6.Determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials by LC-MS/MS.
Hai-hong SI ; Yan-jing LI ; Jia XUE ; Wen-zhe HUANG ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(14):2832-2836
To develop a LC-MS/MS method for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials, the column was Agilent ZORBAX Eclipse plus C18 (3.0 mm x 50 mm, 1.8 µm), and the mobile phase consisted of methanol-water (containing 0.2% formic acid) (95:5) at a flow rate of 0.5 mL · min(-1). The multiple reaction ion monitoring (MRM) with an ESI interface in the negative ion mode was selected. The results showed that the linear ranges of five kinds of ginkgolic acids were in the range of 0.2-36.0 µg · L(-1) (r ≥ 0.999 5). The lowest limit of quantification (LOQ) of ginkgo acid C13: 0, C15:1, C17:2, C15:0 and C17:1 were 0.18, 0.18, 0.21, 0.10 and 0.20 µg · L(-1), respectively. The average recovery was between 73.28% and 87.56%, and the average content of total ginkgolic acids in three batches of samples was in the range of 0.023-0.028 µg · g(-1), which was much lower than 2 µg · g(-1) prescribed in drug registration standards. This method is simple and rapid with high sensitivity, which can be used for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials.
Chromatography, Liquid
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methods
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Ginkgolides
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analysis
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Injections
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Limit of Detection
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Salicylates
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analysis
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Tandem Mass Spectrometry
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methods
7.Epidemiological analysis of human brucellosis in Ningxia from 2004 to 2009
Xiao-jia, SONG ; Feng, JIN ; Xing-zhong, HU ; Tian-bo, MA ; Hai-hong, WANG ; Li, LI
Chinese Journal of Endemiology 2011;30(1):92-95
Objective To learn the prevalence and trends of Brucella disease in Ningxia, in order to provide scientific basis for effective control of the disease. Methods Data of Brucella cases reported through city network from 2004 to 2009 in Yinchuan city, Shizuishan city, Wuzhong city, Guyuan city and Zhongwei city of Ningxia were reviewed and retrospectively analyzed. The data included demographic characteristics, treatment conditions and medical history so on related information. Analytical indicators include reported incidence;with patients' gender, age, regional distribution, urban and rural distribution;become chronic and associated factors;distribution of the cases reporting unit and so on. Results From 2004 to 2009, Ningxia had reported 349 cases of Brucellosis, no deaths, the annual incidence rates reported were 0.017/10 million, 0.543/10 million, 0.151/101 (295/54);The proportion of 34- to 40-year-old age group was higher than other age groups(27.5%, 96/349);Occupational distribution of patients was mainly farmers and herdsmen(70.2% ,245/349), in regional distribution of the patients, the highest percentage was Wuzhong city(61.9%,216/349), followed by Yinchuan city(22.9%,80/349);The proportion of patients in rural areas(97.4% ,340/349) was higher than urban(2.6% ,9/349);the proportion of patients converted to chronic was 11.2% (39/349). With age, the chance of patients converted to chronic was in a decreasing trend(odds ratio was 0.966);cases reported by Centre for Disease Control and Prevention accounted for 74.8%(261/349), by hospital accounted for 25.2%(88/349). Conclusions The reported incidence of Brucellosis in Ningxia is in a rapid upward trend year by year, the patients is mainly young men, the rate of converted to chronic is higher and the ability of hospital in founding and reporting of the cases is weaker.Comprehensive measures should be taken to increase funding, strengthen monitoring, and continuously improve the level of awareness and diagnosis of medical personnel for further strengthen the prevention and control of Brucellosis.
8.Optimization of processing technology for xanthii fructus by UPLC fingerprint technique and contents of toxicity ingredient.
Yan-Quan HAN ; Yan HONG ; Lun-Zhu XIA ; Jia-Rong GAO ; Yong-Zhong WANG ; Yan-Hua SUN ; Jin-Hai YI
China Journal of Chinese Materia Medica 2014;39(7):1248-1254
The experiment's aim was to optimize the processing technology of Xanthii Fructus which through comparing the difference of UPLC fingerprint and contents of toxicity ingredient in water extract of 16 batches of processed sample. The determination condition of UPLC chromatographic and contents of toxicity ingredient were as follows. UPLC chromatographic: ACQUITY BEH C18 column (2.1 mm x 100 mm, 1.7 microm) eluted with the mobile phases of acetonitrile and 0.1% phosphoric acidwater in gradient mode, the flow rate was 0.25 mL x min(-1) and the detection wavelength was set at 327 nm. Contents of toxicity ingredient: Agilent TC-C18 column (4.6 mm x 250 mm, 5 microm), mobile phase was methanol-0.01 mol x L(-1) sodium dihydrogen phosphate (35: 65), flow rate was 1.0 mL x min(-1), and detection wavelength was 203 nm. The chromatographic fingerprints 16 batches of samples were analyzed in using the similarity evaluation system of chromatographic, fingerprint of traditional Chinese medicine, SPSS16.0 and SIMCA13.0 software, respectively. The similarity degrees of the 16 batches samples were more than 0.97, all the samples were classified into four categories, and the PCA showed that the peak area of chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeic acid were significantly effect index in fingerprint of processed Xanthii Fructus sample. The outcome of determination showed that the toxicity ingredient contents of all samples reduced significantly after processing. This method can be used in optimizing the processing technology of Xanthii Fructus.
Caffeic Acids
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analysis
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toxicity
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Chemistry, Pharmaceutical
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Chromatography, High Pressure Liquid
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methods
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Drugs, Chinese Herbal
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analysis
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toxicity
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Quinic Acid
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analogs & derivatives
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analysis
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toxicity
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Xanthium
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chemistry
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classification
9.Garlicin ameliorated pressure overload induced myocardial fibrosis in rats through partial inhibiting TGF-beta1 mediated Smads signal.
Hai-Xiao ZHANG ; Zai-Xiang SHI ; Hai-Zhong JIA
Chinese Journal of Integrated Traditional and Western Medicine 2012;32(5):666-670
OBJECTIVETo observe whether garlicin could ameliorate pressure overload induced myocardial fibrosis in rats through partial inhibiting transforming growth factor beta1 (TGF-beta1) mediated Smads signal.
METHODSForty male SD rats were randomly divided into 4 groups, i. e., the sham-operation group, the model group, the garlicin group, and the Tetramethylpyrazine (TMP) group, 10 in each group. The pressure overload induced myocardial fibrosis rat model was prepared using coarctation of aorta. Three days after modeling 5.0 mg/kg garlicin injection was administered to rats in the garlicin group, 20 mg/kg TMP injection to rats in the TMP group by peritoneal injection, while normal saline was peritoneally injected to rats in the sham-operation group and the model group. Four weeks after medication, the changes of myocardial collagen were observed by picrosirius red staining. The myocardial collagen volume fraction (CVF) and perivascular collagen areas (PVCA) were calculated. The serum transforming growth factor beta1 (TGF-beta1) expression was detected using ELISA. The TGF-beta1 protein expression in the myocardial tissue was observed using immunohistochemical assay. The changes of myocardial Smad2 and Smad7 mRNA expressions were detected using Real-time RT-PCR. The effects of garlicin on TGF-beta1 mediated Smad Signaling through luciferase assay were further verified using Mv1 Lu-(CAGA) 12-Luc cell line response to TGF-beta1.
RESULTSCompared with the sham-operation group, the myocardial levels of CVF and PVCA, the serum TGF-beta1 level, and the TGF-beta1 protein expression in the myocardial tissue obviously increased in the model group (P < 0.05, P < 0.01). Compared with the model group, the PVCA level, the serum TGF-beta1 level, and the TGF-beta1 protein expression in the myocardial tissue of the garlicin group and the TMP group obviously decreased (P < 0.05, P 0O 01). The Smad2 mRNA expression was up-regulated while Smad7 mRNA expression down-regulated in the model group. The Smad2 mRNA expression was obviously down-regulated in the garlicin group and the TMP group (P < 0.05). The Smad7 mRNA expression was obviously up-regulated in the TMP group (P > 0.05). One to 2 microg/mL garlicin could obviously inhibit the luciferase activities of corresponding TGF-beta1, under the stimulation of 2 ng/mL TGF-beta1 (P < 0.05).
CONCLUSIONGarlicin ameliorated pressure overload induced myocardial fibrosis in rats through partial inhibiting TGF-beta-Smads signal pathway.
Allyl Compounds ; pharmacology ; Animals ; Cardiomyopathies ; etiology ; metabolism ; pathology ; Disulfides ; pharmacology ; Fibrosis ; Male ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Smad Proteins ; metabolism ; Transforming Growth Factor beta1 ; metabolism
10.Effects of garlicin on NIH3T3 cell proliferation and collagen synthesis.
Hai-Xiao ZHANG ; Zai-Xiang SHI ; Hai-Zhong JIA
Chinese Journal of Integrated Traditional and Western Medicine 2007;27(5):431-434
OBJECTIVETo investigate the effects of garlicin on fibroblasts proliferation and type I collagen synthesis and explore its anti-fibrosis mechanism.
METHODSGarlicin was added into the culture fluid of NIH3T3 cell, taking Radix Salviae miltiorrhizae as the control medicine. The spiking of H3-thymidine DNA was detected, also the hydroxyproline (HOP) concentration in the culture fluid by alkali digestion method and the protein expression of type I collagen in NIH3T3 cells by immunofluorescent staining.
RESULTSThe NIH3T3 cell growth and proliferation rate were obviously reduced after garlicin treatment concentration-dependently in range of 0.2 - 5 microg/mL; HOP level and protein expression of type I collagen also lowered.
CONCLUSIONGarlicin could inhibit NIH3T3 cell proliferation, reduce the synthesis and protein expression of type I collagen so as to exert the anti-fibrosis effect.
Allyl Compounds ; pharmacology ; Animals ; Cell Proliferation ; drug effects ; Collagen Type I ; biosynthesis ; Disulfides ; pharmacology ; Dose-Response Relationship, Drug ; Garlic ; chemistry ; Hydroxyproline ; analysis ; Mice ; NIH 3T3 Cells