Method of manufacturing agar or agarose beads
The invention discloses method for manufacturing agar or agarose beads, comprising the steps of: a) providing a water phase comprising an aqueous solution of agar or agarose at a temperature of 40-100° C.: b) providing an oil phase comprising a water-immiscible solvent and an emulsifier at a temperature of 40-100° C.; c) emulsifying the water phase in the oil phase to form a water-in-oil emulsion: d) cooling the water-in-oil emulsion to a temperature below a gelation temperature of the agar or agarose to form a dispersion of solidified agar or agarose beads: and e) recovering agar or agarose beads from dispersion, wherein the emulsifier comprises a phosphate ester of an alkoxylated fatty alcohol.
1. A method for manufacturing agar or agarose beads, comprising the steps of:
a) providing a water phase comprising an aqueous solution of agar or agarose at a temperature of 40-100° C.;
b) providing an oil phase comprising a water-immiscible solvent and an emulsifier at a temperature of 40-100° C.;
c) emulsifying said water phase in said oil phase to form a water-in-oil emulsion;
d) cooling said water-in-oil emulsion to a temperature below a gelation temperature of said agar or agarose to form a dispersion of solidified agar or agarose beads; and
e) recovering agar or agarose beads from said dispersion,
wherein said emulsifier comprises a phosphate ester of an alkoxylated fatty alcohol.
2. The method of claim 1 , wherein said water-immiscible solvent comprises toluene or xylene.
3. The method of claim 1 , wherein said oil phase comprises less than 0.1 wt. % of alkyl phenols or alkyl phenol derivatives.
4. The method of claim 1 , wherein said oil phase is devoid of, or essentially devoid of, alkyl phenols and alkyl phenol derivatives.
5. The method of claim 1 , wherein said emulsifier comprises a mixture of phosphate monoester and phosphate diester of said alkoxylated fatty alcohol.
6. The method of claim 1 , wherein said fatty alcohol comprises one or more C 10 -C 20 linear or branched, primary or secondary, alkanols or alkenols.
7. The method of claim 1 , wherein said alkoxylated fatty alcohol has a structure
R 1 —O—(R 2 -O) n —H (I)
wherein:
R 1 is a saturated or unsaturated, linear or branched, aliphatic C 10 -C 20 hydrocarbon,
R 2 is —CH 2 —CH 2 — or a mixture of —CH 2 —CH 2 — and —CH 2 (CH 3 )—CH 2 —, and
n is 2-20.
8. The method of claim 1 , wherein said emulsifier comprises a mixture of
R 1 —O—(R 2 −O) n −P(O)−OH (II)
R 1 —O—(R 2 −O) n −P(O)−(O−R 2 ) n —O−R 1 (III)
wherein:
R 1 is a saturated or unsaturated, linear or branched, aliphatic C 10 -C 20 hydrocarbon,
R 2 is —CH 2 —CH 2 — or a mixture of —CH 2 —CH 2 — and —CH 2 (CH 3 )—CH 2 —, and
n is 2-20.
9. The method of claim 8 , wherein R 1 is a saturated or unsaturated linear aliphatic C 10 -C 18 hydrocarbon.
10. The method of claim 8 , wherein n is 2-10.
11. The method of claim 10 , wherein n is 2-5.
12. The method of claim 1 , wherein said emulsifier comprises a mixture of phosphate monoester and phosphate diester of a mixed ethylene oxide+propylene oxide adduct of a C 10 -C 16 alkanol.
13. The method of claim 1 , wherein said emulsifier comprises oleth-3-phosphate.
14. The method of claim 1 , wherein said emulsifier comprises a C 10 -C 16 ethoxylated/propoxylated phosphate.
15. The method of claim 1 , wherein said emulsifier comprises an oleth-3 phosphate.
16. The method of claim 1 , wherein said aqueous solution of agar or agarose comprises 1-8 wt. % agar or agarose.
17. The method of claim 1 , wherein said oil phase comprises 0.01-2 wt. % of said emulsifier.
18. The method of claim 1 , wherein step c) comprises mixing said water phase and said oil phase in an agitated vessel to form a water-in-oil emulsion.
19. The method of claim 18 , wherein step c) further comprises passing said water-in-oil emulsion through a rotor-stator mixer to reduce a droplet size of said water-in-oil emulsion.
20. The method of claim 18 , wherein step c) further comprises passing said water-in-oil emulsion through a static mixer to reduce a droplet size of said water-in-oil emulsion.
21. The method of claim 18 , wherein step c) further comprises passing said water-in-oil emulsion through a porous membrane to reduce a droplet size of said water-in-oil emulsion.
22. The method of claim 1 , wherein step c) comprises passing said water phase through a porous membrane into said oil phase to form a water-in-oil emulsion.
23. The method of claim 1 , where step d) is performed in an agitated vessel.
24. The method of claim 1 , wherein step d) is performed by passing said water-in-oil emulsion through a conduit with a longitudinally decreasing temperature gradient.
25. The method of claim 1 , wherein step e) comprises adding water or an aqueous solution to said dispersion, decanting said oil phase and recovering said agar or agarose beads as a sediment.
26. The method of claim 1 , wherein step e) further comprises washing said agar or agarose beads with water or an aqueous solution to remove residual emulsifier.
27. The method of claim 1 , further comprising, after step e), a step f) of cross-linking said agar or agarose beads.
28. The method of claim 1 , further comprising, after step e), a step f) of coupling ligands to said agar or agarose beads.