Method for separating and purifying pyrroloquinoline quinone disodium salt based on fermentation liquid composite salting-out
A technology for pyrroloquinoline quinone disodium salt and pyrroloquinoline quinone, which is applied in the fields of biochemical industry and separation and purification, can solve the problems of obtaining pyrroloquinoline quinone disodium salt by separation, and does not involve high-purity separation and extraction of fermentation broth. The effect of shortening the process cycle, short production cycle and reducing waste water discharge
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Embodiment 1
[0074] Preparation of polyvinyl alcohol magnetic microspheres
[0075] Fe 3 o 4 Preparation of magnetic fluid: at 50°C, 0.5mol / L FeCl 2 solution and 0.5mol / L FeCl 3 The solution was added to the three-neck bottle at a volume ratio of 1:2, and while the disperser was stirring (14000r / min), 3mol / L NaOH solution in a molar ratio was added dropwise, and the temperature was raised to 80°C, and stirred for 40min. After the reaction was completed, Separate the precipitate with a magnetic field, wash repeatedly with distilled water until the supernatant is neutral, and dry to obtain black powder Fe 3 o 4 Ferrofluid;
[0076] Preparation of polyvinyl alcohol magnetic microspheres: fully swell PVA with distilled water, stir at 90°C to make a 9% PVA aqueous solution, add an appropriate amount of Fe 3 o 4 Pour the magnetic fluid into the PVA solution, stir and mix well, disperse with a cell pulverizer for 20 minutes, pour it into the dispersion medium (oil: Tween: n-butanol = 2:1:5...
Embodiment 2
[0078] Preparation of pyrroloquinoline quinone disodium salt
[0079] S1: About 21L of pyrroloquinoline quinone fermentation liquid was separated by a ceramic ultrafiltration membrane for the first filtration to obtain a primary filtrate of pyrroloquinoline quinone with a filtration yield of 98.2%;
[0080] S2: Take the primary filtrate obtained in the previous step, adjust the concentration of pyrroloquinoline quinone to about 800-850 mg / L, adjust the pH to 4.3 with sulfuric acid, stir for 1 hour and filter, and pre-coat a layer of mesoporous membrane on the filter before filtering Silica (particle diameter is 600nm, thickness 0.3mm), is filtered to obtain impurity-removing filtrate, and chromatographic purity is 83.6%;
[0081] S3: Add sulfuric acid dropwise to the impurity-removing filtrate obtained in the upward step to make the pH to 2.4, add the polyvinyl alcohol magnetic microspheres prepared in Example 1 to 3.5 wt%, and after 3 hours of adsorption reaction, pyrroloquin...
Embodiment 3
[0088] Preparation of pyrroloquinoline quinone disodium salt
[0089] S1: The pyrroloquinoline quinone fermentation broth was filtered once by ceramic membrane separation technology to obtain the pyrroloquinoline quinone primary filtrate, and the filtration yield was 98%;
[0090] S2: Take the primary filtrate obtained in the previous step, adjust the pH to 4.3 with sulfuric acid, remove impurities, stir for 1 hour and then filter. Before filtering, pre-coat a layer of mesoporous silica (particle size: 0.6-0.7um) on the filter membrane , add mesoporous silica in batches during the filtration process, and add 3 times to the completion of filtration (total consumption is the same as in Example 1), to obtain the impurity-removing filtrate, with a chromatographic purity of 86.1%;
[0091] S3: Add sulfuric acid dropwise to the impurity-removing filtrate obtained in the upward step to make the pH to 2.4, add 5wt% of the polyvinyl alcohol magnetic microspheres prepared in Example 1, ...
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