OBJECTIVETo study the anticoagulant constituents in dried leech (Whitmania pigra Whitman).

METHODThe plasma recalcification time (PRT) as index, the constituents with anticoagulant activity were isolated and purified by anion-exchange chromatography on Sephadex DEAE A-50, gel permeation chromatography on Sephadex G-25 and Sephadex LH-20 columns, and then reversed phase high-performance liquid chromatography successively.

RESULTThree anticoagulant polypeptides were isolated and purified. Compounds 1 and 2 can be translated each other in natural conditions, and their molecular weights are 7100 and 5531, respectively. Compound 3 was identified as a pure polypeptide by HPLC and SDS-PAGE, and its molecular weight was determined as 8 608 by MALDI-TOF-MS. The amino acid composition of compound 3 was also determined.

CONCLUSIONCompound 3 was inferred to be a novel anticoagulant, and named whitmanin.

Animals ; Anticoagulants ; analysis ; chemistry ; isolation & purification ; Leeches ; chemistry ; Molecular Weight

OBJECTIVETo establish the quality control method of the extract enriched hypericins and flavonoids (HFEE) prepared from Hypericum perforatum by means of HPLC fingerprint analysis, and to evaluate the validity, stability of within-day and between-day of the preparing process of HFEE as well as the influence of antioxidant and the herbal source to the quality of HFEE.

METHODHPLC-UV-MS was employed to the qualitative and quantitative analyses of HFEE.

RESULTTen peaks on the HPLC fingerprint of HFEE were indicated, and eight compounds of them had been identified. With respect of the preparing process of HFEE, the validity and stability were significantly observed. There was almost no effect observed on the quality of HFEE whether or not adding the antioxidant during the process. However, different parts of the plant collected as the materials could significantly affect the quality of HFEE.

CONCLUSIONThe method established in the present study is convenient, reliable, and could be used for the quality control of the extract of H. perforatum and for the monitor and control of the preparing process of the extract.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Hypericum ; chemistry ; Perylene ; analogs & derivatives ; chemistry ; isolation & purification ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Quercetin ; chemistry ; isolation & purification ; Rutin ; chemistry ; isolation & purification ; Technology, Pharmaceutical

AIMTo investigate the penetration kinetics of xanthotoxin in human skin and stratum corneum.

METHODSThe penetration experiments were accomplished by the deposit of ethanolic xanthotoxin solution onto human skin and stratum corneum mounted on Franz cells. The diffused xanthotoxin in the receptor solution (1.4% human serum albumin) and the retained amount in the skin and in the stratum corneum after 24 h exposure were quantified by using high performance liquid chromatography.

RESULTSXanthotoxin flux was increased with the concentration deposited onto the human skin, and when the concentration is above 2.5 mg x mL(-1), there is no influence on the xanthotoxin flux. Similar results were obtained from the stratum corneum. And the peak time for the flux in the stratum corneum was preceded about 6 h earlier than that of the whole human skin. The retained xanthotoxin amount after 24 h exposure in the skin and in the stratum corneum increased according to the concentration deposited and has the tendency to saturate. The lag time of ethanolic xanthotoxin solution in the whole human skin is significantly higher than that in the stratum corneum (P < 0.05).

CONCLUSIONThe characteristics of penetration kinetics of xanthotoxin will provide the information for concentration choice of topical formulation and give a reference for ultra violet A (UVA) irradiation time confirmation.

Administration, Cutaneous ; Dose-Response Relationship, Drug ; Epidermis ; metabolism ; Female ; Humans ; In Vitro Techniques ; Methoxsalen ; administration & dosage ; pharmacokinetics ; Middle Aged ; Photosensitizing Agents ; administration & dosage ; pharmacokinetics ; Skin ; metabolism ; Skin Absorption ; Time Factors

Acute Disease ; Adolescent ; Adult ; Aged ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Pesticides ; poisoning ; Poisoning ; epidemiology

OBJECTIVETo establish an advanced purification techniques of the essential oils obtained from Pogostemon cablin.

METHODMolecular distillation (MD) was applied.

RESULTFour distillates were obtained, chemical constituents of which were analyzed with GC-MS. Compared with those in original oils, the contents of active compounds (patchouli alcohol and pogostone) rose by 27%-47% in the distillates II and III.

CONCLUSIONMolecular distillation (MD) effectively raises the contents of patchouli alcohol and pogostone. The work is of great economic and scientific significance for the industrialization of P. cablin and the discovery of new drugs.

Gas Chromatography-Mass Spectrometry ; Lamiaceae ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Sesquiterpenes ; isolation & purification ; Technology, Pharmaceutical ; methods

OBJECTIVEBao-Xie-Ning (BXN), a traditional Chinese herbal medicine (CHM) formula composed of Fructus Evodiae, Flos Caryophylli and Cortex Cinnamomi, and used for the treatment of infant diarrheal illness, was subject to systematic assessment for its putative multiple pharmacodynamic effects and pharmacological antidiarrheal mechanisms.

METHODSHigh-performance liquid chromatography-diode array detector-electrospray ionization-mass spectrometric/mass spectrometry was developed and validated for identification and quantification of the main constituents in different extracts of BXN. Male Kunming mice weighing 20 to 25 g were used for detecting the antidiarrheal activity of the extracts. Ethanolic extract (EE), volatile oil extract (VOE), and aqueous extract (AE) of BXN were respectively subjected to pharmacodynamic and pharmacological comparison in assessing antidiarrheal effects with senna-induced diarrhea, castor oil-induced diarrhea, acetic acid-induced writhing assay, and isolated duodenum test.

RESULTSThe highest yields of three detected components of BXN, rutaecarpine, eugenol and cinnamaldehyde were observed in EE. EE showed the most remarkable antidiarrheal activity in dose-dependent and time-dependent manners in both senna- and castor oil-induced diarrhea models, and presented dose-dependent analgesic activity in acetic acid-induced algesthesia model. In addition, EE extract of BXN also exhibited strong antimobility action on the intestine and strongest depression on spontaneous contraction of isolated duodenum.

CONCLUSIONEthanol extraction is an efficient method to extract the active constituents of BXN. BXN extract demonstrated multiple pharmacological activities affecting the main mechanisms of diarrhea, which validated BXN's usage in the comprehensive clinical treatment of diarrhea.

Animals ; Antidiarrheals ; administration & dosage ; analysis ; Chromatography, High Pressure Liquid ; Diarrhea ; drug therapy ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; Humans ; Male ; Mice ; Plants, Medicinal ; chemistry