A PROCEDURE FOR THE PRODUCTION OF ASTAXANTHIN AND RELATED CAROTENOIDS, ASTAXANTHIN ESTERS, CHITIN, PROTEINS AND MEAT FROM PLANTS, ALGAE, BACTERIA, KRILL, SHRIMPS AND OTHER CRAYFISH AND CRUSTACEA

Abstract: A procedure is described for the production of astaxanthin and related carotenoids, astaxanthin esters, chitin, proteins and meat from plants, algae, bacteria, krill, shrimps and other crayfish and crustacea by extraction in boiling lye. The residue from the extraction is dried and dissolved in anhydride formic acid or another concentrated strong acid, filtered, then the filtrate has water added so that the purified chitin can be precipitated. Astaxanthin and related carotenoids, astaxanthin esters, chitin and proteins are removed from the extract either by acid precipitation or by cooling, removal and subsequent phase separation and purification.

Description: A procedure for the production of astaxanthin and related carotenoids, astaxanthin esters, chitin, proteins and meat from plants, algae, bacteria, krill, shrimps and other crayfish and Crustacea. The present invention consists of a procedure for the production of astaxanthin and related carotenoids, astaxanthin esters, chitin, proteins and meat as according to claim 1. Astaxanthin and related carotenoids are used in the fish farming industry today as dies for colouring the fish meat of species which have red or pale red meat in natural habitats. During cultivation, there is no feed that contains enough dye to give this colour flesh to such fish. These dyes are produced today by fermenting algae and synthesizing from a fermentation mixture. This approach is both expensive and time-rconsuming and necessitates the processing of large volumes of fermentation mixture. So far there has been no product to meet the need for an inexpensive industrial process for producing astaxanthin and related carotenoids for the fish farming industry and other applications for the colouring of foodstuffs. The present invention describes a procedure which enables astaxanthin and related carotenoids to be produced on an industrial basis much cheaper than with present day technology. At the same time, the present invention is based on the exploitation of natural resources that are unexploitable or which represent a waste disposal problem. These include Antarctic krill; plants, bacteria and algae; and waste from the commercial processing of the meat in species such as shrimps, crayfish and Crustacea. The present invention also helps to utilize the above-mentioned natural resources better, by using more of the raw material, either by improving the degree of utilization through the production of the required products, or by utilizing the same raw material to produce other types of products. Thus the present invention can for example be used to produce astaxanthin and related carotenoids astaxanthin esters, chitin, proteins and meat from Antarctic krill in a way that gives an extremely high degree of utilization of this raw 5 material. The shells of Antarctic krill are in addition the greatest unused source of protein in the world. However they cannot be used for feed or food because they contain too many toxic components such as fluorine and toxic heavy 10 metals. With the means of production in the present invention, both the fluorine and the toxic heavy metals can be removed from the shells so that extensive use can be made of the proteins and chitin that can be produced from them. These and other features of the invention become 15 evident from the characterizing part of the Claims of Patent below. The method in accordance with this invention will now be illustrated in greater detail by the following examples. 20 Example 1, Krill When the Antarctic krill is caught, the meat is squeezed out behind the cephalothorax using modified shrimp processing equipment. This extraction of the meat must be done within 3 to 4 hours of the time of catching to avoid 25 contamination from the shell, head and cephalothorax which could affect the meat. As the autolysis of krill occurs very quickly and easily, this is another reason why the meat must be extracted within 4 hours of the time of the catch. After the meat has been squeezed out, it has to be kept at below 30 40 C until it is processed. The contaminants in krill are mainly the fluorine in the shell and toxic heavy metals such as cadmium, mercury, lead and zinc. There are high concentrations of cadmium in the cephalothorax and the content of the head. 35 The further treatment of the rest of the krill after the meat has been extracted, i.e., the shell, cephalothorax .and the head with its content, here termed krill shell, can take alternative forms. Alternative A: The krill shell is boiled in 1.0 N NaOH for 30 min. then the extract is separated from the residue. The residue is then again boiled in 1.0 N NaOH for 30 min. and extract is once more separated from the residue. The residue is dried in concentrated formic acid before being dissolved in anhydride formic acid or another strong concentrated acid, filtrated, then water is added to the filtrate to precipitate the purified chitin. The extracts from the two boilings are mixed and can either: a: have an acid added until all astaxanthin and related carotenoids, astaxanthin esters, proteins are precipitated, whereupon the precipitate is separated from the solution by filtration or a similar process; or b: be removed to cool, after this the mixture is separated into three phases which are divided by floating off the phase with the lowest specific weight and decanting (or the equivalent) the phase with the intermediate specific weight from the phase with the highest specific weight; where the phase with the lowest specific weight is neutralized by the addition of an acid which causes the formation of two liquid phases, which are separated, and the oily phase which consists of astaxanthin esters is taken care of; where the phase with the highest specific weight is neutralized by an acid, and the solids consisting of astaxanthin and related carotenoids are filtered away and taken care of; and where the phase with the intermediate specific weight is neutralized and has marine oil, Ca + -ions, S -ions and (NH. HPO. for the precipitation of the fluorine and the heavy metal contaminants, where this precipitate is removed from the solution by floating or the equivalent, after this the solution is acidified prior to the precipitation of the purified proteins followed by the separation of these proteins from the solution by filtration or an equivalent method. Alternative B: The krill shell is boiled in 0.01 N NaOH for 30 min. then the extract is separated from the residue. The residue is then again boiled in 0.01 N NaOH for 30 min. and the extract is once more separated from the residue. The residue is then again boiled this time in 1.0 N NaOH for 30 min. and the extract is again separated from the residue. The residue is dried in concentrated formic acid before being dissolved in anhydride formic acid or another strong concentrated acid, filtrated, then water is added to the filtrate to precipitate the purified chitin. The extracts from the first two boilings in 0.01 N NaOH are mixed, neutralized and have the addition of marine oil, Ca 2+-ions, S2--ions and (NH 4 Λ )2_HPO4. for the precipitation of the fluorine and the heavy metal contaminants, where this precipitate is removed from the solution by floating or the equivalent, after this the solution is acidified prior to the precipitation of the purified proteins followed by the separation of these proteins from the solution by filtration or an equivalent method. The extracts from the last two boilings in 1.0 N NaOH are mixed and can either: a: have an acid added until all astaxanthin and related carotenoids, astaxanthin esters, proteins are precipitated, whereupon the precipitate is separated from the solution by filtration or a similar process; or b: be placed to cool, after this the mixture is separated into three phases which are divided by floating off the phase with the lowest specific weight and decanting (or the equivalent) the phase with the intermediate specific weight from the phase with the highest specific weight; where the phase with the lowest specific weight is neutralized by the addition of an acid which causes the formation of two liquid phases, which are separated, and the oily phase consisting of astaxanthin esters is taken care of; where the phase with the highest specific weight is neutralized by an acid and solids consisting of astaxanthin and related carotenoids are filtered away and taken care of; and where the phase with the intermediate specific weight is neutralized and followed by the addition of marine oil, Ca 2+-ions, S2--ions and (NH )_HPO. for the precipitation of the fluorine and the heavy metal contaminants, where this precipitate is removed from the solution by floating or the equivalent, after this the solution is acidified prior to the precipitation of the purified proteins followed by the separation of these proteins from the solution by filtration or an equivalent method. Example 2, Plants, algae and bacteria Astaxanthin can be produced from plants, algae and bacteria that contain this die in accordance with the procedure presented in this patent. Plants, algae or bacteria or a mixture of these can be boiled for 30 min. in 1.0 N NaOH, after this the extract is separated from the residue of the plants, algae or bacteria and the extract is removed for cooling. During cooling the astaxanthin is separated and is deposited as sediment in the extract. The sediment is composed of astaxanthin and extract and these are separated by a known method. Example 3, shrimps, crayfish and other Crustacea Shrimps, crayfish and Crustacea differ from krill in that they are boiled before the meat is removed for consumption. The residue from the shrimps, crayfish and Crustacea consists of shell, heads with contents and nonedible components, which can be processed in the same way as krill shell for the production of astaxanthin and related carotenoids, astaxanthin esters, proteins and chitin. The above description only indicates the preferred means of applying the present invention. For specialists in the field it is obvious that various modifications can be made without exceeding the framework of the invention, which is delimited by the following claims of patent. The procedures which are indicated should therefore be considered as illustrative rather than restrictive.

Claims: Claim of Patent: 1. A procedure for the production of astaxanthin and related carotenoids, astaxanthin esters, chitin, proteins and meat from plants, algae, bacteria, krill, shrimps and other crayfish and Crustacea, c h a r a c t e r i z e d by it consisting of the following steps: 1: extraction of proteins, astaxanthin and related carotenoids, and astaxanthin esters by boiling in lye, 2: residue from step 1 is dried in concentrated formic acid then dissolved in anhydride formic acidor another strong acid, filtered, then the filtrate has water added so as to allow the precipitation of the purified chitin, 3: recovery of astaxanthin and related carotenoids, astaxanthin esters, chitin and proteins from the extract in step 1 in one or more product fractions. 2. A procedure in accordance with Claim 1, . c h a r a c t e r i z e d by the above-mentioned extraction occurring in two steps, where the mentioned lye is 1.0 N of a strong base, where the extracts from the two steps have an acid added until all astaxanthin, related carotenoids, astaxanthin esters and proteins are precipitated, after this the unprecipitated compounds are separated from the extracts in a product fraction by decanting and filtration or by equivalent methods or combination of methods. 3. A procedure in accordance with Claim 1, c h a r a c t e r i z e d by the above-mentioned extraction occurring in two steps, where the mentioned lye is 1.0 N of a strong base, where the extracts from the two steps are removed for cooling, whereupon the extracts are separated into three phases which are divided by floating off the phase with the lowest specific weight and decanting (or the equivalent) the phase with the intermediate specific weight from the phase with the highest specific weight; where the phase with the lowest specific weight is neutralized by the addition of an acid which causes the formation of two liquid phases, which are separated, and the oily phase consisting of astaxanthin esters is taken care of; where the phase with the highest specific weight is neutralized by an acid and solids consisting of astaxanthin and related carotenoids are filtered away and taken care of in a known manner; and where the phase with the intermediate specific weight is neutralized and followed by the addition of marine oil, Ca 2+ -ions, S 2~ -ions and (NH 4 ) 2 HP0 4 for the precipitation of F ~ - ions and the heavy metal contaminants, where this precipitate is removed from the solution by floating or the equivalent, after this the solution is acidified prior to the precipitation of the purified proteins followed by the separation of these proteins from the solution by filtration or an equivalent method. 4. A procedure in accordance with Claim 1, c h r a c t e r i z e d by the above-mentioned extraction occurring in four steps, where the mentioned lye in the first two steps is 0.01 N of a strong base, where the mentioned lye in the last two steps is 1.0 N of a strong base; where the extracts from the first two steps are neutralized and this is followed by the addition of marine oil, Ca + -ions, S " -ions and (NH 4 )_HP0 4 for the precipitation of F ~ -ions and the contaminants, where this precipitate is removed from the solution by floating or the equivalent, after this the solution is acidified prior to the precipitation of the purified proteins followed by the separation of these proteins from the solution by filtration or an equivalent method, where the extracts from the last two steps have an acid added until all astaxanthin, related carotenoids, astaxanthin esters and proteins are precipitated, after this the unprecipitated compounds are separated from the extracts in a product fraction by decanting and filtration or by equivalent methods or combination of methods. 5. A procedure in accordance with Claim 1, c h a r a c t e r i z e d by the above-mentioned extraction occurring in four steps, where the mentioned lye in the first two steps is 0.01 N of a strong base, where the mentioned lye in the last two steps is 1.0 N of a strong base; where the extracts from the first two steps are neutralized and this is followed by the addition of marine oil, Ca 2+ -ions, S 2~ -ions and (NH 4 ) 2 HP0 4 for the precipitation of F~-ions and the contaminants, where this precipitate is removed from the solution by floating or the equivalent, after this the solution is acidified prior to the precipitation of the purified proteins followed by the separation of these proteins from the solution by filtration or an equivalent method, where the extracts from the last two steps are removed for cooling, whereupon the extracts are separated into three phases which are divided by floating off the phase with the lowest specific weight and decanting or the equivalent the phase with the intermediate specific weight from the phase with the highest specific weight; where the phase with the lowest specific weight is neutralized by the addition of an acid which causes the formation of two liquid phases, which are separated, and the oily phase consisting of astaxanthin esters is taken care of; where the phase with the highest specific weight is neutralized by an acid and solids consisting of astaxanthin and related carotenoids are filtered away and taken care of in a known manner; and where the phase with the intermediate specific weight is neutralized and followed by the addition of marine oil, Ca 2+-ions, S2--ions and (NH 4 )_HP0 4 for the precipitation of F ~ -ions and the heavy metal contaminants, where this precipitate is removed from the solution by floating or the equivalent, after this the solution is acidified prior to the precipitation of the purified proteins followed by the separation of these proteins from the solution.

Inventors: MIKALSEN GUNNAR, MIKALSEN, GUNNAR +DI
Applicants: MIKALSEN ESTER & HF, MIKALSEN, ESTER +HF
Application year: 1989
Main classification: A23J1
IPC codes: A22C29, A23J1, A23L1, C09B61