A continuous sequential double-column chromatographic separation and purification system and a sophora japonica enzymatic hydrolysis wastewater separation process

Abstract

This invention discloses a continuous sequential dual-column chromatography separation and purification system and a process for separating wastewater from the enzymatic hydrolysis of Sophora japonica, belonging to the field of natural product separation and purification technology. The system consists of two jacketed chromatographic columns, two automatic refractometers, a digital display constant temperature water bath, three flow pumps, and multiple functional storage tanks. Through detector detection and valve switching, the column outlet can selectively connect to different storage tanks, completing the entire process of sample loading, elution, regeneration, and rebalancing. This invention overcomes the technical bottlenecks of traditional single-column chromatography, possessing the advantages of continuous and efficient separation. When applied to the treatment of Sophora japonica enzymatic hydrolysis wastewater, it can achieve efficient separation and enrichment of rutin and D-pinel, with purities reaching over 95% and 90%, respectively, and yields exceeding 90%. The separation effect is comparable to simulated moving bed technology, providing an efficient solution for the resource utilization of Sophora japonica enzymatic hydrolysis wastewater, suitable for large-scale industrial production.

Description

Technical Field

[0001] This invention belongs to the field of natural product separation and purification technology, specifically relating to a continuous sequential dual-column chromatography separation and purification system and a process for separating wastewater from Sophora japonica enzymatic hydrolysis. Background Technology

[0002] Rutinose is a reducing heterodisaccharide composed of α-L-rhamnose and D-glucose linked by α-1,6 glycosidic bonds. Its sugar unit composition and glycosidic bond structure differ fundamentally from common disaccharides such as sucrose, maltose, and lactose. Common disaccharides are mostly formed by the condensation of hexose units with similar configurations; for example, sucrose is composed of α-D-glucose and β-D-fructose, and maltose is composed of two molecules of α-D-glucose linked by glycosidic bonds. In contrast, rutinose is a heterodisaccharide unit composed of 6-deoxyhexose (L-rhamnose) and D-glucose. This unique structure endows rutinose with a unique metabolic pathway and potential physiological activity that distinguish it from conventional disaccharides, and its application potential in functional foods and pharmaceutical intermediates has attracted considerable attention in recent years.

[0003] Currently, rutin sugar is mainly derived from a byproduct of rutin hydrolysis. Existing preparation methods face significant technical bottlenecks: Strong acid hydrolysis easily destroys the glycosidic bonds of rutin, causing it to degrade into monosaccharides, drastically reducing product yield and increasing the difficulty of subsequent separation and purification; enzymatic hydrolysis using exogenous enzyme preparations suffers from high enzyme costs and poor batch-to-batch catalytic efficiency stability, further increasing the recovery cost and scale-up difficulty of rutin sugar. With the deepening research and application of functional sugars in the health industry, the large-scale, low-cost, and stable supply of rutin sugar has become a core bottleneck restricting research on its physiological activity mechanisms and downstream industrial applications.

[0004] Chinese invention patent application 202311264631X reveals that the enzymatic fermentation process of Sophora japonica buds generates a large amount of enzymatic wastewater rich in rutin and D-pinel, providing a novel, low-cost, and renewable raw material source for the preparation of functional sugars. To date, there are no reports, either domestically or internationally, of large-scale processes for the simultaneous and efficient separation and recovery of rutin and D-pinel from Sophora japonica bud enzymatic fermentation wastewater. This type of industrial wastewater, rich in high-value-added functional components, has not yet been developed and utilized for high-value purposes, resulting not only in the waste of valuable biomass resources but also requiring production enterprises to invest in additional wastewater treatment costs, thus imposing a dual burden on both the environment and the economy.

[0005] Simulated moving bed chromatography is a widely used and efficient continuous separation technology in the field of large-scale purification of functional oligosaccharides. However, it generally employs a complex structure of multiple columns in series, which presents challenges such as high equipment investment and maintenance costs, difficulty in coordinating and optimizing multiple operating parameters, and high professional requirements for operators. This makes it difficult to meet the industrialization needs of resource utilization of Sophora japonica processing byproducts. Therefore, developing a continuous chromatographic separation and purification system and supporting processes that are simple in structure, easy to operate, and combine high separation purity with high product yield has significant practical and industrial value. This technology can reduce the treatment cost of Sophora japonica processing wastewater, alleviate the environmental pressure on enterprises, and achieve high-value utilization of industrial waste, aligning with the development concepts of green chemistry and circular economy. Simultaneously, it can fill the technological gap in the low-cost large-scale preparation of rutin sugar and promote the downstream industrial application of functional disaccharides. Summary of the Invention

[0006] In view of this, the purpose of this invention is to provide a continuous sequential dual-column chromatography separation and purification system and a process for separating wastewater from Sophora japonica enzymatic hydrolysis.

[0007] To achieve the above objectives, the present invention provides the following technical solution:

[0008] This invention provides a continuous sequential dual-column chromatography separation and purification system, including a chromatographic separation unit consisting of a first chromatographic column and a second chromatographic column connected in series; An online detection unit is used to monitor the composition of the effluent from the outlet of the first chromatographic column and / or the second chromatographic column; A temperature control unit is used to control the operating temperature of the outlet of the first chromatographic column and / or the second chromatographic column; The control unit is connected to the detection unit by signal and controls the flow path switching according to the detection signal.

[0009] Furthermore, both the first and second chromatographic columns are equipped with inlets and outlets; After the liquid exits from the outlet of the first chromatographic column, the separation waste liquid is detected by the online detection unit, and then the new material to be separated is introduced into the feed port of the second chromatographic column.

[0010] Furthermore, the online detection unit includes an online automatic refractometer.

[0011] Furthermore, the temperature control unit includes a digital display constant temperature water bath for controlling the working temperature of the chromatographic column.

[0012] The present invention also provides a process for purifying rutin and D-pinel from enzymatic hydrolysis wastewater of Sophora japonica, using the above-described system; The pretreated concentrated wastewater from the enzymatic hydrolysis of Sophora japonica flowers was fed into the first jacketed chromatographic column for sample loading, separating the waste liquid from the new analytes to be separated. The new analyte was introduced into the second jacketed column for loading, and elution and separation yielded rutin and D-pinel.

[0013] Furthermore, the pretreated Sophora japonica enzymatic hydrolysis wastewater concentrate is a feed solution pretreated by membrane separation, with a total sugar mass fraction of 20-40%.

[0014] Furthermore, the sample loading volume is 1 / 15 to 1 / 20 of the column volume, and the flow rate of the feed solution is 2-4 mL / min.

[0015] Furthermore, the chromatographic column is operated at 50-75°C.

[0016] Furthermore, the height-to-diameter ratio of the chromatographic column is 15:1-25:1.

[0017] It contains at least the following beneficial technical effects: This invention employs a dual-column tandem continuous sequential separation mode, breaking the limitations of traditional single-column chromatography, such as the limited number of theoretical plates and excessively high bottom pressure under long column conditions. It improves separation accuracy through two-stage tandem separation and achieves continuous separation through a cyclical operation of "product collection and simultaneous second-round sample loading". Compared with the traditional single-column intermittent process, the processing efficiency is improved by more than 50%, with no idle waiting period.

[0018] This system achieves separation results comparable to a simulated moving bed using only two chromatographic columns. Compared to a multi-column simulated moving bed, it reduces equipment investment costs by more than 60%, eliminates the need for complex multi-parameter collaborative optimization, and features convenient operation and low maintenance difficulty, making it perfectly suited for the large-scale industrial production needs of Sophora japonica processing enterprises.

[0019] This invention enables the simultaneous and efficient separation and enrichment of rutin and D-pinel in the enzymatic hydrolysis wastewater of Sophora japonica. The purity of the final rutin can reach over 95%, and the purity of D-pinel can reach over 90%. The yield of both exceeds 90%, and the product indicators are stable, which can directly meet the application requirements of downstream fields such as functional foods and pharmaceutical intermediates.

[0020] Achieving high-value utilization of waste materials aligns with the concept of green development: This invention uses industrial wastewater from the enzymatic hydrolysis of Sophora japonica flowers as raw material, realizing the transformation of waste resources into treasure. It not only fills the technological gap in the low-cost, large-scale preparation of rue sugar, but also significantly reduces the wastewater treatment costs and environmental pressure for enterprises, combining economic value and environmental benefits, and conforming to the development direction of circular economy and green chemical industry. Attached Figure Description

[0021] Figure 1 This is a schematic diagram of the continuous sequential dual-column chromatography separation and purification system of the present invention; Figure 2 The rue candy of this invention1 H-NMR spectrum; Figure 3 The rue candy of this invention 13 C-NMR spectrum; Figure 4 For the present invention, D-pineol 1 H-NMR image Figure 5 For the present invention, D-pineol 13 C-NMR spectrum. Detailed Implementation

[0022] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0023] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0024] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0025] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be obvious to those skilled in the art. This application specification and embodiments are merely exemplary.

[0026] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0027] Unless otherwise specified, "room temperature" and "normal temperature" in this invention refer to 25±2℃.

[0028] Unless otherwise specified, all raw materials or instruments used in the following embodiments of the present invention are commercially available.

[0029] like Figure 1 As shown, a continuous sequential dual-column chromatography separation and purification system includes a chromatography separation unit, an online detection unit, a temperature control unit, a control unit, a fluid delivery unit, a liquid storage unit, and a pipeline valve assembly.

[0030] The chromatographic separation unit includes a first jacketed column (coarse separation column) and a second jacketed column (fine separation column) connected in series. The height-to-diameter ratio of both columns is 15:1-25:1, and the columns are packed with a wet-filled stationary phase, which is a calcium-type or ammonium-type cation exchange resin.

[0031] The online detection unit includes two online automatic refractometers, which are respectively installed in the outlet pipeline of the first jacketed chromatographic column and the outlet pipeline of the second jacketed chromatographic column, for real-time monitoring of the refractive index change of the effluent to identify the material components.

[0032] The temperature control unit includes a digital display constant temperature water bath, whose circulating water path is connected to the jacket cavity of the two jacketed chromatographic columns, and is used to precisely control the working temperature of the chromatographic columns at 50-75℃.

[0033] The fluid transport unit includes three flow pumps: a first feed / elution pump (pump A), a second series transport pump (pump B), and a third elution pump (pump C).

[0034] The liquid storage unit includes multiple functional storage tanks, namely a feed tank for storing raw materials to be separated, an eluent tank for storing the eluent medium (deionized water), a waste liquid tank for collecting waste liquid from the fore- and post-distillate fractions, a rutin product tank for collecting high-purity rutin, and a D-pinel product tank for collecting D-pinel.

[0035] The pipeline valve assembly includes multiple sets of on / off control valves (such as...). Figure 1 Valves 1-6 shown are installed in the corresponding functional pipelines to control the pipeline opening and closing and the fluid flow direction.

[0036] The control unit is connected to the signal output terminals of two online automatic refractometers, the control valves for each on / off switch, and the control terminals of each flow pump. Based on the refractive index signal detected online, the control unit switches the on / off state of the corresponding control valves in real time and controls the start / stop and flow rate of each flow pump. This allows the outlets of the first and second jacketed columns to selectively connect to different functional storage tanks, thereby enabling continuous sequential cycling of sample loading, elution, regeneration, and rebalancing across the dual columns, as well as two-stage tandem separation and purification of materials.

[0037] Example 1 This embodiment provides a process for simultaneously preparing rutin and D-pinel from enzymatic hydrolysis wastewater of Sophora japonica using the system described in the preparation example, as detailed below: Pretreatment of Sophora japonica enzymatic hydrolysis wastewater: Sophora japonica enzymatic hydrolysis wastewater was pretreated using a membrane separation device. The resulting waste mother liquor had a total sugar content of approximately 60% based on its refractive index. This waste mother liquor was diluted with deionized water, resulting in a total sugar content of approximately 30%, which was then used as the feed liquid for separation. Of this, rutin accounted for 50% and D-pinel alcohol accounted for 25% by dry basis mass fraction.

[0038] Continuous sequential dual-column chromatography separation process: Column preparation: Calcium-type cation exchange resin was selected as the stationary phase, with a column height-to-diameter ratio of 20:1. A digital display constant-temperature water bath was started, and 70°C circulating water was supplied to the jackets of both columns to stabilize the column operating temperature at 70°C. Simultaneously, deionized water was circulated at a flow rate of 9.0 mL / min to equilibrate the two columns.

[0039] Feed loading: After equilibration, open valve 1 and start pump A to feed the pretreated feed solution into the first jacketed column at a flow rate of 3.0 mL / min. The loading volume is 1 / 20 of the column volume. During the loading process, open valve 3 to collect the effluent from the first column (mainly containing impurities) into the waste liquid storage tank.

[0040] Coarse separation and tandem fine separation: After sample loading, valve 1 is closed and valve 2 is opened. Deionized water is introduced at a flow rate of 9.0 mL / min to perform adsorption-desorption coarse separation of the material in the first column. The refractive index of the effluent is monitored in real time using an online automatic refractometer at the outlet of the first column. When the target component signal is detected, valve 3 is closed and pump B is started to continuously introduce the effluent from the first column into the second jacketed column for secondary deep fine separation.

[0041] Segmented Product Collection and Synchronous Circulation: The outflow liquid is monitored in real time by an online automatic refractometer at the outlet of the second column. Initially, pumps A and B are shut off, while pump C, valve 6, and valve 3 are opened to collect the foremilk (impurities) into the waste liquid storage tank. When the refractometer detects the target component signal of rutin, the control unit automatically closes valve 3 and opens valve 4 to begin collecting high-purity rutin product into the rutin product storage tank. After the rutin component outflow is complete, the refractometer signal changes, and the control unit closes valve 4 and opens valve 5 to collect D-pineol product into the D-pineol product storage tank. Simultaneously with segmented product collection in the second column, the control unit synchronously initiates the second round of raw material loading (repeating step 2) and coarse separation operation in the first column, achieving parallel processing of "collection" and "loading".

[0042] Continuous cyclic operation: Repeating the above steps completes the full-process separation of the first round of raw materials and the partial separation of the second round of raw materials. Continuous circulation achieves continuous and uninterrupted sequential separation and purification of materials.

[0043] Post-processing: The collected rutin sugar component and D-pineol component are concentrated and spray-dried respectively to obtain high-purity rutin sugar product and D-pineol product.

[0044] Separation results: The yield of the rue sugar product obtained in this example was 92.18%, and the purity was 98.23%; the yield of the D-pineol product was 90.67%, and the purity was 90.42%.

[0045] Example 2 This embodiment is basically the same as that of Embodiment 1, except that the stationary phase of the chromatographic column is replaced with an ammonium cation exchange resin.

[0046] Separation results: After processing according to the process of this embodiment, the yield of the rue sugar product was 91.41% and the purity was 97.34%; the yield of the D-pineol product was 90.35% and the purity was 90.46%.

[0047] Example 3 This embodiment is basically the same as Embodiment 1, except that the total sugar mass fraction of the pretreated feed liquid is adjusted to about 20%, and the feed flow rate is adjusted to 4.0 mL / min.

[0048] Separation results: After processing according to the process of this embodiment, the yield of the rue sugar product was 93.16% and the purity was 98.54%; the yield of the D-pineol product was 91.69% and the purity was 90.89%.

[0049] Example 4 This embodiment is basically the same as Example 1, except that the elution flow rate is adjusted to 8.0 mL / min.

[0050] Separation results: After processing according to the process of this embodiment, the yield of the rue sugar product was 94.59% and the purity was 98.83%; the yield of the D-pineol product was 92.18% and the purity was 91.94%.

[0051] Example 5 This embodiment is basically the same as Example 1, except that the elution flow rate is adjusted to 6.0 mL / min.

[0052] Separation results: After processing according to the process of this embodiment, the yield of the rue sugar product was 95.19% and the purity was 98.98%; the yield of the D-pineol product was 93.54% and the purity was 91.04%.

[0053] Experimental Example 1 Structural confirmation of rutin The organic spectroscopic structure of the rue sugar products prepared in Examples 1-5 was confirmed. All instruments and methods used were conventional organic spectroscopic analysis methods in the field. Specific detection data are as follows: ESI-MS m / z: 349.1105 [M+Na] + Specific curl [α] 22 -4.0° (c 0.40, D2O).

[0054] In the ¹H-NMR spectrum, based on the chemical shifts and integral proportions of the anomeric hydrogens [δα-H-15.24(d,J=4.8Hz,0.4H), δβ-H-14.65(d,J=10.8Hz,0.6H)], the compound was determined to be a mixture of terminal epimers of α-rutose and β-rutose, with α-rutose accounting for 40% and β-rutose accounting for 60%.

[0055] The DEPTQ spectrum revealed 24 carbon signals (with some signal overlap), including 2 methyl signals, 2 methylene signals, and 19 methine signals, among which 4 anomeric carbon signals [δ 100.90, 100.66, 96.05, 92.16]. Preliminary inference suggests that this compound is a mixed disaccharide formed by the condensation of α / β-configured glucose and rhamnose. The splitting peaks at δ 16.70 and 16.64 in the low-field region are characteristic recognition signals of the rhamnose pyranose ring, and this splitting is caused by the configurational difference of the terminal groups of the connected glucose groups. The remaining carbon signals (C2'-C5') of the rhamnose pyranose ring showed no significant shift changes (Δδ < 1 ppm), only minor splitting. The hydrogen signals of H-1', H-2', H-3', H-4', and H-5' in the rhamnose residues were not significantly affected.

[0056] Based on the anomeric hydrogen integral results, the CH coupling constant of the glucose terminal group (3-4 Hz for α-type and 6-8 Hz for β-type, verifying accurate configuration assignment), and the correlation signals of HSQC, HMBC, and ¹H-¹HCOSY two-dimensional spectra, the compound was finally identified as rutinose with the chemical structure α-L-rhamnopyranosyl-(1→6)-α / β-D-glucopyranose, in which α-rutinose accounts for 40% and β-rutinose accounts for 60%.

[0057] Experimental Example 2 Structural confirmation of D-pineol This embodiment confirms the organic spectroscopic structure of the D-pineol products isolated and prepared in Examples 1-5. The detection instruments and methods used are all conventional organic spectroscopic analysis methods in the field. Specific detection data are as follows: ESI-MS m / z: 217.0678 [M+Na]+ The molecular formula is C7H 14 O6; Specific curl [α] 22 +45.2° (c0.30, D2O).

[0058] ¹HNMR(500MHz,D2O)δ:3.92(2H,p,J=4.0Hz,H-1,6),3.72(1H,dd,J=10.0,2.8Hz,H-2),3.67(1H,d d,J=10.0,2.8Hz,H-5),3.56(1H,t,J=9.7Hz,H-4),3.51(3H,s,3-OMe),3.25(1H,t,J=9.7Hz,H-3); ¹³C-NMR (D2O, 125MHz) δ: 72.1 (C-1), 71.6 (C-2), 82.7 (C-3), 70.5 (C-4), 71.4 (C-5), 69.8 (C-6), 59.7 (OMe).

[0059] The above spectral data are in high agreement with the standard data of D-pineol reported in existing literature, therefore the compound is identified as D-pineol.

[0060] Comparative Example 1 Traditional single-column intermittent chromatographic separation process This comparative example uses a conventional single-column intermittent chromatography separation process from existing technology, and is compared in parallel with the continuous sequential dual-column chromatography process of the present invention. The specific operation is as follows: (1) Pretreatment of Sophora japonica enzymatic hydrolysis wastewater: The operation steps and process parameters are completely consistent with those in Example 1.

[0061] (2) Single-column intermittent separation process: The stationary phase of the chromatographic column is calcium-type cation exchange resin, which is packed by wet method. The height-to-diameter ratio and single column volume of the chromatographic column are the same as in Example 1. Before the feeding operation, the digital display constant temperature water bath is started to deliver 70℃ constant temperature circulating water to the chromatographic column jacket and stabilize the working temperature of the chromatographic column at 70℃. At the same time, deionized water is introduced at a flow rate of 9.0 mL / min to equilibrate the chromatographic column.

[0062] After equilibration, feed and sample loading were carried out, with the feed flow rate controlled at 3.0 mL / min and the sample volume at 1 / 20 of the column volume. After sample loading, the elution process was started, with the elution flow rate controlled at 9.0 mL / min. The components of the effluent were monitored in real time using an online automatic refractometer, and the corresponding valves were switched. First, the foremilk impurities such as pigments were collected into the waste liquid storage tank, then the rutin component was collected, followed by the cross-fraction of the two components, which was then collected into the waste liquid storage tank, and finally the D-pinel component was collected.

[0063] (3) Separation results: After processing by the comparative example, the yield of the rue sugar product was 39.59% and the purity was 95.24%, and the yield of the D-pineol product was 32.51% and the purity was 90.23%.

[0064] Comparative Example 2 Traditional single-column intermittent chromatographic separation process The operating steps and process parameters of this comparative example are completely the same as those of Comparative Example 1, except that the stationary phase of the chromatographic column is replaced with an ammonium cation exchange resin.

[0065] Separation results: After processing by the comparative example, the yield of the rue sugar product was 36.43% and the purity was 95.39%, and the yield of the D-pineol product was 30.37% and the purity was 90.28%.

[0066] Therefore, a comparison with Comparative Examples 1 and 2 shows that, under the premise of ensuring comparable product purity (rutin >95%, D-pineol >90%), the system and process described in this invention achieve significantly higher yields of rutin (>90%) and D-pineol (>90%) compared to traditional single-column intermittent chromatography (both yields are below 40%). This indicates that the continuous sequential dual-column system of this invention effectively solves the problems of low separation efficiency and poor yield in traditional single-column chromatography.

[0067] Improved processing efficiency: In Example 1, while the second column was collecting the product, the first column had already begun the next round of sample loading and coarse separation, achieving simultaneous operation of "collection" and "sample loading". In contrast, the single-column process of Comparative Example 1 required waiting for complete elution, regeneration, and equilibration before re-injection, resulting in long equipment downtime. The continuous operation mode of this invention improves processing efficiency by more than 50% compared to Comparative Example 1.

[0068] Optimization of process parameters: Examples 3-5 demonstrate the separation effects under different feed concentrations and elution flow rates. The results show that the process of this invention (total sugar concentration 20-40%, feed flow rate 2-4 mL / min, elution flow rate 5-10 mL / min) can consistently yield products with high purity and high yield, proving that the process has good operational flexibility and stability. Especially at lower elution flow rates (Example 5), although the time may be slightly longer, the yield is further improved.

[0069] In summary, the continuous sequential dual-column chromatography separation and purification system and its process provided by this invention, through a clever design of two columns in series and parallel circulation, combined with online detection and automatic control, successfully achieves efficient, continuous, and high-purity separation of rutin and D-pinel from Sophora japonica enzymatic hydrolysis wastewater. Compared with complex simulated moving bed chromatography, this invention features simpler equipment, lower investment, and easier operation; compared with traditional single-column intermittent chromatography, this invention offers higher separation efficiency and significantly improved product yield, providing a new solution with significant industrial application value for the resource utilization of Sophora japonica enzymatic hydrolysis wastewater.

[0070] The above description is only a preferred embodiment of the present invention. It should be noted that those skilled in the art can make several improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A continuous sequential dual-column chromatographic separation and purification system, characterized in that, It includes a chromatographic separation unit, which consists of a first chromatographic column and a second chromatographic column connected in series; An online detection unit is used to monitor the composition of the effluent from the outlet of the first chromatographic column and / or the second chromatographic column; A temperature control unit is used to control the operating temperature of the outlet of the first chromatographic column and / or the second chromatographic column; The control unit is connected to the detection unit by signal and controls the flow path switching according to the detection signal.

2. The continuous sequential dual-column chromatography separation and purification system according to claim 1, characterized in that, Both the first and second chromatographic columns are equipped with inlets and outlets; After the liquid exits from the outlet of the first chromatographic column, the separation waste liquid is detected by the online detection unit, and then the new material to be separated is introduced into the feed port of the second chromatographic column.

3. The continuous sequential dual-column chromatography separation and purification system according to claim 1, characterized in that, The online detection unit includes an online automatic refractometer.

4. The continuous sequential dual-column chromatography separation and purification system according to claim 1, characterized in that, The temperature control unit includes a digital display constant temperature water bath, used to control the working temperature of the chromatographic column.

5. A process for purifying rutin and D-pinel from enzymatic hydrolysis wastewater of Sophora japonica, characterized in that, The system described in any one of claims 1-4 is used; The pretreated concentrated wastewater from the enzymatic hydrolysis of Sophora japonica flowers was fed into the first jacketed chromatographic column for sample loading, separating the waste liquid from the new analytes to be separated. The new analyte was introduced into the second jacketed column for loading and elution to obtain rutin and D-pinel.

6. The process according to claim 5, characterized in that, The pretreated Sophora japonica enzymatic hydrolysis wastewater concentrate is a feed solution pretreated by membrane separation, with a total sugar mass fraction of 20-40%.

7. The process according to claim 5, characterized in that, The sample loading volume is 1 / 15 to 1 / 20 of the column volume, and the flow rate of the feed solution is 2-4 mL / min.

8. The process according to claim 5, characterized in that, The chromatographic column operates at 50-75℃.

9. The process according to claim 5, characterized in that, The height-to-diameter ratio of the chromatographic column is 15:1-25:1.

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