1.Fingerprint Determination of Panax Notoginseng by LC/MS
Ying WANG ; Xinfa XHEN ; Longsheng SHENG
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(03):-
The fingerprints of Panax Notoginseng medicinal material,intermediate product and injection were detected and 75% ethanol was confirmed as the most suitable extracting solution. Notoginsenoside R 1 and ginsenoside R g1, R f, Rb 1, R d, R e etc. were indentified by LC/MS technigue. The fingerprints of medicinal material, intermediate product and injection were compared with each other.
2.Establishment of the Control of Cyclovirobuxine D
Xinjun XU ; Zhengxing ZHANG ; Dengkui AN ; Longsheng SHENG
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(02):-
Objective To set up the reference standard of cyclovirobuxine D.Methods Thermal analysis,HPLC/MS,HPLC with terminal wavelength,HPLC with fluorescence derivation and with ultraviolet derivation,TLC and nonaqueous titration methods were applied to determine the content of cyclovirobuxine D control.Results Thermal analysis can not be used to analyse the purity of cyclovirobuxine D ,and HPLC/MS,HPLC with terminal wavelength,HPLC with fluorescence derivation and HPLC with ultraviolet derivation can obtain the same purity.Conclusion The methods used for the assay of cyclovirobuxine D control were practical.
3.HIGH PERFORMANCE LIQUID CHROMATOGRAPHY / ELECTROSPRAY IONIZATON MASS SPECTROMETRIC CHARACTERIZATION OF RECOMBINANT L-ASPARAGINASE II
Jun HAN ; Longsheng SHENG ; Zhongyuan YANG ; Bingren XIANG ; Dengkui AN
Acta Pharmaceutica Sinica 2001;36(1):46-50
AIM To characterize the primary structure of recombinant L-asparaginase II product. METHODS The molecular weight of the protein was measured by pneumatically-assisted electrospray ionization mass spectrometry with flow injection mode. Subsequently, tryptic peptide mapping was performed by high performance liquid chromatography on a C8 column with tandem UV and MS detection. An easy-to-use and simple denaturation process with trichloroacetic acid was conducted prior to tryptic digest so as to release the digest resistance from the protein structure. The amino acid sequences of the tryptic peptides were elucidated based on their in-source collision-induced dissociation spectra. RESULTS The measured molecular mass was different from the theoretical value. Three amino acid variations were unambiguously detected along the peptide backbone derived from the gene-encoding sequence. CONCLUSION This paper revealed that LC/ESI/MS had provided a promising and robust technique in primary structure analysis and quality control of DNA-derived recombinant protein pharmaceuticals.
4.Polyphenols from Terminalia chebula
Gang DING ; Yanze LIU ; Maoping SONG ; Dapeng ZOU ; Longsheng SHENG
Journal of China Pharmaceutical University 2001;(3):193-196
AIM The purpose is to examine the chemical constituents in the fruits of Terminalia chebula. METHODS Using combined chromatographies over silica gel, Diaion HP-20, Toyopearl HW-40 and MCI gel CHP -20P to purify the constituents of Terminalia chebula, and identifying their structures on the basis of spectroscopic and chemical evidence were purified. RESULTS Twenty one hydrolyzable tannins and related polyphenols were characterized, here reports eight of them: 2,3-(S)-HHDP-D-glucose, 3,6-di-O-galloyl-D-glucose, 6-O-galloyl-D-glucose, (-)-shikimide 4-O-gallate, (-)-shikimic acid 3-O-gallate+(-)-shikimic acid 5-O-gallate, methyl gallate and 1,2,6-tri-O-galloyl-β-D-glucose were reported. CONCLUSION The above eight polyphenols were obtained from myrobalans for the first time.
5.Study on extraction of Sophora flavescens Ait. by HPLC-MS
Renling MA ; Honghua ZHOU ; Xishui YU ; Longsheng SHENG ; Dengkui AN ;
Chinese Traditional Patent Medicine 1992;0(10):-
Objective:Chemical constituents of Sophora flavescems Ait. were studied by different extraction. Methods: Online HPLC/ESI/MS was used to study the different extraction. Results: numbers of alkaloids and flavone from Sophora flavescens Ait. were 9,8 and 12, respectively by means of chloroform strong aqua , water , and water methanol extraction. Conclusions: For the first time, we studied extraction of Sophora flavescems Ait by HPLC MS. This provided basic for study on fingerprints of Sophora flavescems Ait..
6.Extraction, isolation and purification for ginkgolide B.
Chenfeng ZHANG ; Minghui LI ; Yun TANG ; Yanhai ZHANG ; Min SHI ; Longsheng SHENG
China Journal of Chinese Materia Medica 2010;35(15):1961-1964
OBJECTIVETo establish a simple extraction, isolation and purification method for ginkgolide B from ginkgo leaf.
METHODThe optimum conditions of extraction, isolation and purification were studied by taking the transfer rate of ginkgolide B as index.
RESULTGinkgo leaf was extracted with 70% ethanol for three times, the extracts were concentrated to remove ethanol and diluted by water till the crude drug density reached 0.1 g x mL(-1). The dilution was adsorbed with HPD-450 macroporous resin. The impurities were eluted with 20% ethanol and ginkgolide B was eluted with 80% ethanol. Then the 80% ethanol eluant was concentrated and crystallized. Finally the crude crystals were recrystallized with isopropanol. The purity of the ginkgolide B recrystallization was 95%.
CONCLUSIONThe process was stable and easy to operate, which was suited to industrialized production.
Chemical Fractionation ; methods ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Ginkgo biloba ; chemistry ; Ginkgolides ; analysis ; isolation & purification ; Lactones ; analysis ; isolation & purification ; Plant Leaves ; chemistry
7.Immune intervention with anti-CD80 bivalent antibody in pristane-induced mouse model of lupus nephritis
Hui SHEN ; Han SHENG ; Yuqiang ZHU ; Ying ZHENG ; Huan PAN ; Longsheng XU ; Jianfen SHEN ; Yuhua QIU
Chinese Journal of Microbiology and Immunology 2017;37(9):688-694
Objective To study the therapeutic effect of anti-CD80 bivalent antibody on mouse lu-pus nephritis and to explore the possible molecular mechanism. Methods A mouse model of lupus nephritis was established through intraperitoneal injection of 0. 5 ml of pristine in female C57BL/6J mice. Appearance of urinary protein and significantly increased levels of peripheral antinuclear antibody ( ANA) and anti-doub-le-stranded DNA ( anti-dsDNA) antibody in the fourth month after injection indicated that the mouse model was established successfully. Then the mice were divided into two groups including anti-CD80 bivalent anti-body intervention group (injected with 200μg of anti-CD80 bivalent antibody at day 1, 3, 5, 8 and 15, fol-lowed by three times of injection with an interval of one month) and model group ( injected with the same protein using the same strategy). A normal control group was set up accordingly. Albustix test paper was used to monitor the dynamic changes in mouse urinary protein. Flow cytometry was used to analyze the acti-vation of immune-related cells in spleen. Levels of autoantibodies ( ANA and anti-dsDNA) and levels of IFN-γ and IL-4 in serum were detected by indirect immunofluorescence assay. Renal tissue samples were an-alyzed with hematoxylin and eosin ( HE) staining and immunocomplex ( IC) assay. Results Urinary pro-tein level of the anti-CD80 bivalent antibody intervention group was significantly lower than that of the model group (P<0. 05). Activated macrophages, dendritic cells, neutrophils and B cells in spleen tissues of the anti-CD80 bivalent antibody intervention group were significantly less than those of the model group ( P<0. 05), and the numbers of CD4+ and CD154+ T cells were significantly less than those of the model group (P<0. 05). Positive rates and titers of ANA and dsDNA in serum samples of the intervention group were lower than those of the model group (P<0. 05). Levels of IFN-γand IL-4 in serum samples of the interven-tion group were decreased as compared with those of the model group (P<0. 05). HE staining and immuno-fluorescence assay showed that glomerular inflammatory injury and necrosis were alleviated and kidney im-mune complex deposition was reduced after anti-CD80 bivalent antibody intervention. Conclusion Anti-CD80 bivalent antibody specifically binds to the CD80 molecule on antigen presenting cell surface, blocks the CD80/CD28 co-stimulatory signaling pathway and down-regulates the body′s immune response, which al-leviates and reverses the lupus-like nephritis-induced pathological damages in mice.