Determination of flavonoids constituents in epimedium total flavone capsule and research on its anti-osteoporosis activity / 中国中药杂志

To develop a rapid resolution liquid chromatography (RRLC) method for the simultaneous determination of epimedoside A, epimedin A1, epimedin A, epimedin B, epimedin C, icariin, baohuosideⅡ, icarisideⅠ, sagittatoside B, 2"--rhamnosyl icarisideⅡ, and baohuosideⅠin epimedium total flavone capsule. At the same time, the effects of the above 11 compounds on osteogenic differentiation of MC3T3-E1 cells were investigated by detecting the content of alkaline phosphatase (AKP). The results showed that baohuoside Ⅱ had the highest activities, and both the activities of baohuoside Ⅱ and icariside Ⅰ were stronger than those of icariin.In this study, the content determination method of flavonoid glycosides was established, and the anti-osteoporosis effect of monomers was compared, providing technical support for the study of the pharmacodynamic and mechanism of Epimedium total flavone capsule.

Rapid identification of chemical components in Congrong Zonggan capsule by UPLC-Q-TOF-MS/MS / 中国中药杂志

This study was aimed to qualitatively analyze the chemical components in Congrong Zonggan capsule by using ultra-performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry method (UPLC-Q-TOF-MS/MS). An Agilent SB-C₁₈ Rapid Resolution HD (3.0 mm×100 mm,1.8 μm) was used with acetonitrile (A) - 0.1% formic acid solution (B) as the mobile phase for gradient elution. The flow rate was 0.2 mL•min⁻¹; the detection wavelength was set at 330 nm and the column temperature was maintained at 30 ℃. Electrospray ion (ESI) source was applied for the qualitative analysis under the negative ion mode. Finally, based on comparison with standard samples, database matching analysis and reviewing relevant literature, 41 compounds were identified from Congrong Zonggan capsule. This method could be used to rapidly detect the chemical components in Congrong Zonggan capsule, providing reference for the quality control of Congrong Zonggan capsule and laying a foundation for the further study on active components mechanism.

Rapid analysis on chemical constituents in Yinyanghuo Zonghuangtong capsule by UPLC/Q-TOF-MS/MS / 中国中药杂志

To analyze and identify the chemical constituents from Yinyanghuo Zonghuangtong capsule by using UPLC/Q-TOF-MS/MS. Chromatographic separation was carried out on an Agilent SB-C₁₈ column (2.1 mm×100 mm, 1.8 μm) at the temperature of 30 ℃. The mobile phase was 0.1% formic acid and acetonitrile by gradient elution, with a flow rate at 0.30 mL•min⁻¹, and the injection volume of 2 μL. The MS spectrum was acquired in both negative and positive ion modes using ESI ion source. Based on relative accurate molecular weight, secondary mass spectrometry pyrolysis fragments and chromatographic peak retention time, as well as fragmentation regularity summarized from reference substance and the literature, we could effectively identify the chemical structure of the components under test. As a result, a total of 46 compounds, including flavonoid glycosides, phenolic acids and alkaloids, were successfully identified or tentatively predicted. The results provide technical support for quality control and late-stage clinical application of Yinyanghuo Zonghuangtong capsule, and reference for further expound the pharmacodynamic material basis.

Quality assessment of Shuangyu Granules based on simultaneous determination by HPLC fingerprint and multi-components / 中草药

Objective: To establish an HPLC fingerprint chromatography and determine seven compounds of multi-components in Shuangyu Granules (SG). Methods: The Kromasil C18 (250 mm × 4.6 mm, 3.5 μm) column was used with a mobile phase of acetonitrile-0.05% trifluoroacetic acid gradient elution. The flow rate was 0.8 mL/min, the column temperature was 30℃, and the detection wavelengths were 230 and 327 nm. The common peaks were identified by Q-TOF/MS. Results: The fingerprint chromatography included 17 mutual peaks, and the similarity was more than 0.95. Fourteen common peaks had been identified by LC-Q-TOF/MS, seven of which were unequivocally identified via comparing the retention times and mass spectra data with those of the standard compounds. Then the seven marker components were quantified. The developed quantitative method was validated in terms of accuracy (the recoveries ranged from 97.8% to 101.8% with RSDs less than 2%). Conclusion: The method is rapid, simple, and accurate and can be used for the quality control of SG.

Research on ultrafine grinding technology of improving dissolution rates of effective components in Sanjie Zhentong capsule / 中国中药杂志

The effects of ultrafine grinding on the dissolution rates of the effective components in Sanjie Zhentong capsule (SZC) were studied in this experiment. Fine and ultrafine powder of SZC intermediates were made by ordinary grinding and ultrafine grinding technology, and then granulated by wet granulation. SZC were prepared by fine powder, ultrafine powder and ultrafine granules, respectively. With resveratrol and loureirin B as investigated indexes, dissolution rates of the four intermediates in SZC were determined by cup method and HPLC. The dissolution rates of resveratrol in SZC prepared by fine powder, ultrafine powder and ultrafine granules were 26.11%, 63.27%, 67.49%, respectively; and the dissolution rates of loureirin B were 7.160%, 20.29%, 23.05%, respectively. The dissolution rate of resveratrol and loureirin B in SZC prepared by ultrafine granules was the best. D90 size of ultrafine grinding was 13.221 μm and could improve the dissolution rates of resveratrol and loureirin B in SZC.

Fingerprint analysis of Resina Draconis from different manufactuers by UPLC coupled with chemometrics / 中国中药杂志

This study is to establish an UPLC fingerprint of Resina Draconis from different manufacturers, which can provide a comprehensive evaluation for its quality control. The analysis was performed on a Phenomenex Kinetex 2.6 μ C18 100A column by agradientelution program with acetonitrile-water as mobile phase at a flow rate of 1.7 mL x min(-1). The column temperature was 40 degrees C and the detection wavelengthwas 280 nm. The fingerprints of 18 batches of Draconis Resina were further evaluated by chemometrics methods including similarity analysis (SA), hierarchical clustering analysis (HCA) and principal component analysis (PCA). As a result, there were 15 common peaks, 13 of which had been identified by LC-Q-TOF MS, and the similarity degrees of 15 batches of the samples was more than 0.9, and the samples were divided into 4 clusters by their quality difference. The method is reproducible, simple and reliablethat it can be used for quality control and evaluation of Resina Draconis from different manufacturers.

Isolation and phylogenetic analysis of hemagglutinin gene of H9N2 influenza viruses from chickens in South China from 2012 to 2013

As part of our ongoing influenza surveillance program in South China, 19 field strains of H9N2 subtype avian influenza viruses (AIVs) were isolated from dead or diseased chicken flocks in Guangdong province, South China, between 2012 and 2013. Hemagglutinin (HA) genes of these strains were sequenced and analyzed and phylogenic analysis showed that 12 of the 19 isolates belonged to the lineage h9.4.2.5, while the other seven belonged to h9.4.2.6. Specifically, we found that all of the viruses isolated in 2013 belonged to lineage h9.4.2.5. The lineage h9.4.2.5 viruses contained a PSRSSRdownward arrowGLF motif at HA cleavage site, while the lineage h9.4.2.6 viruses contained a PARSSRdownward arrowGLF at the same position. Most of the isolates in lineage h9.4.2.5 lost one potential glycosylation site at residues 200-202, and had an additional one at residues 295-297 in HA1. Notably, 19 isolates had an amino acid exchange (Q226L) in the receptor binding site, which indicated that the viruses had potential affinity of binding to human like receptor. The present study shows the importance of continuing surveillance of new H9N2 strains to better prepare for the next epidemic or pandemic outbreak of H9N2 AIV infections in chicken flocks.

UPLC-Q-TOF-MS analysis on constituents of Tianshu Capsule absorbed into plasma / 中草药

To study the chemical constituents of Tianshu Capsule in vivo and in vitro. UPLC-Q-TOF-MS was used as the analytic method. The ferulic acid, caffeic acid, ligustrazine hydrochloride, gastrodin, 5-hydroxymethy furfural, and ligustilide were used as the reference compounds. The information on the total ion chromatogram, extraction chromatogram and the mass spectrogram were synthetically analyzed to confirm the constituents of Tianshu Capsule in vivo and in vitro. Twenty-four compounds from Tianshu Capsule were detected among five constituents came from Tianma and 19 constituents came from Chuanxiong. After oral administration of Tianshu Capsule, 13 compounds were absorbed into plasma. The findings obtained from the study can provide the useful information for the determination of bioactive substances and the perfection of quality standard of Tianshu Capsule.

Quality assessment of Longxue Tongluo Capsule based on HPLC fingerprint and multi-components simultaneous determination / 中草药

Objective: To establish an HPLC fingerprint of Longxue Tongluo Capsule (LTC), to determine the multi-components in LTC, and to provide the basis for the evaluation of LTC. Methods: The Kromasil C18 (250 mm×4.6 mm, 5 μm) column was used with a mobile phase of acetonitrile-0.1% formic acid gradient elution, the flow rate was 1.0 mL/min, the column temperature was 35℃, and the detection wavelength was 280 and 325 nm. Ten batches of LTC were detected to eatablish fingerprints, the common peaks were identified by Q-TOF/MS, and some of the characteristic peaks were analyzed. Results: The fingerprint chromatography with good resolution and reproducibility included 18 mutual peaks, and the similarity was more than 0.92.Sixteen common peaks had been identified by LC-Q-TOF/MS, seven of which were unequivocally identified via comparing the retention time, mass spectra, and MS/MS spectra data with those of the standard compounds. Then the seven maker components were identified as resveratrol, 7,4'-dihydroxy flavone, loureirin A, loureirin B, pterostilbene, 2,6-dimethoxy-4,4'-dihydroxydihydrochalcone, and 2-methoxy-4,4'-dihydroxydihydrochalcone. The seven components were quantified, and the recoveries ranged from 98.47% to 101.93% with RSD values of less than 2.27%. Conclusion: The method is rapid, simple, and accurate, and can be used for the quality control of LTC.

Transconjugation and genotyping of the plasmid-mediated AmpC beta-lactamase and extended-spectrum beta-lactamase genes in Klebsiella pneumoniae / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>AmpC beta-lactamases and extended-spectrum beta-lactamases (ESBLs) are becoming predominant causes of resistance to third and forth-generation cephalosporins in Klebsiella pneumoniae (K. pneumoniae). It is very difficult to treat infectious diseases caused by multidrug-resistant K. pneumoniae. The purpose of the present study was to investigate transconjugation and characteristics of beta-lactamase genes in K. pneumoniae producing AmpC beta-lactamases and ESBLs.</p><p><b>METHODS</b>AmpC beta-lactamases were detected by three-dimension test and ESBLs by disc confirmatory test. Minimum inhibitory concentrations (MICs) were determined by agar dilution. Transfer of resistance to EC600 (Rif(r)) was attempted by conjugation in broth and screened on agar containing cefotaxime (2 microg/ml) plus rifampin (1024 microg/ml). The genes encoding AmpC or ESBLs and their transconjugants were detected by PCR and verified by DNA sequencing.</p><p><b>RESULTS</b>The resistant rates to ampicillin and piperacillin were 100% in 18 isolates of K. pneumoniae. However, imipenem was still of great bactericidal activity on K. pneumoniae, and its MIC(50) was 0.5 microg/mL. Eleven beta-lactamase genes, including TEM-1, TEM-11, SHV-13, SHV-28, CTX-M-9, CTX-M-22, CTX-M-55, OXA-1, LEN, OKP-6 and DHA-1, were found from 18 isolates. And at least one beta-lactamase gene occurred in each isolate. To our surprise, there were six beta-lactamase genes in the CZ04 strain. Among 18 isolates of K. pneumoniae, the partial resistant genes in 8 isolates were conjugated successfully, which had 100% homological sequence with donors by sequence analysis. Compared with donors, 8 transconjugants had attained resistance to most beta-lactams, including ampicillin, piperacillin, cefoxitin, cefotaxime and aztreonam, or even amikacin and gentamicin.</p><p><b>CONCLUSIONS</b>R plasmids can be easily transferred between the resistant and sensitive negative bacilli. It is very difficult to block and prevent the spread of antimicrobial resistance. So more attention should be paid to reducing the frequency, times and dosage of antimicrobials, especially third or fourth cephalosporins.</p>