Chemical Fingerprint Analysis and Ultra-Performance Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry-Based Metabolomics Study of the Protective Effect of Buxue Yimu Granule in Medical-Induced Incomplete Abortion Rats.

Medical abortion is a common method to terminate an early pregnancy and often causes serious complications such as abnormal uterine bleeding and endometritis. Buxue Yimu granule (BYG) is a well-known traditional Chinese medicine prescription composed of five kinds of drugs and is widely used in gynecology and obstetrics. The aim of the present study was to establish the quality standard of BYG and investigate its protective effect on incomplete abortion. The chemical fingerprint of BYG was established by high performance liquid chromatography (HPLC). The major compounds of BYG were determined by ultra-performance liquid chromatography with triple quadrupole mass spectrometry. An incomplete abortion rat model was induced by intragastric administration of mifepristone (8.3 mg·kg-1) combined with misoprostol (100.0 µg·kg-1) during early pregnancy. The serum levels of human chorionic gonadotrophin (HCG), estradiol (E2), and progesterone (PG) were determined. The serum endogenous metabolites were analyzed by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Multivariate analysis, including partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA), was employed to analyze the metabolic profiles, and MetaboAnalyst was used to investigate the metabolic pathways. Furthermore, hematoxylin-eosin staining (HE) was used to evaluate the histopathological changes in uterine tissue. The expression levels of VEGFA and NF-κB were detected by immunohistochemistry. The results indicated that HPLC fingerprint analysis can be successfully used to assess the quality of BYG. The medical-induced incomplete abortion rats were clearly separated from control rats, and the biochemical changes were gradually restored to normal after administration of BYG. Moreover, 19 potential biomarkers, including N-lactoylleucine, 2-piperidinone, isobutyryl-l-carnitine, eicosapentaenoylcholine, LysoPC(14:0), LysoPC(20:5), physagulin C, LysoPC(18:3), leukotriene D5, deoxycholic acid 3-glucuronide, glycine, pregnanediol 3-O-glucuronide, LysoPC(18:2), LysoPC(17:0/0:0), N-acetyl-leukotriene E4, LysoPC(18:0), platelet-activating factor, LysoPA(24:1), and LysoPC(18:1), which were mainly related to the amino acids metabolism, lipids metabolism, and bile acid biosynthesis, were identified. Consequently, BYG exerts a potential protective role in the intervention of incomplete abortion by anti-inflammatory, promote endometrial repair, and regulate the metabolic disorders.

[In vivo studies on metabolic network compatibility of herba houttuyniae injection and sodium neohouttuynin injection in rats].

OBJECTIVE: To study metabolic rule in rats for heartleaf houttuynia herb injection and Sodium New Houttuyfonate injection. METHODS: The GC-MS was used to determine metabolites in plasma and urine of rats after intravenous and oral administration of heartleaf houttuynia herb injection as multi-component and Sodium New Houttuyfonate injection as single component. RESULTS: For rats 8 common metabolites were determined in plasma after intravenous administration of heartleaf houttuynia herb injection and Sodium New Houttuyfonate injection, meanwhile 39 in urine. The 16 common metabolites were determined in plasma after oral administration, and 12 in urine. The 9 and 11 of same metabolites were determined in plasma and urine respectively after intravenous and oral administration of heartleaf houttuynia herb injection, 8 and 17 common metabolites for Sodium New Houttuyfonate injection. CONCLUSION: The metabolites of heartleaf houttuynia herb and Sodium New Houttuyfonate injection in rats are the same or similar after intravenous and oral administration, and there are the same structure types between metabolites and original constituents of preparations. The metabolic pathways of the two preparations in rats are the same. There is metabolic Network Compatibility in organism between multi-components, single components and different administration forms for traditional Chinese medicine.

[Theoretical and experimental studies on solubility parameters of multiple components for traditional Chinese materia medica with HPLC].

OBJECTIVE: To establish the theories and methods to determine solubility parameters of multiple components for the traditional Chinese material medica (TCMM) with HPLC. METHOD: The mathematical expresses to determin the solubility parameters were established according to chromatographic and Hildebrand-Scatchard theories, The HPLC experiments were carried out at 40 degrees C on an Alltech Apollo C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetone and water in gradient mode. The flow rate was 1.0 mL min(-1), and the detection wavelength was 254 nm. The injection volume was 1 0 microL. RESULT: The mathematical expresses between the retention time and the solubility parameters were established and used to determin caffeine solubility parameter as 28.31 J(1/2) cm(-3/2) in accordance with 28.84 J(1/2) cm(-3/2) reported by literature, and those of aloe-emodin, rhein, emodin, physcione as 39.70 J(1/2) cm(-3/2), 39.08 J(1/2) cm(-3/2), 38.37 J(1/2) cm(-3/2), 36.42 J(1/2) cm(-3/2) respectively. CONCLUSION: The retention time of HPLC can be used to determine the solubility parameters of multiple componets. The established method is useful for the compatibility rule study of traditional Chinese medicine.

[Fundamentally study on mathematical kinetic model of component extraction from FTCM].

OBJECTIVE: To establish the mathematical kinetic model of the components extracted from the FTMC (formulae of the traditional Chinese medicine) and analyze parameters of the astragaloside IV extracted from the BYHWD (Buyang Huanwu decoction). METHOD: The model, including algebra and differential groups, have been set up according to the FICK discipline and Noyes-whitney soluted theories, as well as two transfer diffusive processes ((1) from protoplasate to apoplasmic, also from material compartment interior cell membrane to outside compartment; (2) apoplasmic to solution, also from outside compartment to solvent compartment) on components extraction from the FTMC. The equation groups, according to laplace transform, have been given a expression as solutions, which indicate the quantitative changes of the component concentration in solvent vs. time. The model kinetic parameters have been analyzed, meanwhile the parameters of the astragaloside IV in the BYHWD under 100 degrees C, extracted by water, have been analyzed by way of this model: RESULT: It has been established a mathematical model that consists of three parts of e exponent. The kinetic parameters: M, alpha, N, beta, L, pi, K, k1', k2', rho1, rho2, tmax, Cmax, AUC, w0, P, D of the BYHWD were respectivelly 0.061 27% , 0.280 2 min(-1), - 1.027% , 0.008 965 min(-1), 1.077%, 0.002 665 min(-1), 3.451 x 10(-3) min(-1), 3.188 x 10(-3) min(-1), 0.375 9 min(-1), 1.420 min, 0.754 7 min, 184.9 min, 0. 0572 1 mg x mL(-1), 289.9 min, 0.070 11%, 46.24%, 22. 35%. CONCLUSION: The kinetic model, applied to isolated system, can have been of the rule of multiplex linear. Each parameters can be analyzed completely.

[Fingerprints of different samples of processed Rhizoma Coptidi by HPLC].

OBJECTIVE: To set up the HPLC fingerprints of different samples of processed Rhizoma Coptidi (RC) in order to offer evidence for identifying different processed RC and preliminary discussion on processing mechanism. METHOD: Seven different samples of processed RC were determined by HPLC and the results were analyzed by Hierarchical clustering and similarity evaluation system for chromatographic fingerprint of TCM. RESULT: The characteristic of common peaks for fingerprints of different samples of processed RC is distinct, but there are differences among various processed RC. The different samples of processed RC were classified into two and the result of hierarchical clustering is correlated with the property of traditional Chinese drug. CONCLUSION: It is proved that the method which is convenient, rapid and repeat well can be used to identify different samples of processed RC. The change of property of different processed RC is related with processing adjuvant, the possible mechanism is that new components are produced or the contents of known components have changed due to the adjuvant during processing.

Cytotoxic withanolides from Physalis angulata L.

Four new withanolides, physagulins L-O (1-4), were isolated from the MeOH extract of the aerial parts of Physalis angulata L. (Solanaceae), together with seven known withanolides, compounds 5-11. Their structures were determined by spectroscopic techniques, including 1H-, 13C-NMR (DEPT), and 2D-NMR (HMBC, HMQC, 1H,1H-COSY, NOESY) experiments, as well as by HR-MS. All eleven compounds were tested for their antiproliferative activities towards human colorectal-carcinoma (HCT-116) and human non-small-cell lung-cancer (NCI-H460) cells. Compound 5 exhibited the highest anticancer activity against the HCT-116 cell line, with an IC50 value of 1.64+/-0.06 microM. Compound 9 exhibited the highest cytotoxicity towards the NCI-H460 cell line, with an IC50 value of 0.43+/-0.02 microM.

[Optimization preparation of chansu-loaded solid lipid nanoparticles by central composite design and response surface method].

OBJECTIVE: To optimize the formulation of chansu-loaded solid lipid nanoparticles (Cs-SLN). METHOD: Cs-SLN was prepared by cold homogenization technique. The effects of influence factors such as the weight of compritol 888 ATO, soybean lecithin and poloxamer 188, on mean diameter, entrapment efficiency, drug loading and overall desirability were investigated by using central composite design and response surface method. The data were imitated using multi-linear equation and second-order polynomial equation. RESULT: The latter was prior to the former considering from multiple correlation coefficients. Under the optimal conditions, the mean diameter, entrapment efficiency, drug loading of the Cs-SLN were 71.5 nm, 92.45% and 5.26%. CONCLUSION: The optimized preparation technique for Cs-SLN is stable, feasible and high inclusion rate. It can be used for production of Cs-SLN.