title: [Content of indole alkaloids and bufadienolides contained in toad medicines].
abstract: OBJECTIVE
  To kinds of establish a HPLC method for determining contents of indole alkaloids and bufadienolides contained in toad medicines, and analyze two kinds of components contained in toad venom, toad skin and toad periostracum.
  
  
  METHODS
  As for alkaloids, Nucleosil C18 column was adopted with acetonitrile and water containing 0.5% potassium dihydrogen phosphate (6: 94, adjust pH to 3.2 with phosphate acid) as the mobile phase. The flow rate was 0.8 mL x min(-1), the detection wavelength was 275 nm, and the column temperature was 30 degrees C. As for bufadienolides, Alltima C18 column was adopted with acetonitrile and water containing 0.3% acetic acid (B) as the mobile phase. The gradient process was as follows: a linear gradient from 28% to 54% acetonitrile in the first 15 min, then kept at 54% for additional 20 min. The flow rate was 0.6 mL x min(-1), the detection wavelength was 296 nm, and the column temperature was 30 degrees C.
  
  
  RESULTS
  The linear ranges were 0.079 6-0.796 microg for serotonin, 0.097 2-1.945 microg for N-methylserotonin, 0.074 4-0.744 microg for N,N-dimethylserotonin, 0.103-2.05 microg for N,N,N-trimethylserotonin, and 0.067 2-0.672 microg for bufothionine, respectively. The average recoveries of serotonin and N-methylserotonin were 98.6% and 91.3%, respectively. The linear ranges of gamabufotalin, bufotalin, bufalin, cinobufagin and resibufogenin were 0.004 83-0.614, 0.007 9-1.006, 0.007 95-1.016, 0.009 7-1.24 and 0.009 6-1.22 microg, respectively, and their average recoveries were 101.6%, 102.5%, 101.0%, 99.1% and 98.9%, respectively.
  
  
  CONCLUSIONS
  Toad venom has the highest contents of indole alkaloids and bufadienolides, followed by toad skin, and toad periostracum showed the lowest contents and even no detection result.
authors: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors: Ting
authors: T
authors: Qu
authors: Hui-Min
authors: HM
authors: Gao
authors: Liang-Mian
authors: LM
authors: Chen
authors: Zhi-Min
authors: ZM
authors: Wang
authors: Qi-Wei
authors: QW
authors: Zhang
authors: Yi-Yu
authors: YY
authors: Cheng
mesh_terms: D000664:Amphibian Venoms / Q000737:chemistry*; D000818:Animals; D002018:Bufanolides / Q000737:chemistry* / Q000302:isolation & purification; D002025:Bufonidae; D002851:Chromatography, High Pressure Liquid; D026121:Indole Alkaloids / Q000737:chemistry* / Q000302:isolation & purification; D012867:Skin / Q000737:chemistry*
chemical_list: D000664:Amphibian Venoms; D002018:Bufanolides; D026121:Indole Alkaloids