A kind of refining method of vasopressin [+gly] impurity
A purification method and vasopressin technology, which are applied in the preparation methods of peptides, oxytocin/vasopressin, chemical instruments and methods, etc., can solve the problem of uneconomical, large amount of hazardous waste liquid, and high waste liquid treatment cost. problems, to achieve the effect of optimizing the production process and reducing the generation of hazardous waste
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[0052] In the following examples, the preparation method of the crude vasopressin [+Gly] impurity solution includes: (1) using solid-phase synthesis to obtain the crude vasopressin [+Gly] impurity; (2) adding the reduced The crude vasopressin [+Gly] impurity is dissolved and diluted to obtain a crude vasopressin [+Gly] impurity solution; (3) the crude vasopressin [+Gly] impurity solution is oxidized to obtain the Vasopressin [+Gly] impurity crude solution.
[0053] The solid-phase synthesis method includes the following steps: using Rink Amide MBHA resin as a starting material, using Fmoc-protected amino acids as monomers, using HOBt / DIC as a condensation agent, and sequentially connecting amino acids one by one; adding peptide cutting reagents to carry out Cut the peptide, add methyl tert-butyl ether for precipitation, and obtain the crude impurity product of reduced vasopressin [+Gly]. Wherein, the peptide cutting reagent is TFA / TIS / H with a volume ratio of 90:7.5:2.5 2 O....
Embodiment 1
[0054] Example 1 Load&Lock4002 preparative column with inner diameter of 50mm
[0055] Using Load&Lock dynamic axial compression and static locking technology, the packing is as described ODS-AQ, pore size 10nm, particle size 10μm, packed to a column bed pressure of 1000psi, Varian chromatography packing system, 300g dry powder as described ODS-AQ super water-resistant filler, 600mL isopropanol is stirred and homogenized, poured into a Load&Lock4002 preparation column with an inner diameter of 50mm, the compression ratio is 1.5:1, and the carrier gas is N 2 , adjust the carrier gas pressure so that the oil pressure gauge pressure is 1500psi, and dynamically axially compress to a column bed height of 25cm, which is used as a preparative column for reverse-phase enrichment, reverse-phase salt conversion and reverse-phase purification schemes.
Embodiment 2
[0056] Example 2 Reverse Phase Enrichment, Reverse Phase Salt Conversion and Reverse Phase Purification of Vasopressin [+Gly] Impurity Crude Solution
[0057] Instrument: Varian SD-1 high pressure liquid phase preparation system
[0058] Chromatographic column: the preparation column Load&Lock400250 * 250mm of embodiment 1, packing is ODS-AQ has a particle size of 10 μm and a pore size of 10 nm.
[0059] The structural formula of the vasopressin [+Gly] impurity is The concentration of the crude product solution of reduced vasopressin [+Gly] impurity is 1 mg / mL, and the solvent of the crude product solution of vasopressin [+Gly] impurity is an aqueous solution containing trifluoroacetic acid and acetic acid.
[0060] Mobile phase A is 0.02% acetic acid / water solution by volume, mobile phase B is 0.02% acetic acid / acetonitrile by volume, and sample C1 is vasopressin [+Gly] impurity crude product solution, according to the HPLC method. The HPLC purity of vasopressin [+Gly] i...
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