An efficient and environmentally-friendly extraction, characterization and activity prediction of polysaccharides from Rhizoma et Radix Notopterygii.

Traditional hot water reflux extraction, ultrasonic-water extraction (UW), ultrasonic-natural deep eutectic solvent (NADES) extraction (U-NADES), ultrasonic-water and enzyme extraction (U-W-E) and ultrasonic-NADES and enzyme extraction (U-NADES-E) are employed for the extraction of Rhizoma et Radix Notopterygii polysaccharides (RNP), in which, the U-NADES-E has being proved as the most effective method. Response Surface Methodology (RSM) was utilized to optimize the conditions for U-NADES-E method. Using the optimal extraction conditions, the yield of RNP can be enhanced by nearly two-fold in comparison to the traditional extraction method, achieving a yield of 7.38 %, with a mere 30-min treatment and low ultrasonic power at 240 W. The RNP's composition included Rhamnose, Arabinose, Galactose, Glucose and Galacturonic Acid by high-performance anion-exchange chromatography. The polysaccharides from two different species of Rhizoma et Radix Notopterygii have also been characterized and identified. Network pharmacology and molecular docking predict that RNP may exert its effects in vivo through binding to PPARA, ACE and REN proteins, thereby potentially impacting diabetes outcomes. This study proposes a new, efficient, energy-saving and environmentally-friendly method for the extraction of RNP.

Nonribosomal antibacterial peptides isolated from Streptomyces agglomeratus 5-1-3 in the Qinghai-Tibet Plateau.

BACKGROUND: New antibiotics are urgently needed in clinical treatment of superdrug-resistant bacteria. Nonribosomal peptides (NRPs) are a major source of antibiotics because they exhibit structural diversity, and unique antibacterial mechanisms and resistance. Analysis of gene clusters of S. agglomeratus 5-1-3 showed that Clusters 3, 6, 12, 21, and 28 were used to synthesize NRPs. Here, we examined secondary metabolites of S. agglomeratus 5-1-3 isolated from soils in the Qinghai-Tibet Plateau, China, for NRPs with antibacterial activity. RESULTS: We isolated a total of 36 Streptomyces strains with distinct colony morphological characteristics from 7 soil samples. We screened 8 Streptomyces strains resistant to methicillin-resistant Staphylococcus aureus (MRSA). We then selected S. agglomeratus 5-1-3 for further study based on results of an antibacterial activity test. Here, we isolated three compounds from S. agglomeratus 5-1-3 and characterized their properties. The crude extract was extracted with ethyl acetate and purified with column chromatography and semipreparative high-performance liquid chromatography (HPLC). We characterized the three compounds using NMR analyses as echinomycin (1), 5,7,4'-trihydroxy-3.3',5'-trimethoxy flavone (2), and 2,6,2', 6'-tetramethoxy-4,4-bis(2,3-epoxy-1-hydroxypropyl)-biphenyl (3). We tested the antibacterial activity of pure compounds from strain 5-1-3 with the Oxford cup method. NRP echinomycin (1) showed excellent anti-MRSA activity with a minimum inhibitory concentration (MIC) of 2.0 µg/mL. Meanwhile, MIC of compound 2 and 3 was 128.0 µg/mL for both. In addition, 203 mg of echinomycin was isolated from 10 L of the crude extract broth of strain 5-1-3. CONCLUSION: In this study, S. agglomeratus 5-1-3 with strong resistance to MRSA was isolated from the soils in the Qinghai-Tibet Plateau. Strain 5-1-3 had a high yield of echinomycin (1) an NRP with a MIC of 2 µg/mL against MRSA. We propose that echinomycin derived from S. agglomeratus 5-1-3 may be a potent antibacterial agent for pharmaceutical use.

Simultaneous Determination of Eight Chemical Components in Angelicae Sinensis Radix and Its Herbal Products by QAMS.

A HPLC method has been developed for simultaneously detecting chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, coniferyl ferulate, senkyunolide A, ligustilide, and levistolide A in Angelicae Sinensis Radix through quantitative analysis of multicomponents by single-marker (QAMS) method with ferulic acid as internal standard substance. The relative analysis correction factors of each component in Angelicae Sinensis Radix have good reproducibility under different chromatography conditions. In addition, no significant difference of results was found between quantitative analysis of multicomponents by single-marker (QAMS) method and external standard method in determining content of these components of different Angelicae Sinensis Radix and its 12 kinds of preparations. As a result, the established QAMS method for Angelicae Sinensis Radix analysis with ferulic acid as internal standard substance is accurate and feasible, which could be used as an effective and economical method to control quality of Angelicae Sinensis Radix and its herbal products.

Multi-Index Quantitative Evaluation of Angelicae sinesis Radix Based on 1H-qNMR.

BACKGROUND: As known to us, HPLC method was often used to determine the contents of Angelicae sinesis Radix. In view of the shortcomings of HPLC method, qNMR has prominent advantages in determining the contents of bioactive components in the quantitative and qualitative analysis of Angelicae sinesis Radix. OBJECTIVE: In this study, a quick, simple, and accurate method was established to determine the components of ferulic acid, coniferyl ferulate, and ligustilide in Angelicae sinesis Radix. METHOD: Using dimethyl sulfoxide-d6(DMSO-d6) as the test solvent and pyrazine as the internal standard substance, 1H-qNMR measurement was performed on a 600 MHz spectrometer. The quantitative resonance peaks of pyrazine, ferulic acid, ligustilide, and coniferyl ferulate were at δ8.66 ppm, δ6.35-6.37 ppm, δ5.53-5.55 ppm, and δ6.50-6.53 ppm, respectively. RESULTS: The linear relationship, limit of detection, limit of quantification, precision, stability, and recovery were verified and the results were good. On the other hand, it was verified by HPLC, and the HPLC used for verification passed the methodological investigation of linearity, precision, repeatability, stability, and recovery, and the results were good. In addition, no significant difference in results was found between the 1H-qNMR and HPLC-UV methods in determining the content of three components in three batches of Angelicae sinesis Radix. CONCLUSIONS: The method can be used for simultaneous determination of three active components, and providing a scientific basis for the overall quality evaluation and quality control of Angelicae sinesis Radix. HIGHTLIGHTS: In this study, 1H-qNMR was used to determine ferulic acid, coniferyl ferulate and ligustilide in Angelicae Sinensis Radix for the first time.

Enhancement of Kidney Invigorating Function in Mouse Model by Cistanches Herba Dried Rapidly at a Medium High Temperature.

Cistanches Herba is a popular Traditional Chinese Medicine and functional food that is used to treat kidney yang deficiency (KYD) in China. In this study, we investigated the effects of different drying methods for Cistanches Herba on kidney invigoration and yang strengthening. We established a mouse model of KYD by intraperitoneal injection of hydrocortisone for 8 days. We dried slices of Cistanches Herba in the sun, in the shade, in a microwave, or in an oven at 40°C, 60°C, 80°C or 100°C, then prepared and administered extracts to the mice by gastric gavage. We measured and evaluated the echinacoside (ECH) and acteoside (ACT) contents of the extracts, as well as the mice's body weight; testicular, epididymal, hepatic, and renal coefficients; and semen quality. All the Cistanches Herba extracts, obtained using different drying techniques, improved symptoms of KYD diagnosis in mice. Among them, treatments with Cistanches Herba dried in a microwave and dried in an oven at 100°C had the best therapeutic effects. Our results suggested that the higher the total content of ECH and ACT in Cistanches Herba extracts, the better the effects of kidney invigoration and yang strengthening. In addition, shorter drying times at higher temperatures lead to the highest recoveries of active components, and Cistanche dried in the sun at a medium-high temperature can improve sperm quality in mice.

[Effects of Different Temperature and Packaging Treatments on Antioxidant Activities of Fresh Cistanche deserticola Fleshy Stem].

Objective: To explore the optimal storage method, antioxidant activities of fresh Cistanche deserticola fleshy stem were studied in response to the types of packaging and storage temperature during storage. Furthermore, this research will provide theoretical guides to preserve Cistanche deserticola fleshy stem. Methods: The fresh fleshy stem of Cistanche deserticola was stored at 4 ℃ and 25 ℃using polyethylene( PE) film, vacuum packaging and vacuum packaging with nitrogen( N2) respectively, and the antioxidant enzyme activities were assayed regularly. Results: The preservation temperature of Cistanche deserticola fleshy stem was better at 4 ℃ than that at25 ℃. Under 4 ℃ condition,vacuum packaging with N2 was conducive to maintaining antioxidant capacity. Conclusion: The combination of vacuum packaging with N2 and lower temperature can keep better quality in fresh Cistanche deserticola fleshy stem during storage.