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Objective:To establish the fingerprints of Microctis Folium by ultra high performance liquid chromatography (UPLC); To determine the contents of three flavonoids in the Microctis Folium; To evaluate the quality difference of Microctis Folium from different producing areas. Methods:The fingerprints were performed on Agilent ZORBAX SB C18 column (2.1 mm×150 mm,1.8 μm). The mobile phase was acetonitrile - 0.1 % acetic acid solution with gradient elution at a flow rate of 0.30 ml/min. The column temperature was 30 ℃ and the detection wavelength was 315 nm. The common fingerprint peaks were identified by UPLC-mass spectrometry, and the identification results were confirmed by comparison of reference materials. Waters Cortecs T3 C18 chromatographic column (2.1 mm × 100 mm,1.6 μm) was used for content determination. The mobile phase was methanol-0.1 % formic acid solution with gradient elution at a flow rate of 0.35 ml/min. The column temperature was 30 ℃ and the detection wavelength was 339 nm. The contents of vitexin, isovitexin and narcissoside in 15 batches of Microctis Folium from different habitats were determine. Results:There were 11 common peaks in the fingerprint of Microctis Folium. Identified by mass spectrometry and confirmed by reference substance,10 chemical components were identified, including caffeic acid, p-hydroxycinnamic acid, ferulic acid, vitexin, isovitexin, kaempferol-3-O-rutoside, astragaloside, narcissoside, isorhamnetin-3-O-glucoside and linden glycoside. The similarity between the fingerprints of 15 batches of Microctis Folium and the control fingerprint was greater than 0.95, indicating that the overall similarity of the fingerprints of Microctis Folium from different producing areas was high. The total contents of three active components were 3.23-5.64 mg/g in Yangjiang City, Guangdong, 3.60-5.78 mg/g in Zhanjiang City, Guangdong, 4.68-5.73 mg/g in Yulin City, Guangxi and 3.87-5.21 mg/g in Wuzhishan City, Hainan . There was no significant difference in the content of three active components in different producing areas. Conclusion:The fingerprints and the determination method established in the study are stable and feasible, which can be used for the quality evaluation of Microctis Folium.
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Objective:To establish the quality evaluation method of Perillae caulis formula granules based on the three kind of quality indexes of standard decoction. Methods:Eighteen batches of Perillae caulis were collected from different habitats according to different technical requirements, eighteen batches of standard decoction and three batches of formula granules were prepared and the paste-forming rates were calculated. The content of Caffeic acid and Rosmarinic acid were determined and calculated by Ultra High Performance Liquid Chromatography (UPLC). Then the fingerprints of standard decoction of and formula granules of Perillae caulis were established by UPLC . The similarity values of fingerprints between formula granules and standard decoction were calculated. Results:The average paste-forming rate of standard decoction was (7.16±1.97)%. The paste-forming rates of three batches of formula granules were 5.52%, 5.25% and 5.34%, respectively. The average content of Caffeic acid and Rosmarinic acid in standard decoction was (12.06±3.37)mg/g. The contents of three batches of formula granules were 5.52, 5.82, 5.77 mg/g, respectively. Seven common fingerprint peaks were identified in the fingerprints of standard decoction and formula granules, three of which were identified as Caffeic acid, N-Feruloyl Octopus amine and Rosmarinic acid by comparison of reference substance. The fingerprints similarity of Perillae caulis dispensing granules and standard decoction were 1.000, 0.995 and 0.997, respectively. Conclusions:The quality indexes of three batches of formulation granules are consistent with standard decoction. This method can provide basis for the establishment of quality standard of Perillae caulis dispensing granules.
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OBJECTIVE：To compare the chemical components in Sinapis alba before and after stir-frying. METHODS ： UPLC-Q-Exactive Obitrap MS was adopted to analyze chemical constituents of S. alba before and after stir-frying. The determination was performed on Waters CORTECS T 3 column with mobile phase consisted of methanol- 0.1％ formic acid solution （gradient elution ）at the flow rate of 0.25 mL/min. The column temperature was 30 ℃ and the sample size was 2 μL. High resolution MS adopted heating electrospray electron source ，positive ion scanning mode ，scanning range m/z 120-1 000. The chemical constituents of S. alba before and after stir-frying were identified by Compound Discover 3.2 software combined with relevant database ，and the content changes of chemical constituents were analyzed by using peak area. Chemometrics analysis was performed for the content changes of chemical constituents using peak area as variable. RESULTS ：A total of 54 chemical components were identified in S. alba ，mainly fatty acids （represented by erucic acid ），alkaloids（represented by sinapine ）， flavonoids. After stir-frying ，the contents of 19 chemical components changed significantly ，of which the contents of 10 components decreased significantly and those of 9 components increased significantly （P＜0.05）. Principal component analysis and orthogonal partial least squares discriminant analysis could clearly distinguish S. alba from stir-fried S. alba . CONCLUSIONS ：The contents of some chemical components of S. alba change significantly after stir-frying ，which may be one of the important reasons for the change of efficacy after stir-frying.
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OBJECTIVE：To compare the c hemical components differences of Inula japonica before and after honey-frying. METHODS：UPLC-MS/MS method was adopted. The determination was performed on Waters ACQUITY UPLC BEH C 18 column with mobile phase consisted of 0.1％ formic acid-acetonitrile （gradient elution ）at the flow rate of 0.3 mL/min. The column temperature was set at 30 ℃，and sample size was 5 µL. The electrospray ion source was scanned by positive ion mode. The first order mass spectrometry scanning range was m/z 70-1 050，the second order mass spectrometry scanning range was m/z 50-1 050， and the normalized collision energy was 40，60 eV ；mass spectrum type was the peak figure ，the flow rate of sheath gas was 35 arb，the auxiliary airflow speed was 10 arb，the spray voltage was 3.80 kV，the S-lens voltage was 50 V，the heating temperature was 350 ℃，and the capillary temperature was 350 ℃. The components were identified by Qual Browser 4.1.39.1 software， referring to the online high-resolution database mzCloud and local database OTCML of high-resolution mass spectrometry of TCM ， and combined with relevant literature. The principal component analysis （PCA）and orthogonal partial least squared-discriminant analysis（OPLS-DA）of I. japonica before and after honey-fried were performed by using SIMCA 14.1 statistical software ，and variable importance projection （VIP）value greater than 1 was used as the standard to screen the differential components before and after honey-frying. RESULTS ：A total of 29 common chemical components were identified from I. japonica and honey-fried I. japonica，including 5 phenolic acids as 1-caffeoylquinic acid ，chlorogenic acid and 3，5-dicaffeoylquinic acid ，12 flavonoids as quercetin，luteolin and evamectin ，as well as 12 sesquiterpene lactones as 1-O-acetylinula diester ，inula bicolor lactone B and 1-O-acetyl-6-O-isobutyryl inulin. The results of PCA showed that I. japonica and honey-fried I. japonica were located on both sides of the score diagram respectively. The results of OPLS-DA showed that the VIP values of 7 components were greater than 1，which were peak 19（britanin），peak 6（quercetagitrin），peak 1（1-caffeoylquinic acid ），peak 21（vitexicarpin），peak 20（tomentosin）， peak 13（spinacetin）and peak 3（daphnetin）. CONCLUSIONS ：After honey-fried ，the content of chemical components of I. japonica changed and decreased to a certain extent. Britanin ，quercetagitrin，1-caffeoylquinic acid ，tomentosin，vitexicarpin， spinacetin and daphnetin may be the differential components of I. japonica and honey-fried I. japonica .
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OBJECTIVE：To esta blish a UPLC fingerprint of Pyrrosia petiolosa from southwest China ，and to determine the contents of 4 kinds of phenolic acids （neochlorogenic acid ，caffeic acid ，chlorogenic acid and cryptochlorogenic acid ）. METHODS：The determination was performed on Waters Cortecs T 3 C18 column（100 mm×2.1 mm，1.6 μm）with mobile phase consisted of methanol- 0.1％ phosphoric acid （gradient elution ）at the flow rate of 0.35 mL/min. The detection wavelength was set at 326 nm. The column temperature was 30 ℃，and injection volume was 1 μL. UPLC method was used to establish the UPLC fingerprint of P. petiolosa in combination with the Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition）. Cluster analysis and principle component analysis （PCA）were performed by using SPSS 20.0 software. The contents of 4 kinds of phenolic acids in 20 batches of P. petiolosa were determined by external standard method. RESULTS ：There were 9 common peaks for the UPLC fingerprint of P. petiolosa . Peaks 1，3，4，5 and 9 were identified as neochlorogenic acid ，caffeic acid，chlorogenic acid ，cryptochlorogenic acid and isochlorogenic acid C ，respectively. RSDs of the relative retention time of each peak in different batches of P. petiolosa were 0-0.68％，and the RSDs of the relative peak area were 0-62.35％. The similarities between the fingerprint of 20 batches of medicinal materials and the control chromatogram were not less than 0.990. The result of cluster analysis showed that P. petiolosa from different regions could be sorted into three species. Results of PCA showed the differences among P. petiolosa from different regions. The linear range of neochlorogenic acid ，caffeic acid ，chlorogenic acid and cryptochlorogenic acid were 0.61-61.41，0.18-17.60，2.00-200.11，0.62-61.51 μ g/mL （R2＞0.999 9）. RSDs of precision ， reproducibility and stability tests were all lower than 2.00％. The recoveries were 96.23％-98.17％（RSD＝0.96％-2.28％， n＝6）. Among 20 batches of samples ，the contents of above 4 kinds of phenolic acids were 0.385 3-1.891 9，0.018 0-0.129 5，2.569 5-10.676 0，0.563.5-1.860 5 mg/g. CONCLUSIONS ： The established UPL C fingerprint could reflect the main chemical constituents of P. pedunculata . Phenolic acids could be used as the main evaluation indexes for the quality of P. petiolosa . The quality order of P. petiolosa from southwest China was Chongqing product＞Sichuan product ＞Guizhou product.