RESUMO
Ultrafine powder and cell wall-broken powder of herbal medicine lack of the morphological characters and microscopic identification features. This makes it hard to identify herb's authenticity with traditional methods. We tested ITS2 sequence as DNA barcode in identification of herbal medicine in ultrafine powder and cell wall-broken powder in this study. We extracted genomic DNAs of 93 samples of 31 representative herbal medicines (28 species), which include whole plant, roots and bulbs, stems, leaves, flowers, fruits and seeds. The ITS2 sequences were amplified and sequenced bidirectionally. The ITS2 sequences were identified using Basic Local Alignment Search Tool (BLAST) method in the GenBank database and DNA barcoding system to identify the herbal medicine. The genetic distance was analyzed using the Kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic tree was constructed using MEGA 6.0. The results showed that DNA can be extracted successfully from 93 samples and high quality ITS2 sequences can be amplified. All 31 herbal medicines can get correct identification via BLAST method. The ITS2 sequences of raw material medicines, ultrafine powder and cell wall-broken powder have same sequence in 26 herbal medicines, while the ITS2 sequences in other 5 herbal medicines exhibited variation. The maximum intraspecific genetic-distances of each species were all less than the minimum interspecific genetic distances. ITS2 sequences of each species are all converged to their standard DNA barcodes using NJ method. Therefore, using ITS2 barcode can accurately and effectively distinguish ultrafine powder and cell wall-broken powder of herbal medicine. It provides a new molecular method to identify ultrafine powder and cell wall-broken powder of herbal medicine in the quality control and market supervision.
RESUMO
This study was aimed to establish an Ultra Fast Liquid Chromatography-Photo Diode Array (UFLC-PDA) method for the simultaneous determination of five chemical components, which included chlorogenic acid, loganin, sweroside, evodia rutaecarpa glycosides and triplostoside A, in Pterocephalus hookeri h eck. Agilent Poroshell 120 SB-C18 (100 mm í 4.6 mm, 2.7 μm) was adopted, with acetonitrile-0.2% phosphoric acid solution in gradient elution as the mobile phase at the flow rate of 1.0 mL·min-1. And the injection volume was 0.4 μL. The detection wavelength was set up at 237 nm and 325 nm. And the column temperature was 30℃. The results showed that the calibration curve was linear within the range of 8.72~218.0, 1.52~38.0, 2.44~61.0, 29.36~734.0, 3.00~75.0μg·mL-1 (r > 0.999 6, n=9) for chlorogenic acid, loganin, sweroside, evodia rutaecarpa glycosides and triplostoside A, respectively. The average recovery rates were 99.46%, 99.41%, 100.14%, 98.89%, and 99.42%, respectively. The RSD was 0.69%, 0.66%, 0.60%, 1.21%, and 0.64%, respectively (n = 9). It was concluded that this method was simple, accurate and reproducible, which can be used for the simultaneous determination of the content of five chemical components in P. hookeri.
RESUMO
The article was aimed to study the influence of low pressure and hypoxia on rat metabolism and evaluate the intervention effect of San-Guo-Tang-San (SGTS) on high altitude polycythemia (HAPC) rats. A total of 25 rats were divided into the plain control group, high altitude model group, Hong-Jing-Tian capsule (Nuo-Di-Kang cap-sule) group, high-dose SGTS group and low-dose SGTS group, with 5 rats in each group. After one week adaptation, rats in the model group and the medication groups were put into the hyperbaric chamber for 40 days (22 h/d) to simulate high altitude environment of 5 000 m. In the end of 40th day, the hemorheology and the dry/wet weight ratio of lung of rats were measured. And plasma samples were derivatized with ECF prior to GC-MS instrumental analysis. Principal component analysis (PCA) was used to find potential biomarkers, and evaluate the intervention effects of SGTS. The results showed that the low pressure and hypoxia changed the hemorheology and dry/wet weight ratio of lung of rats markedly. Metabolomics studies showed that the high altitude model group, high-dose SGTS group, low-dose SGTS group, and Hong-Jing-Tian capsule group can be obviously differentiated. Main markers such as 9-hexylheptadecane, glycine, N-methyl-N-methoxycarbonyl-ethyl ester, 2,4-Di-tert-butylphenol, were found to be the endogenous substances of SGTS which intervening the HAPC rats. It was concluded that SGTS can intervene low pressure and hypoxia induced HAPC.
RESUMO
This study was aimed to find out the optimum parameters to decoction extraction process of Brassicae Radix cream . With methods of integrated score and uniform design , contents of the total extract and the total polysaccharide were set as the target to investigate the optimization process of extraction. The experiment showed that the optimum extraction condition was as follows: ten folds of water, four times decoction at boiling tempera-ture , one hour for each time . It was concluded that the optimum parameters to decoction extraction of Brassicae Radix were stable and feasible, which can be used for the preparation and reference for Brassicae Radix cream in Tibetan clinics .
RESUMO
This study was aimed to establish the quality standard of Pyrethri Tatsienenis Flos. The medical material was identified by the microscopy and the thin layer chromatography ( TLC ) methods . The moisture , total ash , acid-insoluble ash and alcohol-soluble extract were determined according to procedures recorded in the Chi-nese Pharmacopoeia (2010 edition). The content of luteolin was determined by the HPLC method. The results showed a strong characteristic microscopic of Pyrethri Tatsienenis Flos , and its TLC identification had a good resolution with clear spots . The mass fractions of luteolin was 0 . 036%~0 . 104% ( average of 0 . 078%) , moisture was 9 . 32%~15 . 82% ( average of 13 . 11%) , total ash was 6 . 65%~8 . 29% ( average of 7 . 45%) , acid-insoluble ash was 0 . 23%~0 . 59% ( average of 0 . 42%) , and the extraction was 21 . 42%~30 . 15% ( average of 24 . 86%) . It was concluded that this established standard was simple to operate with good stability and reproducibility , which can be used for quality evaluation of Pyrethri Tatsienenis Flos .