CN101158668B - Microwave auxiliary extraction-high speed adverse current chromatograph joint method and device thereof - Google Patents
Microwave auxiliary extraction-high speed adverse current chromatograph joint method and device thereof Download PDFInfo
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- CN101158668B CN101158668B CN200710030959XA CN200710030959A CN101158668B CN 101158668 B CN101158668 B CN 101158668B CN 200710030959X A CN200710030959X A CN 200710030959XA CN 200710030959 A CN200710030959 A CN 200710030959A CN 101158668 B CN101158668 B CN 101158668B
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Abstract
The invention discloses an interconnection method of a microwave assistance extraction -high speed countercurrent chromatography and a device of a microwave assistance extraction-high speed countercurrent chromatography. First, the microwave assistance extraction pattern is adopted to extract the material. Then, the concentration pre-separation of an extract solution is made. Finally, a goal component is acquired through the purification preparation of the high speed countercurrent chromatograph or the goal component in the extract solution of the natural products is analyzed. The microwave assistance extraction, the separation, the purification and the preparation of the high speed countercurrent chromatograph or the integration online of the analysis are realized through a joint and a transformation control. The invention integrates extraction, separation, purification, preparation or analysis. The invention is characterized in quickness and high efficiency and high selectivity. The quick and high efficient online extraction and separation of the natural products, the purification preparation or the analysis are realized. The invention has the widespread application prospect on food, medicine, biology, etc.
Description
Technical field
The present invention relates to a kind of microwave auxiliary extract-high speed adverse current chromatograph joint technology, and the microwave auxiliary extract-high speed adverse current chromatograph joint instrument of implementing this technology, be applicable to separation, purifying, preparation or the analysis of target component in the fields such as food, medicine and biology.
Background technology
Traditional natural product extraction and separation and purification process are separate, and the running time is long, cause the target component extraction ratio and the recovery low.Microwave-assisted extracts difference that (MAE) technology utilizes material to absorb the microwave ability in microwave field makes in the base matter some regional or some component be heated by selectivity, thereby extraction of substance is separated from matrix fast.It has rapidly and efficiently, saves the strong and favorable reproducibility of solvent, selectivity, characteristics such as applied widely, and the extraction that is suitable for natural materials separates.
High speed adverse current chromatogram (HSCCC) is a kind of liquid luquid partition chromatography separation, analysis and technology of preparing of continuous high-efficient, realization separates according to the partition factor of material in the immiscible liquid of two-phase is different for it, do not need solid carrier, avoided because of problems such as separated material loss that irreversible adsorption causes, inactivation, sex change, sample is all reclaimed, and the sample of recovery more can reflect the characteristic that it is original; And, make the preparation amount of sample improve greatly owing to can fully contact between separated material and the liquid stationary phase.According to the effective difference of separating column volume, high speed adverse current chromatogram mainly comprises two kinds on analytic type and preparation type, and wherein, it is fast that analytic type has the target components velocity of separation, the characteristics that separation purity is high; The preparation type then has advantages such as the purifying ability is strong, preparation amount is big, the recovery is high, applied widely.
Use although these two kinds of technology of microwave current assisted extraction and high speed adverse current chromatogram have more widely in natural products, food, medicine and other fields, all be in relatively independent research state, quality is respectively arranged.
Summary of the invention
For addressing the above problem, the object of the present invention is to provide a kind of microwave auxiliary extract-high speed adverse current chromatograph joint method that integrates extraction separation, purifying preparation or purity analysis rapidly and efficiently, can effectively reduce sample loss, shorten extraction time, improve product recovery rate.
Another object of the present invention provides a kind of microwave auxiliary extract-high speed adverse current chromatograph joint instrument.
In order to achieve the above object, the technical solution used in the present invention is: a kind of microwave auxiliary extract-high speed adverse current chromatograph joint method is characterized in that may further comprise the steps: (1) adopts microwave-assisted extraction pattern to extract material; (2) extract concentrates pre-separation; (3) prepare target components by the countercurrent chromatography purifying, or adopt the target components in the analytic type high speed adverse current chromatogram analysis natural product extraction liquid; Above-mentioned steps realizes the integrated online of microwave-assisted extraction, separation, purifying, high speed adverse current chromatogram preparation or analysis by interface and conversion and control.
Described microwave-assisted extraction pattern comprises that normal pressure extracts and low temperature extracts.
A kind of microwave auxiliary extract-high speed adverse current chromatograph joint device is characterized in that comprising :～microwave-assisted extracts separation module and microwave extraction tank, is used to extract material;～concentrate module, pre-separation module, extract separation module with microwave-assisted and microwave extraction tank is connected, extract is carried out the online pre-separation of online concentrated and object and matrix;～high performance countercurrent chromatography purifying prepares module, is connected the target components in preparation or the analysis extract with concentrated module, pre-separation module;～interface and conversion and control module become one the online connection of above-mentioned each module.Microwave extraction tank in the described extraction separation module is normal pressure microwave extraction tank or low-temperature microwave extractor.
Described extraction separation module, concentrate module, pre-separation module and purifying and prepare module by teflon or the polypropylene connecting pipe is organic is connected, the switching by operation valve of interface and conversion and control module is to realize the full-automatic or semi-automatic operation of complete machine.
The present invention in conjunction with microwave-assisted extract rapidly and efficiently, the advantage of the high efficiency separation purifying of advantage that selective extraction separates and high speed adverse current chromatogram, develop a kind of microwave auxiliary extract-high speed adverse current chromatograph joint method, can effectively reduce target components in the materials such as natural products and prepare sample loss the overall process from extracting, separate, analyze purifying, shorten extraction time, improve product recovery rate.Simultaneously, can be according to the difference of goal in research, target components in preparation of ON-LINE SEPARATION purifying or the ON-LINE SEPARATION analytic sample, parameters such as sample size, extraction pattern, Microwave Extraction condition (as heating-up temperature, extraction time etc.), concentrated pre-separation speed, purifying preparation amount can be controlled as required.
Description of drawings
Fig. 1 is a MAE-HSCCC combined apparatus module frame chart of the present invention;
Fig. 2 is that MAE-HSCCC combined apparatus complete machine of the present invention connects the control schematic block diagram;
Fig. 3 is that MAE-of the present invention prepares the high speed adverse current chromatogram figure that type HSCCC separates the preparation giant knotweed;
Fig. 4 is the high-efficient liquid phase chromatogram (B) of peak VII among giant knotweed extract (A) and the HSCCC;
Fig. 5 is the high speed adverse current chromatogram figure of MAE-analytic type HSCCC compartment analysis Glabrous Sarcandra Herb of the present invention;
Fig. 6 A and 6B are respectively the high-efficient liquid phase chromatograms (B) of peak III among Glabrous Sarcandra Herb extract (A) and the analytic type HSCCC.
Embodiment
Further the present invention will be described with reference to the accompanying drawings, but the present invention is not limited to this specific examples.
The present invention is a kind of microwave auxiliary extract-high speed adverse current chromatograph joint method, it is characterized in that may further comprise the steps: (1) adopts microwave-assisted extraction pattern to extract material; (2) extract concentrates and pre-separation; (3) prepare target components by the countercurrent chromatography purifying, or adopt the target components in the analytic type high speed adverse current chromatogram analysis natural product extraction liquid; Above-mentioned steps is extracted microwave-assisted and the high speed adverse current chromatogram on-line coupling by coupling technique, realizes the integrated online of extraction, separation, purifying, preparation or analysis.Described coupling technique is by interface and conversion and control microwave auxiliary extraction technology and high-speed countercurrent chromatography on-line coupling to be realized the extraction separation and purification preparation of target components or extracted compartment analysis.Described microwave-assisted extraction pattern comprises that normal pressure extracts and low temperature extracts.The controllable parameters such as single purifying preparation amount of extracted amount, pre-separation speed and the high speed adverse current chromatogram of extraction pattern, Microwave Extraction condition (as heating-up temperature, extraction time etc.), single sample are determined according to different objects, do not limit at this.
As shown in Figure 1, 2, microwave auxiliary extract-high speed adverse current chromatograph joint device of the present invention includes: microwave-assisted extracts separation module and microwave extraction tank, is used to extract material; Concentrate module, pre-separation module, extract separation module with microwave-assisted and microwave extraction tank is connected, extract is carried out the online pre-separation of online concentrated and object and matrix; The high performance countercurrent chromatography purifying prepares module, is connected the target components in preparation or the analysis extract with concentrated module, pre-separation module; Interface and conversion and control module realize the online connection of above-mentioned each module and integrated by connecting pipe, transfer valve or reversal valve and peristaltic pump, and circulation or the order of controlling solvent between each module flow.Microwave extraction tank in the described extraction separation module is normal pressure microwave extraction tank or low-temperature microwave extractor.Described high speed adverse current chromatogram comprises preparation type and analytic type.Described extraction separation module, concentrate module, pre-separation module and purifying and prepare module by teflon or the polypropylene connecting pipe is organic is connected, interface and conversion and control module are made up of multidirectional transfer valve (as six-way valve V1 V2 V3, solenoid controlled directional control valve V4 V5 V6), peristaltic pump (or ram pump) P1 P2 P3 P4 P5 P6 and connecting pipe etc.Switch the full-automatic or semi-automatic operation that realizes complete machine by valve.Connecting pipe internal diameter size is adjustable according to the applied sample amount of solvent flow rate and high speed adverse current chromatogram, and diameter is 0.8-3.0mm.The high-speed counter-current chromatograph of purifying module can be selected according to the demand of object.
Embodiment one (separation and purification of employing microwave-assisted extraction-countercurrent chromatography coupling technique prepares the resveratrol in the giant knotweed, obtains highly purified resveratrol)
At first adopt the resveratrol in the microwave-assisted extraction giant knotweed.Its concrete operations are: after the giant knotweed sample is crushed to the suitable particles size, take by weighing 2.0g giant knotweed sample in 200mL normal pressure microwave extraction tank, the methyl alcohol that adds 40mL extracts 10min at 60 ℃ of following microwave-assisted, and extract obtains medicinal extract through pre-separate device, online the concentrating of enrichment facility.Giant knotweed medicinal extract is directly used the preparation of high speed adverse current chromatogram moving phase and stationary phase (1: 1) solvent 2mL dissolving carrying out purifying, at 800rmp with the complete sample introduction of flow velocity 2.0mL/min.The countercurrent chromatography condition is: column volume is 240mL, and dicyandiamide solution is a chloroform: methyl alcohol: water=4: 3: 2 (volume ratio); Above phase methanol is a stationary phase, and following phase methyl alcohol/chloroform is a moving phase; The stationary phase retention rate is about 70%, ultraviolet detection wavelength 254nm.
Fig. 3 prepares the high speed adverse current chromatogram figure of giant knotweed for the microwave auxiliary extract-high speed adverse current chromatographic resolution; Fig. 4 is the high-efficient liquid phase chromatogram of the resveratrol of giant knotweed extract and high speed adverse current chromatogram figure superiors VII correspondence.The giant knotweed sample is after the microwave-assisted extraction directly enters the high speed adverse current chromatogram separation and purification, and the liquid chromatographic detection purity of resveratrol is greater than 95%, and the recovery of resveratrol is not less than 95% in the medicinal extract.
Embodiment two (adopting the isofraxidin in the high speed adverse current chromatograph joint used compartment analysis Glabrous Sarcandra Herb of microwave-assisted extraction-analytic type)
At first adopt the isofraxidin in the microwave-assisted extraction Glabrous Sarcandra Herb.Its concrete operations are: after the Glabrous Sarcandra Herb sample is crushed to the suitable particles size, take by weighing 2.0g Glabrous Sarcandra Herb sample in 100mL normal pressure microwave extraction tank, 60% ethanolic solution that adds 20mL, at 65 ℃ of following Microwave Extraction 20min, extract obtains medicinal extract through pre-separate device, online the concentrating of enrichment facility.Medicinal extract is directly with the dissolving of 5.0mL high speed adverse current chromatogram moving phase, at 1600rmp with flow velocity 1.0mL/min sample introduction 1.0mL.Analytic type high speed adverse current chromatogram condition is: column volume is 43mL, and dicyandiamide solution is a normal hexane: ethyl acetate: methyl alcohol: water=1: 2: 1: 2 (volume ratios); Above phase n-hexane/ethyl acetate is a stationary phase, and following phase methanol is a moving phase; The stationary phase retention rate is about 60%, ultraviolet detection wavelength 254nm.
Fig. 5 is that Glabrous Sarcandra Herb medicinal extract directly carries out the analytic type chromatogram that the high speed adverse current chromatogram separation obtains; Fig. 6 A, Fig. 6 B are respectively the high-efficient liquid phase chromatograms of the isofraxidin of Glabrous Sarcandra Herb extract and analytic type high speed adverse current chromatogram figure superiors III correspondence.The Glabrous Sarcandra Herb sample is after the microwave-assisted extraction directly enters analytic type high speed adverse current chromatogram compartment analysis, and the liquid chromatographic detection purity of isofraxidin is greater than 99%.
Clearly, after adopting microwave auxiliary extract-high speed adverse current chromatograph joint of the present invention with instrument, both can reach and directly from giant knotweed, obtain purity greater than 95% effective constituent resveratrol, resveratrol only need separate with a high speed adverse current chromatogram through microwave-assisted extraction fast, concentrated pre-separation, and the recovery of resveratrol can reach more than 95%; Rapid extraction, separation and the high speed adverse current chromatogram compartment analysis of isofraxidin in also can the canbe used on line Glabrous Sarcandra Herb.Rapidly and efficiently and the optionally advantage that this coupling instrument combines that microwave-assisted extract to separate and the advantage of high speed adverse current chromatogram efficiently purifying preparation, effectively reduce target components extraction in the samples such as natural products, separation, purifying and prepared the sample loss in the overall process, shorten extraction time, improved product recovery rate.
In addition, in above-mentioned example, adopt in the giant knotweed that isofraxidin illustrates as object in the resveratrol and Glabrous Sarcandra Herb, but the present invention is not only applicable to extraction separation, the purifying preparation of effective constituent in the natural products or analyzes, and can be applied to the separating at line drawing fast of target components in other complex objects such as food, medicine, biology, purifying preparation yet or analyzes.
By above explanation as can be known, microwave auxiliary extract-high speed adverse current chromatograph joint of the present invention with instrument have adapt to wide, rapidly and efficiently, strong, the recovery advantages of higher of purifying preparative capacibility, can be widely used in extraction separation, purifying, preparation and the analysis of target components in the complex objects such as food, medicine, biology.
Claims (6)
1. a microwave auxiliary extract-high speed adverse current chromatograph joint method is characterized in that may further comprise the steps: (1) employing normal pressure and low-temperature microwave assisted extraction pattern extraction material; (2) extract concentrates pre-separation; (3) prepare target components by the countercurrent chromatography purifying, or adopt the target components in the analytic type high speed adverse current chromatogram analysis natural product extraction liquid; Above-mentioned steps realizes the integrated online of microwave-assisted extraction, separation, purifying, high speed adverse current chromatogram preparation or analysis by interface and conversion and control.
2. microwave auxiliary extract-high speed adverse current chromatograph joint device is characterized in that comprising:
～microwave-assisted extracts separation module and microwave extraction tank, is used to extract material, and described microwave extraction tank is normal pressure microwave extraction tank or low-temperature microwave extractor;
～concentrate module, pre-separation module, extract separation module with microwave-assisted and microwave extraction tank is connected, extract is carried out the online pre-separation of online concentrated and object and matrix;
～high performance countercurrent chromatography purifying prepares module, is connected the target components in preparation or the analysis extract with concentrated module, pre-separation module;
～interface and conversion and control module connect above-mentioned each module, become one.
3. microwave auxiliary extract-high speed adverse current chromatograph joint device according to claim 2 is characterized in that: it is countercurrent chromatography or analytic type high speed adverse current chromatogram that described high speed adverse current chromatogram purifying prepares module.
4. microwave auxiliary extract-high speed adverse current chromatograph joint device according to claim 2 is characterized in that: described interface and conversion and control module comprise transfer valve, peristaltic pump/ram pump and connecting pipe.
5. microwave auxiliary extract-high speed adverse current chromatograph joint device according to claim 4, it is characterized in that: described extraction separation module, concentrate module, pre-separation module and purifying and prepare module by teflon or the polypropylene connecting pipe is organic is connected, the switching by the valve on the control linkage pipe of interface and conversion and control module is to realize the full-automatic or semi-automatic operation of complete machine.
6. microwave auxiliary extract-high speed adverse current chromatograph joint device according to claim 4 is characterized in that: described connecting pipe internal diameter size is adjustable according to the applied sample amount of solvent flow rate and high speed adverse current chromatogram, and diameter is 0.8-3.0mm.
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| CN103913533B (en) * | 2013-01-09 | 2016-08-24 | 丽珠集团利民制药厂 | For evaluating method and the application thereof of ginseng and astragalus injection for strengthening body chemical composition |
| CN104645667B (en) * | 2013-11-18 | 2017-02-08 | 华东理工大学 | Expanded bed chromatography and counter-current chromatography on-line combination method, application and apparatus thereof |
| CN107151203A (en) * | 2016-03-03 | 2017-09-12 | 复旦大学 | Separate the method for preparing natural naphthoquinone compound |
| CN109187162A (en) * | 2018-09-30 | 2019-01-11 | 镇江华智睿安物联科技有限公司 | A kind of on-line monitoring Microwave Extraction device |
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| CN1434019A (en) * | 2003-03-03 | 2003-08-06 | 清华大学 | Process for on-line extracting and concentrating organic acid in organic acid fermentation |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1434019A (en) * | 2003-03-03 | 2003-08-06 | 清华大学 | Process for on-line extracting and concentrating organic acid in organic acid fermentation |
| CN1490319A (en) * | 2003-07-28 | 2004-04-21 | 浙江一新制药股份有限公司 | Method for separating high-purity galanthamine from short-tube lycoris crude extract |
| CN1763029A (en) * | 2005-09-06 | 2006-04-26 | 浙江大学 | Method for separating rubescensinA from rabdosia |
Non-Patent Citations (8)
| Title |
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| Chen, LG |
| Chen, LG;Jin, HY;Ding, L, et al..On-line coupling of dynamic microwave-assisted extraction with high-performance liquid chromatography for determination of andrographolide and dehydroandrographolide in Andrographis paniculata Nees.《JOURNAL OF CHROMATOGRAPHY A》.2007,第1140卷(第1-2期),71-77. * |
| Ding, L, et al..On-line coupling of dynamic microwave-assisted extraction with high-performance liquid chromatography for determination of andrographolide and dehydroandrographolide in Andrographis paniculata Nees.《JOURNAL OF CHROMATOGRAPHY A》.2007,第1140卷(第1-2期),71-77. |
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| Liu, ZL;Wang, J;Shen, PN, et al..Microwave-assisted extraction and high-speed counter-current chromatography purification of ferulic acid from Radix Angelicae sinensis.《SEPARATION AND PURIFICATION TECHNOLOGY》.2006,第52卷(第1期),18-21. * |
| Shen, PN, et al..Microwave-assisted extraction and high-speed counter-current chromatography purification of ferulic acid from Radix Angelicae sinensis.《SEPARATION AND PURIFICATION TECHNOLOGY》.2006,第52卷(第1期),18-21. |
| Wang, J |
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