CN104017059A - Method for extracting microcystic toxins - Google Patents
Method for extracting microcystic toxins Download PDFInfo
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- CN104017059A CN104017059A CN201410274127.2A CN201410274127A CN104017059A CN 104017059 A CN104017059 A CN 104017059A CN 201410274127 A CN201410274127 A CN 201410274127A CN 104017059 A CN104017059 A CN 104017059A
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- microcystin
- ethanol
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Abstract
The invention discloses a method for extracting microcystic toxins. The method comprises the following steps: by taking fresh blue-green algae of natural blooms as raw materials, performing coarse extraction through ethanol, and separating and purifying MCs through a secondary macroporous resin column. The MCs purified by the method has the advantages of low cost, safety in operation, intuitiveness, simplicity, convenience and the like.
Description
Technical field
The present invention relates to the biochemical technical field separating, specifically, relate to a kind of extracting method of Microcystin.
Background technology
In recent years, the blue-green alga bloom being caused by body eutrophication pollutes and has caused global concern.It is poisonous blue-green algae to the Microcystin (mi-crocystis that discharges number of different types in water body that blue-green alga bloom pollutes the main harm of bringing, MCs), wherein microcapsule phycotoxin MC-LR and MC-RR are topmost two kinds of algae toxin in China's eutrophication water.MCs is a kind of generally acknowledged hepatotoxin and carcinogenic promoting agent, and the water that long-term drinking contains MCs can cause damage to people's liver, and under extreme case, entail dangers to is to people's life.
Along with the further investigation of water bloom pollution problem, the environmental behaviours such as distribution, migration, accumulation and the home to return to of MCs in environment, and aspect environmental pollution improvement, be subject to efforts at environmental protection person's special concern.Up to the present, do a lot of work in the aspects such as distribution, Structural Identification, content detection, toxicity assessment and toxicity mechanism to MCs, but the lacking of MCs sterling, becomes one of bottleneck problem that puzzlement researchist furthers investigate MCs, therefore in the urgent need to studying the method for effective extraction, purifying MCs a kind of.
Summary of the invention
The object of this invention is to provide one taking the fresh blue-green algae of wawter bloom as raw material, extract Microcystin by extraction and macroporous resin chromatography, method is simple, easy and simple to handle, with low cost.
For achieving the above object, the invention provides following technical scheme:
An extracting method for Microcystin, comprises the following steps:
Step 1: taking the fresh blue-green algae of natural bloom as raw material, the methanol solution that volume ratio is 65-75% is extraction agent, and magnetic agitation 1.5-2 hour is after ultrasonic 20-30 minute, centrifugal, takes out supernatant liquor;
Step 2: filter residue is repeated to step above, merge the supernatant liquor of twice, remove residue;
Step 3: regulate PH to filter after 7.5-8 crude extract, steam methyl alcohol at the temperature of 35-40 DEG C;
Step 4: concentrated crude extract, by the microfiltration membrane of 0.45 μ m, is obtained to Microcystin crude product;
Step 5: regulating the pH value of Microcystin crude product is 3-4, and crude product liquid is crossed to D101 macroporous adsorptive resins, and flow velocity is 3mL/ min, then the ethanol elution that is 45% by volume fraction;
Step 6: collect elutriant, evaporate after ethanol, then to regulate pH value be 3-4, cross D101 macroporous adsorptive resins, flow velocity is 3mL/ min, then the ethanol elution that is 55% by volume fraction;
Step 7: collect MC-RR peak in the time of 6.892min, after 15.282min collection MC-LR collects, collect respectively the MC-RR and the MC-LR elutriant that obtain, use Rotary Evaporators evaporate to dryness, then add dissolve with ethanol;
Step 8: by the MC-RR obtaining and MC-LR ethanolic soln, solvent evaporated, obtains product.
Further, the volume ratio of the blue-green algae in step 1 and methanol solution is 1:10-1:20.
Further, the rotating speed of the magnetic agitation in step 1 is: 2 000 r/min-2500 r/min.
Further, the centrifugal condition in step 1 is: rotating speed 10000r/min-20000r/min, time 5-10 minute.
Beneficial effect:
In the technical scheme of the inventive method, the blue-green algae producing when the natural environmental water grey hair that raw material adopts is given birth to, algae powder treating processes is simple, and raw materials cost is negligible, the operation adopting in operating process is relatively simple, and the pre-treatment of algae powder requires lower to experimenter;
With traditional anti-phase C18 filler as the Solid-Phase Extraction purifying MCs that is separated, although can obtain the MCs that purity is higher, but because the restriction of upper column quantity and flow velocity cannot obtain a large amount of MCs that purify for, MCs after 2 purification with macroreticular resin, although purity is between 70%～85%, but can obtain a large amount of MCs, thereby meet the needs of the various environmental chemistry behavioral experiments of MCs;
Because purification amount is large, the methyl alcohol of comparing, adopts different concentration ethanol safer in concrete operations as elutriant, convenient; Methods For Purification MCs of the present invention has that cost is low, operational safety, the advantage such as directly perceived easy.
Embodiment
Below in conjunction with embodiment, the present invention is further detailed explanation.
Embodiment
An extracting method for Microcystin, comprises the following steps:
Step 1: taking the fresh blue-green algae of natural bloom as raw material, the methanol solution that volume ratio is 65-75% is extraction agent, and magnetic agitation 1.5-2 hour is after ultrasonic 20-30 minute, centrifugal, takes out supernatant liquor;
Step 2: filter residue is repeated to step above, merge the supernatant liquor of twice, remove residue;
Step 3: regulate PH to filter after 7.5-8 crude extract, steam methyl alcohol at the temperature of 35-40 DEG C;
Step 4: concentrated crude extract, by the microfiltration membrane of 0.45 μ m, is obtained to Microcystin crude product;
Step 5: regulating the pH value of Microcystin crude product is 3-4, and crude product liquid is crossed to D101 macroporous adsorptive resins, and flow velocity is 3mL/ min, then the ethanol elution that is 45% by volume fraction;
Step 6: collect elutriant, evaporate after ethanol, then to regulate pH value be 3-4, cross D101 macroporous adsorptive resins, flow velocity is 3mL/ min, then the ethanol elution that is 55% by volume fraction;
Step 7: collect MC-RR peak in the time of 6.892min, after 15.282min collection MC-LR collects, collect respectively the MC-RR and the MC-LR elutriant that obtain, use Rotary Evaporators evaporate to dryness, then add dissolve with ethanol;
Step 8: by the MC-RR obtaining and MC-LR ethanolic soln, solvent evaporated, obtains product.
Further, the volume ratio of the blue-green algae in step 1 and methanol solution is 1:10-1:20.
Further, the rotating speed of the magnetic agitation in step 1 is: 2 000 r/min-2500 r/min.
Further, the centrifugal condition in step 1 is: rotating speed 10000r/min-20000r/min, time 5-10 minute.
Claims (4)
1. an extracting method for Microcystin, is characterized in that: comprise the following steps:
Step 1: taking the fresh blue-green algae of natural bloom as raw material, the ethanolic soln that volume ratio is 65-75% is extraction agent, and magnetic agitation 1.5-2 hour is after ultrasonic 20-30 minute, centrifugal, takes out supernatant liquor;
Step 2: filter residue is repeated to step above, merge the supernatant liquor of twice, remove residue;
Step 3: regulate PH to filter after 7.5-8 crude extract, steam methyl alcohol at the temperature of 35-40 DEG C;
Step 4: concentrated crude extract, by the microfiltration membrane of 0.45 μ m, is obtained to Microcystin crude product;
Step 5: regulating the pH value of Microcystin crude product is 3-4, and crude product liquid is crossed to D101 macroporous adsorptive resins, and flow velocity is 3mL/ min, then the ethanol elution that is 45% by volume fraction;
Step 6: collect elutriant, evaporate after ethanol, then to regulate pH value be 3-4, cross D101 macroporous adsorptive resins, flow velocity is 3mL/ min, then the ethanol elution that is 55% by volume fraction;
Step 7: collect MC-RR peak in the time of 6.892min, after 15.282min collection MC-LR collects, collect respectively the MC-RR and the MC-LR elutriant that obtain, use Rotary Evaporators evaporate to dryness, then add dissolve with ethanol;
Step 8: by the MC-RR obtaining and MC-LR ethanolic soln, solvent evaporated, obtains product.
2. the extracting method of a kind of Microcystin as claimed in claim 1, is characterized in that: the blue-green algae in step 1 and the volume ratio of ethanolic soln are 1:10-1:20.
3. the extracting method of a kind of Microcystin as claimed in claim 1, is characterized in that: the rotating speed of the magnetic agitation in step 1 is: 2 000 r/min-2500 r/min.
4. the extracting method of a kind of Microcystin as claimed in claim 1, is characterized in that: the centrifugal condition in step 1 is: rotating speed 10000r/min-20000r/min, time 5-10 minute.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410274127.2A CN104017059A (en) | 2014-06-19 | 2014-06-19 | Method for extracting microcystic toxins |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410274127.2A CN104017059A (en) | 2014-06-19 | 2014-06-19 | Method for extracting microcystic toxins |
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| CN104017059A true CN104017059A (en) | 2014-09-03 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201410274127.2A Pending CN104017059A (en) | 2014-06-19 | 2014-06-19 | Method for extracting microcystic toxins |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106943437A (en) * | 2017-03-06 | 2017-07-14 | 广东药科大学 | A kind of Microcystis aeruginosa ethanol extract and preparation method and application |
| CN108752440A (en) * | 2018-06-21 | 2018-11-06 | 水利部中国科学院水工程生态研究所 | A kind of other extensive method for extraction and purification of the gram-grade of Microcystin |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1546991A (en) * | 2003-12-05 | 2004-11-17 | 浙江大学 | Detection method of trace microcapsule algal toxin in water |
| CN1563081A (en) * | 2004-03-27 | 2005-01-12 | 中国科学院水生生物研究所 | Method for separating and purifying toxin of microcapsule alga |
| CN101168561A (en) * | 2007-11-27 | 2008-04-30 | 江南大学 | Method for separating and purifying microcystin |
| CN101955517A (en) * | 2010-10-29 | 2011-01-26 | 中国科学院水生生物研究所 | Method for extracting and purifying microcystin |
-
2014
- 2014-06-19 CN CN201410274127.2A patent/CN104017059A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1546991A (en) * | 2003-12-05 | 2004-11-17 | 浙江大学 | Detection method of trace microcapsule algal toxin in water |
| CN1563081A (en) * | 2004-03-27 | 2005-01-12 | 中国科学院水生生物研究所 | Method for separating and purifying toxin of microcapsule alga |
| CN101168561A (en) * | 2007-11-27 | 2008-04-30 | 江南大学 | Method for separating and purifying microcystin |
| CN101955517A (en) * | 2010-10-29 | 2011-01-26 | 中国科学院水生生物研究所 | Method for extracting and purifying microcystin |
Non-Patent Citations (3)
| Title |
|---|
| 向伦辉等: "微囊藻毒素与饮用水有机提取物联合致HepG2细胞DNA损伤的研究", 《环境与健康杂志》 * |
| 樊洁等: "大孔树脂去除水中微囊藻毒素初步试验", 《环境研究与检测》 * |
| 王中婵等: "大孔吸附树脂分离纯化微囊藻毒素", 《环境科学学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106943437A (en) * | 2017-03-06 | 2017-07-14 | 广东药科大学 | A kind of Microcystis aeruginosa ethanol extract and preparation method and application |
| CN106943437B (en) * | 2017-03-06 | 2020-01-03 | 广东药科大学 | Ethanol extract of microcystis and preparation method and application thereof |
| CN108752440A (en) * | 2018-06-21 | 2018-11-06 | 水利部中国科学院水工程生态研究所 | A kind of other extensive method for extraction and purification of the gram-grade of Microcystin |
| CN108752440B (en) * | 2018-06-21 | 2022-08-26 | 水利部中国科学院水工程生态研究所 | Gram-level large-scale extraction and purification method of microcystin |
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