ObjectiveThis study aims to systematically analyze the blood-absorbed components and metabolic profiles of Xiebaisan（XBS） in normal and chronic bronchitis （CB） mice using ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS）， while comparing differences between the two states. MethodsThirty female BABL/c mice were randomly divided into the normal group， the normal drug administration group， the CB group， the CB drug administration group and the dexamethasone group， with 6 mice in each group. The CB mouse model was established by inducing with ovalbumin （OVA）. The mice in the normal drug administration group and the CB drug administration group started to be gavaged with XBS（13.2 g·kg^-1） from the 21^st day， and the dexamethasone group mice were simultaneously gavaged with dexamethasone （0.5 mg·kg^-1） until the end of the 35^th day of the experiment. Subsequently， serum samples were collected and evaluated for their efficacy， based on the pharmacological evaluation indicators， to determine the efficacy of XBS in treating CB. Then the UPLC-Q-Exactive Orbitrap MS was employed to identify and analyze the chemical constituents， blood-absorbed components， and metabolites of XBS. Chemometric analysis was conducted to reveal metabolic profile differences under "dual states". Concurrently， Real-time PCR technology was utilized to detect the expression levels of key liver metabolic enzymes CYP2E1， CYP3A1， UGT1A1， and UGT1A6. ResultsA total of 28 prototype components and 158 metabolites （including 48 phase Ⅰ metabolites and 110 phase Ⅱ metabolites） of XBS were unambiguously identified in the serum of normal mice. Additionally， a comprehensive characterization was performed on a total of 32 prototype components and 178 metabolites （including 50 phase Ⅰ metabolites and 128 phase Ⅱ metabolites） of XBS in the serum of CB mice. Among them， 27 prototype components were detected in both states， including 12 flavonoids， 2 alkaloids， 3 triterpenes， 4 organic acids， 3 amides， 1 stilbene and 2 other compounds. The chemometrics analysis revealed no significant difference in the prototype components and metabolites of XBS between normal and CB mice； however， there was a significant increase in the in-vivo exposure of XBS in CB mice. Compared to normal mice， the levels of phase Ⅰ metabolites such as oxidation， reduction and methylation of blood components of XBS as well as phase Ⅱ metabolites of glucuronidation showed significant changes in CB mice. Real-time PCR further confirmed that these alterations were attributed to the upregulation of CYP2E1 （P<0.05）， CYP3A1 （P>0.05）， UGT1A1 （P<0.01） and UGT1A6 （P<0.01） enzymes expression in the liver of CB mice. ConclusionThis study elucidated the disparities in the levels of the blood-absorbed components and metabolic profiles of XBS in normal and CB mice， especially in oxidation， reduction， methylation in phase Ⅰ metabolism and glucoaldehyde acidification in phase Ⅱ metabolism. And there are related to the differences in the expression levels of phase Ⅰ and phase Ⅱ metabolic enzymes CYP2E1， CYP3A1， UGT1A1 and UGT1A6 in the liver.

Editor's note: The research on effective constituents of traditional Chinese medicine is one of the crucial areas of basic research of traditional Chinese medicine. It is also a research hotspot which gains widespread attention and active application of projects of national natural science funds in the discipline of traditional Chinese medicine. In 2013, the National Natural Science Foundation of Chinese traditional medicine in the application code H2803 in the research field of effective constituents of traditional Chinese medicine , the direction of gener-al project , Youth Science Foundation and the regional Science Foundation project application 449 , which accept-ed 425 . In this paper , the general situation of the application and approved projects of national natural science funds in the research field of effective constituents of traditional Chinese medicine in 2013 has been introduced. The research strategies and plans of the approved projects have been summarized , and the problems of the ap-plications have been also analyzed .

A HPLC-ELSD method was developed and validated for simultaneous determination of five Hetisane-type diterpenoid alkaloids in a Tibetan traditional herbal medicine, “Gebu Dilu” (Herba Delphinii), using a Kromasil C18 column (250 mm ? 4.6 mm, 5μm) with the mobile phase consisting of acetonitrile and 0.1% triethylamine in gradient (detected by evaporative light scattering detector). The linear ranges of five compounds were determined and method validation was evaluated completely. The established method is rapid and accurate with high repeatability, and can be applied for the quality control of Herba Delphinii.

OBJECTIVETo study the conjugation reaction characteristics of caffeic acid micromolecule cistanoside F and bovine serum albumin.

METHODThe interaction between bovine serum albumin (BSA) and cistanoside F that was separated from Callicarpa plant for the first time and abbreviated CF was detected by fluorescence (FS), UV-vis absorbance and circular dichroism (CD) under simulative physiological conditions.

RESULTCF-BSA's static apparent binding constant (K(a)), number of binding sites (n), efficiency of energy transfer (E), spatial distance (r), thermodynamic parameters deltaG, deltaH and deltaS and changes in alpha-helical structure content in BSA before and after CF's effect were calculated to define the binding site of CF in BSA and analyze the impact of several common metal ions on the interaction of CF and BSA.

CONCLUSIONGround state compounds formed by CF and BSA could cause intrinsic fluorescence quenching. Their binding constant K(a) of cistanoside F with BSA was 4.36 x 10(4) L x mol at 25 degrees C, the number of binding site n was 1, and the spatial distance r was 3.09 nm. The results indicated that the hydrogen bond played a major role in cistanoside F-BSA association. The displacement experiments confirmed that cistanoside F can bind to site I of BSA. In addition, the binding constant of cistanoside F with BSA was enhanced after the addition of some common metal ions Mg2+, Fe3+, Cu2+ and Zn2+. The intrinsic fluorescence of BSA was quenched by cistanoside F via forming cistanoside F-BSA complex and non-radiation energy transfer. CD spectra showed that the binding of cistanoside F with BSA induced conformational changes in BSA.

Animals ; Caffeic Acids ; chemistry ; Catechols ; chemistry ; Cattle ; Circular Dichroism ; Glycosides ; chemistry ; Serum Albumin, Bovine ; chemistry ; Spectrometry, Fluorescence ; Spectrophotometry, Ultraviolet ; Thermodynamics

A new triterpenoid saponin and fourteen known triterpenoids were isolated from the methanol extract of the stems and leaves of Callicarpa integerrima Champ, which is used in Chinese folk medicine for stopping bleeding, expelling the wind, dissipating stagnation, and treating scrofula, by using various chromatographies, such as silica gel, Sephadex LH-20 and RP-C18 column chromatography. Their structures were identified as a new compound 2alpha, 3beta, 19alpha, 23-tetrahydroxy-olean-12-en-28-oic acid-28-O-beta-D-glucopyranosyl-(1 --> 4)-beta-D-glucopyranoside (1), together with fourteen known compounds: oleanolic acid (2), 3-acetyl oleanolic acid (3), 3beta-O-acetyl ursolic acid (4), 2alpha-hydroxy-ursolic acid (5), 2alpha, 3beta, 19alpha, 23-tetrahydroxy-urs-12-en-28-oic acid (6), alpha-amyrin-3-O-beta-D-glucopyranoside (7), pomolic acid (8), betulinic acid (9), ursolic acid (10), 2alpha, 3beta, 19alpha, 23-tetrahydroxy-olean-12-en-28-oic acid (arjungenin) (11), 2alpha-hydroxy-oleanolic acid (12), hederagenin (13), 2alpha, 19alpha-dihydroxy-ursolic acid (14) and pruvuloside A (15), by the spectroscopic techniques of NMR, HMBC, IR and MS, separately. All these compounds were obtained from this plant for the first time, and compounds 3, 4 and 15 were isolated from genus Callicarpa L. for the first time.

To study the chemical constituents of Picrasma quassioides. The chemical constituents were isolated and purified by chromatographic methods over Sephadex LH-20 and silica gel column, and structurally elucidated by spectral analysis, including UV, IR, MS, 1H-NMR, 13C-NMR. Fourteen compounds were obtained and identified as trifolirhizin(1), maackiain(2), 3', 7-dihydroxy-4'-methoxylisoflavone(3), umbelliferone(4), emodin(5), nigakilactone F(6), picrasin B(7),picraqualide B (8),4-methoxy-5-hydroxycanthin-6-one(9), 4,5-dimethoxycanthin-6-one (10),5-methoxycanthin-6-one(11), 11-hydroxycanthin-6-one(12) , 1-methoxycarbonyl-beta-carboline(13), 1-hydroxymethyl-beta-carboline(14). Compounds 1-5 are reported from the first time for the genus Pricrasma.
Organic Chemicals ; analysis ; isolation & purification ; Picrasma ; chemistry ; Spectrum Analysis

The study is to develop an HPLC method for simultaneous determination of rhamnazin (1), rhamnocitrin (2), rhamnetin (3), rhamnazin-3-O-beta-D-glucopyranoside (4), rhamnazin-3-O-beta-D-xylopyranosyl-(1-->4)-beta-D-glucopyranoside (5), rhamnazin-3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranoside (6), and rhamnocitrin-3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranoside (7) in Nervilia fordii. The separation was performed on a Kromasil C18 column (250 mm x 4.6 mm, 5 microm) with 0.4% phosphoric acid-acetonitrile as the mobile phase in a gradient elution at a flow rate of 1.0 mL x min(-1). The detect wavelength was set at 256 nm, and the column temperature was set at 40 degrees C. There were good linear relationships between the logarithm values of concentrations and those of the peak areas of seven flavonoids (1-7) in the range of 0.55-70.00 microg x mL(-1) (r = 0.9997), 0.86-110.00 microg x mL(-1) (r = 0.9997), 0.39-50.00 microg x mL(-1) (r = 0.999 7), 0.55-70.00 microg x mL(-1) (r = 0.999 5), 1.33-170.00 microg x mL(-1) (r = 0.9998), 1.33-170.00 microg x mL(-1) (r = 0.9998), 0.16-20.00 microg x mL(-1) (r = 0.9995), respectively. The recoveries of the seven flavonoids were between 97.19%-99.45%, the relative standard deviations (RSDs) were between 0.91%-2.69%. The established method is rapid, accurate with high repeatability, which could provide scientific evidence for the quality control of Nervilia fordii.

Objective To establish a new method for the identification of Cortex Phellodendri. Methods A HPLC fingerprint at 220 nm has been established for identification of Cortex Phellodendri, using a Merck-Lichrospher RP-C_(18) column(4.6?250mm, 5?m) and ACN(A) with the buffer of 0.3% H_3PO_4-NH(CH_2CH_3)_2 (B) in gradient as the mobile phase. The results were compared with the chromatograms of three species of Rbizoma Coptidis under the same conditions. Results HPLC fingerprint of Cortex Phellodendri at 220nm consists of 14 specific peaks. The steady appearance of the peaks and their relative contents were considered as important signs for the identification of this crude drug. The chromatograms of Rhizoma Coptidis were obviously different from the fingerprint of Cortex Phellodendri. Conclusion This method has been proven to be feasible for the identification of Cortex Phellodendri.

Objective To study the protective effects of aqueous extract and alcohol-soluble extract of Isodon lophanthoides var.gerardianus (Benth.)Hara (ILVG) on immunity hepatic injury induced by concanavalin A (Con A) in mice.Methods One hundred and eight NIH mice were allocated into normal control group,model group,ILVG groups of aqueous extract and alcohol-soluble extract (low-,middle-and high-dosage),and bifendate group randomly.In the experimental groups,mice received either ILVG aqueous extract or alcohol-soluble extract (18.20 g?kg-1,9.10 g?kg-1,4.55 g?kg-1 respectively) or Bifendate (45 mg?kg-1) by gastric perfusion daily for consecutive 5 days.In the 5th day,Con A (20 mg?kg-1) was injected into mice via the tail vein 4h after administration.And then the blood was obtained by picking out the eyeball and the serum was separated after 8-hour fasting.The serum levels of ALT and AST were analyzed,the body weight as well as the weight of liver,spleen and thymus were measured,and pathological features of hepatic tissue were observed.Results ILVG can decrease the ALT and AST,restrain the enlargement of liver and the shrinkage of thymus and reduce the necrosis of hepatic tissue.Conclusion Aqueous extract and alcohol-soluble extract of ILVG possess the effects of protecting liver from immunity injury induced by Con A in NIH mice.

Objective To establish a HPLC method for the assay of psoralen and isopsoralen in different effective extracts of Fructus Psoraleae. Methods HPLC was carried out on the column of Kromasil RP-C18. The mobile phase was methanol -water(65 ∶35). The flow rate was 1.0 mL/min and the UV detection wavelength was 245 nm. Results Good linearity of psoralen was showed within the range of 10.5 ng～525 ng(r= 0.999 3)and isopsoralen within the range of 9 ng～450 ng (r= 0. 999 9). The content of psoralen and isopsoralen differed in different extractions of Fructus Psoraleae. Among them,the extract C (extracted by ethyl acetate ) contained the highest contents of psoralen and isopsoralen,while the contents of psoralen and isopsoralen were very low in the extract D (extracted by n-butyl alcohol) and E (supernatant of water extract). Conclusion The method is simple,accurate and reproducible. The anti-asthma effect and the dose-effect relationship of the different effective extracts of Fructus Psoralea need further pharmacodynamics study.