ABSTRACT
Objective To establish HPLC characteristic chromatogram ofKanglao Qingfei Granules.Methods HPLC analysis of samples was performed on Kromasil C18 column (4.6 mm × 250 mm, 5μm), with acetonitrile-1% glacial acetic acid as the mobile phase of gradient elution (0-50 min, 5%→15% acetonitrile;50-70 min, 15%→25% acetonitrile;70-80 min, 25%→40% acetonitrile;80-90 min, 40%→65% acetonitrile, 90-120 min, 65%→95% acetonitrile);the volume flow rate was 1.0 mL/min;detection wavelength was set at 290 nm;column temperature was 30℃. Chromatographic peaks were identified by HPLC-MS/MS method.Results The similarity degrees of 10 batches of samples were all greater than 0.995, and 13 chromatographic peaks were determined as common characteristic peaks, of which 10 peaks were confirmed in the source attribution and 8 peaks were identified in chemical component.Conclusion The established HPLC characteristic chromatogram can be used for the quality control ofKanglao Qingfei Granules.
ABSTRACT
Objective To optimize the extraction process of Kanglao Qingfei granules. Methods Orthogonal test design was used to study the influence of soaking time, water content, extraction time on extraction technology with bergenin's extraction rate, total polysaccharides's extraction rate and extract yield as evaluation indexes. Results The optimum extraction process conditions were as follows:the raw materials were immersed in water for 1.0 h and extracted two times under reflux, and the ratio of water volume to raw materials weight was 10 and 8 (mL/g), and extraction time was 2.5 h and 2.0 h, respectively. Conclusion The optimized extraction process was simple and stable, which can provide foundation for industrial production of Kanglao Qingfei granules.
ABSTRACT
The corona-like spikes or peplomers on the surface of the virion under electronic microscope are the most striking features of coronaviruses. The S (spike) protein is the largest structural protein, with 1,255 amino acids, in the viral genome. Its structure can be divided into three regions: a long N-terminal region in the exterior, a characteristic transmembrane (TM) region, and a short C-terminus in the interior of a virion. We detected fifteen substitutions of nucleotides by comparisons with the seventeen published SARS-CoV genome sequences, eight (53.3%) of which are non-synonymous mutations leading to amino acid alternations with predicted physiochemical changes. The possible antigenic determinants of the S protein are predicted, and the result is confirmed by ELISA (enzyme-linked immunosorbent assay) with synthesized peptides. Another profound finding is that three disulfide bonds are defined at the C-terminus with the N-terminus of the E (envelope) protein, based on the typical sequence and positions, thus establishing the structural connection with these two important structural proteins, if confirmed. Phylogenetic analysis reveals several conserved regions that might be potent drug targets.