Extraction and isolation of flavonoid glycosides from Flos Sophorae Immaturus using ultrasonic‐assisted extraction followed by high‐speed countercurrent chromatography

A method of ultrasonic‐assisted extraction followed by high‐speed countercurrent chromatography was established for the extraction and isolation of three flavonoid glycosides, i.e. rutin, narcissin, and nicotiflorin from Flos Sophorae Immaturus. The effects of ultrasonic‐assisted extraction factors for the main flavonoid compound (rutin) from Flos Sophorae Immaturus were optimized using Box–Behnken design combined with response surface methodology. The optimum conditions were determined as ultrasonic power 83% (600 W), solvent‐to‐material ratio 56:1, methanol concentration 82% v/v, and extraction time 60 min. Three bioactive flavonol glucosides, rutin, narcissin, and nicotiflorin were isolated from Flos Sophorae Immaturus using high‐speed countercurrent chromatography. The separation was performed with a two‐phase solvent system containing ethyl acetate/n‐butanol/methanol/water (4:0.9:0.2:5, v/v). Amounts of 87 mg of rutin, 10.8 mg of narcissin, and 1.8 mg of nicotiflorin were isolated from 302 mg of crude extract of Flos Sophorae Immaturus in a one‐step separation within 160 min with purities of 99.3, 98.0, and 95.1%, respectively, as determined by HPLC with diode array detection. Their structures were characterized by UV, MS, and NMR spectroscopy. It was demonstrated that the established method was simple, fast, and convenient, which was feasible to extract and isolate active flavonoid glycosides from Flos Sophorae Immaturus.     

补骨脂及其混伪品蛋白质电泳鉴别

为鉴别补骨脂与其混伪品,对两种种子的可溶性蛋白质进行了聚丙烯酞胺凝胶电泳分析.结果表明,补骨脂及其混伪品的电泳图谱存在显著差异,电泳图谱可作为补骨脂及其混伪品的鉴别依据.     

Carbon nanotube/polystyrene composite electrode for microchip electrophoretic determination of rutin and quercetin in Flos Sophorae Immaturus

In this report, carbon nanotube/polystyrene (CNT/PS) composite electrodes have been fabricated as sensitive amperometric detectors of microchip capillary electrophoresis (CE) for the determination of rutin and quercetin in Flos Sophorae Immaturus. The composite electrode was fabricated on the basis of the in situ polymerization of a mixture of CNT and styrene in the microchannel of a piece of fused silica capillary under heat. The surface morphologies of the composite in the electrodes were observed by using a scanning electron microscope. The performance of this unique system has been demonstrated by separating and detecting rutin and quercetin. The new CNT-based CE detector offered significantly lower detection potentials, yielded substantially enhanced signal-to-noise characteristics, and exhibited resistance to surface fouling and hence enhanced stability. It demonstrated long-term stability and reproducibility with a relative standard deviation of less than 5% for the peak current (n = 20) and should also find a wide range of applications in conventional CE, flowing injection analysis, and other microfluidic analysis systems.     

高效液相色谱-电喷雾质谱法测定枳壳中黄酮苷类化合物

利用高效液相色谱与电喷雾质谱联用技术研究了枳壳中的黄酮苷类化合物。实验采用反相C18色谱柱，二元线性梯度洗脱，分离并检测了枳壳中的6种黄酮苷类化合物；它们分别是新圣草苷（neoeriocitrin）、异柚皮苷（isonaringin）、柚皮苷（naringin）、橙皮苷（hesperidin）、新橙皮苷（neohesperidin）和新枸橘苷（neoponcirin）；通过与电喷雾质谱联用获得了这6种黄酮苷的准分子离子峰（[M＋H]^＋）及分子加钠峰（[M＋Na]^＋），利用质谱的碰撞诱导解离技术获得了碎片裂解信息。通过这此质谱信息并结合文献，对这6种化合物进行了结构鉴定。     

A simple liquid chromatography coupled with tandem mass spectrometry approach for the simultaneous quantification of thirteen compounds in rats following oral administration of raw and processed Fructus Xanthii: Application in a comparative pharmacokinetic study

A simple and sensitive analysis using ultra high performance liquid chromatography with a tandem mass spectrometric system operated in selected reaction monitoring mode was developed for the determination of 11 phenolic acids, atractyloside, and carboxyatractyloside in rat plasma. The two classes of analytes were then separated on a Waters ACQUITY? UPLC HSS T3 column (50 mm × 2.1 mm, 1.8 µm) using gradient elution with a mobile phase of 0.2% formic acid in water containing 10 mM ammonium acetate and methanol. Detection was accomplished by selected reaction monitoring scanning via an electrospray source operating in negative ionization mode. The calibration curve was linear (R² = 0.990) over a concentration range of 1.20–3500 ng/mL, while the validated lower limit of quantification was 1.20 ng/mL. The precision varied from 0.84 to 4.62%, and the accuracy varied within ±5%. The method proved robust with sample freezing and thawing and with short‐ and long‐term sample storage. The established method was used for simultaneous quantification and was successfully used for the first time for the pharmacokinetic evaluation of 13 compounds after the intragastric administration of raw and processed Fructus Xanthii in rats. The results indicated that processing affects the absorption and metabolism of Fructus Xanthii extract. Importantly, the results also indicated the importance of processing for the clinical application of traditional Chinese medicine.     

Classification of Aurantii Fructus samples by multivariate analysis

Twenty commercial samples of Aurantii Fructus Immaturus (Poncitrus trifoliata) and 30 of Aurantii Fructus Maturus (Citrus aurantium and C. wilsonii) were collected from the Taiwan and China herbal markets. The contents of 12 constituents in these samples were determined by HPLC and were used to assess the potential relationships with their plant origins. Multivariate analysis including principal component analysis (PCA), cluster analysis (CA), and linear discriminant analysis (LDA) were used as classification procedures. Natural groupings of the samples divided into three sets successfully, 20 P. trifoliata, 15 C. aurantium, and 15 C. wilsonii, were observed by using PCA and CA. The application of LDA gave correct assignation percentages of 100.0% for all three groups.     

Association between chemical and genetic variation of Vitex rotundifolia populations from different locations in China: its implication for quality control of medicinal plants

Vitex rotundifolia is a widely distributed plant species that has been extensively used in traditional Chinese medicine. Its fruits, Fructus Viticis, are recorded as Manjingzi in the Pharmacopoeia of the People's Republic of China. For the effective quality control of its medicinal values reflected by chemical variation patterns, in addition to the relationship with genetic diversity, analyses based on high-performance liquid chromatographic (HPLC) fingerprinting and inter simple sequence repeat (ISSR) molecular markers were carried out, involving 14 V. rotundifolia populations from different locations in China. The HPLC data showed considerable variation of chemical constituents among the V. rotundifolia populations. The hierarchical clustering analysis further revealed four major groups based on their chemotype variation. Abundant genetic diversity was detected among the V. rotundifolia populations that also were clustered into four groups based on their ISSR data. It is important to point out that the genetic variation pattern revealed by molecular markers was closely associated with that indicated by chemical constitutes in the fruits of V. rotundifolia. This finding provides a solid basis for the combined use of chemical and genetic fingerprints in efficiently evaluating qualities and choosing favourable chemotypes with appropriate pharmacological properties of V. rotundifolia, in addition to establishing good agricultural practices for medicinal plants.     

药材瓜蒌不同部位的红外光谱分析

采用傅立叶红外光谱和二阶导数谱分析方法,对药材瓜蒌不同部位(皮、瓤、子)的红外光谱进行分析.红外谱图显示,瓜蒌各部位均含糖及苷;瓜蒌皮和瓜蒌子中含黄酮及酯类成分;瓜蒌瓤和瓜蒌皮中含相当量的淀粉.二阶导数谱中,在1800～700 cm-1区域内,瓜蒌子的谱峰最简单,次之为瓜蒌瓤,全瓜蒌和瓜蒌皮的峰数较多,峰形和峰强比较相...     

Metabolomics analysis of Ligustri Lucidi Fructus at different harvest times during the whole growing period based on ultra-high-performance liquid chromatography with mass spectrometry

After medicinal market research, it was found that the harvest time of Ligustri Lucidi Fructus (LLF) was chaotic in practice. In order to determine the optimal harvest period of LLF to ensure its pharmacological activity, metabolomics analysis of LLF at different harvest times based on ultra-high-performance liquid chromatography-triple quadrupole-(linear ion trap)-tandem mass spectrometry was established. In this study, 166 differential metabolites (DMs) in 448 metabolites at different harvest times were screened out based on variable importance in projection value, and among them, 94 DMs with regular trends of change in relative content (59 increased and 35 decreased with the growth period) were chosen to further research. The result of the multivariate statistical analysis showed that November was the optimal harvest period of LLF. Additionally, 10-hydroxyligustroside, oleoside 11-methyl ester, and salidroside were screened out to be used as the evaluation indicators of immature LLF, while specnuezhenide, nuezhenoside G13, and neonuezhenide were the evaluation indicators of mature LLF. This study provides fundamental insight for metabolite identification and proposes the best harvest period of LLF to avoid confusion in the medicinal market.     

Anti-rheumatoid arthritis effects of Xanthii Fructus by affecting the PI3K-AKT signaling pathway based on TMT-labeled quantitative proteomics

Rheumatoid arthritis is a systemic autoimmune disease characterized by chronic symmetrical multiple arthritis. Current traditional counter-therapies are expensive and have side effects. Xanthii Fructus has effects in expelling wind and cold, draining the nasal orifice, and removing wind and dampness. However, its mechanism of action against rheumatoid arthritis is unknown. In this paper, the mechanism of the anti- rheumatoid arthritis effect of Xanthii Fructus is studied by proteomics. The experimental results show that it could significantly reduce serum inflammatory factor levels, alleviate joint edema, improve vasodilation and congestion, and significantly reduce the number of inflammatory cells. Proteomics results show that the PI3K-AKT signaling pathway is the key pathway for Xanthii Fructus to treat rheumatoid arthritis. In this study, we obtained a new understanding of the mechanism of Xanthii Fructus in the treatment of rheumatoid arthritis, which provided a theoretical basis for its prevention and treatment and laid the foundation for further research.