ABSTRACT
Objective:Chemical constituents in hypoglycemic effective fractions of Longan Folium were isolated and identified by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） to clarify the hypoglycemic substance basis of Longan Folium. Method:Chemical constituents in hypoglycemic effective fractions of Longan Folium were isolated on a Thermo Hypersil GOLD C<sub>18</sub> column （2.1 mm×100 mm， 1.9 μm）， the mobile phase was 0.1% formic acid acetonitrile solution and 0.1% formic acid solution （containing and 10 mmol ammonium acetate） for gradient elution. HRMS was operated in the positive and negative ion modes with the scanning range of <italic>m</italic>/<italic>z</italic> 100-1 500. Result:The secondary fragment ion information of target compounds was selected and compared with the compounds reported in the databases and related literature to further confirm these compounds. Nine compounds were identified in the ethanol fraction of Longan Folium， including cynaroside， kaempferol， quercitrin， luteolin， shikimic acid， citric acid， <italic>L</italic>-tyrosine， adenosine and nicotinamide. A total of 11 compounds were determined in the ethyl acetate fraction （cynaroside， quercitrin， kaempferol， luteolin， shikimic acid， gallic acid， protocatechuic acid， adenosine， nicotinamide， <italic>L</italic>-phenylalanine and scopoletin）， and 10 compounds were identified in the <italic>n</italic>-butanol fraction （cynaroside， kaempferol-3-<italic>O</italic>-rutinoside， kaempferol， astragalin， luteolin， citric acid， gallic acid， adenosine， nicotinamide and 5-hydroxymethylfurfural）. And five common compounds were identified in these three hypoglycemic effective fractions. Conclusion:The established UPLC-Q-Orbitrap HRMS can quickly identify chemical constituents in three hypoglycemic effective fractions of Longan Folium， their main chemical constituents are flavonoids and their glycosides， organic acids and nitrogen-containing compounds， which provides technical support and scientific evidence for the study on pharmacodynamic material basis and quality control of Longan Folium.
ABSTRACT
Objective:The aim of this study was to research the relationship between HPLC fingerprint and anti-inflammatory effect of Zhideke granules， and the substance basis of its anti-inflammatory effect was preliminary explored. Method:The fingerprint of 10 batches of Zhideke granules were determined by HPLC， the mobile phase was consisted of methanol-0.2% phosphoric acid solution for gradient elution with a detection wavelength of 254 nm. Similarity analysis， hierarchical cluster analysis （HCA）， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were used to evaluate the quality difference between batches of Zhideke granules. The correlation analysis between the common peaks and the inhibition rates of Zhideke granules on ear swelling and cotton ball granuloma in mice was carried out by partial least squares （PLS）， and the peaks greatly contributing to the anti-inflammatory effect were screened out. Result:There were 31 common peaks in the HPLC fingerprint of Zhideke granules. The similarities of 10 batches samples were ≥0.992. The HCA and PCA analysis results were consistent， and the samples were divided into 3 categories. Combined with the OPLS-DA results， 15 components were the main markers affecting the differences of different batches of samples. Different batches of Zhideke granules differed in anti-inflammatory effect. The chromatographic peaks being positively correlated with the anti-inflammatory effect were mainly from Belamcandae Rhizoma and Scutellariae Radix， Chromatographic peaks 3， 6， 19， 27-30 had significant contribution to anti-inflammatory effect， of which peaks 28 and 30 were irisflorentin and wogonin. Conclusion:HPLC fingerprint combined with chemical pattern recognition method can provide a reference for systematic evaluation of the overall quality of Zhideke granules. Zhideke granules has a certain inhibitory effect on acute and chronic inflammation in mice， and the anti-inflammatory effect is the result of the combined action of various ingredients， while Belamcandae Rhizoma and Scutellariae Radix have significant significance for the anti-inflammatory effect.
ABSTRACT
Objective: To summarize and evaluate the existing animal models of pharyngitis, in order to provide a theoretical basis for its treatment and research of traditional Chinese medicine (TCM) for throat impediment. Method: Nearly 5 years of literatures were retrieved form databases of Chinese national knowledge infrastructure (CNKI), SinoMed and Pubmed, with throat impediment, pharyngitis, and animal model as medicine subject headings terms. Result: At present, rats, rabbits, guinea pigs and macaques are available as model animals. The main methods for reproduction of pharyngitis animal model were direct ammonia molding, ammonia and turpentine combined molding and bacterial injection molding. The rarely used methods, such as pyridine induction method, capsaicin induction method and sand dust induction method, have their own characteristics. The pharyngitis animal models with the combination of TCM syndrome include YIN-deficiency type and stagnation of liver-Qi with deficiency of spleen type. These models evaluation indexes were diverse, including apparent behavior, histopathology, biochemical indicator and hemorheology. The problems of these modeling methods included the differences in etiology and mechanism between the methods of modeling and the formation of clinical patients, the lack of molecular level of pathological mechanism elaboration, the non-uniform standard for concentration, frequency and cycle of drug administration during modeling, the subjectivity in judging partial evaluation indexes and the deficiency of the pharyngitis animal model with the combination of TCM syndrome. Conclusion: Firstly, the existing models needed to be further improved by such means as the exploration of the correlation or consistency between the molecular pathology mechanism and the clinical pathogenesis, and the concentration and frequency of uniform model administration and uniform standard for evaluation indexes. Secondly, the pharyngitis animal model with the combination of TCM syndrome and modern medicine disease was urgently needed to be explored, in order to facilitate the development of basic research in TCM.