CN1931835A - Process of extracting citrulline from water melon - Google Patents
Process of extracting citrulline from water melon Download PDFInfo
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- CN1931835A CN1931835A CNA2006101135105A CN200610113510A CN1931835A CN 1931835 A CN1931835 A CN 1931835A CN A2006101135105 A CNA2006101135105 A CN A2006101135105A CN 200610113510 A CN200610113510 A CN 200610113510A CN 1931835 A CN1931835 A CN 1931835A
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- citrulline
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- ultrafiltrated
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Abstract
The process of extracting citrulline from water melon belongs to the field of food and medicine industry technology. The process includes the following steps: stirring water melon juice or water melon peel, stem or leaf leaching liquor and centrifuging to obtain supernatant; filtering with ultrafilter film, regulating pH to 3-5 and cationic exchange resin column separating; linear gradient eluting with phase A of ammonia water and phase B of deionized water in the initial eluent concentration of 0 % and final concentration of 0.5-2.0 %; merging and collecting the eluted liquid near the eluting peaks, vacuum distilling to concentrate and freeze drying to obtain the citrulline product. The citrulline product contains no harmful matter, and may be used in medicine production and as food additive.
Description
Technical field
The invention belongs to food and medicine field, particularly the citrulline technology of preparing.
Background technology
Citrulline is a kind of nonprotein amino acid, has a lot of important physical functions, in medicine and field of food important effect is arranged.
The method of producing citrulline at present mainly is fermentation method, enzyme process and synthesis method.Need to add other chemical additives in the process of these methods production citrulline, the final product that obtains is unsuitable for being directly used in pharmaceutical production and food adds.
Agnes M.Rimando, the method that Penelope M.Per kins-Veazie mentions in article, earlier with the watermelon sample dissolution in 5mL80% methyl alcohol, again through ultrasonication 20 minutes, filtering the back washs with 5mL80% methyl alcohol (MeOH), obtain the citrulline sample and test (A.M.R imando, P.M.Perkins-Veazie.Jornal of Chromatography is (2005) 196-200 A.1078).This method only is used for the usefulness of the test analysis of a small amount of test sample, does not reach industrial level.
Summary of the invention
The object of the present invention is to provide a kind of commercial run that directly extracts citrulline from watermelon, the citrulline product of acquisition does not contain hazardous substance, and can be directly used in pharmaceutical production or as foodstuff additive.
The present invention includes following steps:
(1) Pulp Citrulli juice that will squeeze from flesh of Pulp Citrulli is fully stirring the centrifugal supernatant liquor that gets in back; (2) with the gained supernatant liquor be through molecular weight cut-off after the ultra-filtration membrane ultrafiltration of 5000～8000 (preferred 6000) ultrafiltrated; (3) carry out separation and purification with ion exchange method, the gained ultrafiltrated is regulated pH value to 3～5 (preferred 4) with hydrochloric acid, with 4～8BV/h (resin bed volume/hour) flow velocity upper prop, through cation exchange resin column separation and purification (optimization styrene sulfonic resin); (4) carry out the straight line gradient elution, eluent A is ammoniacal liquor mutually, and B is deionized water mutually, elution flow rate 1BV/h, and eluent initial concentration 0%, stopping concentration is 0.5～2.0%; (5) draw elution curve, merge the eluent of collecting the elution peak place; (6) underpressure distillation concentrates, and the lyophilize of gained concentrated solution is got citrulline (powder).
In above-mentioned (2) step, the tangential flow flow velocity of ultrafiltration is 15～150mL/min, and filter pressure is 0.01～0.10MPa.
In above-mentioned (3) step, the pH value of ion exchange column resin is 4.
In above-mentioned (5) step, after merging the elutriant of collecting the elution peak place, available low-pole macroporous resin dynamically decolours.
In above-mentioned (6) step, the spissated temperature of underpressure distillation can be 40 ℃.
In above-mentioned (6) step, after underpressure distillation concentrates, the gained concentrated solution can be added dehydrated alcohol, the freezing citrulline crystal of separating out.
In (1) step, also can use watermelon stem, skin, leaf to make raw material, raw material is clean, pulverizing adds deionized water and stirring, and the vat liquor that obtains after the lixiviate, obtains supernatant liquor in abundant stirring back centrifugation at 2 hours extraction times.The solid-liquid ratio of leaching process can select 1: 4～8.
In above-mentioned (2) step, before ultrafiltration, also can carry out vacuum filtration and handle, with further removal impurity to supernatant liquor.
Method used in the present invention utilizes ultrafiltration and chromatography method directly to extract citrulline from watermelon, and technical process is short, and energy consumption is low, is suitable for suitability for industrialized production.The citrulline product that obtains does not contain hazardous substance, and can be directly used in pharmaceutical production or as foodstuff additive, and the melon leaf in the agriculture production, melon skin the like waste can be utilized again, produces the high citrulline product of added value, good economy performance.
In the present invention, the supernatant liquor that extracts is filtered, can obviously improve resin isolation efficient, alleviate pollution, prolong the resin life cycle, reduce investment outlay, and can improve product purity resin with ultra-filtration membrane.Adopt the straight line gradient elution, can significantly improve the purity of final citrulline product.
As dynamically decolouring with the low-pole macroporous resin, the elutriant color is obviously shoaled, improve the quality of final citrulline product.
Description of drawings
Accompanying drawing 1 is the process flow sheet of present method.
Embodiment
Embodiment 1:
(1) squeeze the juice: with juice extractor flesh of Pulp Citrulli is pressed into juice, get under the 500ml room temperature and stirred 2 hours, centrifugal 15 minutes of whizzer 5000rpm gets supernatant liquor, vacuum filtration.
(2) ultrafiltration: supernatant liquor is crossed the hollow cellulose film, and (a kind of ultra-filtration membrane, molecular weight cut-off are 6000, model: UEOS503, Tianjin MoTian Membrane Engineering Technology Co., Ltd produces), the tangential flow flow velocity 150mL/min of charging, filter pressure is 0.05MPa, collects ultrafiltrated 460ml.
(3) upper prop: 732 (styrene sulfonic acid type) Zeo-karb is carried out pre-treatment, be treated to the H type, pH value is about 4.Regulating ultrafiltrated PH is 3, with 4BV/h flow velocity upper prop (internal diameter: 19 millimeters, length: 950 millimeters), and the positive upper prop that stops of triketohydrindene hydrate color reaction, finally going up column volume is 380ml.
(4) wash-out: upper prop finishes back top water 40ml, with concentration is that (initial concentration is 0% to 0～0.5% ammoniacal liquor straight line gradient elution, stopping concentration is 0.5%), flow velocity 1BV/h collects elutriant with partly collecting instrument automatically, every 5ml one test tube, collect 80 pipes altogether, utilize the HPLC standard measure to detect citrulline content, draw elution curve, the elutriant at citrulline elution peak place is merged, altogether 100ml.
(5) decolouring: (internal diameter: 13mm height: 250mm) dynamically decolour, flow velocity is 1.0ml/min with the pillar of the elutriant that the merges low-pole macroporous resin by AB-8 (production of resin processing plant of Nankai University) is housed.
(6) concentrate: 40 ℃ of underpressure distillation of the elutriant after will decolouring concentrate, and with enriched material vacuum lyophilization 48h, obtain the faint yellow citrulline powder of 1.69g, yield 75.6%, purity 37.7%.
Embodiment 2:
(1) squeeze the juice: with juice extractor flesh of Pulp Citrulli is pressed into juice, get under the 500ml room temperature and stirred 3 hours, the centrifugal 15min of 5000rpm gets supernatant liquor, vacuum filtration.
(2) ultrafiltration: supernatant liquor is crossed hollow cellulose film (a kind of ultra-filtration membrane, molecular weight cut-off is 6000, model: UEOS503 Tianjin MoTian Membrane Engineering Technology Co., Ltd), and the tangential flow flow velocity 150mL/min of charging, filter pressure is 0.05MPa, collects ultrafiltrated 460ml.
(3) upper prop: 732 (styrene sulfonic acid type) Zeo-karb is carried out pre-treatment, be treated to the H type, pH value is about 4.Regulating ultrafiltrated PH is 4, with 6BV/h flow velocity upper prop (internal diameter: 19 millimeters, length: 950 millimeters), and the positive upper prop that stops of triketohydrindene hydrate color reaction, finally going up column volume is 380ml.
(4) wash-out: upper prop finishes back top water 40ml, with concentration is 0～1.0% ammoniacal liquor straight line gradient elution (initial concentration 0%, stopping concentration is 1.0%), flow velocity 1BV/h collects elutriant with partly collecting instrument automatically, every 5ml one test tube, collect 80 pipes altogether, utilize the HPLC standard measure to detect citrulline content, draw elution curve, the elutriant at citrulline elution peak place is merged, altogether 85ml.
(5) decolouring: (internal diameter: 13mm height: 250mm) dynamically decolour, flow velocity is 1.0ml/min with the pillar of the elutriant that the merges low-pole macroporous resin by AB-8 (production of resin processing plant of Nankai University) is housed.
(6) concentrate: 40 ℃ of underpressure distillation of the elutriant after will decolouring concentrate, and with enriched material vacuum lyophilization 48h, obtain the faint yellow citrulline powder of 1.78g, yield 81.0%, purity 41.0%.Adding the dehydrated alcohol freezing and crystallizing, once can to obtain 0.41g purity be 65% citrulline crystal.
Embodiment 3:
(1) squeeze the juice: with juice extractor flesh of Pulp Citrulli is pressed into juice, get under the 500ml room temperature and stirred 4 hours, the centrifugal 15min of 5000rpm gets supernatant liquor, vacuum filtration.
(2) ultrafiltration: supernatant liquor is crossed the hollow cellulose film, and (a kind of ultra-filtration membrane, molecular weight cut-off are 6000, model: UEOS503, Tianjin MoTian Membrane Engineering Technology Co., Ltd produces), the tangential flow flow velocity 150mL/min of charging, filter pressure is 0.05MPa, collects ultrafiltrated 460ml.
(3) upper prop: 732 (styrene sulfonic acid type) Zeo-karb is carried out pre-treatment, be treated to the H type, pH value is about 4.Regulating ultrafiltrated PH is 5, with 8BV/h flow velocity upper prop (internal diameter: 19 millimeters, length: 950 millimeters), and the positive upper prop that stops of triketohydrindene hydrate color reaction, finally going up column volume is 360ml.
(4) wash-out: upper prop finishes back top water 40ml, with concentration is 0～2.0% ammoniacal liquor straight line gradient elution (initial concentration 0%, stopping concentration is 2.0%), flow velocity 1BV/h collects elutriant with partly collecting instrument automatically, every 5ml one test tube, collect 80 pipes altogether, utilize the HPLC standard measure to detect citrulline content, draw elution curve, the elutriant at citrulline elution peak place is merged, altogether 80ml.
(5) decolouring: (internal diameter: 13mm height: 250mm) dynamically decolour, flow velocity is 1.0ml/min with the pillar of the elutriant that the merges low-pole macroporous resin by AB-8 (production of resin processing plant of Nankai University) is housed.
(6) concentrate: 40 ℃ of underpressure distillation of the elutriant after will decolouring concentrate, and with enriched material vacuum lyophilization 48h, obtain the faint yellow citrulline powder of 1.72g, yield 77.5%, purity 35.5%.Adding the dehydrated alcohol freezing and crystallizing, once can to obtain 0.43g purity be 57% citrulline crystal.
Claims (10)
1. a method of extracting citrulline from watermelon is characterized in that: comprise the steps that the Pulp Citrulli juice that (1) will squeeze is fully stirring the centrifugal supernatant liquor that gets in back from flesh of Pulp Citrulli; (2) with the gained supernatant liquor be through molecular weight cut-off after 5000～8000 the ultra-filtration membrane ultrafiltration ultrafiltrated; (3) the gained ultrafiltrated is regulated pH value to 3～5 with hydrochloric acid, with 4～8BV/h flow velocity upper prop, through the cation exchange resin column separation and purification; (4) carry out the straight line gradient elution, eluent A is ammoniacal liquor mutually, and B is deionized water mutually, elution flow rate 1BV/h, and eluent initial concentration 0%, stopping concentration is 0.5～2.0%; (5) merge the elutriant of collecting the elution peak place; (6) underpressure distillation concentrates, and the lyophilize of gained concentrated solution is got citrulline.
2. method according to claim 1 is characterized in that: in (2) step, the molecular weight cut-off of described ultra-filtration membrane is 6000, and the tangential flow flow velocity of ultrafiltration is 15～150mL/min, and filter pressure is 0.01～0.10MPa.
3. method according to claim 1 and 2 is characterized in that: in (3) step, be the H type with the pre-treatment of ion exchange column resin, pH value is 4.
4. according to claim 1 or 2 or 3 described methods, it is characterized in that: in (3) step, the gained ultrafiltrated is regulated pH value to 4 with hydrochloric acid.
5. according to the arbitrary described method of claim 1～4, it is characterized in that: the Zeo-karb described in (3) step is a styrene sulfonic acid type resin.
6. according to the arbitrary described method of claim 1～5, it is characterized in that: (5) step dynamically decoloured with the low-pole macroporous resin after merging the elutriant of collecting the elution peak place.
7. according to the arbitrary described method of claim 1～6, it is characterized in that: in (6) step, the spissated temperature of underpressure distillation is 40 ℃.
8. according to the arbitrary described method of claim 1～7, it is characterized in that: in (6) step, after underpressure distillation concentrates, the gained concentrated solution is added dehydrated alcohol, the freezing citrulline crystal of separating out.
9. according to the arbitrary described method of claim 1～8, it is characterized in that: in (1) step, described supernatant liquor is to obtain fully stirring the back centrifugation by watermelon stem, skin, leaf being pulverized the vat liquor that adds gained after the deionized water and stirring lixiviate.
10. method according to claim 9 is characterized in that, in (1) step, the solid-liquid ratio of leaching process is 1: 4～8.
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| Application Number | Priority Date | Filing Date | Title |
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| CNA2006101135105A CN1931835A (en) | 2006-09-29 | 2006-09-29 | Process of extracting citrulline from water melon |
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| CNA2006101135105A CN1931835A (en) | 2006-09-29 | 2006-09-29 | Process of extracting citrulline from water melon |
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| CN1931835A true CN1931835A (en) | 2007-03-21 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101372465B (en) * | 2007-08-24 | 2011-11-16 | 北京健健康康生物技术有限公司 | Industrial method for extracting natural L-citrulline from water melon or smacking watermelon and other plant tissues |
| CN103063762A (en) * | 2012-12-21 | 2013-04-24 | 青岛谱尼测试有限公司 | Method for measuring citrulline contents in watermelon by utilizing high performance liquid chromatography |
| CN103360283A (en) * | 2012-04-11 | 2013-10-23 | 周学义 | Method for extracting citrulline from selenium-contained watermelon plant tissues |
| CN104418774A (en) * | 2013-09-11 | 2015-03-18 | 安阳天尊生物工程有限公司 | Method for extracting L-citrulline employing microbial fermentation of trichosanthes kirilowii maxim pulp |
| CN108822001A (en) * | 2018-08-30 | 2018-11-16 | 中国农业科学院郑州果树研究所 | A method of extracting citrulling from watermelon |
-
2006
- 2006-09-29 CN CNA2006101135105A patent/CN1931835A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101372465B (en) * | 2007-08-24 | 2011-11-16 | 北京健健康康生物技术有限公司 | Industrial method for extracting natural L-citrulline from water melon or smacking watermelon and other plant tissues |
| CN103360283A (en) * | 2012-04-11 | 2013-10-23 | 周学义 | Method for extracting citrulline from selenium-contained watermelon plant tissues |
| CN103360283B (en) * | 2012-04-11 | 2014-12-31 | 周学义 | Method for extracting citrulline from selenium-contained watermelon plant tissues |
| CN103063762A (en) * | 2012-12-21 | 2013-04-24 | 青岛谱尼测试有限公司 | Method for measuring citrulline contents in watermelon by utilizing high performance liquid chromatography |
| CN104418774A (en) * | 2013-09-11 | 2015-03-18 | 安阳天尊生物工程有限公司 | Method for extracting L-citrulline employing microbial fermentation of trichosanthes kirilowii maxim pulp |
| CN108822001A (en) * | 2018-08-30 | 2018-11-16 | 中国农业科学院郑州果树研究所 | A method of extracting citrulling from watermelon |
| CN108822001B (en) * | 2018-08-30 | 2019-08-16 | 中国农业科学院郑州果树研究所 | A method of extracting citrulling from watermelon |
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