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  • Chen SUN, Zhe-yu XIE, Yong-jiang WU
    Acta Pharmaceutica Sinica. 2019, 54(11): 2059-2063.

    An HPLC fingerprint and multi-component determination method of Leonurus japonicus was established for comprehensive evaluation and quality control of Leonurus japonicus. The sample was incubated in 70% ethanol in a water bath for 2 h, and the extract was analyzed by HPLC using Kromasil C18 column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-0.1% formic acid with gradient elution. The flow rate was 1.0 mL·min-1. The temperature of column was 30℃. The detection wavelength was 280 nm. HPLC fingerprint of characteristic components of Leonurus japonicus was established. There were 12 common peaks among 25 batches of samples, and 5 of them were identified and determined. Syringic acid, leonurine hydrochloride, rutin, hyperoside or isoquercitrin showed a good linearity in the ranges of 0.426 1-85.22 ng (r=0.999 9), 7.948-1 590 ng (r=0.999 3), 10.20-2 040 ng (r=1.000 0), 2.018-403.6 ng (r=0.999 9), or 8.704-1 741 ng (r=0.999 9), respectively. The average recoveries were 99.0%, 97.6%, 97.4%, 96.9% and 98.5% with RSD of 1.1%, 1.8%, 1.4%, 1.5% and 1.3%, respectively. The HPLC characteristic fingerprint of Leonurus japonicus was specific, and this method can simultaneously determine the content of 5 components.

  • En-ying WANG, Cheng-ting JIN, Jun-jun WANG, Yong CHEN, Feng-mei HAN
    Acta Pharmaceutica Sinica. 2019, 54(11): 2064-2068.

    Vinpocetine (VP) has been widely used to treat cerebrovascular disorders and nerve injury. Borneol (BN), as an important traditional Chinese medicine, is commonly used to promote the absorption and distribution of central nervous system drugs. In this work, a LC-MS/MS method was developed to determine the level of VP in rat plasma and tissues, and to evaluate the effect of co-administration of BN with VP by oral gavage on the absorption and tissue distribution of VP in rats. Rats were divided into VP (10 mg·kg-1), VP (10 mg·kg-1) + BN (75 mg·kg-1) and VP (10 mg·kg-1) + BN (150 mg·kg-1) groups for pharmacokinetic study, and divided into VP (10 mg·kg-1) and VP (10 mg·kg-1) + BN (150 mg·kg-1) groups for tissue distribution study. The animal experiment was approved by Ethics Committee of Hubei University, and complied with the guideline for caring and using of laboratory animals. Compared to VP group, the AUC0-∞, MRT0-∞ and t1/2z of VP + BN (150 mg·kg-1) group increased significantly, by 1.98-, 1.22- and 1.42-fold respectively, and the exposure in plasma, liver, kidney and brain increased by 2-, 1.5-, 1.5- and 1.3-fold respectively. The pharmacokinetic results suggested that co-administration of BN with VP is beneficial for overcoming the undesirable pharmacokinetic characteristics of VP, such as short residence time, low oral bioavailability and brain exposure in clinical usage.

  • Yu-xuan MA, Na WANG, Su-yun DAI, Bing WANG, Gang LI
    Acta Pharmaceutica Sinica. 2019, 54(11): 2069-2073.

    The preparation of polymorphic forms of rivaroxaban was carried out using a recrystallization method based on that for crystal form-Ⅰ. Preparation methods were developed for two crystal forms-Ⅱ (medicinal crystal form) and five crystal forms-Ⅳ and the crystals were then characterized. The crystalline form was identified by applying modern analytical means including X-ray powder diffraction (PXRD), differential scanning calorimetry (DSC), element analysis (EA), mass spectrometry (MS), and infrared spectrum (IR), and the morphology of different crystal forms was observed by scanning electron microscopy (SEM). The results show that the PXRD and DSC characteristics of prepared crystal forms-Ⅱ and Ⅳ are consistent with those described in patents at home and abroad, and the test results with EA, MS and IR are in accordance with the chemical structure of rivaroxaban. The crystal form-Ⅰ is lamellar, the crystal form-Ⅱ is linear and crystalline form-Ⅳ is striped as determined by SEM. In summary, the methods for preparing crystal form-Ⅱ and form-Ⅳ are reliable, the required reagents are easily available, the experimental conditions are easy to implement and the preparation process is simple. Our study provides a new reference for the production and application of rivaroxaban.

  • Xiao-ru HU, Hai-rong ZHENG, Hai-xia DANG, Qin HU, Shuai KANG, Zhong DAI, Shuang-cheng MA
    Acta Pharmaceutica Sinica. 2019, 54(11): 2074-2082.

    We determined a component-target-disease network for Carthamus tinctorius L. and the key compounds, identified by topological analysis, were related to vasculitis, coronary heart and cerebrovascular disease. Based on these compounds, the chromatographic fingerprint of Carthamus tinctorius L. was established. Firstly, 132 compounds were obtained from TCMID and TCMSP databases. Their targets were predicted in the PharmMapp and HemMapper databases. CardioGenBase, Therapeutic Target Database and DisGeNET databases were used to collect targets of vasculitis, coronary heart disease and cerebrovascular disease. The corresponding relationships between component and target protein were established by mapping. Finally, the " component-target-disease" network was built with Cytoscape software. The core network and key nodes were analyzed with the Cytohubba plug-in. The results showed that the 24 key compounds were alpha-tocopherol, adenosine, quinone chalcone pigments such as hydroxysafflor yellow A, safflower yellow, quercetin, kaempferol and other flavonoids, organic acids such as stearic acid, linolenic acid, coumaric acid and cinnamic acid. This resulting chromatographic fingerprint of Carthamus tinctorius L. showed good consistency, and the core chemical compounds obtained by topological analysis of the network of " component-target-disease" , could be used as quality control markers. Our research provides a new approach for the identification of quality control indicators in Chinese medicinal materials.

  • Fei-fei WANG, Jie LIU, Feng-yan HE, Nan-ping ZHANG, Yi HE, Jing LI, Yu-mei ZHANG, Jian ZHENG, Shuang-cheng MA
    Acta Pharmaceutica Sinica. 2019, 54(11): 2083-2088.

    The morphological identification, chemical component analysis, and DNA barcode determination were investigated on Genus Mentha (including Mentha haplocalyx Briq., Mentha piperita Linn., Mentha spicata Linn. and Mentha cultivated) in order to reveal the origin of Herba Mentha as a drug, and ensure the accuracy in clinic application. The morphological characters, chemical composition analysis by GC-MS/MS and DNA content measure by polymerase chain reaction (PCR) were reported in this study for inter- or intra- species divergence. Based on the morphology, axillary verticillasters was recognized as the typical character for Mentha haplocalyx Briq. Carvone was used as an index component for chemical composition analysis of Mentha spicata Linn. Gene clustering analysis divided 22 batches of samples into two molecular groups. Mentha haplocalyx Briq. is distinguishably different from Mentha spicata Linn. Mentha piperita Linn. and other cultivated plants were distributed between these two species. The results obtained by morphological identification, chemical composition analysis, and DNA barcode determination show good correlations, but each identification method has its limit. In view of the fact that hybridization of the plants in Genus Mentha is common, identification relying on only one method is not recommended.

  • Jie GAO, Xian-cheng ZHAN
    Acta Pharmaceutica Sinica. 2019, 54(11): 2089-2092.

    The methods for determination of freezing point include cooling curve cryoscopy and air humidity cryoscopy. These methods are usually influenced by many factors, such as instrumentation, environment and operators. Despite the numerous experimental methods, precise freezing point values are challenging to obtain due to time-consuming procedures, limited sample size and extensive manual work. In this study, a semi-empirical hydration model (SEHM) was developed to calculate freezing point of NaH2PO4-K2HPO4 mixed electrolyte solution. According to SEHM, the water activity of mixed electrolyte solution was calculated by molality of solutes. Then the freezing point of solution was calculated. The calculated results were compared with those obtained by cooling curve cryoscopy and air humidity cryoscopy. The results indicate that the SEHM calculations are comparable to the measurements of cooling curve cryoscopy and air humidity cryoscopy.

  • Lei LI, Lan SHEN, Xiao-ning WANG, Xiao LIN, Yan-long HONG, Yi FENG
    Acta Pharmaceutica Sinica. 2019, 54(11): 2093-2099.

    Bioadhesive preparation can be attached to specific sites to control drug release rate, increase drug concentration and increase efficacy, which is based on natural or synthetic polymer material. In this paper, based on the physical properties of wet mass, a method for screening adhesion formulation was proposed, which was different from conventional way of screening optimal formulation, and astragalosides loaded bioadhesive pellets were prepared by extrusion-spheronization method (extrusion speed 30 r·min-1, spheronization speed 808 r·min-1, spheronization time 7.5 min) based on this formulation screening method, small living animal imaging technology and mucin from porcine stomach model were used to evaluate the in vivo and in vitro adhesiveness behaviour of the pellets. According to the relationship between the physical properties of wet mass and the formability and adhesiveness of bioadhesive pellets, five key physical properties hardness (Ha), adhesiveness (Ad), springiness (Sp), cohesiveness (Co), chewiness (Ch) were selected as the index of screening optimal formulation, therefore a comprehensive evaluation model was established, which based on principal component analysis, to did digital ranking for these proposed adhesion formulation, the optimal formulation was determined:microcrystalline cellulose:(chitosan:Carbomer 940=2:1), the adhesive material dosage accounted for 20% of the excipient dosage, and the ratio of drugs to excipients was 1:4. All animal experiments have been approved by Ethics Committee of Shanghai University of Traditional Chinese Medicine. The in vivo and in vitro adhesive evaluation results showed the pellets had a clear advantage in intestinal adhesion over normal pellets, its also proved the scientificity and reliability of the method of screening bioadhesive formulation.

  • Ting LI, Dan JIANG, Xiao-song HU, Xiao-xi CHANG, Zhen XU, Ying GONG, Guo-dong HUA, Chun-sheng LIU
    Acta Pharmaceutica Sinica. 2019, 54(11): 2100-2105.

    The structural composition of the surface fungal community of commercially Platycladi semen was analyzed to reveal the surface fungal biodiversity and structural differences. Platycladi semen was collected from Henan, Shandong and Hong Kong, their DNA was extracted, ITS fragments in DNA were amplified by PCR. Miseq was sequenced on Illumina Hiseq 2500 platform after the PCR products were qualified for quality inspection. The sequence OTU cluster was obtained and bioinformatics analysis was carried out. Microbial communities were not observed in the eyes of the Platycladi semen in the three regions. Sequencing results showed that the surface microbial community had high biodiversity, but there were significant differences in species composition. Seven samples o Platycladi semen obtained 345 947 valid sequences, which were divided into 267 OTUs, 3 phylums. 18 classes, 40 orders, 82 families, 120 genus, 191 species fungi. At the genus level, Aspergillus is the dominant species, accounting for the highest proportion, reaching (93.36 ±6.01)%. Seven samples were contaminated by Aspergillus flavus, and the pollution levels were 14.58%, 15.98%, 17.64%, 16.44%, 0.97%, 23.39% and 18.86%. Except sample No. 5, Aspergillus cibarius was the most abundant, the other six samples were Aspergillus niger, Aspergillus fumigatus and Aspergillus flavus as the core microflora. By analyzing the diversity of fungi distribution in different habitats, we can fully understand the fungi on the surface of Platycladi semen, lay a foundation for early risk warning of Aspergillus flavus contamination and its aflatoxin contamination, and provide a theoretical basis for the quality and safety of Platycladi semen.

  • Yong-gang QIAO, Jia-xin HE, Yong-fei WANG, Ya-ping CAO, Meng-jun JIA, Xin-rui ZHANG, Jian-ping LIANG, Yun SONG
    Acta Pharmaceutica Sinica. 2019, 54(11): 2106-2112.

    In order to explore the chloroplast genome characteristics of Sophora flavescens and the phylogenetic relationship of the genus, this study used high-throughput sequencing technology to sequence and functionally annotate the chloroplast genome of Sophora flavescens. The results showed that the full length 154 165 bp of Sophora flavescens chloroplast genome showed a typical four-stage structure. The chloroplast genome contains 123 genes, including 77 protein-coding genes, 38 tRNA genes and 8 rRNA genes. Sequence analysis revealed 104 SSR loci, most of which consisted of A and T. In addition, the chloroplast genome codon preference is weak, and the coding region is biased towards the use of A and T bases. A comparative analysis of two different regions of Sophora flavescens chloroplast genome revealed four differential genes. Based on the maximum likelihood method (ML) for phylogenetic analysis of Sophora flavescens and 16 other leguminous, it was found that the relationship between Sophora flavescens and the genus Sophora alopecuroides is the closest. This study provides an important theoretical basis for the genetic variation, breeding and phylogenetic analysis of Sophora flavescens, and has certain reference value.

  • Lian-quan QI, Yu BAI, Jian-hui LUO
    Acta Pharmaceutica Sinica. 2019, 54(11): 2113-2117.

    Sequence analysis of DNA, mRNA and protein is an essential component of biologics or bioprocess development. Analysis of sequences at the DNA, mRNA, and protein levels after the transfer of the gene of interest into a host cell is an important part of quality control. This article reviews the application of new technologies such as next-generation sequencing and LC-MS/MS in biological drug development such as monoclonal antibodies. These techniques have different requirements in term of cost, handling time and expertise. Selecting an appropriate technique with a sound rationale at different stages of drug development will add to the success rate of research and development, and ensure product quality, thus ensuring the clinical efficacy and safety.