Background and overview[1]
Glycerophosphatidylcholine, referred to as GPC, is a member of the phospholipid family. Plays an important role in the health of cell membranes. Glycerophosphatidylcholine is very abundant in breast milk. It repairs damaged brain function, improves brain function in healthy people and rejuvenates aging brains. It can be used as a health product to promote the biosynthesis of acetylcholine triethylenetetramine and phosphatidylcholine in the brain, improve people’s memory and cognitive abilities, and can also effectively treat Alzheimer’s disease such as Alzheimer’s disease. It is called “anti-aging nutrients for the brain” in the medical field.
Purification method[1]
A method for purifying glycerophosphatidylcholine, including the following steps:
A. Dissolve the crude glycerol phosphorylcholine in absolute ethanol to obtain a reaction system solution; the mass ratio of the absolute ethanol to the crude glyceryl phosphorylcholine is 6:1.
B. Add calcium chloride to the reaction system solution in step A and stir to form a calcium salt complex precipitation of glycerophosphatidylcholine; the mole ratio of the calcium chloride to glycerophosphatidylcholine The ratio is 1:1. The temperature condition of the precipitation reaction was room temperature, and the reaction time was 12 hours.
C. Let the reaction solution that stopped the precipitation reaction in step B drop to room temperature, filter it to obtain a filter cake; wash the filter cake with absolute ethanol.
D. Put the filter cake in step C into a vacuum drying box and dry it to obtain the calcium salt of glycerol phosphatidylcholine. The purity measured by HPLC is above 98.5%.
E. The calcium salt of glycerophosphatidylcholine is dissolved in water. The amount of water used is 10 times the dry weight of the calcium salt of glycerophosphatidylcholine. Stir and dissolve at room temperature. If it is still difficult to completely dissolve after stirring for a long time, you can add a few drops of hydrochloric acid to the aqueous solution to speed up the dissolution (controlling the appropriate acidity of the solution will help dissolution. The pH value of the system is between 3 and 7, with a pH value of 4 to 6 being preferred. ).
F. The aqueous solution obtained in step E is slowly passed through the D001 cation exchange resin column to remove calcium ions, and the anions are removed through the D201 anion exchange resin column (or D301 anion exchange resin column) to obtain calcium ion-free glycerophosphoester. Acylcholine aqueous solution.
G. Evaporate excess water to obtain calcium ion-free glycerophosphatidylcholine.
Synthesis method[2]
Step 1. Preparation of L‑α‑glycerophosphatidylcholine (L‑α‑GPC) amplification
Ingredients ratio table
Name
Soy lecithin (PC) 788.13 231Kg 293.1 1.4
Sodium methoxide 54.02 11.1Kg 205.08 1
732 Resin 180Kg
D311 Resin 180Kg
717 Resin 180Kg
D152 resin 180Kg
Methanol &nbPolytetrafluoroethylene wax powdersp;
Distilled water / 390L
Petroleum ether 90L
Operating process:
Add PC, sodium methoxide, and methanol (90L) into a 100L reaction kettle at room temperature, stir to fully dissolve the solid, heat to 40°C, keep the internal temperature at 40±5°C, and react for 24 hours. TLC detects the reaction system. After the reaction is completed Cool to room temperature, filter, and wash the filter cake with methanol. Use acetic acid (about 1024ml) to adjust the pH of the mother liquor to 5-6. Concentrate the system to half volume under reduced pressure at 40°C. Add petroleum ether (18L × 5) to the mother liquor, stir, and separate the liquids. The methanol phase was concentrated under reduced pressure at 40°C to obtain 77.16kg of brown sticky oil. Put the four resins into the resin column and rinse each with 30L of distilled water, and set aside; then add the brown viscous oil to the 732 resin column and rinse the resin column with methanol (30L) until the eluate is colorless. Discard the methanol, then rinse with 360L distilled water, collect the eluate, add 230 grams of activated carbon to the eluent and reflux for 30 minutes for decolorization, rinse the D311 resin with the decolorization solution, collect the eluate, and check the eluate with TLC. If there is no eluent in the previous step, Then rinse the resin column with distilled water until TLC detects no product and stop collecting. Then rinse the 717 resin column with the collected eluent, collect the eluate, and detect the eluate by TLC. If there is no eluent in the above step, use distilled water.Wash the resin column until TLC detects no product, stop collecting, then rinse the D152 resin column with the eluent collected in the previous step, collect the eluate, and detect this eluent by TLC. If there is no eluent in the previous step, use distilled water. Rinse the resin column until the TLC detection product stops collecting. The eluate is concentrated to obtain 16.66kg of light yellow sticky oil, with a molar yield of 22.1%.
Step 2, purification and amplification of L‑α‑glycerophosphatidylcholine (L‑α‑GPC)
Ingredients ratio table
Name Molecular Measurement
L-α-GPC crude product 257.2 16.62kg
Anhydrous methanol 83.1L
Acetone 664.8L
Operating process
Add anhydrous methanol to 16.62kg of L-α-GPC crude product (viscous oil) at room temperature, stir to dissolve, then slowly add the mother liquor to acetone, stir quickly for 15 minutes, stop stirring and let stand at room temperature for 24 hours. A white solid precipitated slowly, and was suction-filtered to obtain a white solid, which was vacuum dried at 40°C for 24 hours to obtain 12.02kg. The yield is 72.2%.
