ABSTRACT
OBJECTIVE: To determine the contents of hirudin's hydrolysates in processed leeches, set up a new evaluating method of dot blotting to evaluate the qualities of those processed Chinese medicines as leeches. METHOD: Contents of hirudin's hydrolysates in processed leeches were determined by dot blotting with rat antibody of anti-hirudin as the first antibody. Blotting signal was analyzed by software of Quantity One. RESULT: Contents of hirudin's hydrolysates in four batches of processed leeches were 296.51, 165.47, 95.58, and 298.05 microg g(-1), respectively. CONCLUSION: Difference among four batches of processed leeches was significant in the content of hirudin's hydrolysates. Dot blotting, as a convenient and accurate method can be broadly used for evaluating processed products of Chinese crude drugs similar to leeches.
Subject(s)
Hirudins/metabolism , Leeches/chemistry , Animals , Drugs, Chinese Herbal/chemistry , Hirudins/immunology , Immunoblotting/methods , Reproducibility of ResultsABSTRACT
OBJECTIVE: To determine the cocoonase content in maxilla of four silkmoth breeds of Bombyx mori and set up a dot blotting method to determine the content of the drug action components in Chinese medicines. METHODS: The contents of cocoonase in maxilla of four silkworm breed were determined by dot blotting with rabbit antibody of anti-cocoonase as the first antibody. Blotting signals were analyzed by software of Quantity one. RESULTS: Contents of cocoonase in maxilla of four silkmoth breeds were 6.41, 8.09, 7.62, and 1.88 microg/moth, respectively. CONCLUSION: The difference among four silkmoth breeds was significant in the content of maxilla cocoonase. Dot blotting, as a convenient and accurate method could be used for determination of the contents of drug action components in Chinese crude drugs similar to silkworm.
Subject(s)
Bombyx , Immunoblotting/methods , Materia Medica/chemistry , Serine Endopeptidases/metabolism , Animals , Antibody Formation , Bombyx/chemistry , Bombyx/classification , Bombyx/metabolism , Enzyme Precursors/metabolism , Male , Quality Control , Rabbits , Reproducibility of Results , Serine Endopeptidases/immunologyABSTRACT
OBJECTIVE: To research the effect of cryogenic processing technology of leech on the components of water-solubility proteins and the fibrinolytic activity. METHODS: Leech was respectively processed by traditional processing technology (TPT) and cryogenic processing technology (CPT). The former was done according to the method described in 2005 pharmacopeia of our country, and the latter adopted the quick-liquid-nitrogen-frozen technology and the freeze-drying technology. The difference of the components of water-solubility proteins in leech between the two processing technology was qualitatively and quantitatively determined by SDS-PAGE and Scion Image software. Fibrin plate method and thrombolysis test in vitro were used to qualitatively and quantitatively determine the difference between the two processing technologies in the fibrinolytic and thrombolysis activity. RESULTS: Water-solubility proteins in leech processed by CPT were more abundant than those of TPT. Not only the number and concentration of protein bands in SDS-PAGE gel, but also the fibrinolytic and thrombolysis activity of CPT, were markedly higher than those of TPT. CONCLUSION: CPT is advantaged to TPT in assuring activity of peptides and proteins of leech. CPT can be theoretically applied into the processing of other animal Chinese traditional medicine in which the main active components are peptides or proteins.