CN105330668A - Method for separating and enriching sodium chlorophyllin - Google Patents
Method for separating and enriching sodium chlorophyllin Download PDFInfo
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Abstract
The invention relates to a method for separating and enriching sodium chlorophyllin, in particular to a method for carrying out flotation on sodium chlorophyllin in a bamboo leaf saponified solution through aqueous two-phase gas flotation solvents, and belongs to the technical field of natural product extraction separation. The method includes the steps that fresh bamboo leaves are collected and pretreated, and the bamboo leaf saponified solution is obtained; a tripotassium phosphate solution and the bamboo leaf saponified solution are mixed in a colorimetric tube, oscillated, mixed to be even and then transferred into a flotation column, ethyl alcohol is added from the top end of the column, and nitrogen is introduced for flotation; after flotation is completed, the absorbancy of the upper-phase sodium chlorophyllin, the absorbancy of the lower-phase sodium chlorophyllin and the absorbancy of upper-phase sugar are measured, and the recovery rate of the sodium chlorophyllin and the removing rate of the sugar are obtained; upper-phase ethyl alcohol and lower-phase tripotassium phosphate are recycled. By means of the method, the defects that in the extraction separation process of the sodium chlorophyllin serving as the intermediate of sodium copper chlorophyllin, the technology is complex, energy consumption is high, and purity is low are overcome, and the ethyl alcohol-tripotassium phosphate aqueous two-phase gas flotation solvent flotation method which is easy to operate, low in cost, environmentally friendly and efficient is provided for separating/enriching the sodium chlorophyllin.
Description
Technical field
The present invention relates to a kind of method of separation and concentration CHLOROPHYLLINE sodium, particularly relate to the method for CHLOROPHYLLINE sodium in a kind of double water-phase air-float solvent floatation leaf of bamboo saponification liquor, belong to natural product extraction and separation technology field.
Background technology
Chlorophyll is a kind of natural green pigment, is extensively present in natural green plants and algae.But chlorophyll extremely unstable, runs into acid, alkali, light, heat and enzyme etc. and all easily decompose, and chlorophyll is difficult to water-soluble, so its application is very limited.Sodium copper chlorophyllin is chlorophyllous synthesis of derivatives, good water solubility and very stable, is widely used as foodstuff additive, cosmetics additive and tinting material etc.Because sodium copper chlorophyllin has higher value, so mainly chlorophyll is made sodium copper chlorophyllin to use to chlorophyllous exploitation.Therefore, the preparation method of sodium copper chlorophyllin is subject to paying close attention to widely both at home and abroad.
Tradition prepares the technological process of sodium copper chlorophyllin mainly: organic solvent lixiviate is concentrated obtains chlorophyll paste, CHLOROPHYLLINE sodium is prepared in chlorophyll paste saponification, acidifying is put copper and is obtained Chlorophyll Cu acid crude, and alkali is for salify and removal of impurities obtains final product sodium copper chlorophyllin.In this series of process process, CHLOROPHYLLINE sodium is the important intermediate preparing sodium copper chlorophyllin, and its purity directly affects yield and the quality product of sodium copper chlorophyllin.Traditional technology is when preparing CHLOROPHYLLINE sodium, not only to consume a large amount of organic solvent and the energy, and be also extracted out together with chlorophyll due to water-soluble and oil-soluble impurities in organic solvent leaching process, the CHLOROPHYLLINE sodium causing saponification to obtain also adulterates a large amount of impurity, and purity is very low.Therefore, productive rate and the quality of product sodium copper chlorophyllin be improved, need to set up a kind of efficient, extracting and developing of green non-pollution and the method for enrichment CHLOROPHYLLINE sodium.
Conventional extracting substances separation method mainly contains solvent extraction, aqueous two-phase extraction and solvent signal suppression etc.And when using solvent extraction, need to consume a large amount of organic solvent, and in the extraction and isolation of some valuable biologically active substances, solvent extraction can cause distortion and the inactivation of biologically active substance.Aqueous two-phase extraction operational condition is gentle, and technique is simple, can solve the problem that solvent extraction destroys biologically active substance, but the enrichment times of aqueous two-phase extraction to target compound is lower.Although solvent floatation can solve target compound enrichment times and be limited to the shortcoming compared, usually can use some toxic organic solvents, cause environmental pollution and threaten human health.
Double water-phase air-float solvent floatation is a kind of New Pre enrichment and the isolation technique having future occurred in recent years, it integrates solvent floatation technology and aqueous two phase extraction technique, by the mutual supplement with each other's advantages of the two, there is lot of advantages, comprise that separation efficiency is high, enrichment times is large, operational condition is gentle, operating process is simple and organic solvent consumption little etc.And double water-phase air-float solvent floatation can scale up test parameter, is highly suitable for industrial production.Especially adopt small molecular alcohol-inorganic salt double water-phase air-float solvent floatation method, not only have the advantage that the low and phase composite composition of environmental protection, cost is easy to reclaim simultaneously, and can effectively be separated from mixture by target substance, enrichment times is large.Therefore, this small molecular alcohol-inorganic salt double water-phase air-float solvent floatation method is with a wide range of applications in the separation and concentration field of natural product.
Summary of the invention
The object of the invention is to overcome complex process in sodium copper chlorophyllin intermediate CHLOROPHYLLINE sodium extraction and isolation process, energy consumption is high, purity is low defect, provide a kind of simple to operate, low cost, environmental protection and efficiently the method for ethanol-Tripotassium phosphate double water-phase air-float solvent floatation be separated/enrichment leaf of bamboo saponification liquor in CHLOROPHYLLINE sodium.
For reaching above technical purpose, the present invention adopts following technique means:
A method for separation and concentration CHLOROPHYLLINE sodium, described method double-aqueous phase system and air-float solvent is floated to combine, and selects the method for CHLOROPHYLLINE sodium in leaf of bamboo saponification liquor, carry out according to following step:
(1) gather the fresh leaf of bamboo and carry out pre-treatment and obtain leaf of bamboo saponification liquor, being placed in 4 DEG C of refrigerators and preserving, within every 5 days, again preparing a storing solution;
(2) certain density Tripotassium phosphate solution and leaf of bamboo saponification liquor are mixed in colorimetric cylinder, are transferred in flotation column after concussion mixes, then add the ethanol of certain volume from capital end, pass into nitrogen and carry out flotation;
(3), after flotation terminates, phase ethanol phase is pipetted out with dropper, lower to the outflow of flotation column bottom switch, measure the absorbancy of phase CHLOROPHYLLINE sodium and upper sugar mutually up and down with ultraviolet visible spectrophotometry, thus draw the rate of recovery of CHLOROPHYLLINE sodium and the clearance of sugar;
(4) upper phase ethanol and the recovery of lower phase Tripotassium phosphate are used for setting up secondary flotation;
Wherein, the Tripotassium phosphate strength of solution described in step (2) is 0.46-0.62g/mL;
The volume ratio of described Tripotassium phosphate solution and leaf of bamboo saponification liquor is 45:5;
Described ethanol contend is 4-5.5mL; Described nitrogen flow rate is 10-30mL/min; Described flotation time is 15-35min.
beneficial effect of the present invention is as follows:
(1) the present invention adopts double water-phase air-float solvent floatation method; double-aqueous phase system and solvent floatation are combined; both solve target compound enrichment times in double-aqueous phase system and be limited to the shortcoming compared, turn avoid the defect that solvent floatation usually can use some toxic organic solvents.
(2) the present invention adopts double water-phase air-float solvent floatation method flotation CHLOROPHYLLINE sodium first.CHLOROPHYLLINE sodium molecule has extraordinary surfactivity, can be adsorbed on bubble surface flotation in a large number to organic solvent phase.Therefore, CHLOROPHYLLINE sodium is highly suitable for double water-phase air-float solvent floatation compared with other materials.
(3) prepare with tradition compared with CHLOROPHYLLINE sodium process, the present invention eliminates extraction and concentrated chlorophyllous step in separation and concentration CHLOROPHYLLINE sodium process, avoid the use of a large amount of organic solvent, save the energy and time, and solve the problem that a large amount of impurity occurs to degrade and be mixed with leaching process Determination of Chlorophyll.
(4) the present invention adopts ethanol-Tripotassium phosphate double water-phase air-float solvent floatation system.Ethanol is nontoxic, cost is low and be easy to reclaim; Tripotassium phosphate belongs to basic salt, can not destroy the CHLOROPHYLLINE sodium in leaf of bamboo saponification liquor, and the double water-phase air-float solvent floatation system formed with ethanol can the more CHLOROPHYLLINE sodium of separation and concentration and to remove a large amount of water-soluble impurities sugared.The CHLOROPHYLLINE sodium enrichment times obtained like this is large, and purity is high, prepares sodium copper chlorophyllin as raw material, is very beneficial for the yield and the quality that improve product.
Accompanying drawing explanation
Fig. 1 is (a) and the bubbling behaviour figure containing (b) in CHLOROPHYLLINE sodium deionized water in deionized water in embodiment 7.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in further detail.Should understand; described embodiment is preferred embodiment of the present invention; but the present invention is not limited to above-mentioned embodiment; when not deviating from flesh and blood of the present invention, any apparent improvement that those skilled in the art can make is replaced or modification all belongs to protection scope of the present invention.
embodiment 1:
A method for CHLOROPHYLLINE sodium in double water-phase air-float solvent floatation leaf of bamboo saponification liquor, concrete steps are as follows:
(1) gather the fresh leaf of bamboo and shred.By the broken leaf of bamboo and NaOH solution (10%, w/w) proportionally 1:16(w/v) mix and be placed in 65 DEG C of waters bath with thermostatic control, react 2h under constantly stirring.Juice after saponification is obtained leaf of bamboo saponification liquor and be placed in 4 DEG C of refrigerators preserving through cloth and filter paper filtering successively, within every 5 days, again prepares a storing solution.
(2) 0.50g/mL Tripotassium phosphate solution and 5mL leaf of bamboo saponification liquor are mixed in 50mL colorimetric cylinder, make mixed solution cumulative volume be 50mL.Being transferred to by mixed solution in flotation column and adding the ethanol of 4mL from capital end, logical nitrogen flotation 15min, nitrogen flow rate is 10mL/min.
(3), after flotation terminates, measure phase CHLOROPHYLLINE sodium up and down respectively and, in the absorbancy at 639nm place, measure up and down the absorbancy of sugar at 490nm place mutually respectively by phenol-sulfuric acid process.Show that the rate of recovery of CHLOROPHYLLINE sodium is 40.62%, enrichment times is 3.28, and the clearance of impurity sugar is 96.14%.
(4) Tripotassium phosphate is reclaimed by alcohol precipitation, filtration, evaporation; Ethanol is reclaimed by rotary evaporation.The ethanol of recovery and Tripotassium phosphate are used for setting up secondary flotation.
embodiment 2:
A method for CHLOROPHYLLINE sodium in double water-phase air-float solvent floatation leaf of bamboo saponification liquor, concrete steps are as follows:
(1) gather the fresh leaf of bamboo and shred.By the broken leaf of bamboo and NaOH solution (10%, w/w) proportionally 1:16(w/v) mix and be placed in 65 DEG C of waters bath with thermostatic control, react 2h under constantly stirring.Juice after saponification is obtained leaf of bamboo saponification liquor and be placed in 4 DEG C of refrigerators preserving through cloth and filter paper filtering successively, within every 5 days, again prepares a storing solution.
(2) 0.54g/mL Tripotassium phosphate solution and 5mL leaf of bamboo saponification liquor are mixed in 50mL colorimetric cylinder, make mixed solution cumulative volume be 50mL.Being transferred to by mixed solution in flotation column and adding the ethanol of 5mL from capital end, logical nitrogen flotation 20min, nitrogen flow rate is 15mL/min.
(3), after flotation terminates, measure phase CHLOROPHYLLINE sodium up and down respectively and, in the absorbancy at 639nm place, measure up and down the absorbancy of sugar at 490nm place mutually respectively by phenol-sulfuric acid process.Show that the rate of recovery of CHLOROPHYLLINE sodium is 52.56%, enrichment times is 4.24, and the clearance of impurity sugar is 96.93%.
(4) Tripotassium phosphate is reclaimed by alcohol precipitation, filtration, evaporation; Ethanol is reclaimed by rotary evaporation.The ethanol of recovery and Tripotassium phosphate are used for setting up secondary flotation.
embodiment 3:
A method for CHLOROPHYLLINE sodium in double water-phase air-float solvent floatation leaf of bamboo saponification liquor, concrete steps are as follows:
(1) gather the fresh leaf of bamboo and shred.By the broken leaf of bamboo and NaOH solution (10%, w/w) proportionally 1:16(w/v) mix and be placed in 65 DEG C of waters bath with thermostatic control, react 2h under constantly stirring.Juice after saponification is obtained leaf of bamboo saponification liquor and be placed in 4 DEG C of refrigerators preserving through cloth and filter paper filtering successively, within every 5 days, again prepares a storing solution.
(2) 0.58g/mL Tripotassium phosphate solution and 5mL leaf of bamboo saponification liquor are mixed in 50mL colorimetric cylinder, make mixed solution cumulative volume be 50mL.Being transferred to by mixed solution in flotation column and adding the ethanol of 5.5mL from capital end, logical nitrogen flotation 25min, nitrogen flow rate is 20mL/min.
(3), after flotation terminates, measure phase CHLOROPHYLLINE sodium up and down respectively and, in the absorbancy at 639nm place, measure up and down the absorbancy of sugar at 490nm place mutually respectively by phenol-sulfuric acid process.Show that the rate of recovery of CHLOROPHYLLINE sodium is 78.63%, enrichment times is 6.45, and the clearance of impurity sugar is 96.20%.
(4) Tripotassium phosphate is reclaimed by alcohol precipitation, filtration, evaporation; Ethanol is reclaimed by rotary evaporation.The ethanol of recovery and Tripotassium phosphate are used for setting up secondary flotation.
embodiment 4:
A method for CHLOROPHYLLINE sodium in double water-phase air-float solvent floatation leaf of bamboo saponification liquor, concrete steps are as follows:
(1) gather the fresh leaf of bamboo and shred.By the broken leaf of bamboo and NaOH solution (10%, w/w) proportionally 1:16(w/v) mix and be placed in 65 DEG C of waters bath with thermostatic control, react 2h under constantly stirring.Juice after saponification is obtained leaf of bamboo saponification liquor and be placed in 4 DEG C of refrigerators preserving through cloth and filter paper filtering successively, within every 5 days, again prepares a storing solution.
(2) 0.46g/mL Tripotassium phosphate solution and 5mL leaf of bamboo saponification liquor are mixed in 50mL colorimetric cylinder, make mixed solution cumulative volume be 50mL.Being transferred to by mixed solution in flotation column and adding the ethanol of 4.5mL from capital end, logical nitrogen flotation 30min, nitrogen flow rate is 15mL/min.
(3), after flotation terminates, measure phase CHLOROPHYLLINE sodium up and down respectively and, in the absorbancy at 639nm place, measure up and down the absorbancy of sugar at 490nm place mutually respectively by phenol-sulfuric acid process.Show that the rate of recovery of CHLOROPHYLLINE sodium is 46.59%, enrichment times is 3.76, and the clearance of impurity sugar is 95.90%.
(4) Tripotassium phosphate is reclaimed by alcohol precipitation, filtration, evaporation; Ethanol is reclaimed by rotary evaporation.The ethanol of recovery and Tripotassium phosphate are used for setting up secondary flotation.
embodiment 5:
A method for CHLOROPHYLLINE sodium in double water-phase air-float solvent floatation leaf of bamboo saponification liquor, concrete steps are as follows:
(1) gather the fresh leaf of bamboo and shred.By the broken leaf of bamboo and NaOH solution (10%, w/w) proportionally 1:16(w/v) mix and be placed in 65 DEG C of waters bath with thermostatic control, react 2h under constantly stirring.Juice after saponification is obtained leaf of bamboo saponification liquor and be placed in 4 DEG C of refrigerators preserving through cloth and filter paper filtering successively, within every 5 days, again prepares a storing solution.
(2) 0.62g/mL Tripotassium phosphate solution and 5mL leaf of bamboo saponification liquor are mixed in 50mL colorimetric cylinder, make mixed solution cumulative volume be 50mL.Being transferred to by mixed solution in flotation column and adding the ethanol of 5mL from capital end, logical nitrogen flotation 35min, nitrogen flow rate is 15mL/min.
(3), after flotation terminates, measure phase CHLOROPHYLLINE sodium up and down respectively and, in the absorbancy at 639nm place, measure up and down the absorbancy of sugar at 490nm place mutually respectively by phenol-sulfuric acid process.Show that the rate of recovery of CHLOROPHYLLINE sodium is 48.19%, enrichment times is 3.95, and the clearance of impurity sugar is 95.53%.
(4) Tripotassium phosphate is reclaimed by alcohol precipitation, filtration, evaporation; Ethanol is reclaimed by rotary evaporation.The ethanol of recovery and Tripotassium phosphate are used for setting up secondary flotation.
embodiment 6:
A method for CHLOROPHYLLINE sodium in double water-phase air-float solvent floatation leaf of bamboo saponification liquor, concrete steps are as follows:
(1) gather the fresh leaf of bamboo and shred.By the broken leaf of bamboo and NaOH solution (10%, w/w) proportionally 1:16(w/v) mix and be placed in 65 DEG C of waters bath with thermostatic control, react 2h under constantly stirring.Juice after saponification is obtained leaf of bamboo saponification liquor and be placed in 4 DEG C of refrigerators preserving through cloth and filter paper filtering successively, within every 5 days, again prepares a storing solution.
(2) 0.54g/mL Tripotassium phosphate solution and 5mL leaf of bamboo saponification liquor are mixed in 50mL colorimetric cylinder, make mixed solution cumulative volume be 50mL.Being transferred to by mixed solution in flotation column and adding the ethanol of 5mL from capital end, logical nitrogen flotation 20min, nitrogen flow rate is 30mL/min.
(3), after flotation terminates, measure phase CHLOROPHYLLINE sodium up and down respectively and, in the absorbancy at 639nm place, measure up and down the absorbancy of sugar at 490nm place mutually respectively by phenol-sulfuric acid process.Show that the rate of recovery of CHLOROPHYLLINE sodium is 77.53%, enrichment times is 6.25, and the clearance of impurity sugar is 95.61%.
(4) Tripotassium phosphate is reclaimed by alcohol precipitation, filtration, evaporation; Ethanol is reclaimed by rotary evaporation.The ethanol of recovery and Tripotassium phosphate are used for setting up secondary flotation.
embodiment 7:
The surface-active discussion of CHLOROPHYLLINE sodium molecule:
First 50mL deionized water is proceeded in flotation column and ventilate, observe bubble situation; Then 5mL CHLOROPHYLLINE sodium is joined in deionized water and make mixed solution cumulative volume be also 50mL, proceed in flotation column and ventilate, make nitrogen flow rate identical with first time, observe bubble situation.Two kinds of situation comparing result displays, almost do not have bubble formation in deionized water; And in the deionized water containing CHLOROPHYLLINE sodium, there is a large amount of bubble and bubble stabilizes existence.This is because CHLOROPHYLLINE sodium image surface promoting agent equally has surfactivity, it can reduce solution surface tension, improves liquid film viscosity, is so just conducive to bubbling and steady bubble.Therefore, target compound CHLOROPHYLLINE sodium is suitable for double water-phase air-float solvent floatation very much.
embodiment 8:
The contrast of double water-phase air-float solvent floatation and aqueous two-phase extraction:
Double water-phase air-float solvent floatation CHLOROPHYLLINE sodium and aqueous two-phase extraction CHLOROPHYLLINE sodium are contrasted.The condition of double water-phase air-float solvent floatation is: Tripotassium phosphate concentration is 0.55g/mL, and upper phase ethanol original volume is 5mL, and nitrogen flow rate is 25mL/min, and flotation time is 26min, and adding the thick liquid of CHLOROPHYLLINE sodium is 5mL; The condition of aqueous two-phase extraction is: Tripotassium phosphate concentration is 0.4g/mL, and upper phase ethanol original volume is 20mL, and extraction time is 26min, and adding the thick liquid of CHLOROPHYLLINE sodium is 5mL.Comparing result is as shown in table 1.The rate of recovery of double water-phase air-float solvent floatation CHLOROPHYLLINE sodium is slightly lower than aqueous two-phase extraction, but the partition ratio of double water-phase air-float solvent floatation and enrichment times are all apparently higher than aqueous two-phase extraction.Another advantage of double water-phase air-float solvent floatation is that sugared clearance is higher than aqueous two-phase extraction a lot, more effectively can remove impurity.
Table 1 double water-phase air-float solvent floatation and aqueous two-phase extraction comparing result
Claims (7)
1. a method for separation and concentration CHLOROPHYLLINE sodium, is characterized in that, described method is combined at double-aqueous phase system and solvent floatation.
2. the method for a kind of separation and concentration CHLOROPHYLLINE sodium according to claim 1, is characterized in that, the source of described CHLOROPHYLLINE sodium is the leaf of bamboo.
3. the method for a kind of separation and concentration CHLOROPHYLLINE sodium according to claim 1, is characterized in that, carry out according to following step:
(1) gather the fresh leaf of bamboo and carry out pre-treatment and obtain leaf of bamboo saponification liquor, being placed in refrigerator and saving backup, within every 5 days, again preparing a storing solution;
(2) certain density Tripotassium phosphate solution and leaf of bamboo saponification liquor are mixed in colorimetric cylinder, are transferred in flotation column after concussion mixes, then add the ethanol of certain volume from capital end, pass into nitrogen and carry out flotation;
(3) after flotation terminates, take out upper and lower phase respectively, measure up and down the absorbancy of middle CHLOROPHYLLINE sodium and upper sugar mutually mutually with ultraviolet visible spectrophotometry, thus draw the rate of recovery of CHLOROPHYLLINE sodium and the clearance of sugar;
(4) upper phase ethanol and the recovery of lower phase Tripotassium phosphate are used for setting up secondary flotation.
4. the method for a kind of separation and concentration CHLOROPHYLLINE sodium according to claim 1, it is characterized in that, it is characterized in that, the Tripotassium phosphate strength of solution described in step (2) is 0.46-0.62g/mL.
5. the method for a kind of separation and concentration CHLOROPHYLLINE sodium according to claim 1, it is characterized in that, described in step (2), the volume ratio of Tripotassium phosphate solution and leaf of bamboo saponification liquor is 45:5.
6. the method for a kind of separation and concentration CHLOROPHYLLINE sodium according to claim 1, it is characterized in that, described in step (2), ethanol contend is 4-5.5mL.
7. the method for a kind of separation and concentration CHLOROPHYLLINE sodium according to claim 1, it is characterized in that, described in step (2), nitrogen flow rate is 10-30mL/min; Described flotation time is 15-35min.
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