. The inter-day and intra-day precisions (RSD) were <15%, and the accuracy (RE) was from −13.84% to 11.16%. The overall matrix effect was within an acceptable range, from 87.79% to 111.71%. Morris water maze (MWM) test was performed to evaluate the differences of cognitive abilities among normal control, diabetes mellitus (DM) model and Rhizoma coptidis (R. coptidis)-treated DM group rats. Moreover, the developed method was applied for the further exploration of the R. coptidis efficacies on the eight analytes changes in the hippocampus of diabetic rats with mild cognitive impairment. It was found that an improvement in neurochemicals imbalance was observed in diabetic rats after treatment with R. coptidis. In conclusion, the developed online analysis method is selective, sensitive and accurate, which is suitable for in vivo neurochemicals monitoring and could be extended to pharmacology studies.

Ginsenosides attenuate d-galactose- and AlCl3-inducedspatial memory impairment by restoring the dysfunction of the neurotransmitter systems in the rat model of Alzheimer's disease.

ETHNOPHARMACOLOGICAL RELEVANCE Panax ginseng C.A.Mey. is a traditional Chinese herbal medicine, which has been used to treat Alzheimers disease (AD) for thousands of years. Ginsenoside is one of the major compounds found in P. ginseng. This study aimed to explore the attenuation of spatial memory impairment by ginsenosides and its correlation with restoring the dysfunction of the neurotransmitter systems in AD model rats to understand the mechanism underlying the anti-AD effect of P. ginseng. MATERIALS AND METHODS In this study, the AD model was established by combining d-galactose (d-gal) with AlCl3 (Al) for 60 days. From day 30, the ginsenosides group was intragastrically administered with ginsenosides for 30 days. The ethology of rats was tested through the Morris water maze test(MWM). Histopathological changes in the hippocampus of rats were observed through hematoxylin and eosin staining. The expressions of amyloid β peptide (Aβ) and phospho-tau (p-tau) in the hippocampus and cortex of rats were detected by immunohistochemistry. A liquid chromatography-mass spectrometry assay was used to measure neurotransmitter concentrations in the hippocampus, cortex, and blood. RESULTS Ginsenosides could significantly decrease the escape latency time and the average latency time in the place navigation test and increase the times of crossing the platform area, the percentage of residence time, and the distance in the original platform quadrant in the spatial probe test. Ginsenosides could repair the damage of the hippocampus and reduce the expressions of Aβ and p-tau. Ginsenosides could also increase γ-aminobutyric acid, acetylcholine, and dopamine levels and decrease glutamate and aspartic acid levels in the hippocampus and cortex and increase glycine and serotonin levels in the blood. CONCLUSIONS After effectively administrated, ginsenosides attenuate d-gal- and Al-induced spatial memory impairment. The possible mechanism of the beneficial effect is restoring the dysfunction of various neurotransmitters.