Objective To optimize ethanol extraction technologyDingtongning Granule.Methods With yield of extract and content of tanshinoneⅡA as comprehensive evaluation indexes, the concentration of ethanol, the amount of ethanol, soaking time, extraction time and extraction times were selected as observing factors. Orthogonal design test L18(37) was used to optimize ethanol extraction technology ofDingtongning Granule.Results Optimum extraction conditions were as follows: soaking medicine for 0 hour; 2 times with 6 times of the amount of 80% ethanolre; flux extracted 3 hours each time.Conclusion This optimal extraction technology is stable and reliable, which can provide basis for industrial production ofDingtongning Granule.

OBJECTIVETo establish an HPLC method for simultaneous determination of nuzhenide, specnuezhenide, wedelolactone and oleanic acid in Erzhiwan.

METHODThe DIKMA C18 (4.6 mm x 200 mm, 5 microm) column was adopted with acetonitrile and 0.1% phosphoric acid solution as the mobile phase and gradient elution. The flow rate was 1.0 mL x min(-1) and the volume of injection was 20 microL. The column temperature was maintained at 30 degrees C and the detective wavelength was set at 215 nm.

RESULTThere were good linear relationships between the peak area and concentration at the range of 2.008-80.32 (r = 0.999 6), 5.872-234.88 (r = 0.999 7) , 0.9-36 (r = 0.999 9), 13.24-529.6 mg x L(-1) (r = 0.999 6) for nuzhenide, specnuezhenide, wedelolactone and oleanic acid, respectively. The average recovery rates of nuzhenide, specnuezhenide, wedelolactone and oleanic acid were 99.25%, 98.70%, 96.23% and 101.5%, respectively, with RSD of less than 3%.

CONCLUSIONThe established method was so easy, rapid and accurate that it can be used as an effective way for quality control of Erzhiwan.

Chromatography, High Pressure Liquid ; methods ; Coumarins ; analysis ; Drugs, Chinese Herbal ; analysis ; Glucosides ; analysis ; Pyrans ; analysis

OBJECTIVETo study the absorption kinetic characteristics of mollugin and purpurin in each intestinal segment of rats.

METHODThe in situ single-way perfusion rat model was established to study absorption characteristics of mollugin and purpurin in each intestinal segment of rats. The volume of recirculation fluid was regulated by phenol red.

RESULTDifferent quality concentrations (12.33, 24.66, 49.32 mg x L(-1)) of mollugin and (8.455, 16.91, 33.82 mg x L(-1)) purpurin showed a concentration gradient of absorption dose in each intestinal segment, with the osmotic coefficient increasing to more than 0.2 x 10(-4) cm x s(-1). In the same concentration, mollugin and purpurin showed an identical trend of P(eff) in each intestinal segment in the order of colon > duodenum > ileum > jejunum, with a significant difference (P < 0.05).

CONCLUSIONMollugin and purpurin are highly permeable in rat intestinal segments, with absorption in each segment, while the specific absorption existed in the colon segment.

Animals ; Anthraquinones ; metabolism ; Female ; Intestinal Absorption ; Male ; Pyrans ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results

OBJECTIVETo compare the toxicity on normal liver cells LO2 before and after Kansui Radix stir-baked with vinegar, and make a preliminary study on the mechanism of detoxication of Kansui Radix stir-baked with vinegar.

METHODThe MTT method was adopted to detect the cell activity, with normal liver cells LO2 as the study object. The morphology of cells were observed, and the level or content of AST, ALT, LDH, SOD, Na+-K+-ATPase, Ca2+-Mg2+ -ATPase, GSH and MDA were determined in cell culture supernatant and splitting supernatant.

RESULTCompared with the control group, Kansui can obviously inhibit the cell activity (P < 0.01) and morphology, and increase the levels of ALT, AST, and LDH (P < 0.01) in the supernatant fluid of cell incubation, and decrease the level of SOD and the content of GSH (P < 0.01). Besides, it significantly increased the content of MDA (P < 0.01) and significantly decreased the level of Na+-K+-ATPase and Ca2+-Mg2+ -ATPase (P < 0.01) in the supernatant fluid of cell dissociation. Compared with Kansui group of various doses, Kansui Radix stir-baked with vinegar can significantly decrease the cell proliferation inhibition and the trend of morphological variation, and obviously decrease the levels of ALT, AST, and LDH (P < 0.01) in the supernatant fluid of cell incubation, and significantly increase the level of SOD and the content of GSH (P < 0.01), and significantly decrease the content of MDA (P < 0.01). Additionally, it significantly increased the level of Na+-K+-ATPase and Ca2+-Mg2+ ATPase (P < 0.01) in the supernatant fluid of cell dissociation, and showed a certain dose-effect relationship.

CONCLUSIONStir-baking with rice vinegar can release the hepatotoxicity of Kansui Radix. Its possible mechanism was that Kansui Radix stir-baked with vinegar can decrease the influence of Kansui Radix on the permeability of liver cells LO2 membrane and oxidative damage, in order to provide basis for further exploration of the detoxication mechanism of Kansui Radix stir-baked with vinegar.

Acetic Acid ; chemistry ; Cell Line ; Cell Proliferation ; drug effects ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Euphorbia ; chemistry ; Humans ; Liver ; cytology ; Oxidative Stress ; drug effects ; Plant Roots ; chemistry ; Sodium-Potassium-Exchanging ATPase ; metabolism

OBJECTIVETo study the active ingredients in liver protection from Erzhi Wan (AIEP) on acute hepatic injury induced by carbon tetrachloride (CCl4) in mice.

METHODSixty Kunming mice were randomly divided into six groups: the normal group, the model group, bifendate group (150 mg x kg(-1)), high AIEP group (19.8 g x kg(-1)), middle AIEP group (13.2 g x kg(-1)) and low AIEP group (6.6 g x kg(-1)). The treatment groups were orally administered once per day for 7 d separately, whereas the normal and model groups were orally administered with saline. Except normal rats, all the other rats were injected intraperitoneally CCl4 20 mL x kg(-1) once. The rats were sacrificed 16 h after CCl4 administration. Serum and liver samples were collected for analysis. The acute hepatic injury model was prepared by CCl4 injected intraperitoneally. Then, the therapeutic effects of AIEP on the model were evaluated by the activity determination of serum alanine aminotransferase and aspirate aminotransferase (ALT and AST), superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in liver,and the hepatic pathohistological changes following the treatment.

RESULTThe activities of ALT and AST and the MDA content in liver was significantly increased and the activity of SOD was largely inhibited in the animals of modeling group. Following the treatment with AIEP, ALT and AST activities and MDA content were significantly reduced and SOD activity was obviously increased in the mice of treatment group. Furthermore, AIEP could ameliorate the hepatic pathological changes.

CONCLUSIONAIEP have protective effects on acute hepatic injury induced by CCL4 in mice, and are the effect of the liver protecting active sites.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; physiology ; Carbon Tetrachloride Poisoning ; drug therapy ; Chemical and Drug Induced Liver Injury ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Liver ; drug effects ; injuries ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mice

OBJECTIVETo make a qualitative analysis on chemical components in Ligustrum lucidum by UPLC-ESI-Q-TOF-MS.

METHODACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 microm) column was adopted, with methyl cyanides-0.1% formic acid as the mobile phase for gradient elution; ESI ion source was used for mass spectra, and data were collected in positive and negative mode.

RESULTFourteen compounds of L. lucidum were identified by analyzing positive and negative ion mass spectra information and element composition and comparing controls with data from relevant literature.

CONCLUSIONAfter the separation by ultra high performance liquid chromatography, relative molecular mass was determined by mass spectra, and chemical compounds in L. lucidum were determined by information search through relevant literature data, in order to provide powerful therapeutic material basis for L. lucidum.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ligustrum ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; methods

This study is to investigate the effects of phenytoin sodium, lidocaine (sodium channel blockers), propranolol (beta-adrenergic receptor antagonist), amiodarone (drugs prolonging the action potential duration) and verapamil (calcium channel blockers) on arrhythmia of mice induced by Bufonis Venenum (Chansu) and isolated mouse hearts lethal dose of Chansu. Arrhythmia of mice were induced by Chansu and then electrocardiograms (ECGs) were recorded. The changes of P-R interval, QRS complex, Q-T interval, T wave amplitude, heart rate (HR) were observed. Moreover, arrhythmia rate, survival rate and arrhythmia score were counted. Isolated mouse hearts were prefused, and the lethal dose of Chansu was recorded. Compared with control group, after pretreatment with phenytoin sodium, broadening of QRS complex and HR were inhibited, and the incidence of ventricular arrhythmia was reduced dramatically, while survival rate was improved; the isolated mouse hearts lethal dose of Chansu was increased significantly. After pretreatment with lidocaine, the prolongation of P-R interval and broadening of QRS complex were inhibited, and the incidences of ventricular arrhythmia were reduced dramatically, while survival rate was improved; the isolated mouse hearts lethal dose of Chansu was increased significantly. After pretreatment with propranolol, prolongation of P-R interval, broadening of QRS complex, prolongation of Q-T interval and HR were inhibited, and the incidences of both supraventricular and ventricular arrhythmias were reduced dramatically, while survival rate was improved. After pretreatment with amiodarone, HR was inhibited, the incidences of ventricular tachycardia were reduced dramatically. Lastly, after pretreatment with verapamil, the prolongation of P-R interval and Q-T interval were inhibited and the incidences of both supraventricular and ventricular arrhythmias were reduced dramatically; the isolated mouse hearts lethal dose of Chansu was reduced significantly. In in vivo experiments, phenytoin sodium was most effective against the mice arrhythmias induced by Chansu while cautious use of verapamil for Chansu inducing arrhythmia should be noted. It is also concluded that mice ventricular arrhythmias induced by Chansu might be most closely related to sodium channel, supraventricular arrhythmias might be related to beta-adrenergic receptor, and calcium channel plays an important role in conduction block. In in vitro experiments, phenytoin sodium was most effective, followed by lidocaine and propranolol, and amiodarone had no obvious effect and verapamil reduced the lethal dose of Chansu.

OBJECTIVETo study the bioaffinity between 8 bufadienolides(Bu) and tumor cells and analyze the correlation between the bioaffinity and the anti-tumor activities of Bu.

METHODMix and cultivate the chloroform extract of Chansu and MGC-803. Measure the content of 8 Bu in supernatant and cells using HPLC and calculate their affinity rate.

RESULTThe coefficient correlation between the decrease of Bu in cell supernatant after affinity and its MGC-803 restrictive activities, and between the cotent percentage of the free Bu in free cells with its MGC-803 restrictive activities, and between the difference between the decrease and the percentage and its MGC-803 restrictive activities is r = 0.82 (P < 0.05), r = -0.04 and r = 0.83 (P < 0.05) respectively.

CONCLUSIONEight Bu have different levels of affinity with MGC-803 which correlate with their anti-tumor activities.

Amphibian Venoms ; chemistry ; Animals ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Anura ; Bufanolides ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Chromatography, High Pressure Liquid ; methods ; Neoplasms ; drug therapy

OBJECTIVETo investigate the flavonoids in Platycladi Cacumen.

METHODThe constituents in Platycladi Cacumen were determined by UPLC-MS. A Waters BEH C18 column (2.1 mm x 150 mm, 1.7 microm) was used with a gradient elution of methanol-water containing 0.2% formic acid. The mass spectrometer equipped with electrospay ionization source was used as defector and operated in data was collected under the negative ion modes.

RESULTEleven constituents were identified.

CONCLUSIONIn this study, the main flavonoids in Platycladi Cacumen were separated by UPLC, and identified through the information of mass number and UV spectra. With the information of MS/MS from flavonoids, the modes of breaking up flavonoids were discussed. It is an accurate and effective method which can be applied for the constituent identification of Platycladi Cacumen.

Chromatography, Liquid ; methods ; Cupressaceae ; chemistry ; Flavonoids ; chemistry ; Mass Spectrometry ; methods ; Plant Extracts ; chemistry

OBJECTIVETo analyze and identify the chemical ingredients in Sini San by LC-MS.

METHODThese compounds were analyzed with an ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 microm) by gradient elution of methyl cyanides and 0.1% formic acid. ESI source was used and data were collected in positive and negative mode.

RESULTSeventeen compounds of Sini San were identified by direct comparison in both positive and negative modes of Q-TOF-MS, the elemental compositions analysis, the standard substance and the data of the literature.

CONCLUSIONIt is a rapid and accurate method that the composition of compound prescription of traditional Chinese medicine can be identified in terms of the separation of ultra high performance liquid chromatography, the accurate molecurlar weights measured by MS and other information, which can trace to the source of compounds in Sinisan.

Chromatography, Liquid ; methods ; Medicine, Chinese Traditional ; Spectrometry, Mass, Electrospray Ionization ; methods