2.Analysis of Mass Isotopomer Distributions of Proteingenic Amino Acids Used GC-MS during ~(13)CMFA
Jian WANG ; Ze-Jian WANG ; Ming-Zhi HUANG ; Jiang-Chao QIAN ; Ju CHU ; Si-Liang ZHANG ;
China Biotechnology 2006;0(07):-
13C metabolic flux analysis(13CMFA)have been the research hotspots of metabolic engineering internationally due to its accuracy and applicability.It is vital that the measurement of 13C labelling pattern of proteinogenic amino acids for 13C metabolic flux analysis.To acquire 13C-labelling proteinogenic amino acids,Pseudomonas denitrifican which products Vitamin B12 was firstly fed with mimimal culture medium contained 20% U-13C and 80% natural glucose,after the culture reached a steady state,then about 20 mg biomass was hydrolyzed by 1 ml of 6 mol/L hydrochloric acid for 24h at 110℃.Then amino acids was separated,concentrated,evaporated in a vacuum,and derivatized with MBDSTFA,TBDMS-derivatized amino acids can be observed by GC-MS last we get 13C labelling pattern of fifteen aminio acids through mass spectrum.The experimental methods and sample preparation offers referential value for the development of 13C metabolic flux analysis in our courtry.
3.Identification of bufadienolides profiling in cinobufacino by HPLC-DAD-FT-ICR-MS method.
Jun-Qiu LIU ; Nan SI ; Jian YANG ; Hai-Yu ZHAO ; Bao-Lin BIAN ; Hong-Jie WANG
Acta Pharmaceutica Sinica 2014;49(2):244-248
Cinobufacino injection is a significant anti-tumor medicine for the treatment of various tumors in clinic, which was made from water extraction of the skin of Bufo bufo gargarizans. In present paper, HPLC-DAD-FT-ICR-MS method was used to identify the major bufadienolides in cinobufacino for the first time. Solid-phase extraction with dichloromethane and silica was used to enrich the total bufadienolides in cinobufacino. Based on the UV and high resolution MS/MS data, 33 bufadienolides were analyzed and characterized. Among them, eight compounds were identified by comparing with standard references unambiguously. This study elucidated the major bufadienolides in cinobufacino, which provided material foundation of cinobufacino and will be benefit for the further pharmacological research.
Amphibian Venoms
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chemistry
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Animals
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Bufanolides
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analysis
;
chemistry
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Bufo bufo
;
Chromatography, High Pressure Liquid
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Molecular Structure
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Spectrometry, Mass, Electrospray Ionization
;
Tandem Mass Spectrometry
4.LC-FT-ICR-MS analysis of the prototypes and metabolites in rat plasma after administration of huang-lian-jie-du decoction.
Ran ZUO ; Hong-Jie WANG ; Nan SI ; Hai-Yu ZHAO ; Jian YANG ; Bao-Lin BIAN
Acta Pharmaceutica Sinica 2014;49(2):237-243
To obtain chemical constituent information of rat plasma after oral administration of Huang-Lian-Jie-Du Decoction (HLJDD), a LC-FT-ICR-MS method has been established, and both positive and negative ions scan modes were include in the analysis. By comparing their retention time, high resolution mass data of HLJDD extracts, blank plasma and dosed plasma, 38 constituents, including 22 prototype compounds and 16 metabolites, were detected in rat plasma after oral administration of HLJDD. In the 22 prototype compounds, 16 constituents were determined unambiguously by comparing with references. In the analysis of metabolites, phase II reactions like glucuronidation and sulfation were the major biotransformation pathways of HLJDD. M11 was observed as the only phase I metabolite in present experiment. The results will be beneficial for the further pharmacokinetics and pharmacological evaluations of HLJDD.
Administration, Oral
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Alkaloids
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blood
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Animals
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Biotransformation
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Chromatography, High Pressure Liquid
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Drug Combinations
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Drugs, Chinese Herbal
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administration & dosage
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isolation & purification
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metabolism
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Flavonoids
;
blood
;
Iridoids
;
blood
;
Male
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Plants, Medicinal
;
chemistry
;
Rats
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Rats, Sprague-Dawley
;
Spectrometry, Mass, Electrospray Ionization
;
Tandem Mass Spectrometry
5.In vivo comparison analysis of scandoside methyl ester metabolites in four kinds of liver microsomes using ultra-performance liquid chromatography combined with electrospray ionization tandem orbitrap mass spectrometry.
Yan LI ; Hongjie WANG ; Jing HE ; Nan SI ; Jian YANG ; Haiyu ZHAO ; Baolin BIAN
Acta Pharmaceutica Sinica 2014;49(9):1315-9
In order to clarify the metabolism pathways of scandoside methyl ester, the analysis of metabolites profiling in four kinds of liver microsomes was performed by using an ultra-performance liquid chromatography/ electrospray-tandem mass spectrometry (UPLC-ESI-MS). The data obtained from the 0 h-incubation and the 2 h-incubation were compared and analyzed. After incubation, 5 metabolites of scandoside methyl ester were found in rat, Beagles, rhesus monkey and human liver microsome. The results showed that scandoside methyl ester's major metabolic pathway in the liver microsomes is hydrolysis, oxidation and reduction reactions, and there are certain kinds differences between species. The study provides a research base for further research about iridoid compounds in vivo metabolic pathways.
6.Chemical constituents of bufadienolides in cinobufacino injection.
Lingyu HAN ; Nan SI ; Junqiu LIU ; Haiyu ZHAO ; Jian YANG ; Baolin BIAN ; Hongjie WANG
Acta Pharmaceutica Sinica 2014;49(11):1574-7
Cinobufacino injection is purified from water extraction of the skin of Bufo bufo gargarizans, which has been widely used for various cancers in clinic with significant anti-tumor effects. Bufadienolides were regarded as the main active constituents of cinobufacino injection in previous reports. In present study, 6 bufadienolides were isolated and purified from Cinobufacino injection. Their structures were identified as 3-epi-ψ-bufarenogin (1), ψ-bufarenogin (2), 3-epi-arenobufagin (3), arenobufagin (4), 3-epi-gamabufotalin (5), and 3-oxo-arenobufagin (6), separately. Among them, 1 and 3 were new compounds, 5 and 6 were new natural products. Compounds 1, 2 and compounds 3, 4 were two pairs configuration isomers at C-3, separately.
7.LC-FT-ICR-MS analysis of the prototypes and metabolites in rat plasma after administration of huang-lian-jie-du decoction.
Ran ZUO ; Hongjie WANG ; Nan SI ; Haiyu ZHAO ; Jian YANG ; Baolin BIAN
Acta Pharmaceutica Sinica 2014;49(2):237-43
To obtain chemical constituent information of rat plasma after oral administration of Huang-Lian-Jie-Du Decoction (HLJDD), a LC-FT-ICR-MS method has been established, and both positive and negative ions scan modes were include in the analysis. By comparing their retention time, high resolution mass data of HLJDD extracts, blank plasma and dosed plasma, 38 constituents, including 22 prototype compounds and 16 metabolites, were detected in rat plasma after oral administration of HLJDD. In the 22 prototype compounds, 16 constituents were determined unambiguously by comparing with references. In the analysis of metabolites, phase II reactions like glucuronidation and sulfation were the major biotransformation pathways of HLJDD. M11 was observed as the only phase I metabolite in present experiment. The results will be beneficial for the further pharmacokinetics and pharmacological evaluations of HLJDD.
8.Identification of bufadienolides profiling in cinobufacino by HPLC-DAD-FT-ICR-MS method.
Junqiu LIU ; Nan SI ; Jian YANG ; Haiyu ZHAO ; Baolin BIAN ; Hongjie WANG
Acta Pharmaceutica Sinica 2014;49(2):244-8
Cinobufacino injection is a significant anti-tumor medicine for the treatment of various tumors in clinic, which was made from water extraction of the skin of Bufo bufo gargarizans. In present paper, HPLC-DAD-FT-ICR-MS method was used to identify the major bufadienolides in cinobufacino for the first time. Solid-phase extraction with dichloromethane and silica was used to enrich the total bufadienolides in cinobufacino. Based on the UV and high resolution MS/MS data, 33 bufadienolides were analyzed and characterized. Among them, eight compounds were identified by comparing with standard references unambiguously. This study elucidated the major bufadienolides in cinobufacino, which provided material foundation of cinobufacino and will be benefit for the further pharmacological research.
9.Mechanism of combined treatment with Shenkangling Decoction and prednisone in rats with adriamycin-induced nephropathy.
Qing LIN ; Jian ZHENG ; Si AI ; Lixing LIN ; Yuxin CHEN ; Fang WANG ; Ling ZHU
Journal of Integrative Medicine 2009;7(7):661-6
To study the mechanism of Shenkangling (SKL), a compound traditional Chinese herbal medicine, combined with prednisone in treating adriamycin-induced nephropathy in rats.
10.99Tcm-MIBI myocardial perfusion imaging for evaluation of the myocardial blood supply in patients with metabolic syndrome
Xiao-shan, GUO ; Zhi-fang, WU ; Jian-zhong, LIU ; Guang, HU ; Jin, WANG ; Si-jin, LI
Chinese Journal of Nuclear Medicine 2011;31(3):174-177
Objective To evaluate the myocardial blood supply in patients with metabolic syndrome (MS) using 99Tcm-MIBI SPECT MPI. Methods A total of 342 patients were divided into four groups according to the number of abnormal metabolic indices: no abnormal metabolic index (Group 1), one abnormal index (Group 2), two abnormal indices (Group 3), three or more abnormal indices (Group 4). Each patient underwent two-day protocol of gated stress and rest 99Tcm-MIBI MPI. One hundred and three of the 342 patients were clinically diagnosed as MS and underwent CAG within 1 month after MPI. χ2test was used to evaluate the difference among the four groups and Kappa test to analyze the correlation between MPI and CAG. Results Compared with CAG, the diagnostic sensitivity, specificity, positive and negative predictive values by 99Tcm-MIBI SPECT MPI for coronary artery diseases (CAD) in 103 MS patients were 80.5% (33/41), 85.5% (53/62), 78.6% (33/42) and 86.9% (53/61), respectively. The correlation coefficient between MPI and CAG was 0.657 (P<0.001). The abnormal MPI rates in group 1, 2, 3 and 4 were 23.3% (10/43), 32.9% (26/79), 54.4% (56/103), and 57.3% (67/117), respectively (χ2=23.22, P<0.001). Conclusions In MS patients,99Tcm-MIBI SPECT MPI can be useful for evaluating myocardial blood supply and the myocardial ischemia rates may correlate positively with the number of abnormal metabolic indices.