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CN104262315A - Method for separating and purifying tocotrienols - Google Patents

Method for separating and purifying tocotrienols Download PDF

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Publication number
CN104262315A
CN104262315A CN201410453257.2A CN201410453257A CN104262315A CN 104262315 A CN104262315 A CN 104262315A CN 201410453257 A CN201410453257 A CN 201410453257A CN 104262315 A CN104262315 A CN 104262315A
Authority
CN
China
Prior art keywords
polar solvent
tocotrienols
silica gel
separating
silver nitrate
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201410453257.2A
Other languages
Chinese (zh)
Inventor
张源仁
王嘉磊
甄洪峰
魏国华
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
NINGBO DAHONGYING BIO-ENGINEERING Co Ltd
Original Assignee
NINGBO DAHONGYING BIO-ENGINEERING Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by NINGBO DAHONGYING BIO-ENGINEERING Co Ltd filed Critical NINGBO DAHONGYING BIO-ENGINEERING Co Ltd
Priority to CN201410453257.2A priority Critical patent/CN104262315A/en
Publication of CN104262315A publication Critical patent/CN104262315A/en
Pending legal-status Critical Current

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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07DHETEROCYCLIC COMPOUNDS
    • C07D311/00Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/58Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
    • C07D311/70Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with two hydrocarbon radicals attached in position 2 and elements other than carbon and hydrogen in position 6
    • C07D311/723,4-Dihydro derivatives having in position 2 at least one methyl radical and in position 6 one oxygen atom, e.g. tocopherols
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01DSEPARATION
    • B01D15/00Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08Selective adsorption, e.g. chromatography
    • B01D15/10Selective adsorption, e.g. chromatography characterised by constructional or operational features
    • B01D15/20Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the sorbent material
    • B01D15/206Packing or coating
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01JCHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/02Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material
    • B01J20/10Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising silica or silicate
    • B01J20/103Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising silica or silicate comprising silica

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Inorganic Chemistry (AREA)
  • Pyrane Compounds (AREA)

Abstract

The invention discloses a method for separating and purifying tocotrienols. The method comprises the following steps: (1) preparing filler for chromatography; (2) carrying out column chromatography; and (3) fractionally collecting eluent, combining the eluent with same components according to TLC detection and carrying out concentration to obtain tocotrienols concentrate, wherein the filler for column chromatography is silver nitrate silica gel, in the preparation process of the silver nitrate silica gel, the proportion of the silver nitrate to the silica gel, the proportion of the added water to the ethyl alcohol and the parameters such as the column amount of the column chromatography and the drying time are adjusted, and a plurality of solvent systems are used for sequential elution, so that the elution of the active constituents are more complete and the yield of the tocotrienols is greatly improved. The method is simple in process, low in equipment requirement and high in separation efficiency.

Description

A kind of separating and purifying method of tocotrienols
Technical field
The invention belongs to field of chemical engineering, be specifically related to a kind of separating and purifying method of tocotrienols.
Background technology
Vitamin-E is the general name of tocopherol and tocotrienols and the derivative (as ester class etc.) with d-alpha-tocopherol physiologically active.From plant resources, 4 kinds of tocopherols (α, β, γ, δ) and 4 kinds of tocotrienolss (α, β, γ, δ) are isolated at present.Tocotrienols and the structural key distinction of tocopherol are that isoprenoid base side chain degree of saturation is different.Why tocotrienols has carried no weight two reasons: one is in various tocopherol and tocotrienols, and the physiologically active of alpha-tocopherol is the highest, preferentially absorbed and utilize, and the physiologically active of tocotrienols is relatively low in human body; Two is that the distribution of tocotrienols is wide, only exist with monocotyledons (Gramineae and Palmae) and minority dicotyledons in find.
At present, the extraction purification research report about tocotrienols is little, and the difficulty of its extraction and isolation is: easily oxidational losses; Tocopherol and tocotrienols structure and character close, not easily separated.Its purification technique comprises: saponification is to remove the Ester in vegetables oil, but saponification reaction needs comparatively high temps and carries out in the basic conditions, the very easily oxidized loss of tocotrienols, therefore needs to add antioxidant; Supercritical CO 2extraction, the method effectively can overcome the shortcomings such as oxidizable, the dissolvent residual in tocotrienols sepn process, but required equipment cost is higher; Column chromatography chromatogram is separated can carry out roughing out to tocopherol and tocotrienols, makes most of tocopherol and tocotrienols carry out enrichment, but tocopherol cannot be made effectively to be separated with tocotrienols; Preparative liquid chromatography can obtain highly purified tocotrienols, but the method cost is high, equipment requirements is high, output is little, is not suitable for industrialization.
Summary of the invention
In order to overcome the deficiency of above-mentioned prior art, the invention provides a kind of separating and purifying method of tocotrienols, the method technique is simple, equipment requirements is low, separation efficiency is high.
The technical solution adopted in the present invention is, a kind of separating and purifying method of tocotrienols, specifically comprises the following steps:
(1) prepared by chromatography filler: in silica gel: the ratio mixing of Silver Nitrate=2.5 ~ 3.5:1 (m/m), add silica gel quality 1:1 ~ 2 (m/v) distilled water and 1:1.5 ~ 3 (m/v) ethanol, be uniformly mixed, underpressure distillation desolvation, the sample powder that must flow dries 16 ~ 24h in 100-120 DEG C;
(2) column chromatography: adopt wet method dress post, chromatography column is proceeded to by disposable after the immersion of Silver Nitrate silica gel non-polar solvent, with described non-polar solvent as eluent repeatedly wash-out filler make Silver Nitrate silica gel deposit compacting, by the upper column quantity of Silver Nitrate silica gel quality 5% ~ 30%, sample is transferred to silica gel top, then 0.5-1BV (column volume) non-polar solvent is pressed, the mixing elutriant of 0.5-1BV (column volume) 5% ~ 10% polar solvent-non-polar solvent, the sequentially eluting sample of the mixing elutriant of 0.5-1BV (column volume) 50% ~ 100% polar solvent-non-polar solvent,
(3) Fractional Collections elutriant, according to TLC combining data detection same composition elutriant, obtains tocotrienols concentrated solution after concentrated.
Preferably, in step (1), in silica gel: the ratio mixing of Silver Nitrate=2.5:1 (m/m).
Preferably, the non-polar solvent in step (2) is normal hexane, normal heptane or sherwood oil.
Preferably, the polar solvent in step (2) is ethyl acetate, tetrahydrofuran (THF) or chloroform.
Preferably, the non-polar solvent in step (2) is normal hexane, and polar solvent is ethyl acetate.
Preferably, by the upper column quantity of Silver Nitrate silica gel quality 5%, sample is transferred to silica gel top in step (2).
Preferably, the mixing elutriant of the polar solvent-non-polar solvent in step (2) is ethyl acetate-hexane.
Preferably, in step (2), by the sequentially eluting sample of the mixing elutriant of 1BV (column volume) non-polar solvent, 1BV (column volume) 10% polar solvent-non-polar solvent, the mixing elutriant of 1BV (column volume) 50% polar solvent-non-polar solvent.
Preferably, the simmer down to molecular distillation in step (3) concentrates.
The sample source that the present invention is separated is extensive, both plam oil and Rice pollard oil can be derived from, also can derive from the tankage of cereal processing and contain than more rich tocotrienols, the sample to be separated containing certain density tocotrienols is all suitable for method of the present invention.Sample source also can be obtained by purchase, the tocotrienols raw material produced as Malaysian Carotech company (wherein mixes tocotrienols content 25.32%, alpha-tocopherol content 6.9%, comprises squalene, plant sterol, Coenzyme Q10 99.0 and mixed type carotenoid etc. containing other compositions in addition).
Compared to existing technology, the present invention has following advantage:
1, column chromatography filler adopts Silver Nitrate silica gel, in the preparation process of Silver Nitrate silica gel, comprising the parameter such as ratio and drying time of Silver Nitrate and silica gel ratio, the water added and ethanol, is all the optimum proportioning that applicant gropes out through long-term creative work; The use successively of the upper column quantity that column chromatography adopts and multi-solvents system, namely the mixing elution of the polar solvent-non-polar solvent increased from non-polar solvent to concentration is adopted, make the wash-out of active principle more complete, greatly improve the yield of tocotrienols;
2, method technique of the present invention is simple, equipment requirements is low, separation efficiency is high, is suitable for industrialization.
Embodiment
Below in conjunction with embodiment, the invention will be further described:
Embodiment 1
(1) prepared by chromatography filler: get silica gel 100g, Silver Nitrate 40g, adds 100ml distilled water and 150ml ethanol, is uniformly mixed, underpressure distillation desolvation, and the sample powder that must flow dries 20h in 120 DEG C;
(2) column chromatography: adopt wet method dress post, chromatography column is proceeded to by disposable after the immersion of Silver Nitrate silica gel normal hexane, with normal hexane as eluent repeatedly wash-out filler make Silver Nitrate silica gel deposit compacting, get 7g sample and be transferred to silica gel top, then press the eluent sequentially eluting sample of normal hexane 1BV, 10% ethyl acetate-hexane 1BV, 50% ethyl acetate-hexane 1BV; Sample source is obtained by purchase, the tocotrienols raw material (wherein mixing tocotrienols content 25.32%, alpha-tocopherol content 6.9%) that Malaysian Carotech company produces;
(3) Fractional Collections elutriant, according to TLC combining data detection same composition elutriant, obtains tocotrienols concentrated solution 1.99g, wherein tocotrienols content 85.57%, yield 81.0% after concentrated.
Above embodiment display and describe ultimate principle of the present invention and principal character and advantage of the present invention.The technician of the industry should understand; the present invention is not restricted to the described embodiments; what describe in above-described embodiment and specification sheets just illustrates principle of the present invention; instead of limit the scope of the invention by any way; without departing from the scope of the invention; the present invention also has various changes and modifications, and these changes and improvements all fall in claimed scope.

Claims (8)

1. a separating and purifying method for tocotrienols, specifically comprises the following steps:
(1) prepared by chromatography filler: in silica gel: the ratio mixing of Silver Nitrate=2.5 ~ 3.5:1 (m/m), add silica gel quality 1:1 ~ 2 (m/v) distilled water and 1:1.5 ~ 3 (m/v) ethanol, be uniformly mixed, underpressure distillation desolvation, the sample powder that must flow dries 16 ~ 24h in 100-120 DEG C;
(2) column chromatography: adopt wet method dress post, chromatography column is proceeded to by disposable after the immersion of Silver Nitrate silica gel non-polar solvent, with described non-polar solvent as eluent repeatedly wash-out filler make Silver Nitrate silica gel deposit compacting, by the upper column quantity of Silver Nitrate silica gel quality 5% ~ 30%, sample is transferred to silica gel top, then 0.5-1BV (column volume) non-polar solvent is pressed, the mixing elutriant of 0.5-1BV (column volume) 5% ~ 10% polar solvent-non-polar solvent, the sequentially eluting sample of the mixing elutriant of 0.5-1BV (column volume) 50% ~ 100% polar solvent-non-polar solvent,
(3) Fractional Collections elutriant, according to TLC combining data detection same composition elutriant, obtains tocotrienols concentrated solution after concentrated.
2. the separating and purifying method of a kind of tocotrienols according to claim 1, is characterized in that, in step (1), in silica gel: the ratio mixing of Silver Nitrate=2.5:1 (m/m).
3. the separating and purifying method of a kind of tocotrienols according to claim 1, is characterized in that, the non-polar solvent in step (2) is normal hexane, normal heptane or sherwood oil.
4. the separating and purifying method of a kind of tocotrienols according to claim 1, is characterized in that, the polar solvent in step (2) is ethyl acetate, tetrahydrofuran (THF) or chloroform.
5. the separating and purifying method of a kind of tocotrienols according to claim 1, is characterized in that, the non-polar solvent in step (2) is normal hexane, and polar solvent is ethyl acetate.
6. the separating and purifying method of a kind of tocotrienols according to claim 1, is characterized in that, by the upper column quantity of Silver Nitrate silica gel quality 5%, sample is transferred to silica gel top in step (2).
7. the separating and purifying method of a kind of tocotrienols according to claim 1, it is characterized in that, in step (2), by the sequentially eluting sample of the mixing elutriant of 1BV (column volume) non-polar solvent, 1BV (column volume) 10% polar solvent-non-polar solvent, the mixing elutriant of 1BV (column volume) 50% polar solvent-non-polar solvent.
8. the separating and purifying method of a kind of tocotrienols according to claim 1, is characterized in that, the simmer down to molecular distillation in step (3) concentrates.
CN201410453257.2A 2014-09-05 2014-09-05 Method for separating and purifying tocotrienols Pending CN104262315A (en)

Priority Applications (1)

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CN201410453257.2A CN104262315A (en) 2014-09-05 2014-09-05 Method for separating and purifying tocotrienols

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201410453257.2A CN104262315A (en) 2014-09-05 2014-09-05 Method for separating and purifying tocotrienols

Publications (1)

Publication Number Publication Date
CN104262315A true CN104262315A (en) 2015-01-07

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Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1171106A (en) * 1994-11-07 1998-01-21 伊斯曼化学公司 Process for production of tocopherol and tocopherol/tocotrienol concentrates
US6395915B1 (en) * 1999-09-10 2002-05-28 Technikrom, Inc. Method for producing purified tocotrienols and tocopherols using liquid chromatography
US20040026323A1 (en) * 2000-11-21 2004-02-12 Kikuzo Kaneko Method for chromatographic preparation of tocotrienol
JP2004305155A (en) * 2003-04-09 2004-11-04 Tama Seikagaku Kk Method for producing tocopherol and tocotrienol
CN101445498A (en) * 2008-11-14 2009-06-03 天津大学 Production method of high purity natural vitamin E and separation method of tocopherol homologues
JP2009143896A (en) * 2007-11-19 2009-07-02 Nisshin Pharma Inc Method for concentrating fat-soluble vitamins
JP2009190989A (en) * 2008-02-13 2009-08-27 Tohoku Univ Method for simultaneous production of tocotrienol and biodiesel fuel from fats and oils
CN102264730A (en) * 2008-10-28 2011-11-30 爱迪生药物公司 Process for the production of tocotrienols and derivatives

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1171106A (en) * 1994-11-07 1998-01-21 伊斯曼化学公司 Process for production of tocopherol and tocopherol/tocotrienol concentrates
US6395915B1 (en) * 1999-09-10 2002-05-28 Technikrom, Inc. Method for producing purified tocotrienols and tocopherols using liquid chromatography
US20040026323A1 (en) * 2000-11-21 2004-02-12 Kikuzo Kaneko Method for chromatographic preparation of tocotrienol
JP2004305155A (en) * 2003-04-09 2004-11-04 Tama Seikagaku Kk Method for producing tocopherol and tocotrienol
JP2009143896A (en) * 2007-11-19 2009-07-02 Nisshin Pharma Inc Method for concentrating fat-soluble vitamins
JP2009190989A (en) * 2008-02-13 2009-08-27 Tohoku Univ Method for simultaneous production of tocotrienol and biodiesel fuel from fats and oils
CN102264730A (en) * 2008-10-28 2011-11-30 爱迪生药物公司 Process for the production of tocotrienols and derivatives
CN101445498A (en) * 2008-11-14 2009-06-03 天津大学 Production method of high purity natural vitamin E and separation method of tocopherol homologues

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
万建春 等: "天然VE色谱法分离与纯化的研究进展", 《农产品加工》, no. 12, 31 December 2007 (2007-12-31), pages 65 - 70 *
代志凯 等: "生育三烯酚分离纯化研究进展", 《中国油脂》, vol. 39, no. 1, 20 January 2014 (2014-01-20), pages 79 - 83 *
夏向东 等: "硅胶和硝酸银硅胶柱层析分离生育酚与生育三烯酚", 《中国粮油学报》, vol. 19, no. 3, 30 June 2004 (2004-06-30) *

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Application publication date: 20150107