[Advance on quality control of toad venom and its key influence factors].

Toad venom (Chansu) is prepared from the dried secretion of Bufo bufo gargarizans or B. melanostictus. It is not only one of the famous and expensive traditional Chinese medicines(TCMs) from animal origin, but also one of 28 kinds of toxic TCMs to be required for special management issued by the State Council of the People's Republic of China. Chansu contains the rich bufadienolides and indole alkaloids, and displays various bioactivity including cardiotonic, anti-tumor, analgesia, and local anesthesia. Based on the published references in the recent years, the advance on the identification of adulterants and quality evaluation as well as the influence factors on the quality of toad venom was summarized to improve the quality standards and promote the level of quality control of toad venom and its preparations.

[Research on therapeutic effects of bufonis venenum on L7212 leukemia and its mechanism].

OBJECTIVE: To study the therapeutic effects of Bufonis Venenum on L7212 leukemia and the potential mechanism. METHODS: L7212 leukemia model mice were randomly divided into four groups: model group, low and high dose Bufonis Venenum groups and chemotherapy group. Normal mice were treated as control group. Mice were injected intraperitoneally for 10 days continually. The body weight, survival time, peripheral blood leukocyte, hepatic and splenic indexes, bone marrow leukocyte and T lymphocyte were observed and determined. RESULTS: Body weight of L7212 leukemia model group mice were decreased significantly. Compared with other groups, high dose Bufonis Venenum group's weight loss was the least. Bufonis Venenum groups survived longer than L7212 model group. Compared with model group, high dose Bufonis Venenum group's liver index was higher (P < 0.05). After inoculation for 1 day, leukocyte count as well as percentage of leukemic cells within five groups had no significant difference (P > 0.05). Compared with the model group, after inoculation for 10 days, leukocyte count in Bufonis Venenum groups and the chemotherapy group were significantly reduced (P < 0.05). Percentage of leukemia cells in blood and bone marrow in high dose Bufonis Venenum group was significantly decreased (P < 0.05). Compared with model group, CD3+ and CD4+ in Bufonis Venenum groups and the chemotherapy group were increased, CD8+ was decreased, but had no significant difference (P > 0.05). CONCLUSION: Bufonis Venenum has therapeutic effects on the L7212 leukemia by inducing apoptosis and improving immune system.

[Study on the HPLC fingerprint of toad skin from different regions].

OBJECTIVE: To study the HPLC fingerprint of toad skin and provide a reliable method for quality control and identification. METHODS: It used HPLC for detection and computer aided similarity evaluation system for processing and analysing HPLC fingerprint. RESULTS: The common pattern of HPLC fingerprint of toad skin was astablished, 29 peaks were identified as the characteristic fingerprints, in which 9 peaks corresponded to 9 bufogenins. (2) Each samples' similarity of relative retention time was all above 0.99, but the similarity of relative peak areas was low. CONCLUSION: (1) The method is accurate and with good reproducibility. The fingerprints can be used for the identification and quality control of toad skin. (2) The toad skin from different regions are stable in composition, but the contents of the components are different.

Treatment of toad venom poisoning with digoxin-specific Fab fragments.

Toxicity from toad venom poisoning is similar to digoxin toxicity and carries a high mortality rate. We report on six previously healthy men who developed vomiting and bradycardia after ingesting a purported topical aphrodisiac. Each patient had positive apparent digoxin levels and the first four patients died of cardiac dysrhythmias. The last two patients recovered following treatment with digoxin Fab fragments. We analyzed samples of the purported aphrodisiac and found that it was identical to Chan Su, a Chinese medication made from toad venom. To our knowledge, this is the first reported use of digoxin Fab fragments to treat toad venom poisoning.

[Dynamic effect of low frequency complex impulse current on transdermal absorption of secretio bufonis].

OBJECTIVE: To explore the regulation of transdermal absorption of Secretio Bufonis (SB) and the effect of low frequency complex impulse current (LFCIC) on it. METHODS: By modifying three-chamber flow diffusion pool to develop a prototype LFCIC device for transdermal delivery, using high performance liquid chromatograph (HPLC) to determine the quantitative transdermal absorption of the amount of ingredients of SB, including bufalin, cinobufagin and resibufogenin, etc. and the transdermal absorption velocity was calculated. RESULTS: The chief ingredients of SB could be absorbed through skin, but the volume was low. Additional application of LFCIC could enhance the cumulative infiltration volume and velocity of transdermal diffusion. Difference appeared 2 hrs after and significant difference appeared 4 hrs after the application, and 13.8 Hz showed the optimal effect of transdermal delivery. CONCLUSION: Chief ingredients of SB could be absorbed through transdermal medication, and LFCIC can evidently enhance the amount and velocity of transdermal absorption of SB.

[Determination of resibufogenin and cinobufagin in venenum Bufonis by HPLC].

OBJECTIVE: To establish a HPLC method to determine the contents of active constituents in Venenum Bufonis. METHOD: In determining the contents of resibufogenin and cinobufagin in the traditional Chinese medicine Venenum Bufonis, the mobile phase was acetonitrile-0.5% KH2PO4 solution(50:50)(pH adjusted with value phosphoric acid to 3.25 +/- 0.02). RESULT: The constituents thus determined have good linearity and separation. The average recovery of resibufogenin was 100.35%, RSD 1.86%; the average recovery of cinobufagin was 100.38%, RSD 2.09%. CONCLUSION: The method was convenient, rapid, accurate and practicable.

[Determination of resibufogenin in xinli pills by RP-HPLC].

A method has been developed for the quantitative determination of Resibufogenin in Xinli pills by RP-HPLC. The stainless steel column(200 x 4.6 mm) was packed with ODS-3 (particle size 5 microns). The mobile phase was a mixture of methanol-water (60:40). The flow rate was 1 ml/min. The column temperature was 30 degrees C. The detection wavelength was 299 nm. The retention time for recibufogenin was 5.6 min. The average recovery for resibufogenin was 99.9%. The linear calibration curve for resibufogenin was obtained in the range 53.0-159.0 micrograms with the correlation coefficient 0.9998.

Nux vomica 30 prepared with and without succession shows antialcoholic effect on toads and distinctive molecular association.

Adult toads, Bufo melanostictus, were administered Nux vomica (Nux v) 30 prepared with and without succussion on the tongue. The drug was mixed with sterile distilled water at the rate 0.05ml/ml water and given orally 0.05ml/individual. The control consisted of blank ethanol solution. Seeds of Strychnos nuxvomica were ground and extracted with 90% ethanol in the laboratory. Nux v 30 was prepared by successive dilution and succussion in 30 steps, Nux v 30 u was prepared by successive dilution only. Four hours after treatment, toads were given 25% ethanol i.p. at 8g/kg body weight. The duration of ethanol induced sleep time was recorded for each toad. Both Nux v 30 and Nux v 30 u significantly reduced ethanol induced sleep time in toads as compared to their respective controls. Electronic, infra red and nuclear magnetic resonance spectra of Nux v 30, Nux v 30 u and their diluent medium (90% ethanol) show marked differences from each other. These dilutions and ethanol 30 and ethanol 30 u show marked differences from each other with respect to spin-lattice relaxation time (T1) and chemical shift. The difference has been attributed to the variation in intra and inter-molecular association of ethanol and water.