Chemical profiling of Qixue Shuangbu Tincture by ultra-performance liquid chromatography with electrospray ionization quadrupole-time-of-flight high-definition mass spectrometry (UPLC-QTOF/MS)

The present study was designed to develop and validate a sensitive and reliable ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS) method to separate and identify the chemical constituents of Qixue Shuangbu Tincture(QXSBT), a classic traditional Chinese medicine(TCM) prescription. Under the optimized UPLC and QTOF/MS conditions, 56 components in QXSBT, including chalcones, triterpenoids, protopanaxatriol, flavones and flavanones were identified and tentatively characterized within a running time of 42 min. The components were identified by comparing the retention times, accurate mass, and mass spectrometric fragmentation characteristic ions, and matching empirical molecular formula with that of the published compounds. In conclusion, the established UPLC-QTOF/MS method was reliable for a rapid identification of complicated components in the TCM prescriptions.

Optimization of the processing technology of Fructus Arctii by response surface methodology

The present study was designed to optimize the processing of Fructus Arctii by response surface methodology(RSM).Based on single factor studies,a three-variable,three-level Box-Behnken design(BBD) was used to monitor the effects of independent variables,including processing temperature and time,on the dependent variables.Response surfaces and contour plots of the contents of total lignans,chlorogenic acid,arctiin,and arctigenin were obtained through ultraviolet and visible(UV-Vis) monitoring and high performance liquid chromatography(HPLC).Fructus Arctii should be processed under heating in a pot at 311℃,medicine at 119 °C for 123 s with flipping frequently.The experimental values under the optimized processing technology were consistent with the predicted values.In conclusion,RSM is an effective method to optimize the processing of traditional Chinese medicine(TCM).

Identification and differentiation of major components in three different “Sheng-ma” crude drug species by UPLC/Q-TOF-MS

Cimicifugae Rhizoma(Sheng ma) is a Ranunculaceae herb belonging to a composite family and well known in China,has been widely used in traditional Chinese medicine.The Pharmacopoeia of the People's Republic of China contains three varieties(Cimicifuga dahurica(Turcz.),Cimicifuga foetida L.and Cimicifuga heracleifolia Kom.) which have been used clinically as "Sheng-ma".However,the chemical constituents of three components of "Sheng-ma" have never been documented.In this study,a rapid method for the analysis of the main components of "Sheng-ma" was developed using ultra-high performance liquid chromatography with quadrupole-time-of-flight mass spectrometry(UPLC/Q-TOF-MS).The present study reveals the major common and distinct chemical constituents of C.dahurica,C.foetida and C.heracleifolia and also reports principal component and statistical analyses of these results.The components were identified by comparing the retention time,accurate mass,mass spectrometric fragmentation characteristic ions and matching empirical molecular formula with that of the published compounds.A total of 32 common components and 8 markers for different "Sheng-ma" components were identified.These findings provide an important basis for the further study and clinical utilities of the three "Sheng-ma" varieties.

Development and validation of a HPLC-UV-ESI-MS method for the simultaneous quantitation of ten bioactive compounds in Dahuang Fuzi Tang

AIM: To develop a high performance liquid chromatography(HPLC) coupled with electrospray ionization mass spectrometry(ESI-MS) and ultraviolet(UV) detector method for the acid-alkaline simultaneous determination of ten bioactive compounds, and analyze the effect of compatible medicinal plants on the concentration of components in Dahuang Fuzi Tang(DFT). METHOD: The chromatographic separation was performed on a Hypersil BDS C18 analytical column by gradient elution with acetonitrile and formate buffer(containing 0.15% formic acid, V/V) at 25 °C with a flow rate of 1.0 mL ·min–1 and UV detection at 280 nm. Four of the ten compounds in DFT were identified and their MS fragments were elucidated by HPLC-ESI-MS, and the contents of the six compounds were determined by HPLC-UV. RESULTS: All calibration curves showed good linear regression(r2 ≥ 0.9990). The limits of detection and limits of quantification were 0.021–0.155 μg·mL –1 and 0.076–0.520 μg·mL –1, respectively. Overall precision RSD(intra-day and inter-day) were less than 2.96%, and the average recoveries were 98.35%–101.45%, with RSD ranging from 1.54% to 3.01% for the analytes. CONCLUSION: The developed method can be applied for the quality control and provide analytical evidence on the chemical basis and combinational principles of DFT.

Development of liquiritigenin-phospholipid complex with the enhanced oral bioavailability

In the present study,liquiritigenin-phospholipid complex(LPC)was developed and evaluated to increase the oral bioavailability of liquiritigenin.A single-factor test methodology was applied to optimize the formulation and process for preparing LPC.The effects of solvent,drug concentration,reaction time,temperature and drug-to-phospholipid ratio on encapsulation efficiency were investigated.LPCs were characterized by UV-visible spectroscopy,differential scanning calorimetry(DSC),fourier transform infrared spectroscopy(FTIR),and powder X-ray diffractometry(PXRD).The apparent solubility and n-octanol/water partition coefficient were tested.The pharmacokinetic characteristics and bioavailability of the LPC were investigated after oral administration in rats in comparison with liquiritigenin alone.An LPC was successfully prepared.The optimum level of various parameters for liquiritigeninphospholipid complex was obtained at the drug concentration of 8 mg·mL-1,reaction time for 15 min,reaction temperature of 30℃,a ratio of 1∶4.5(W/W)drug-to-phospholipid and anhydrous ethanol as reaction solvent.Compared to liquiritigenin,the AUC0-t of the LPC was increased by 239%.The liquiritigenin-phospholipid complex significantly increase the lipid solubility and bioavailability of liquiritigenin,suggesting that it is an effective formulation for further development and clinical applications.