Method 1. Extraction method (the method currently used in my country has low purity but low cost) for the extraction of oxytocin 10 solution. Take 100 g of dry powder of the posterior leaf and 30 g of quartz powder into a ball mill, add distilled water, and extract four times with the same method. The quantity of distilled water added was 1.4 L two times as well as 1.3 L twice. Each extraction was performed for 45 min, centrifuged, the liquid was gathered, and the residue was removed again. The 4 essences were incorporated to acquire oxytocin service. Back fallen leave completely dry powder [water, sphere mill] → oxytocin removal fluid chromatographic splitting up Add 1500g of pre-treated artificial zeolite, add 20L of 0.25% acetic acid solution, put it right into the exchange column after stirring, wait on the acetic acid remedy to drop to above the surface of the zeolite When it is 2-3cm, the drawing out option is added, and the white turbid solution is accumulated at a proper circulation price (primarily including oxytocin, since its pI7.7, as well as vasopressin pI10.9, which is easy to end up being a favorable ion and be adsorbed). When the liquid level of the remove goes down to the surface of the zeolite, include distilled water instantly, and also remain to accumulate the turbid fluid until it goes out. The turbid solution was adapted to pH 3.5 with glacial acetic acid, swiftly heated up to 95 ° C in a water bathroom for 3 minutes, swiftly cooled down and also refrigerated over night. Oxytocin extract [HAc, man-made zeolite column] → turbid fluid [HAc, heating] → splitting up liquid adsorption, elution The next day, the refrigerated fluid was filtered, and also 10% bentonite slurry was included in the filtrate under stirring (include 3ml per 100ml ), mixed for 1 h, adsorbed, and centrifuged. The supernatant was contacted sulfosalicylic acid test option, and also the adsorption was full if no rainfall took place. If it is not complete, the adsorption is performed once more, and also the two bentonite slurry speeds up are incorporated. The bentonite was eluted four times with 1% acetic acid (1.6 L, 1.4 L, 1.2 L, and 0.8 L specifically). When the eluent was heated to 80 ° C, chlorobutanol was added, and then the temperature level was increased to 95 ° C., quick-cooled to 25 ℃, filteringed system or centrifuged, combined 4 filtrates, and also tested the titer and also vasopressin restriction. Chilled solution [purification] → filtrate [bentonite slurry] → adsorbate [1%HAc] → eluent Preparation of oxytocin injection Water down the determined qualified oxytocin solution to 5U/ml or 10U/ml, infiltrated a No. 4 or No. 5 vertical melting channel, potted, and also disinfected by flowing vapor at 100 ° C for thirty minutes to acquire the completed product. Oxytocin solution (qualified for determination) [filtered, potted, disinfected] → oxytocin 10 shot. Regrowth of man-made zeolite The man-made zeolite was cleaned twice with distilled water, after that 1.5 L of water (calculated according to 500g of zeolite) and 150g of sodium chloride were included and mixed for 2h, washed to get rid of vasopressin, and after that washed with distilled water till there was no chloride ion. Add an ideal amount of water, change the pH to 9 with sodium hydroxide, mix for half a hr, pour off the supernatant, wash with pure water, change the pH to 5.5-6 with sulfuric acid, clean with water up until almost neutral, filter completely dry, and cook at 105 ° C Dry extra. Include suitable amount of water before usage, change with glacial acetic acid, mix and soak. The preparation of 10% bentonite slurry was mixed in a ratio of bentonite mass to water quantity ratio of 1:10, ground for about 1 hr, and also the pH was gotten used to 3.5 with acetic acid during grinding, and positioned in a fridge for later use.
Technique 2. Chemical synthesis approach Initially, 7-peptide amide (paragraph 3-9) was manufactured from benzyloxycarbonylleucine-p-nitrophenyl ester, and then benzyloxycarbonyl-S-benzylcysteine · tyrosylazide was synthesized (paragraph 1-2), the 3rd action is to synthesize oxytocin 10.