UFLC-Q-TOF-MS fingerprints of rhizome of Curcuma phaeocaulis and its vinegar processed products and inhibitory effect on thrombosis / 中国中药杂志

Both raw and vinegar products of the rhizome of Curcuma phaeocaulis are common drugs for promoting blood circulation and removing blood stasis in traditional Chinese medicine,which could be reflected in the inhibition of tail thrombosis in mice. As the traditional processing theory instructs,vinegar tastes sour and bitter,but can activate blood circulation and remove stasis after being infiltrated into the rhizome of C. phaeocaulis as an excipient. In this study,under the help of the ultrafast liquid chromatography-quadrupole time-offlight mass spectrometry( UFLC-Q-TOF-MS),the spectrum-effect relationship between the inhibition of tail thrombosis in mice and the rhizome of C. phaeocaulis both before and after the vinegar processing,were established to explore the functional changes of blood circulation and stasis after vinegar process. Based on the peak area from the fingerprint of UFLC-Q-TOF-MS of the alcohol extracts from the raw and vinegar-processed rhizome of C. phaeocaulis and their efficacy for inhibiting tail thrombosis,the correlation between the chromatography of UFLC-Q-TOF-MS and the inhibition of tail thrombosis in mice were analyzed by orthogonal partial least squares discriminant analysis( OPLS-DA) method. The results,produced by Simca-P software,showed that effective components consisted of eight peaks 16,24( aromadendrene oxide),3,11,22( dehydro-α-curcumene),19[( R)-(-)-α-curcumene],23 and 10 from the fingerprint,making great contribution to distinguish C. phaeocaulis raw products and the corresponding vinegar processed products. Therefore,from the perspective of inhibiting the formation of tail thrombosis in mice,the marker components could be found through the spectrum-effect relationship to distinguish C.phaeocaulis raw and vinegar products. This study provided new basis to explain the difference between the raw and the processed products of traditional Chinese medicine in the functional change of promoting blood circulation and removing blood stasis.

Qualitative evaluation of Forsythia suspensa by HPLC-PDA fingerprint combined with UFLC-Q-TOF-MS qualitative identification / 中国中药杂志

The analysis of Forsythia suspensa was performed on Waters Symmetry C18 column( 4. 6 mm×250 mm,5 μm) and mobile phase was methanol( A)-0. 1% formic acid aqueous solution( B) with the elution gradient. Column temperature was maintained at 30℃,and the flow rate was 1. 0 m L·min-1 with detection wavelength 265 nm. The HPLC-PDA fingerprint of F. suspensa was optimized.Chemical constituents in F. suspensa were analyzed by UFLC-Q-TOF-MS in positive and negative ion mode. The quality of 48 batches of F. suspensa from different habitats,processing methods and specifications was evaluated by similarity evaluation and cluster analysis.The 18 common peaks were confirmed. The similarity of F. suspensa from different habitats was more than 0. 98,and 56 chemical constituents were identified. Different processing methods had great influence on the quality of F. suspensa. Compared with boiled and direct drying,the quality of F. suspensa processed by sun-drying was obviously decreased. The similarity was about 0. 58. Different specifications of F. suspensa also had obvious distinction,and the similarity was about 0. 78. The effective components of grown F. suspensa,such as forsythoside A and phillyrin,were significantly reduced. The results of cluster analysis were basically consistent with the results of similarity evaluation. The establishment of fingerprint and the recognition of chemical pattern of F. suspensa can provide a more comprehensive reference for the quality control of herbs.

Comparative study on acute toxicity of different active ingredient fractions from Kansui stir-baked with vinegar on zebrafish embryos / 中国中药杂志

The 24 h normal developing zebrafish embryos were used to evaluate the acute toxicity and the compounds of respective fractions were analyzed by UFLC-Q-TOF-MS simultaneously. Nine concentration groups with respective concentration and a blank control group were designed for each fraction to investigate their effect on survival rates of zebrafish embryos 96 h after drug administration, and calculate the median lethal concentration (LC₅₀) of different fractions to zebrafish embryos. The results showed that all of the fractions had acute toxicity to zebrafish embryos except VEKD, and the order was as follows: VEKB, VEKC, VEKA and VEKD. According to the results of UFLC-Q-TOF-MS, the chemical ingredients contained in VEKB and VEKC were mainly composed of ingenane-type and japhane-type diterpenoids, respectively. It could be speculated that japhane-type diterpenoids might be the active compounds with lower toxicity associated with the results of toxicity study, providing some references for the further research on effective material basis of Kansui stir-baked with vinegar according to the principle of "drastic medicine, no death risks".

Qualitative evaluation of Atractylodis Macrocephalae Rhizoma from different habitats by HPLC-PDA fingerprint combined with UFLC-Q-TOF/MS qualitative identification / 中草药

Objective: To establish and identify the HPLC-PDA fingerprint of Atractylodis Macrocephalae Rhizoma (AMR) and provide a reference for the comprehensive control of the quality of AMR. Methods: AMR was extracted with 70% methanol by sonicating for 60 min. The analysis of AMR extract was performed on Inertsil® ODS-SP column (150 mm × 4.6 mm, 5 μm), column temperature was maintained at 40 ℃, flow rate was 1.0 mL/min, and detector was Waters 2998 UV detector with detection wavelength 235 nm. Mobile phase was acetonitrile (B)-water (A) with the elution gradient 0 -10 min, 30%-45% B, 10-25 min, 45% B, 25-50 min, 45%-70% B, 50-55 min, 70% B, 55-62 min, 70%-30% B, 62-75 min, 30% B. Time-of-flight mass spectrometer (TOF/MS) and electro-spray ion (ESI) source were used for the qualitative analysis in a positive ion mode, and mass scan range was m/z 50-1 500. Results: Comparing and fitting the peaks of AMR from different habitats (Zhejiang, Anhui, and Hunan Provinces), the HPLC-PDA fingerprint was set up with six common peaks, and they were identified by UFLC-Q-TOF/MS as 5-(hydroxymethyl)-2-furaldehyde, atractylenolide III, atractylenolide I, atractylenolide II, atractylenolide VI, and biatractylenolide. System suitability, extraction, and chromatographic conditions of AMR were optimized. RSD of accuracy, stability and repeatability was all less than 2%. Measuring ten batches and fitting fingerprint similarity, the values were all greater than 0.95. Conclusion: The HPLC fingerprint can be used as standard uniformity and stability of quality control methods for AMR slice.

Chemical constituents in Akebiae Fructus based on UFLC-Q-TOF/MS / 中草药

Objective: To identify the chemical constituents in Akebiae Fructus by UFLC-Q-TOF/MS method. Methods: The separation was performed on an Acquity UPLC BEH C18 Column (100 mm × 2.1 mm, 1.7 μm), with a mobile phase using 0.1% formic acid-acetonitrile and water containing 0.1% formic acid (B) for gradient elution. The flow rate was 0.3 mL/min, the temperature of column was 40°C with injection volume of 1 μL. TOF/MS and electrospray ion (ESI) source were applied for the qualitative analysis under the negative ion mode, and the full mass scan range was m/z 100-1500. Results: According to MS principle, twenty-five triterpenoids were identified from the methanol extract of Akebiae Fructus and the chemical structures of other nine unknown compounds were deduced. Conclusion: UFLC-Q-TOF/MS method could identify the main chemical constituents in Akebiae Fructus rapidly and accurately, which lays a foundation for the quality control of Akebiae Fructus.