Method 1. Removal approach
Saturating, alkali refining, acidification Put about 2-3 tons of faucet water in the algae washing storage tank, put in 120kg of algae, and also after the algae broaden, carefully clean the Mannitol Pulver on the algae right into the water, and also the washed seaweed is made use of for removing sodium alginate. Clean the second set of seaweed with the washing fluid, and also clean about four batches like this. Include 300g/L (30%) sodium hydroxide option to the above cleaning remedy, Ph10-11, and also allow it stand for 8 hours until the fucoidan service, starch as well as other organic thick materials are fully coagulated and also sped up. Siphon the supernatant, acidify it with sulfuric acid (1:1), readjust it to pH 6-7, better get rid of the jelly, and obtain a neutral supernatant.
Seaweed or kelp [faucet water] → lotion [NaOH] → [pH10-11, 8h] supernatant [H2SO4] → [pH6-7] neutral clear liquid focused, alcohol clean the above neutral clear liquid
Warmth it with direct fire or heavy steam up until it steams as well as evaporates, the temperature is 110-115 ° C, a huge amount of sodium chloride speeds up, and also the salt and also adhesive dirt are continuously removed till it becomes a concentrated solution, take a little sample and pour it on the ground, as well as it should strengthen after a little air conditioning The feeding material contains greater than 30% Mannitol Pulver and also regarding 10% moisture. Trendy the concentrated remedy to 60-70 ° C, include 95% ethanol (2:1) while it is hot, keep mixing, and also progressively trendy to room temperature, after that centrifuge to completely dry to remove periodontal, and also acquire off-white loosened matter.
Neutral clear liquid [110-115 ° C] → concentrate [ethanol] → [60-70 ° C] loosened issue extraction Consider the loosened issue, put it into a removal pot furnished with a reflux condenser, add 94% of 8 times the quantity Ethanol, stirred, gradually heated up, steamed as well as refluxed for 30 minutes to discharge, cooled in running water for 8 hrs, left for a day and night, centrifuged and also dried out to get white loosened crude mannitol, containing 70% -80% mannitol. The same procedure as above, ethanol recrystallization once, to obtain Mannitol Pulver for commercial usage, the content is more than 90%, and the Cl- web content is <0.5%.
Loose matter [94% ethanol] → [reflux for 30 minutes, cooling for 8 hours] refine unrefined mannitol, include industrial mannitol and also pure water to liquify, after that add 1/10 -1/ 8 medicinal triggered carbon, keep stirring, maintain warm at 80 ℃ for half hours, filter while warm, clean the turned on carbon twice with a little water, integrate the cleaned filtrate, as well as concentrate up until the mannitol gets to 70%, if there is turbidity, re-filter. Great to room temperature level under stirring, take shape, filter with suction, clean the crystals, as well as filter with suction to obtain crystalline mannitol.
Unrefined mannitol [pure water, triggered carbon] → [80 ° C, 30min] focus the filtrate → crystallize mannitol as well as dry the above crystallized mannitol, after the Cl- is qualified (Cl-< 0.007%), completely dry it with vapor at 105-110 ° C, transformed frequently, and also obtained in 4 hours, it is the completed item of medical mannitol. Web content 98% -102%, melting point 166-168 ° C, [α] 20D +23 °- +24 °. Crystalline mannitol [105-110 ° C, 4h] → Preparation of ended up medicinal mannitol injection The ended up mannitol is dissolved in water for injection according to the dose to create an option, and also mannitol injection is acquired after sanitation. Mannitol completed item.
[Water for Shot] → Mannitol Injection.
Method 2, straight fermentation method.
Breeding strains Aspergillus Oryzae3.409 was hair transplanted right into a test tube loaded with 10ml slant tool (2.1% agar was included in the wort) and cultured at 31 ° C for 4 days. Get slant microorganisms. The stress conservation system embraces the low-temperature preservation method, put the stress on a slant in a fridge at 4 ° C, and shop it for 2-3 months for passage as soon as. Prior to use, it needs to be re-transplanted as well as turned on to ensure fermentation.
Amitramycin [stress angle medium] → [31 ° C, 4d] slant strain.
For seed society, take 2 wort slant planes of stress cultured for 4 days, and also transplant them right into 17.5 L seed society tool. Prep work of seed tool: salt nitrate 0.3%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.05%, potassium chloride 0.05%, ferrous sulfate 0.001%, corn high alcohol 0.5%, starch hydrolyzed sugar 2%, corn flour 2%, initial pH6-7. Ventilation 1: 0.5 V/( V · min), temperature 31 ° C ± 1 ° C, stirring at 350r/min, tank stress 98.1 kPa (1kgf/cm2) as well as culture for 20-24h to obtain seed society solution.
Incline pressures [seed medium] → [pH6-7, 31 ° C, 20-24h] seed culture service.
Prep work of fermentation medium: salt nitrate 0.3%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.05%, ferrous sulfate 0.001%, corn steep liquor 3%, corn flour 3.5%, starch hydrolysis sugar 12.5%, preliminary pH 6-7.
Take 350L of fermentation medium as well as placed it right into a 600L fermentation tank, heavy steam sterilize it at 146.15 kPa (1.5 kgf/cm2) for 30min, move it right into the seed culture remedy, the inoculum dimension is 5%, the storage tank pressure is 98.1 kPa (1kgf/cm2), the air flow quantity 1: 0.3 V/( V minutes), after 20 hours of fermentation, modification to 1: 0.4 V/( V minutes), temperature 30-32 ° C, stirring 230r/min, fermentation cycle 4-5d. In order to protect against frothing, include 200ml soybean oil when making ingredients. The obtained fermented fluid consists of greater than 5% of mannitol, as well as the conversion price is 40%.
Seed society fluid [steam] → [pH6-7, 30-32 ° C, 4-5d] fermentation liquid.
Coagulation, concentration, crystallization, decolorization, desalination Heat the fermentation brew at 100 ° C for 5 mins, add 1% triggered carbon at 80-85 ° C for 30 minutes, filter or tremble completely dry to obtain a clear fluid. After that the clear filtrate was concentrated in vacuum at 50-60 ° C to 31 ° Be, left at area temperature level for 24h, and dried to get Mannitol Pulver crystals. Liquify the crystals in 0.7 times water, add 2% triggered carbon, heat at 70 ° C for 30 minutes to decolorize, and also filter. The filtrate is desalted by 717 strong basic anion exchange material (OH-type) as well as solid acidic cation exchange material (H+ kind) 2:1 blended bed material, and the effluent is looked for no chloride ion reaction.
Fermentation broth [1% activated carbon] → [100 ° C, 5min; 80-85 ° C, 30min] cleared up filtrate [55-60 ° C] → [decompression] crude item [water, 2% turned on carbon] → [70 ° C, 30min] Filtrate [yin and yang combined bed material] → effluent.
Concentrate, take shape, and dry the effluent with vapor at 55-60 ° C, 80kPa (600mmHg) under decreased stress to 25 ° Be, allow the focused solution cool to area temperature for 24h to crystallize, tremble completely dry, dry at 105-110 ° C, as well as pulverize to obtain Medicinal completed item, return 60%, web content 98.83% -100.9%.
Effluent [55-60 ° C] → [Decompression] Concentrate [Area Temperature level] → [24h] Formation [105-110 ° C] → API.
For the prep work, take an appropriate quantity of water for injection, warmth it to regarding 90 ° C, evaluate Mannitol Pulver at a content of 20%, dissolve it, add turned on carbon in a water bath and also warm for 5 minutes to decolorize, infiltrate a Buchner channel, and afterwards include water for injection fully quantity. After the resolution of web content as well as pH is certified, filter it with G3 vertically merged glass filter ball till it is clear, and distribute it in 50ml, 100ml or 250ml ampoules or mixture bottles, and also disinfect it with 98.1 kPa (1kgf/cm2) hot-pressed vapor for 40min to get manna Alcohol Shot.
