Rapid flavonoid-focused sub-chemome characterization of Draconis Sanguis using UPLC-Q-TOF-MS in combination with molecular weight imprinting and mass defect filtering / 中国中药杂志

Draconis Sanguis is a precious Chinese medicinal material for activating blood and resolving stasis, and its effective components are flavonoids. However, the structural diversity of flavonoids in Draconis Sanguis brings great challenges to the in-depth chara-cterization of its chemical composition profiles. To clarify the substance basis of Draconis Sanguis, ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used in this study to acquire MS data of Draconis Sanguis. The molecular weight imprinting(MWI) and mass defect filtering(MDF) were developed for rapid screening of flavonoids in Draconis Sanguis. Full-scan MS and MS~2 were recorded within the mass range m/z 100-1 000 in positive ion mode. Accor-ding to previous literature, MWI was employed to hunt for reported flavonoids in Draconis Sanguis, and the mass tolerance range of [M+H]~+ was set as ±10×10~(-3). A five-point MDF screening frame was further constructed to narrow the screening range of flavonoids from Draconis Sanguis. Combined with diagnostic fragment ions(DFI) and neutral loss(NL) as well as mass fragmentation pathways, 70 compounds were preliminarily identified from the extract of Draconis Sanguis, including 5 flavan oxidized congeners, 12 flavans, 1 dihydrochalcones, 49 flavonoids dimers, 1 flavonoids trimer and 2 flavonoid derivatives. This study clarified the chemical composition of flavonoids in Draconis Sanguis. Moreover, it also showed that high-resolution MS combined with data post-processing methods such as MWI and MDF could achieve rapid characterization of the chemical composition in Chinese medicinal materials.

Anti-tumor Effect of Draconis Sanguis Petroleum Ether Fraction on Human Gastric Cancer HGC-27 and MGC-803 Cells / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Draconis Sanguis petroleum ether fraction （DSPEF） on the proliferation， apoptosis， migration， and autophagy of human gastric cancer HGC-27 and MGC-803 cells， and preliminarily elucidate its molecular mechanism. MethodCell counting kit-8 （CCK-8） assay was used to detect the effect of DSPEF at different concentrations （0， 20， 40， 60， 80 mg·L-1） on the proliferation of HGC-27 and MGC-803 cells after 24， 48， 72 h. Hoechst staining and flow cytometry were used to explore the effects of DSPEF at different concentrations on the apoptosis and apoptosis rate of HGC-27 and MGC-803 cells after 48 h treatment， respectively. The wound healing assay and acridine orange staining were used to investigate the effects of DSPEF on the migration and autophagy of HGC-27 and MGC-803 cells， respectively. Western blot was used to detect the expression levels of signaling pathway-related proteins in HGC-27 and MGC-803 cells treated with DSPEF for 48 h. ResultCompared with the control group， DSPEF（30 mg·L-1） inhibited the proliferation and migration of HGC-27 and MGC-803 cells in a concentration- and time-dependent manner （P<0.05）， and induced the apoptosis （P<0.01） and autophagy of HGC-27 and MGC-803 cells. DSPEF （60 mg·L-1） down-regulated the protein levels of phosphorylated mammalian target of rapamycin （p-mTOR） （P<0.05， P<0.01） and down-regulated phospho-signal transducer and activator of transcription 3 （p-STAT3） in HGC-27 and MGC-803 cells （P<0.01）， suggesting that DSPEF presumedly inhibited the proliferation and migration of human gastric cancer HGC-27 and MGC-803 cells and induced their apoptosis and autophagy by inhibiting the mTOR/STAT3 signaling pathway. ConclusionThe down-regulation of the mTOR/STAT3 signaling pathway may be involved in the anti-gastric cancer effect of DSPEF. This study is expected to provide a reference for the investigation of the anti-tumor effect of Draconis Sanguis.

New Quality Problems and Control Methods for Imported Draconis Sanguis / 中国药学杂志

OBJECTIVE: To explore the new quality problems of Draconis Sanguis imported from Indonesia. METHODS: According to the quality standard of imported medicinal materials and Ch.P method, exploratory analysis was carried out on 10 batches of imported Draconis Sanguis and three batches of Draconis Sanguis fruits. RESULTS: The dracohodin content in three batches of Draconis Sanguis fruits was 1.2%-2.4%. Three batches of imported Draconis Sanguis were obtained by directly pulverization of Draconis Sanguis fruits, and loureirin A and loureirin B were detected in one batch of Draconis Sanguis, suggesting that dracaena cochinchinensis was added. CONCLUSION: Among the 10 batches of imported Draconis Sanguis, three batches are determined to be unqualified due to character inconformity, and the other batch is judged to be unqualified due to the addition of dracaena cochinchinensis.

DNA barcodes identification of one rare traditional Chinese medicine Draconis Sanguis / 中国药学杂志

OBJECTIVE: To choose suitable barcodes for Draconis Sanguis and find the new method to identify Draconis Sanguis. METHODS: The genomic DNAs from 41 samples were extracted. Draconis Sanguis samples were amplified and sequenced based on seven candidate DNA barcodes [ITS, ITS2, psbA-trnH, rbcL, matK, trnh (UAA) intron, P6 loop of trnh (UAA) intron]. The obtained sequences were assembled using the CodonCode Aligner V 4.2. The Kimura 2-Parameter (K2P) distances were calculated using MEGA 5.0. The neighbor-joining (NJ) phylogenetic trees were constructed. RESULTS: Six out of the seven candidates [ITS, ITS2, psbA-trnH, rbcL, matK, trnh (UAA) intron] were not able to be amplified, while P6 loop of trnh (UAA) intron could be used for identification. The length of the P6 loop of trnh( UAA) intron sequence of Daemonorops draco was 45 bp. The maximum K2P intra-spe-cific genetic distances of Daemonorops draco were much smaller than the minimum inter-specific genetic distances between Daemonorops draco and its adulterants. The NJ tree showed that Draconis Sanguis differed from its adulterants obviously. CONCLUSION: The P6 loop of trnL (UAA) intron can be used as DNA barcode for the identification of Draconis Sanguis.