NO122123B - - Google Patents
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- Publication number
- NO122123B NO122123B NO16730967A NO16730967A NO122123B NO 122123 B NO122123 B NO 122123B NO 16730967 A NO16730967 A NO 16730967A NO 16730967 A NO16730967 A NO 16730967A NO 122123 B NO122123 B NO 122123B
- Authority
- NO
- Norway
- Prior art keywords
- nucleate
- alkali
- nucleic acid
- alcohol
- precipitated
- Prior art date
Links
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 15
- 102000039446 nucleic acids Human genes 0.000 claims description 15
- 108020004707 nucleic acids Proteins 0.000 claims description 15
- 150000007523 nucleic acids Chemical class 0.000 claims description 15
- 239000003513 alkali Substances 0.000 claims description 12
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 7
- 239000011734 sodium Substances 0.000 claims description 7
- 229910052708 sodium Inorganic materials 0.000 claims description 7
- 241000251468 Actinopterygii Species 0.000 claims description 6
- 230000001476 alcoholic effect Effects 0.000 claims description 6
- 239000008267 milk Substances 0.000 claims description 6
- 210000004080 milk Anatomy 0.000 claims description 6
- 235000013336 milk Nutrition 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 150000003839 salts Chemical class 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 5
- 241001465754 Metazoa Species 0.000 claims description 4
- 238000010438 heat treatment Methods 0.000 claims description 4
- 229910052751 metal Inorganic materials 0.000 claims description 4
- 239000002184 metal Substances 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 239000000725 suspension Substances 0.000 claims description 3
- 235000013311 vegetables Nutrition 0.000 claims description 2
- 235000011132 calcium sulphate Nutrition 0.000 claims 2
- 239000001175 calcium sulphate Substances 0.000 claims 1
- 239000007788 liquid Substances 0.000 description 11
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 10
- 239000002244 precipitate Substances 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 235000011121 sodium hydroxide Nutrition 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 2
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 229910052802 copper Inorganic materials 0.000 description 2
- 239000010949 copper Substances 0.000 description 2
- 238000010908 decantation Methods 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 239000011777 magnesium Substances 0.000 description 2
- 229910052749 magnesium Inorganic materials 0.000 description 2
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000011707 mineral Substances 0.000 description 2
- 235000010755 mineral Nutrition 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 235000008733 Citrus aurantifolia Nutrition 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 239000005905 Hydrolysed protein Substances 0.000 description 1
- CPLXHLVBOLITMK-UHFFFAOYSA-N Magnesium oxide Chemical group [Mg]=O CPLXHLVBOLITMK-UHFFFAOYSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 235000011941 Tilia x europaea Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000012213 gelatinous substance Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000004571 lime Substances 0.000 description 1
- VTHJTEIRLNZDEV-UHFFFAOYSA-L magnesium dihydroxide Chemical compound [OH-].[OH-].[Mg+2] VTHJTEIRLNZDEV-UHFFFAOYSA-L 0.000 description 1
- 239000000347 magnesium hydroxide Substances 0.000 description 1
- 229910001862 magnesium hydroxide Inorganic materials 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 150000003467 sulfuric acid derivatives Chemical class 0.000 description 1
- 230000007306 turnover Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/56—Ring systems containing three or more rings
- C07D209/80—[b, c]- or [b, d]-condensed
- C07D209/82—Carbazoles; Hydrogenated carbazoles
- C07D209/88—Carbazoles; Hydrogenated carbazoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the ring system
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Indole Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Meat, Egg Or Seafood Products (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Nitrogen And Oxygen As The Only Ring Hetero Atoms (AREA)
Description
Fremgangsmåte til utvinning av ren nukleinsyre. Method for extracting pure nucleic acid.
Foreliggende oppfinnelse angår en fremgangsmåte for fra animalske eller ve-getabilske råmaterialer, og spesielt fra kold fiskemelke, å utvinne ren nukleinsyre. The present invention relates to a method for extracting pure nucleic acid from animal or vegetable raw materials, and in particular from cold fish milk.
Det kjennes fremgangsmåter til behandling av forskjellige animalske stoffer for å utvinne nukleinsyre, særlig ved behandling av animalske materialer med natronlut og utfelling av nukleat med alkohol. Disse fremgangsmåter gir forurensede nuk-leinsyrer. Hvis man ønsker å få ren nukleinsyre, må man utsette materialet for en behandling for å befri det for de organiske og mineralske forurensninger, og utføre ekstrahering og utfelling først med tem-melig rene produkter. Tysk patent nr. 304.003 beskriver således en fremgangsmåte for rensning av hydrolyserte proteiner ifølge hvilken hydrolyseoppløsningene behandles med kalsium- eller magnesium-hydroksyd eller en blanding av disse for å eliminere visse forurensninger som gela-tinøse stoffer eller stoffer som med kalk eller magnesia danner uoppløselige salter. Selve proteinstoffene påvirkes ikke av denne behandling. Methods are known for treating various animal substances to extract nucleic acid, in particular by treating animal materials with caustic soda and precipitation of nucleate with alcohol. These methods give contaminated nucleic acids. If one wishes to obtain pure nucleic acid, one must subject the material to a treatment to rid it of organic and mineral contamination, and carry out extraction and precipitation first with relatively clean products. German patent no. 304,003 thus describes a method for the purification of hydrolysed proteins according to which the hydrolysis solutions are treated with calcium or magnesium hydroxide or a mixture of these to eliminate certain impurities such as gelatinous substances or substances which with lime or magnesia form insoluble salts . The protein substances themselves are not affected by this treatment.
Fremgangsmåten etter oppfinnelsen, The method according to the invention,
hvor ren nukleinsyre utvinnes fra de nevnte råstoffer, og hvor råstoffer, f. eks. på forhånd opphakket fersk fiskemelke behandles med alkalilut og alkalinukleat felles ut med alkohol, karakterisert ved at det utfelte alkalinukleat renses ved behandling under oppvarmning med kalsiumsulfat i nærvær av vann og ved at det fra den rensede oppløsning av alkalinukleat felles ut praktisk talt ren nukleinsyre ved hjelp av en blanding av alkohol og saltsyre, hvor- where pure nucleic acid is extracted from the aforementioned raw materials, and where raw materials, e.g. previously chopped fresh fish milk is treated with alkali lye and alkali nucleate is precipitated with alcohol, characterized in that the precipitated alkali nucleate is purified by treatment while heating with calcium sulfate in the presence of water and that practically pure nucleic acid is precipitated from the purified solution of alkali nucleate using of a mixture of alcohol and hydrochloric acid, where-
etter, om ønsket, den erholdte nukleinsyre i alkoholisk suspensjon behandles med en alkoholisk oppløsning av natrium eller av et metallsalt for å danne ved dobbelt omsetning det tilsvarende nukleat. after, if desired, the obtained nucleic acid in alcoholic suspension is treated with an alcoholic solution of sodium or of a metal salt to form by double reaction the corresponding nucleate.
For å utføre fremgangsmåten ifølge foreliggende oppfinnelse må fersk fiskemelke opphakkes slik at det dannes en flytende velling. Denne velling behandles med konsentrert, varm alkalilut. Blandingen blir langsomt flytende, og hydrolysen stop-per når det hele danner en farget væske. In order to carry out the method according to the present invention, fresh fish milk must be chopped up so that a liquid gruel is formed. This gruel is treated with concentrated, hot alkaline lye. The mixture slowly becomes liquid, and the hydrolysis stops when the whole thing forms a colored liquid.
Det avkjøles til ca. 40°, og rå alkalinukleat utfelles med alkohol, det vaskes ved dekantering, og nukleatet fraskilles ved hjelp av kjente fysikalske metoder, f. eks. ved sentrifugering eller filtrering under trykk. It cools to approx. 40°, and crude alkali nucleate is precipitated with alcohol, it is washed by decantation, and the nucleate is separated by means of known physical methods, e.g. by centrifugation or filtration under pressure.
Den ikke tørkede utfelling renses ved innvirkning av kalsiumsulfat under oppvarmning i nærvær av vann. En første filtrering utført i varm tilstand for å hin-dre tilstopping av filtrene etterfølges av en annen filtrering av saften i kold tilstand for å fjerne alt av proteiner som er holdt tilbake av alkalinukleatet. Det fåes en klar, lett farvet oppløsning, som inneholder alkalinukleatet i praktisk talt ren tilstand. The undried precipitate is purified by the action of calcium sulfate during heating in the presence of water. A first filtration carried out in a hot state to prevent clogging of the filters is followed by a second filtration of the juice in a cold state to remove all proteins retained by the alkali nucleate. A clear, slightly colored solution is obtained, which contains the alkali nucleate in a practically pure state.
Nukleinsyre utfelles fra denne rensede oppløsning på hvilken som helst kjent måte, f. eks. ved hjelp av saltsur alkohol. Den fraskilles ved væskeatskillelse eller sentrifugering og vaskes med alkohol og aceton. Der fåes en praktisk talt ren tymonukleinsyre eller desoksyribonukleinsyre. Nucleic acid is precipitated from this purified solution in any known manner, e.g. using hydrochloric acid alcohol. It is separated by liquid separation or centrifugation and washed with alcohol and acetone. A practically pure thymonucleic acid or deoxyribonucleic acid is obtained.
For å fremstille salter av nukleinsyre tilsettes til en suspensjon av nukleinsyre i alkohol en alkoholisk oppløsning av natrium eller av et salt av det metall som skal danne nukleatet. Egnede salter er angitt under avsnitt II senere i denne be-skrivelse/ Metallnukleatet dannes ved dobbelt omsetning og kan fraskilles ved hvilken som helst kjent metode. To prepare salts of nucleic acid, an alcoholic solution of sodium or of a salt of the metal that is to form the nucleate is added to a suspension of nucleic acid in alcohol. Suitable salts are indicated under section II later in this description/ The metal nucleate is formed by double reaction and can be separated by any known method.
Eksempel. Example.
500 kg fersk fiskemelke hakkes opp inn-til der fåes en flytende velling. Denne velling behandles med 85 liter natronlut ved 36° Bé i en time ved 80° C. Det røres om under oppvarmningen. Den seige masse som til å begynne med dannes ved opp-tagelsen av natrium blir langsomt flytende, og der dannes etter en time en brunaktig væske som kjøles til 40° C. 500 kg of fresh fish milk is chopped up until a liquid gruel is obtained. This gruel is treated with 85 liters of caustic soda at 36° B for one hour at 80° C. It is stirred during the heating. The tough mass which is initially formed by the absorption of sodium slowly liquefies, and after an hour a brownish liquid is formed which cools to 40°C.
Hydrolysevæsken helles så i 650 liter alkohol av 80° G.L. Urent natriumnukleat felles ut og avsettes hurtig Væsken dekan-teres, og etter å ha vasket utfellingen tre ganger med alkohol ved 80° G.L. samles utfellingen opp etter å ha atskilt væsken i størst mulig utstrekning. The hydrolysis liquid is then poured into 650 liters of alcohol of 80° G.L. Impure sodium nucleate precipitates out and settles quickly. The liquid is decanted, and after washing the precipitate three times with alcohol at 80° G.L. the precipitate is collected after having separated the liquid to the greatest possible extent.
Utfellingen som er befridd for væske men ikke tørket, blandes med en pasta dannet av 97 liter vann og 5.25 kg kalsiumsulfat for 75 kg av utfellingen. Det blandes energisk og opphetes til 80° C. Blandingen filtreres i varm tilstand, filtratet avkjøles og filtreres påny. Det fåes mellom 85 og 90 liter av en klar, gylden saft. 8 volumdeler av denne saft helles i en lunken blanding av 16 volumdeler alkohol av 75/80° G.L. og 1.8 volumdeler saltsyre. Etter energisk omrøring avsetter nukleinsyre seg i form av et hvitt bunnfall. Dette oppsamles, vaskes med alkohol ved dekantering, væsken skilles fra eller den sentri-fugeres og det vaskes påny tre ganger med alkohol og en gang med aceton. The precipitate, which is freed from liquid but not dried, is mixed with a paste formed from 97 liters of water and 5.25 kg of calcium sulfate for 75 kg of the precipitate. It is mixed vigorously and heated to 80° C. The mixture is filtered while hot, the filtrate is cooled and filtered again. Between 85 and 90 liters of a clear, golden juice is obtained. 8 parts by volume of this juice are poured into a lukewarm mixture of 16 parts by volume of alcohol of 75/80° G.L. and 1.8 parts by volume of hydrochloric acid. After vigorous stirring, nucleic acid settles in the form of a white precipitate. This is collected, washed with alcohol by decantation, the liquid is separated or it is centrifuged and it is washed again three times with alcohol and once with acetone.
Der fåes et hvitt, lett og meget fint pulver av tymonukleinsyre som inneholder omtrent: A white, light and very fine powder of thymonucleic acid is obtained which contains approximately:
Denne syre gir med natriumacetat en stabil oppløsning som ikke inneholder organiske eller mineralske stoffer. II. En del av tymonukleinsyre utvun-net i eksempel 1 utrøres med alkohol av 85° G.L. Det tilsettes en alkoholisk oppløs-ning av natrium, eller av sulfater av mangan, kopper, jern, kalsium-, magnesium-eller koboltklorid etc. for å få de tilsvarende tymonukleater av natrium, mangan, kopper, jern, magnesium, kobolt etc. som dannes ved dobbelt omsetning. With sodium acetate, this acid gives a stable solution that does not contain organic or mineral substances. II. A portion of thymonucleic acid extracted in example 1 is stirred with alcohol of 85° G.L. An alcoholic solution of sodium, or of sulphates of manganese, copper, iron, calcium, magnesium or cobalt chloride etc. is added to obtain the corresponding thymonucleates of sodium, manganese, copper, iron, magnesium, cobalt etc. which formed by double turnover.
Claims (2)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US53729366A | 1966-03-25 | 1966-03-25 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| NO122123B true NO122123B (en) | 1971-05-24 |
Family
ID=24142044
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| NO16730967A NO122123B (en) | 1966-03-25 | 1967-03-16 |
Country Status (12)
| Country | Link |
|---|---|
| AT (1) | AT272335B (en) |
| BE (1) | BE696061A (en) |
| CH (1) | CH511842A (en) |
| DE (1) | DE1695399B2 (en) |
| DK (1) | DK114623B (en) |
| ES (1) | ES338416A1 (en) |
| FI (1) | FI48732C (en) |
| FR (1) | FR1515629A (en) |
| GB (1) | GB1148123A (en) |
| NL (1) | NL151073B (en) |
| NO (1) | NO122123B (en) |
| SE (1) | SE327400B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE60135230D1 (en) | 2000-08-16 | 2008-09-18 | Hoffmann La Roche | Aminocyclohexane derivatives |
| US7012077B2 (en) | 2001-12-20 | 2006-03-14 | Hoffmann-La Roche Inc. | Substituted cyclohexane derivatives |
-
1967
- 1967-03-16 NO NO16730967A patent/NO122123B/no unknown
- 1967-03-17 GB GB1257767A patent/GB1148123A/en not_active Expired
- 1967-03-20 CH CH395567A patent/CH511842A/en not_active IP Right Cessation
- 1967-03-21 SE SE395667A patent/SE327400B/xx unknown
- 1967-03-21 DK DK147767A patent/DK114623B/en unknown
- 1967-03-21 NL NL6704176A patent/NL151073B/en unknown
- 1967-03-21 FI FI83567A patent/FI48732C/en active
- 1967-03-23 DE DE1967M0073307 patent/DE1695399B2/en active Granted
- 1967-03-23 ES ES338416A patent/ES338416A1/en not_active Expired
- 1967-03-23 AT AT285267A patent/AT272335B/en active
- 1967-03-24 FR FR100340A patent/FR1515629A/en not_active Expired
- 1967-03-24 BE BE696061D patent/BE696061A/xx unknown
Also Published As
| Publication number | Publication date |
|---|---|
| NL151073B (en) | 1976-10-15 |
| GB1148123A (en) | 1969-04-10 |
| ES338416A1 (en) | 1968-04-01 |
| FR1515629A (en) | 1968-03-01 |
| CH511842A (en) | 1971-08-31 |
| AT272335B (en) | 1969-07-10 |
| NL6704176A (en) | 1967-09-26 |
| DK114623B (en) | 1969-07-21 |
| SE327400B (en) | 1970-08-24 |
| FI48732B (en) | 1974-09-02 |
| DE1695399B2 (en) | 1976-09-30 |
| DE1695399A1 (en) | 1971-03-18 |
| BE696061A (en) | 1967-09-01 |
| FI48732C (en) | 1974-12-10 |
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