OBJECTIVE: To establish a liquid chromatography-tandem mass chromatography (LC-MS/MS) method for the simultaneous screening for 22 poisonous alkaloids in blood. METHODS: This method involves a liquid-liquid extraction (LLE) followed by liquid chromatography-tandem mass spectrometry with multi-ple-reaction monitoring (MRM). After blood was extracted with buprenorphine as the internal standard, the target compounds were analyzed with LC-MS/MS-ESI in the positive ionization mode. RESULTS: Identification was based on the compound's retention time and two precursor-to-product ion transitions. The limits of detection ranged from 0.1 ng/mL to 20 ng/mL in blood. CONCLUSION The method was sufficiently selective and sensitive to detect poisonous alkaloids and can be applied in forensic and clinical toxicology.
Aconitine/blood* ; Alkaloids/chemistry* ; Chromatography, Liquid/methods* ; Colchicine/blood* ; Forensic Medicine ; Humans ; Sensitivity and Specificity ; Strychnine/blood* ; Tandem Mass Spectrometry/methods*

Effects of beta-adrenergic blockers and related agents were investigated on experimental convulsions of chicks induced with strychnine, pentylenetetrazol or electroshock and on thiopental sleeping time of rabbits. Convulsions of chicks due to strychnine were significantly inhibited by all beta-adrenergic blockers except dichloroisopreterenol. Propranolol inhibited electroshock convulsion as well, but none of the blockers inhibited pentylenetetrazol convulsion. Furthermore, the mortality of chicks due to large dose of pentylenetetrazol was greatly increased by treatment of beta-adrenergic blockers. Pindolol alone showed diazepam-like anticonvulsive effect against low doses of pentylenetetrazol. Pretreatment with beta-adrenergic blockers caused a marked increase in thiopental sleeping time in rabbits. Prolongation of thiopental sleep due to propranolol was abolished by premedication of animals with reserpine or tranylcypromine. Thiopental sleeping time was prolonged by Zizyphus extract, though less effective than beta-adrenergic blockers. It is felt that the anticonvulsive or sleep enhancing effect of beta-adrenergic blocking agents has an intimate relationship with endogenous adrenergic amines and the receptors.
Adrenergic beta-Antagonists/pharmacology* ; Anesthesia ; Animal ; Anticonvulsants* ; Blood Pressure/drug effects ; Chickens ; Convulsions/chemically induced ; Heart Rate/drug effects ; Male ; Propranolol/pharmacology* ; Rabbits ; Strychnine/antagonists & inhibitors ; Thiopental

The HPLC method for determining plasma concentration of brucine was optimized during the study on the effect of the extraction reagent, the extraction frequency and the volume of extraction solvent on the extraction recovery of brucine. The optimum sample treatment method was obtained in the study. Specifically, ammonia water was added, 4 mL extraction solvent (N-hexane-methylene chloride-isopropyl alcohol 65:30:5) were adopted to extract brucine for twice. The method to determine plasma concentration of brucine was applied in pharmacokinetic study to compare pharmacokinetic properties of intravenous injection (5 mg x kg(-1)) and transdermal administration (40 mg x kg(-1)) of brucine aqueous alkali. The results showed that both pharmacokinetic parameters of brucine after intravenous injection and transdermal administration were in conformity with the two-compartment model. After transdermal administration, the absolute bioavailability was calculated to be 18.72%. The optimized HPLC method can satisfy the demands of the pharmacokinetic study on brucine.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Strychnine ; administration & dosage ; analogs & derivatives ; blood ; pharmacokinetics

OBJECTIVETo study the effects of brucine on vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) in a nude mouse model of bone metastasis due to breast cancer, and to assess the possible antitumor mechanism of brucine.

METHODSA syringe needle was used to directly inject 0.2 mL monoplast suspension (with 2×10(5) human breast cancer cells contained) into the bony femoral cortex of the right hind leg for modeling. Twenty-five nude mice were randomized into five groups and administered with an intraperitoneal injection of saline or drug for 8 consecutive days: model group (0.2 mL normal saline), low-dose brucine group (1.73 mg·kg(-1)), medium-dose brucine group (3.45 mg·kg(-1)), high-dose brucine group (6.90 mg·kg(-1)), and thalidomide group (200 mg·kg(-1)). Diet and activity were recorded, and the tumors were harvested 5 weeks later. The percentage of VEGF-positive cells was determined with hematoxylin and eosin staining and immunohistochemical staining, and MVD expression was determined by optical microscopy.

RESULTSThe VEGF expressions in brucine- or thalidomide-treated mice were significantly reduced as compared with mice in the model group (P <0.01). There were no significant difference between the high-dose brucine group and the thalidomide group (P >0.05). Significant difference was between the high- and low-dose brucine group P<0.05). Further, VEGF expression was significantly increased in the low- and medium-dose brucine groups compared with the thalidomide group (P <0.05). The MVD values in the three brucine and thalidomide groups were significantly lower than that in the model group (P <0.01). The MVD values in the medium- and high-dose brucine groups were not significantly different from those in the thalidomide group (P >0.05), while the MVD value showed a significant increase in the low-dose group compared with the thalidomide group (P <0.05).

CONCLUSIONBrucine could inhibit the growth of breast cancer to bone metastases, possibly by inhibiting tumor angiogenesis.

Animals ; Bone Neoplasms ; blood supply ; metabolism ; secondary ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Humans ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microvessels ; drug effects ; pathology ; Strychnine ; analogs & derivatives ; pharmacology ; therapeutic use ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays