OBJECTIVE To evaluate the inter-batch quality consistency of 12 batches of Huobahuagen tablets using UPLC fingerprints, and to identify the main chemical constituents for clarifying their substance bases. METHODS Chromatographic separation was performed on a Waters CORTECS T₃ column (2.1 mm×100 mm, 1.6 μm). Acetonitrile-0.1% formic acid aqueous solution was adopted as mobile phase for gradient elution. The flow rate was set at 0.4 mL·min^-1. The detection wavelength was set at 260 nm. The column temperature was maintained at 35 ℃. The similarity of fingerprint was evaluated by the “Similarity Evaluatio System of Chromatographic Fingerprint of Traditional Chinese Medicine” (2012 edition). At the same time, the main chemical constituents of Huobahuagen tablets were identified by UPLC-Q-TOF-MS method. RESULTS In the established fingerprint, a total of 27 common peaks were calibrated. The similarity of the tested samples was 0.992-0.999. Eighteen main constituents were identified, most of which were phenolic glycosides and flavonoids. CONCLUSION Huobahuagen tablets have good inter-batch quality consistency. The main components contained in this preparation have relatively high polarity, which is due to the production process of water extraction and alcohol precipitation. The results presented above can serve as a foundation for a later quality control study of this preparation.

OBJECTIVE To establish a method for characterization and evaluation of the O-glycosaminoglycan in ulinastatin. METHODS The research released and separated the O-glycosaminoglycan through the chemical degrading and salt deposition, then acquired the mass data of the O-glycosaminoglycan via the UHPLC-Orbitrap-HRMS. RESULTS The result showed that the products derived from different companies are consistent in their O-glycosaminoglycan by the analysis and calculation of the molecular weight and molecular ion peaks. CONCLUSION The method for characterization of the O-glycosaminoglycan in ulinastatin in this research is precise and reliable which could not only realize to evaluate the quality of ulinastatin, but also could apply in other extracted glycoprotein drugs.

OBJECTIVE To establish an ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of 15 active components in Tripterygium hypoglaucum tablets. METHODS Chromatographic separation was performed on a Waters Cortecs T₃ column (2.1 mm × 100 mm, 1.6 μm) at a column temperature of 35 ℃. The 0.1% formic acid acetonitrile solution (A)-0.1% formic acid aqueous solution (B) was adopted as mobile phase for gradient elution. The flow rate was set at 0.3 mL·min^-1. The detection was performed in multiple reaction monitoring (MRM) mode by positive and negative ion switching scan for 15 active components, including two flavonoids (catechin and epicatechin), nine diterpenoids (triptriolide, triptolidenol, tripdiolide, triptolide, tripchlorolide, triptonide, triptophenolide, triptoquinone A, and triptoquinone B), and four triterpenoids (demethylzeylasteral, celastrol, wilforlide A, and pristimerin). RESULTS The 15 investigated components showed good linearity (r²≥0.999 1) in their respective ranges, and the recoveries were in the range of 88.36%-115.91% (RSD≤4.94%). The quantitative results showed that the content of the active components in Tripterygium hypoglaucum tablets from different manufacturers varied significantly, which may have an impact on the clinical efficacy and safety of the preparations. CONCLUSION The established method is sensitive and accurate, which can be used for the determination of the active components in Tripterygium hypoglaucum tablets. The results can provide a reference and basis for the quality evaluation of this preparation.

OBJECTIVE To evaluate the quality of 15 batches of Curcuma phaeocaulis polysaccharide (CPP) from different areas by the combination of HPLC fingerprint, antioxidant activity and stoichiometric analysis. METHODS With glucose as the control substance, the content of total sugar was determined by phenol-sulfuric acid method, CPP was hydrolyzed by trifluoroacetic acid, and the fingerprint of polysaccharide was established by PMP-HPLC, and the similarity of fingerprint, hierarchical cluster analysis and partial least square-discriminant analysis were evaluated. The antioxidant activity of CPP was evaluated by using the DPPH free radical scavenging ability, hydroxyl radical scavenging ability and total Fe³⁺ reducing ability as indexes. RESULTS Six common peaks were identified by the fingerprint of CPP, which were mannose, rhamnose, galacturonic acid, glucose, galactose and arabinose. Based on similarity evaluation, cluster analysis and partial least square-discriminant analysis, mannose, glucose and galacturonic acid were selected as potential chemical markers, and the antioxidant activity corresponding to DPPH free radical scavenging activity could be used as candidate detection indexes for auxiliary identification. CONCLUSION The establishment of PMP-HPLC fingerprint of CPP combined with antioxidant activity can provide a more comprehensive reference for the quality control of Rhizoma Curcumae polysaccharide.

OBJECTIVE To establish the first batch of national reference substance (RS) for the first-class new drug sodium oligomannate in China, and confirm the structure of the RS. METHODS Structure of the candidate was confirmed by liquid chromatography-mass spectrometry, nuclear magnetic resonance and other methods. GE Superdex 30 Increase 10/300GL column was used to separate oligosaccharides by liquid chromatography, and negative ion mode for mass spectrometry detection was used; one-dimensional and two-dimensional nuclear magnetic resonance techniques were used to analyze the structures of polymannuronic acid, polyguluronic acid and the sodium oligomannate RS . RESULTS Through the analysis of mass spectrometry and spectrum, it was confirmed that the sodium oligomannate RS is an oligosaccharide mixture composed of 2-10 saccharides, and the molecular backbone is mainly mannuronic acid, which is consistent with the theoretical structure. CONCLUSIONS In this study, the structure confirmation methods of sodium oligomannate are established, and the molecular weight and molecular structure of the first batch of national RS of sodium oligomannate are confirmed.

OBJECTIVE To establish the UPLC-Q-Exactive Orbitrap-MS fingerprint of Rosa odorata Sweet var. gigantean root and study the spectrum-effect relationship of its free radical scavenging activity. METHODS The fingerprint of Rosa odorata Sweet var. gigantean root was established by UPLC-Q-Exactive Orbitrap-MS method, the similarity of the fingerprint was evaluated, and 19 common peaks were evaluated by heat map analysis. The antioxidant activity of Rosa odorata Sweet var. gigantean root was evaluated by1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) method, and the spectrum-effect relationship was studied by grey correlation analysis (grey relational analysis, GRA). RESULTS The UPLC-Q-Exactive Orbitrap-MS fingerprint of Rosa odorata Sweet var. gigantean root was established, and 19 common chromatographic peaks with Area %>1% were selected, and the chromatographic peaks were identified by reference method, database and reference. According to the heat map analysis of 19 peaks and the grey correlation degree of DPPH oxidation resistance, the heat map analysis of 19 peaks showed that the peaks with large area color changes were 2, 4, 8, 14, 15, 17. The spectrum-effect relationship showed that peaks 11, 18, 17, 19, 8, 6, 16, 12, 1, 5, 7 and 15 were positively correlated with antioxidant activity. The following nine peaks were selected from the top five as quality evaluation indexes, Peak 2 (catechin), Peak 4 (epicatechin), Peak 8 [(+)-fisetinidol-(4β, 8)-epicatechin], Peak 11 (ellagic acid), Peak 14 (rosamultin), Peak 15 (kaji-ichigoside F1), Peak 17 (2α, 3β, 19α-trihydroxyurs-12-en-28-oic acid), Peak 18 (euscaphic acid), Peak 19 (quillaic acid). CONCLUSION The antioxidant activity of Rosa odorata Sweet var. gigantean root is the result of the combined effect of multi-components such as phenols and triterpenoid saponins, which provides a more comprehensive reference for the quality control of medicinal materials.