The active peptides purified by chromatographic separation were used.
Ion exchange chromatography, gel filter chromatography, liquid chromatography a series mass spectrometry is commonly used. Separation purification of snake venom analgesic peptides: the dissolved CM-SephadexC-25 is loaded into the column (2.6cm x 60cm) and balanced with 0.01mol/L phosphate buffer (pH 6.0). 1000.0mg coarse venom is dissolved in the same buffer, the above sample. The first 2 peaks are washed out with a balance buffer, then the first 2 peaks are washed with a straight-line gradient (0.00 to 0.65mol/LNaCl) at a flow rate of 36 ml/h, 6 tubes/h. The collected active components were freeze-dried, desalted and then concentrated, further purified with Sephadex G-50 column (1.6cm x 100cm), elution of 0.15mol/LNH4Ac buffer (pH 7.0), flow rate of 16ml/h, 4 tube/h. The resulting active components were eluded by a CM-Sepha-dexC-25 bar (1.6cm x 120cm) balanced with 0.15mol/LNH4Ac, with a straight gradient (0.15 to 0.60mol/LNH4Ac) at a flow rate of 16 ml/h, 4 tubes/h. Collected active components frozen dry, snake venom analgesic peptide single component. The product reaches the color spectrum purity level.