JPS60207567A - Production of lipid containing carotinoids from algae - Google Patents

Abstract

PURPOSE:Carbon dioxide is brought into contact with algae under supercritical conditions to extract lipids containing carotinoids such as beta-carotin in such a state as chlorophyll is selectively excluded to enable the extraction of lipids free from chlorophyll. CONSTITUTION:Carbon dioxide is used as an extractant and pressurized with compressor 3, heated with heat-exchanging type heater 4 to prepare carbon dioxide gas under supercritical conditions (over 75.2kg/cm<2> of the critical pressure, over 31.1 deg.C of the critical point) and the gas is fed into the extraction tank 2. The carbon dioxide under the supercritical conditions is brought into contact with a powder of spirulina in the extraction tank 2 to extract lipids free from chlorophyll. The carbon dioxide containing lipids are reduced in pressure by passing through pressure-controlling valve 6, then the lipids 7 is collected in the tank 5 by feeding the carbon dioxide reduced in pressure continuously into the recovery tank 5.

Description

[Detailed Description of the Invention] The present invention relates to a method for producing carotenoid-containing lipids from algae such as spirulina and chlorella, and a method for quickly and easily producing lipids from algae while selectively excluding chlorophyll. Provide a method to extract.

Algae, which is the object of the present invention, is a general term for lower plants that grow in water, have assimilable pigments, and carry out photosynthesis, and are taxonomically classified as follows. However, the numbers in parentheses indicate specific examples.

1. Blue-green algae (Spirulina, Agaricella) 2. Green algae (Aonori, Mil, Chlorella, Scenedesmus) 3. I-Algae M (Kelp, Wakame, Hijiki, Arame) 4. Red Algae (Amanita, Amanita, Funori) 5. Diatom M1
6, Brassinophyceae (
Pyramimonas, Platymonas) 7.1 Colored Algae M (7 Usenmo, 7 Sinasimidoro) 8. Golden Algae (Hikarimo) 9. Cryptoalgae (Chloomonas) 10. Green Flagellate (Chatonella) Hydinoflagellate M (Seratium, Goniolax) ) 12, Euglena algae (Euglena, Colacium) 13, Ring algae M
Among the algae mentioned above, kelp, seaweed, hijiki, chinensis, and amanita are familiar to the Japanese diet, but this pond also has spirulina (in the formal system of plant taxonomy, Blue-green algae - order Nostocales - suborder Nostocales -
Yuremo Jinkoma, Chlorella (Green Algae - Chlorococcumales - Chlorellaceae) and Scenedesmus (Green Algae - Chlorococcales - Coelastorum Family) etc. have a protein content of more than 60% per dry algal body, so initially It was expected to have potential as a new protein resource.

However, centrifugation or over-cropping is required when harvesting from the culture solution, and the processing capacity is naturally limited and the production price is high (for example, the production price of spirulina is higher than that of soybeans and fishmeal for feed). It is about 20 times that of
In addition, Chlorella has a tough cell wall and has problems with digestibility, so it is currently used as a processed health food or food additive rather than as a food resource. be.

In other words, chlorella, spirulina, etc. are made into tablets as health medicines, and the latter in particular is used in some fish feed as a coloring agent for Nishikigoi, goldfish, and prawns. The blue pigment contained in it, called phycocyanin, is commercialized as a natural coloring agent.

In general, the lipids contained in algae consist of neutral lipids and complex lipids, the former containing triglycerides, carotenoids, hydrocarbons, free fatty acids, etc., and the latter containing phospholipids, glycolipids, sulfur-containing lipids, etc.; Contains abundant carotenoids.

For example, 100 g of spirulina contains 650 mg of carotenoids, and the main components of these carotenoids are β-carotene, zeaxanthin, and sugar carotenoids, and β-carotene is absorbed in the intestines and becomes vitamin A (
That is, β-carotene and vitamin A are nutritionally important because they are precursors of vitamin A), and both β-carotene and vitamin A have recently attracted attention for their cancer-suppressing effects.

The yellow carotenoid zeaxanthin is used as a color enhancer for carp and goldfish, and the red sugar carotenoid may have antibacterial activity, so it is expected to be used as a drug.

Furthermore, the glyceroglycolipids contained in the complex lipids of Spirulina may have antitumor effects, and are expected to be used as medicines.

However, at present, research on algae is often focused on proteins and polysaccharides, and there are few attempts to actively extract lipids from algae and utilize them. For example, in the production of the protein pigment phycocyanin from Spirulina mentioned above, phycocyanin is recovered by elution into water, but the residue containing lipids is separated and removed by centrifugation and then discarded, or at most It is only used as feed.

However, as mentioned above, the lipids contained in algae are rich in useful components, including carotenoids such as β-carotene, which have anticancer effects. It is possible to produce high value-added products such as foods, pigment oils, and medicines.

In this way, the extraction of lipids from algae opens the door to processed foods and nutritionally enriched feed, but on the other hand, algae that carry out photosynthesis naturally contain a large amount of chlorophyll (for example, spirulina). (25,000 mg of chlorophyll is contained per 1,008 mg of chlorophyll), and it is difficult to avoid the contamination of this large amount of chlorophyll at the same time during lipid extraction.

For example, Japanese Patent Publication No. 48-16189 discloses a method for extracting lycopene, a hydrocarbon carotenoid, from fungi classified as algae using n-hexane or petroleum ether. As can be seen, the method for extracting carotenoid-containing lipids from algae is a multistage solvent extraction method using common organic solvents such as ethanol, acetone, ether, chloroform, methylene chloride, or n-hexane. However, since chlorophyll is easily soluble in the above-mentioned organic solvents, chlorophyll must also be extracted at the same time when extracting lipids, and the reality is that there is no solvent extraction method that can selectively remove only chlorophyll.

By the way, the chlorophyll contained in this algae causes photosensitivity, and for example, cases of damage to livestock due to feeding have been known for a long time, and it has recently been reported that eating too many chlorella tablets can cause damage to humans. It became widely known.

This is directly caused by 7eo7olbide a produced by the decomposition of chlorophyll, which causes photosensitivity such as sun-induced dermatitis due to a prior sensitization reaction, but this pheophorbite a The results of a phototoxicity test (after oral administration, irradiation with light for 60 minutes at an illuminance of 20,000 lux the next day) showed that LDl was 45.5 + 100% body weight.
g or more, MLD50 is said to have a value of 12 mg/100 g or more (Japanese Society of Agricultural Chemistry, 1980, V
ol, 54, No. 9.721-726).

Moreover, chlorophyll tends to be acidic, and when organic solvents coexist with it, it becomes easier to produce 7eo7olbide a, so from an overall perspective, lipids extracted from algae can be used as they are as health foods or pigment oils. Problems remain.

The present invention relates to lipid extraction from algae, which has not been actively attempted in the past, and aims to extract lipids while excluding chlorophyll, which could not be achieved with the above-mentioned organic solvent extraction method, and in order to achieve this purpose, Carbon dioxide is used as an extractant, and the critical point of carbon dioxide is 75.2 kg/cm2・31
, carbon dioxide is produced in the supercritical gas range exceeding 1°C, and this carbonic acid is brought into contact with algae to selectively eliminate chlorophyll, and lipids containing carotenoids such as β-carotene are carbonated from within the algae. It is configured to extract into the gas.

That is, the present inventors have newly discovered the fact that chlorophyll is not eluted into carbon dioxide gas in the supercritical gas region A shown in FIG. 2 due to its affinity, and based on this discovery, the present invention has been developed. This is the completed version.

Hereinafter, the various steps for practically implementing this extraction method will be explained in detail in (E) using Spirulina as an example, with reference to FIG.

(B) Pre-treatment process Spirulina has the property of floating and clumping into blocks, so the algae are harvested by a filtration method, and then dried using a low-temperature spray dryer or freeze-drying method to form a blue-black powder. do.

This powder is supplied as a raw material to the following extraction process.

(b) Raw material storage step The powder obtained in the pretreatment step is stored in the extraction tank 2 as the raw material 1.

(c) Extractant supply step Carbon dioxide gas is used as the extractant, and it is pressurized with a pressurizer (compressor or pump, etc.) 3 and heated with a heat exchange type heater 4 to reach a critical pressure of 5.2 kg/am. ”(Pl)
At a supercritical pressure exceeding , and at a critical temperature of 31.1°C (
A supercritical gas region A where the supercritical temperature exceeds T,
The carbon dioxide (see Figure 2) is supplied to the extraction tank 2.

(d) Lipid extraction step Supercritical carbon dioxide gas is brought into contact with the Spirulina powder 1 in the extraction tank 2, and the lipids contained in the powder are extracted into the supercritical carbon dioxide gas with chlorophyll removed.

(e) Lipid recovery process Carbon dioxide in a supercritical state containing lipids is reduced in pressure by passing through the pressure regulating valve 6 for setting the primary pressure from the extraction tank 2, and then continuously injected into the recovery tank 5. Pressure 5,
Bring the pressure below 2 kg/cm2.

Carbon dioxide gas below this critical pressure loses its affinity for a single lipid, so it easily separates the lipids, and the recovery tank 5 contains a rich amount of υtinoids mainly composed of β-carotene, and at the same time contains chlorophyll. The lipids 7 that do not contain the lipids 7 can be quickly recovered.

In the figure, numeral 10 is a filter, 11 is a discharge valve, 12 is a check valve, 9 is a carbon dioxide gas supply source, and 13 is a carbon dioxide gas supply valve.

Incidentally, in order to perform each of the above steps (a) to (e) more efficiently, at least one of the following steps (v) to (v) may be employed.

(f) The cell membrane of Spirulina is thin and soft, and cell components can be easily eluted. After the lipids are eluted, the residual matter that is insoluble in water is centrifuged and subjected to a drying process such as low temperature or freezing.
It is then passed through a pressurizer 3 and a heater 4 to be recirculated to the extraction tank 2.

(h) The carbon dioxide gas after lipid separation is circulated from the filter 10 to the pressurizer (compressor) 3 without the heat exchanger 8.

(S) Supplying liquefied carbon dioxide gas from the supply source 9 to the pump 3,
The gaseous carbon dioxide gas is discharged from the collection tank 5 to the outside through the gaseous carbon dioxide discharge valve 11 and is not circulated to the pump 3.

(v) The carbon dioxide gas containing lipids is intermittently transferred from the extraction tank 2 to the recovery tank 5.

Therefore, the results of applying the method of the present invention, which is a combination of the above-mentioned steps, to freeze-dried products or low-temperature-dried products of Spirulina and Chlorella will be sequentially described below.

<Example 1> Freeze-dried spirulina was placed in an extraction tank of 75 brass, and each sample was placed in a supercritical gas region at an average flow rate of 1.0 kg/br and temperature and pressure as shown in Table I. 3 using carbon dioxide gas
Lipid extraction was performed over time, and the extracted lipid amount and residual lipid amount obtained from each sample are summarized in Table 1.

Two extraction experiments were conducted under each extraction condition to confirm the reproducibility of the extraction amount.

Here, the total lipid amount is determined by quantifying the crude fat contained per 100 g of sample using the Bligh-Dyer method, and naturally shows the same value for each sample.

Furthermore, the amount of extracted lipids is determined by determining the amount of lipids in the extracted portion recovered using supercritical carbon dioxide gas, and the amount of residual lipids is determined by quantifying the amount of lipids remaining in the prepared sample after extraction using the Bligh-Diameter method, each converted to 1008. This is what I did.

According to the above extraction example, the pressure is kept constant (e.g.
250kg/c+a2 constant), and when the temperature is increased (4
0 → 60 → 80℃), the total extraction amount increases, but even if the temperature is kept constant (for example, 40℃ constant) and the pressure is increased (
250 → 400 kg/c wheel 2), it can be seen that there is no significant increase in the total extraction amount.

Therefore, when extracting lipids from frozen Spirulina crystals using carbon dioxide gas in the supercritical gas region, the economic efficiency of the extraction can be increased by increasing the temperature and lowering the pressure.

<Example 2> A low-temperature dried product of Spirulina, which was dried so that the components would not undergo thermal denaturation, was placed in an extraction tank 41, and sample HL1A 91
cm+ Sako16'J 0 and If 4tJA111. −
When extraction was carried out for 1.5 hours at an average flow rate of 7°0 kg/br using carbon dioxide gas in the supercritical gas range of /-2, the following values were obtained.

Total lipid amount 7.5g/sample 100g Extracted lipid amount 0.4g/sample 100g Residual lipid amount 4.7g/sample 100g From the results of Examples 1 and 2, calculate the extracted lipid amount per unit weight of carbon dioxide gas and compare. (Both are compared at a temperature of 40°C and a pressure of 400kg/c+e2 extraction), and the average flow rate for the freeze-dried product is 1. 100 samples after flowing at Okg/br for 3 hours
Since 1.29-1.49g can be extracted per g, 1.29-1.49g/l, Okg/brX 31+r
= 0.43 to 0.50g/CO21kg, whereas the low temperature dry product shows an average flow rate of 7.0 and 1 at 8/hr.
．． Since 0.4g can be extracted per 100g of sample after 5 hours of flow, 0.4g/7.0kg/brX 1.5br=0.04
The numerical value is g/CO21kg.

<Example 3> A freeze-dried product of chlorella was stored in an extraction tank of 75 mρ, and each sample amount was taken at an average flow rate of 1.1 kg/I+r and the temperature and pressure were adjusted to the values shown in Table ■ to extract carbon dioxide gas in the supercritical gas range. using 3
Lipid extraction was carried out over a period of time, and the extracted lipid amount and residual lipid amount obtained from each sample are summarized in Table 3.

In order to confirm the reproducibility of the extraction amount under each extraction condition, two extraction experiments were conducted.

The total lipid amount and extracted lipid amount in the above table are the same as in Example 1.

In addition, in the above example, as in Example 1, the amount of light oil output increases when the pressure is held constant and the temperature is increased, but even when the temperature is held constant and the pressure is increased, the amount of light oil output increases significantly. There isn't.

<Example 4> Chlorella was dried to 120-130% using a spray dryer.
The high-temperature dry product dried at ℃ was placed in a 4ρ extraction tank, and a sample amount of 978.3g was placed at a temperature of 40℃ and a pressure of 400kg/am2.
Using carbon dioxide gas in the supercritical gas region, the average flow rate is 3. Okg
/l+r for 7 hours, the following values were obtained.

Total lipid amount 2.5 g/sample 100g Extracted lipid amount 0.95 g/sample 100g Residual lipid amount 1
．． 5 g/100g of sample The total lipid content of the above high temperature dried product is 10.4g/100g of sample for the freeze-dried chlorella product.
Compared to
This can be presumed to be due to heat denaturation and decomposition of a considerable portion of the lipids contained in the raw material chlorella.

Therefore, the amount of lipid extracted per unit weight of carbon dioxide is also 0,
05 g/C021kg (0.95g/3.0 to 8/
Calculated from hr
It remains at a considerably lower value compared to .

As described above, according to various examples, lipids can be quickly and easily extracted from dried products of Spirulina and Chlorella using carbon dioxide gas in the supercritical gas region, but the composition of the extracted lipids is It was necessary to analyze this and confirm the presence of chlorophyll in the lipid, and for this confirmation, tests (1) to (3) were conducted.

(1) Quantitative test of neutral lipids and complex lipids in extracted lipids <Test Example 1> Lipids extracted from low-temperature dried Spirulina (extraction amount of Example 2) 1 g of 75II was added to silica gel (Wokogel).
The lipids were sequentially eluted using a column (2X4cI11) filled with C-200) and 50d of chloroform and 50d of methanol as developing solutions, and the neutral lipid fraction (reaa form fraction) and complex lipid fraction (methanol fraction) were separated. The following results were obtained by calculating the quantitative ratio of each fraction by gravimetric method.

Spirulina low temperature dried product extracted lipids (total amount 75B) Neutral lipids 71+f1g (weight ratio 94.6%) Complex lipids 3m
g (weight ratio 4.0%) According to the above test, most of the extracted lipids are neutral lipids consisting of glycerides, carotenoids, etc., and only a small amount of complex lipids consisting of glycolipids, etc. are included. can be confirmed.

<Test Example 2> 78II 1 g of lipid extracted from a high-temperature dried product of Chlorella (extracted amount in Example 4) was subjected to column chromatography in the same manner as in Test Example 1, and the following results were obtained.

Chlorella high temperature dried product extracted lipids (total amount 78cm) Neutral lipids 65,111g (weight ratio 83.5%) Complex lipids 4.
6 mg (weight ratio 5.9%) According to the above component ratio, the amount of neutral lipids is slightly smaller than that of Spirulina, and the amount of complex lipids is correspondingly larger.

(2) Qualitative test of lipid composition <Test Example 3> Among the lyophilized Spirulina products used in Example 1, 40
For each of the total lipids, residual fats, and carbon dioxide gas-extracted lipids obtained from the extraction conditions of 0 kg/cu2/40°C,
A thin layer of commercially available silica gel 60F254 (Merck) was used and petroleum ether/ether/acetic acid (90:10
The resulting product was subjected to thin layer chromatography using the developing solvent of 1) to obtain Figure 3 (A).

In addition, since the freeze-dried product and the low-temperature-dried product of Spirulina both showed similar results, the above-mentioned extract was used as a representative example of the extraction test.

According to the above diagram A, the lipids that make up Spirulina are β-
It consists of carotene, chlorophyll, yellow carotenoid, triglyceride, etc. (see the development state of total lipid amount). When extracted with supercritical carbon dioxide, β-carotene, yellow carotenoid, triglyceride, cholesterol, etc. are extracted from the total lipids in carbon dioxide gas. However, it can be seen that chlorophyll becomes residual lipid and remains at the origin.

Therefore, in order to confirm the detailed composition of the lipids that remain near the origin in the thin layer chromatogram above, this carbon dioxide extract was further extracted using chloroform/methanol/water (chloroform/methanol/water).
65:25:4) as a developing solvent to obtain Figure 3(B).

According to the above diagram B, in addition to those shown in diagram A, the components contained in the total lipids include red sugar carotenoids such as myxoxanthophyll and osilaxanthin, and glyceroglycolipids such as monoglycosyl diglyceride, monoglycosyl diglyceride, and diglycosyl monoglyceride. can be mentioned.

In addition, near the development tip of the thin-layer chromatogram, overlapping development spots of chlorophyll and yellow carotenoid appear in the case of total lipid content, but only yellow development spot of yellow carotenoid appears in the case of supercritical carbon dioxide extraction. It can be reconfirmed that chlorophyll remains in the residual lipids and does not transfer to the extracted lipids.

Therefore, in lipid extraction using supercritical carbon dioxide gas, neutral lipids, especially carotenoids such as β-carotene and yellow carotenoids, can be recovered in high yield, and at the same time, chlorophyll can be removed and retained as a residue. can.

In addition to chlorophyll, the residual lipids are concentrated in glyceroglycolipids and red sugar carotenoids, and the former may have antitumor activity, and the latter may have antibacterial activity, and both may be used as drugs. You can expect it.

<Test Example 4> Chlorella freeze-dried product used in Example 3 (250 kg/
For each of the residual lipids obtained from am2・40℃ extraction) and the carbon dioxide extracted lipids, the silica gel 60
A thin plate of F 254 (Merck) was used and two developing solvents were used: petroleum ether/ether/acetic acid (90%
:10:1) and chloroform/methanol/water (65
:25:4) to obtain a thin layer chromatogram.

In addition, since chlorella freeze-dried products and high-temperature dried products both showed similar results, the above-mentioned extract was used as a representative example of the extraction test.

Figure 4 (A) shows the result developed in a petroleum ether/ether/acetic acid system; β-carotene, triglycerides, yellow carotenoids, hydrocarbons, cholesterol esters, etc. are transferred to the carbon dioxide extract, and almost no residual lipids are present. On the other hand, chlorophyll remains in the residual lipid fraction and does not transfer to the extracted lipid fraction.

Figure 4 (B) shows a further development test in a chloroform/methanol/water system in order to clarify the lipid components that remain near the origin in Figure A above, and shows that chlorophyll does not migrate to the extracted lipid fraction. can be confirmed.

In addition, the neutral lipids contained in chlorella contain more hydrocarbons, free fatty acids, and carotenoids such as lutein than those in spirulina, while the complex lipids contain many types of glyceroglycolipids, sulfur-containing lipids, etc. It can be confirmed that

(3) Qualitative test of carotenoids contained in extracted lipids According to Test Example 2 (Figure B in Figure 3), the pigment components contained in the total lipids consist of yellow carotenoids, chlorophyll, myxoxanthophyll, and osillaxanthin. However, we can mainly recover yellow carotenoids.

Test Example 5> Therefore, 400 kg/cn+ of lyophilized Spirulina product
For extracted lipids mainly composed of yellow carotenoids obtained from 2.40℃, acetone/petroleum ether (3:17)
Using the developing solvent of silica gel 60F254 (M
When the mixture was developed on a thin layer plate prepared using erck), the thin layer chromatogram shown in FIG. 5 was obtained.

From FIG. 5, it can be seen that the extracted carotenoids are mainly composed of β-carotene, zeaxanthin, echinenone, 3'-hydroxyechinenone, β-cryptoxanthin, etc.

As mentioned above, taking Spirulina and Chlorella as an example,
If lipids are extracted from these with carbon dioxide gas in the supercritical gas region, chlorophyll can be removed and lipids rich in carotenoids can be obtained. As such, the present invention can be applied to all the algae mentioned above, which contain a large amount of chlorophyll in the algal bodies because they photosynthesize in water, and the effects of the present invention when applied to algae in general are as follows.

(1) Taking advantage of the property that carbon dioxide in a supercritical state does not elute chlorophyll due to its affinity,
By bringing supercritical carbon dioxide into contact with algae, lipids can be extracted from algae with chlorophyll removed, so by ingesting the extracted lipids as in the past, the decomposition products produced from chlorophyll can be absorbed into the human body. It eliminates the stain that causes photosensitivity when taken into the skin.

Moreover, this selective removal of chlorophyll from lipids could not be achieved using conventional solvent extraction methods.
If the present invention is applied to algae, only useful lipids including β-carotene etc. can be easily and safely extracted from algae, and this can be used as the main health food or pigment oil.

Furthermore, if conventional solvent extraction is performed on the extraction residue after carbon dioxide gas extraction of the present invention is applied to algae, a chlorophyll concentrate can be obtained. It can be used as

Moreover, while chlorophyll is an indirect cause of the aforementioned photosensitivity, it also has the pharmacological effect of lowering cholesterol in the body if taken into the human body in an appropriate amount, so it can be used as a drug.

(2) The lipid extracted from algae according to the present invention is β-
Contains a large amount of carotene and other carotenoids, so it can be used as a medicinal nutritional supplement or pigmented feed. It can be used as an improver for chicken meat color and egg color.

Masuko, the residue Φ after extraction is enriched with chlorophyll, sugars, and glycolipids1), and is expected to have various medicinal effects, so the residue can also be used. , thereby further increasing the added value of the extraction operation of the present invention.

(3) Selectively remove chlorophyll from algae with a single step of pressurizing and heating liquefied carbon dioxide gas to a supercritical state that exceeds the critical point, bringing it into contact with algae, and then removing carbon dioxide gas. It is possible to extract and separate the lipids in this state and manufacture them directly.

Furthermore, since the extractant is carbon dioxide gas in a supercritical state, separation from the desired lipid can be carried out very smoothly.

That is, in conventional solvent extraction methods, in order to selectively obtain lipids from algae, the extraction must be carried out in multiple stages using different types of solvents, whereas the present invention requires only one stage operation.

Therefore, the labor required for lipid production can be minimized, the production cost can be reduced, and the entire equipment can be largely integrated into Funbaku V.

(4) Although carbon dioxide in a supercritical state is gaseous, it has a very high density close to that of a liquid, and also has a high diffusion coefficient, which is an attribute of gas, so it It can spread widely to every nook and corner, increase both the extraction surface area and the contact flow rate, and has a high elution power like a liquid.

Therefore, the extraction efficiency is high and the amount extracted can be increased.

(5) Since carbon dioxide gas is used as the extractant, the lipids are kept in an inert atmosphere of carbon dioxide gas from the beginning to the end of the extraction operation, so there is no rancidity or deterioration, and the quality of extracted lipids, especially carotenoids, is improved. Can be maintained well.

Inert carbon dioxide gas also poses no risk of flame or explosion compared to traditional solvent extraction methods, making it extremely safe to operate.

Furthermore, even if carbon dioxide gas is discharged or leaked into the atmosphere, there is no risk of polluting the atmosphere or the environment.

(6) In conventional solvent extraction methods, there is a risk that organic solvents harmful to the human body, such as chloroform and methylene chloride, may remain in the lipids, but in the method of extraction with carbon dioxide gas as in the present invention, Since carbon dioxide gas is a gas, it is likely that this gas will remain in the product (even if a small amount remains, it is highly safe), making lipid products safe and non-toxic as health foods or pigment oils. It can be used in

(7) Since the present invention can be applied to algae in general, with Spirulina and Chlorella as representatives, there is a possibility of application to plants that carry out photosynthesis and therefore have chlorophyll. It is expected that useful products can be created by extracting the

[Brief explanation of the drawing]

The drawings relate to the present invention; Figure 1 is a flowchart of the present invention, and Figure 2 shows the pressure and pressure of carbon dioxide gas used as an extractant.
Temperature state diagram, Figure 3 (A) (B), Figure 4 (A) (B)
Figures 4 and 45 are thin layer chromatograms of lipids tested using various developing solvents. 1... Algae, 2... Extraction tank, 3... Pressurizer, 4...
... Heater, 5 ... Recovery tank, 6 ... Pressure regulating valve, PI.
...Critical pressure, T1...Critical temperature, A...Supercritical gas region. Patent applicant: Iwatani Sangyo Co., Ltd. Osaka Hydrogen Industry Co., Ltd. Japan Spirulina Co., Ltd. Yaeyama Shokusan Co., Ltd. Figure 1 Figure 2 Figure 4 (A) Figure 4 (B) Continued from page 1 @ Inventor Murai Katsuyuki 0 Inventor Tetsuo Kaji Yama 15-101 Suita Nagano Higashi 6-4 Shimanokaku, Aramaki, Itami City Procedural amendment (spontaneous) Day 1, Indication of case Patent Application No. 62595 of 1982 2, Invention Name of method for producing lipids containing carotenoids from algae 3. Person making the amendment Related to the case Patent applicant: 18 Iwatani Sangyo Co., Ltd. (and 8 others) 4. Agent: 5. Date of amendment order: Showa year Sent on Monday → 1. The scope of claims will be amended as follows. Note 1. As an extractant, gas is pressurized and heated to produce carbon dioxide gas in the supercritical gas range with a critical pressure of 5.2 kg/d and a critical temperature of 31.1°C, and this carbon dioxide gas is brought into contact with algae. , Method 2 for producing lipids containing carotenoids from algae, characterized by extracting lipids containing carotenoids from algae into carbon dioxide gas while selectively excluding chlorophyll, page 4, line 14 of the specification. Correct "β-carotene" written from line 15 to "β-carotene". 3. "Phycocyanian" listed on page 5, line 9 of the specification will be corrected to "phycocyanin." 4. Change “use” written in line 12 of page 5 of the specification to “
Correct to "Use". 5. "Carbon dioxide" described on page 8, line 10 of the specification is "
Correct to "carbon dioxide gas". 6. Amend "to administer" written in line 4 on page 11 of the specification to "apply." 7. “Lyophilized product 75” described on page 12, line 1 of the specification
Correct "Ill" to "75mj!j for lyophilized product." 8. Correct "light oil yield" listed in lines 9 and 11 of page 13 of the specification to "extracted lipid amount." 9. rWokoge described on page 17, line 16 of the specification
Correct lJ to rWakogelJ. 10. Correct the "total lipid amount" listed on page 19, line 14 of the specification to "total lipid." 11. The "yellow carotenoid" listed in the second line of page 23 of the specification is corrected to "the yellow carotenoid when extracted with supercritical gas." 12. "Flame" described on page 27, line 16 of the specification is replaced with "
Fire” will be corrected. that's all

Claims (1)

[Claims] 1. Use carbon dioxide gas as an extractant, pressurize and heat the carbon dioxide gas to reach critical pressure. 5.
kg/cI112, carbon dioxide in the supercritical gas range with a critical temperature of over 31.1°C is brought into contact with the algae, and while chlorophyll is selectively removed, lipids containing carotenoids are carbonated from within the algae. A method for producing lipids containing carotenoids from algae, which is characterized by extraction into gas.