A method for preparing a gentiana scabra extract

Abstract

The application discloses a preparation method of gentiana scabra extract and relates to the technical field of plant extract. The preparation method comprises the following steps: S1, ultrasonic extraction is conducted on gentiana scabra by using a first ethanol aqueous solution, solid-liquid separation is conducted, and a clear liquid is obtained; S2, the clear liquid is sequentially subjected to decoloring treatment and ultrafiltration, and a filtrate is obtained; S3, column chromatography is conducted on the filtrate by using a macroporous resin, water, a second ethanol aqueous solution and a third ethanol aqueous solution are sequentially used for elution, the second ethanol aqueous solution eluent is collected, and a loganin acid product is obtained; the third ethanol aqueous solution eluent is collected, and a gentiopicroside product is obtained; the volume percentage of ethanol in the second ethanol aqueous solution is 20% to 40%; and the volume percentage of ethanol in the third ethanol aqueous solution is 60% to 80%. The process is simple, the production efficiency is high, the two products of gentiopicroside and loganin acid can be effectively separated at the same time, the method is suitable for industrial production, and the method is environment-friendly.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of plant extracts, and particularly relates to a preparation method of gentiana scabra extract. BACKGROUND

[0002] Gentiana scabra Bunge is a perennial herb of Gentianaceae and Gentiana. The main active chemical components of gentiana are iridoid glycosides, including gentiopicroside, loganic acid, 6'-O-β-D-glucosyl gentiopicroside, swertiamarin and sweroside. Among them, gentiopicroside and loganic acid have the highest content and have the functions of protecting liver, promoting bile, resisting inflammation, protecting nervous system and resisting cancer.

[0003] In related art, a new preparation method of gentiana effective part and its application (application number: 200410042874.X) provides a preparation method. The crude powder of gentiana medicinal material is extracted by hot reflux with 30% to 90% methanol, ethanol or acetone. The extract is concentrated under reduced pressure to 1g to 5g of medicinal material per milliliter of concentrated solution, and is refrigerated for more than 24 hours. The precipitate is removed by centrifugation or filtration to obtain the extract. The extract is purified by column chromatography with DIAION HP20 as the filler. 0.5mL to 2.5mL of liquid medicine is added to each gram of filler. The filler is first washed with deionized water, and then eluted with 40% to 70% methanol or ethanol. The alcohol eluate is collected, dried under reduced pressure, and vacuum dried to obtain the gentiana effective part mainly composed of loganic acid and gentiopicroside (the content of loganic acid and gentiopicroside is 50% to 95%). The process does not effectively separate loganic acid and gentiopicroside, and only obtains a mixture of the two.

[0004] A method for preparing high-purity gentiopicroside (application number: 200610016839.X) selects chloroform-methanol-n-butanol-water as the solvent system, and adopts small-scale preparation and semi-preparation high-speed counter-current chromatography. Once separation, gentiopicroside can be obtained. The process uses toxic and harmful organic solvents such as chloroform, methanol and n-butanol, which is not conducive to the environment, health and safety. In addition, the production capacity of the preparation chromatography is low, which is not conducive to industrial production.

[0005] A preparation method of a gentian extract, gentiopicroside, and application thereof (application number: 202111436362.1), the gentian root and stem are crushed, then pure water is added for extraction, then the extract is filtered and concentrated to obtain a concentrated solution; the concentrated solution is filtered, then the filtrate is poured into a macroporous resin, then water is added for elution, the eluate is discharged, then ethanol is poured for elution, the eluate is collected, and the ethanol is recovered, then a extract is obtained; the extract is added to silica gel, stirred uniformly, and then subjected to silica gel column chromatography, eluted with a mixed mobile phase of ethyl acetate, ethanol and water, eluted until no gentiopicroside is detected by thin layer chromatography, the eluate is collected, concentrated to 1 / 3-1 / 2 of the eluate volume, allowed to stand for crystallization, filtered, the filter cake is washed with ethanol, and dried to obtain gentiopicroside. The process does not identify the content and yield of gentiopicroside, and the separation method of silica gel column chromatography not only requires the use of a large amount of organic solvent (ethyl acetate), but also has low production capacity.

[0006] In summary, it is necessary to provide a method which is environmentally friendly, has high production efficiency, and can effectively separate gentiopicroside and loganic acid. SUMMARY

[0007] The present application aims to at least solve one of the technical problems existing in the prior art. To this end, the present application provides a preparation method of a gentian extract, which is simple, has high production efficiency, can effectively separate gentiopicroside and loganic acid at the same time, is suitable for industrial production, and only uses organic solvent ethanol in the preparation process, which is environmentally friendly.

[0008] The present application also discloses a gentian extract prepared by the preparation method.

[0009] The present application also discloses application of the gentian extract in preparation of cosmetics, food, health products or medicines.

[0010] According to the preparation method of the gentian extract according to the first aspect of the present application, the following steps are included:

[0011] S1, ultrasonic extraction of gentian with a first ethanol aqueous solution, solid-liquid separation, to obtain a clear liquid;

[0012] S2, the clear liquid is sequentially subjected to decolorization treatment and ultrafiltration to obtain a filtrate;

[0013] S3, column chromatography of the filtrate by using a macroporous resin, elution with water, a second ethanol aqueous solution and a third ethanol aqueous solution in sequence, collection of the second ethanol aqueous solution eluate, to obtain a loganic acid product; and collection of the third ethanol aqueous solution eluate, to obtain a gentiopicroside product;

[0014] The volume percentage of ethanol in the second ethanol aqueous solution is 20%-40%;

[0015] The volume percentage of ethanol in the third ethanol aqueous solution is 60% to 80%.

[0016] According to the preparation method of the embodiment of the present application, at least the following beneficial effects are achieved:

[0017] The preparation method of the embodiment has simple process, high production efficiency, and high raw material utilization efficiency, can effectively reduce the production cost, and is suitable for industrial production. The high content of gentiopicroside product and loganic acid product can be obtained at the same time, and the resource utilization efficiency is improved. Moreover, only the ethanol aqueous solution is used as the solvent, and the production process is environmentally friendly, safe and healthy.

[0018] According to some embodiments of the present application, the volume percentage of ethanol in the first ethanol aqueous solution is 30% to 50%. In this way, the solvent polarity can be close to the target component, thereby ensuring high target component extraction efficiency and reducing the extraction rate of impurity components.

[0019] According to some embodiments of the present application, the solid-liquid ratio of the gentian and the first ethanol aqueous solution is 1 kg: 15 L to 25 L.

[0020] According to some embodiments of the present application, the ultrasonic frequency of the ultrasonic extraction is 25 KHz to 40 KHz. For example, it can be specifically 30 KHz.

[0021] According to some embodiments of the present application, the ultrasonic temperature of the ultrasonic extraction is 25±5℃.

[0022] According to some embodiments of the present application, the ultrasonic extraction time is 30 min to 60 min.

[0023] According to some embodiments of the present application, the solid-liquid separation is selected from centrifugation or suction filtration.

[0024] According to some embodiments of the present application, the resin used for the decolorization treatment includes at least one of LSA-900E and LSA-960. In this way, decolorization and impurity removal can be performed to perform preliminary purification.

[0025] According to some embodiments of the present application, the molecular weight cut-off of the ultrafiltration is 500 to 1000. Too large molecular weight cut-off can easily lead to a decrease in the filtered impurities, affecting the purification effect; too small molecular weight cut-off can filter out the target components, affecting the product yield.

[0026] According to some embodiments of the present application, the filtrate needs to be subjected to concentration pretreatment before the column chromatography. The concentration pretreatment can be performed by a reverse osmosis membrane. In this way, the solvent can be recovered, and the loading concentration of the macroporous resin can be improved.

[0027] According to some embodiments of the present application, the temperature of the concentrated pretreatment is 25±5℃. In this way, the thermal degradation of the target component can be reduced.

[0028] According to some embodiments of the present application, the filtrate is concentrated to a soluble solid content of 5% to 10% by the pretreatment.

[0029] According to some embodiments of the present application, in step S3, the type of the macroporous resin includes at least one of LSA-10, LX-12, LX-38, and AB-8.

[0030] According to some embodiments of the present application, in step S3, the amount of water used is 1BV to 3BV.

[0031] According to some embodiments of the present application, in step S3, the amount of the second ethanol aqueous solution used is 1BV to 3BV.

[0032] According to some embodiments of the present application, in step S3, the amount of the third ethanol aqueous solution used is 1BV to 3BV.

[0033] According to some embodiments of the present application, the preparation method further comprises concentrating and drying the second ethanol aqueous solution eluate and / or the third ethanol aqueous solution eluate.

[0034] According to some embodiments of the present application, the concentration can be reduced pressure concentration. The temperature is not higher than 65℃. The single concentration time is controlled within 3h.

[0035] According to some embodiments of the present application, the drying is selected from microwave drying or freeze drying.

[0036] According to some embodiments of the present application, in the gentiopicroside product, the content of gentiopicroside is not less than 90%. Further, not less than 94%.

[0037] According to some embodiments of the present application, in the loganic acid product, the content of loganic acid is not less than 90%.

[0038] According to some embodiments of the present application, the transfer rate of gentiopicroside is not less than 80%.

[0039] According to some embodiments of the present application, the transfer rate of loganic acid is not less than 80%. Further, not less than 82%.

[0040] The gentiana extract prepared by the above preparation method according to the second aspect of embodiments of the present application.

[0041] According to some embodiments of the present application, the gentiana extract includes at least one of the loganic acid product and the gentiopicroside product.

[0042] Use of the above-mentioned gentian extract according to the third aspect of the present application in the preparation of cosmetics, food, health products or pharmaceuticals.

[0043] Other features and advantages of the present application will be set forth in the following description, and in part will become apparent to those skilled in the art from the description, or can be learned by practice of the present application. BRIEF DESCRIPTION OF DRAWINGS

[0044] Figure 1 is a liquid chromatogram of a control solution (strychnine acid standard and gentiopicroside standard) ;

[0045] Figure 2 is a liquid chromatogram of a strychnine acid product of Example 1 of the present application;

[0046] Figure 3 is a liquid chromatogram of a gentiopicroside product of Example 1 of the present application;

[0047] Figure 4 is a liquid chromatogram of a strychnine acid product of Comparative Example 1 of the present application;

[0048] Figure 5 is a liquid chromatogram of a gentiopicroside product of Comparative Example 1 of the present application;

[0049] Figure 6 is a liquid chromatogram of a strychnine acid product of Comparative Example 2 of the present application;

[0050] Figure 7 is a liquid chromatogram of a gentiopicroside product of Comparative Example 2 of the present application;

[0051] Figure 8 is a liquid chromatogram of a strychnine acid product of Comparative Example 3 of the present application;

[0052] Figure 9 is a liquid chromatogram of a gentiopicroside product of Comparative Example 3 of the present application. DETAILED DESCRIPTION

[0053] The idea and the technical effects of the present application will be described in detail below with reference to the embodiments, so as to fully understand the purpose, features and effects of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments of the present application, other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the present application.

[0054] The specific conditions not mentioned in the embodiments are carried out according to the conventional conditions or the conditions suggested by the manufacturers. The reagents or instruments not mentioned by the manufacturers are all conventional products that can be obtained by market purchase.

[0055] In the description of the present application, if there is described first, second, third, etc. only for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of indicated technical features or implicitly indicating the sequence of the indicated technical features.

[0056] In the description of the present application, the terms "include" and "have" and any variations thereof are intended to cover non-exclusive inclusion, for example, a process, method, system, product or device including a series of steps or units does not have to be limited to those steps or units clearly listed, but can include other steps or units not clearly listed or inherent to these processes, methods, products or devices.

[0057] Unless otherwise specified, "not less than" in the present application means greater than and greater than or equal to.

[0058] Unless otherwise specified, "room temperature" in the present application refers to 25℃±5℃.

[0059] In the embodiments of the present application, the content of gentiopicroside in the gentiana crude material is 5.02%, and the content of loganic acid is 1.15%.

[0060] In the embodiments of the present application, the gentiopicroside standard (purity: 99.77%) and the loganic acid standard (purity: 99.38%) are purchased from Chengdu Man Sit Biotechnology Co., Ltd.

[0061] In the embodiments of the present application, the liquid chromatography conditions of the loganic acid product and the gentiopicroside product are as follows:

[0062] Chromatographic column: C18 chromatographic column (250mm×4.6mm, 5μm);

[0063] Mobile phase: 0.4% phosphoric acid aqueous solution (A)-methanol (B); gradient elution (0-10min, 10%B→28%B; 10-30min, 28%B→29%B);

[0064] Equilibrium time: 5min;

[0065] Flow rate: 1mL / min;

[0066] Detection wavelength: 240nm;

[0067] Column temperature: 35℃.

[0068] The calculation formula of the product content is as follows:

[0069]

[0070] A 样The peak area of gentiopicroside or loganic acid in the sample solution; A 对 The peak area of gentiopicroside or loganic acid in the control solution; W 样 The sample weight (mg); W 对 The sample weight (mg); V 样 The sample solution volume (mL); V 对 The control solution volume (mL); K is the purity of the standard.

[0071] The preparation method of the control solution is as follows:

[0072] A certain amount of gentiopicroside standard and loganic acid standard were precisely weighed and placed in a volumetric flask, 80% (V / V) methanol solution was added, and a mixed standard solution with a gentiopicroside concentration of 0.4 mg / mL and a loganic acid concentration of 0.5 mg / mL was prepared.

[0073] The preparation method of the sample solution is as follows:

[0074] A certain amount of loganic acid product and gentiopicroside product were precisely weighed and placed in a volumetric flask, 80% (V / V) methanol solution was added, and the solution was ultrasonically dissolved. Then, the solution was diluted to the mark with 80% methanol solution, filtered through a 0.22 μm microporous filter, and used.

[0075] The calculation formula of the product transfer rate is as follows:

[0076] Transfer rate % = (product ingredient content * product weight) / (gentian raw material weight * gentian raw material ingredient content) * 100%;

[0077] Wherein, the ingredient refers to gentiopicroside or loganic acid.

[0078] Example 1

[0079] The present embodiment provides a preparation method of gentian extract, and the specific process is as follows:

[0080] S1, 10 kg of dried gentian raw material was weighed and placed in an ultrasonic extraction tank, 150 L of 50% (V / V) ethanol solution was added, and ultrasonic extraction was carried out once at room temperature, with an ultrasonic frequency of 30 KHz and an extraction time of 60 min / time, to obtain the extraction liquid. The extraction liquid was filtered through a double filter and then centrifuged in a butterfly centrifuge to obtain the clear liquid.

[0081] S2, the clear liquid was decolorized by LSA-900E resin, and 131 L of effluent was collected. The effluent was further filtered through an ultrafiltration membrane (molecular weight 1000), and the filtrate and the retentate were collected respectively.

[0082] S3, under normal temperature, the filtrate was concentrated by reverse osmosis membrane, and 15 L of concentrated solution (soluble solid content concentration of 6.21%) was obtained; the concentrated solution was continuously passed through LSA-10 macroporous resin, the loading flow rate was 1 BV / h, then 2 BV of deionized water, 2 BV of 30% (V / V) ethanol aqueous solution and 2 BV of 70% (V / V) ethanol aqueous solution were used for elution in sequence, and the elution flow rate was 1 BV / h. The eluate of 30% ethanol aqueous solution and the eluate of 70% ethanol aqueous solution were collected respectively, and were concentrated under reduced pressure and dried by microwave to obtain 0.105 kg of loganic acid product and 0.431 kg of gentiopicroside product respectively.

[0083] It was detected that the content of loganic acid in the loganic acid product was 91.09%, and the transfer rate was 83.17%; the content of gentiopicroside in the gentiopicroside product was 94.58%, and the transfer rate was 81.20%.

[0084] Example 2

[0085] The present example provides a preparation method of gentian extract, and the specific process is as follows:

[0086] S1, 10 kg of dried gentian raw material was weighed and placed in an ultrasonic extraction tank, 200 L of 40% (V / V) ethanol aqueous solution was added, and ultrasonic extraction was carried out once at room temperature, the ultrasonic frequency was 30 KHz, and the extraction time was 30 min / time, to obtain the extraction solution. The extraction solution was filtered by double filtration, and then was centrifuged in a butterfly centrifuge to obtain the clear solution.

[0087] S2, the clear solution was decolorized by LSA-900E resin, and 182 L of effluent was collected. The effluent was continuously passed through an ultrafiltration membrane (molecular weight of 500), and the filtrate and the retentate were collected respectively.

[0088] S3, under normal temperature, the filtrate was concentrated by reverse osmosis membrane, and 16 L of concentrated solution (soluble solid content concentration of 7.06%) was obtained; the concentrated solution was continuously passed through LX-12 macroporous resin, the loading flow rate was 1 BV / h, then 2 BV of deionized water, 2 BV of 20% (V / V) ethanol aqueous solution and 2 BV of 60% (V / V) ethanol aqueous solution were used for elution in sequence, and the elution flow rate was 1 BV / h. The eluate of 20% ethanol aqueous solution and the eluate of 60% ethanol aqueous solution were collected respectively, and were concentrated under reduced pressure and dried by microwave to obtain 0.107 kg of loganic acid product and 0.439 kg of gentiopicroside product respectively.

[0089] It was detected that the content of loganic acid in the loganic acid product was 90.98%, and the transfer rate was 84.65%; the content of gentiopicroside in the gentiopicroside product was 94.36%, and the transfer rate was 82.52%.

[0090] Example 3

[0091] The embodiment provides a preparation method of gentiana scabra extract, and the specific process is as follows:

[0092] S1, 10 kg of dried gentiana scabra raw material is weighed and placed in an ultrasonic extraction tank, 250 L of 30% (V / V) ethanol aqueous solution is added, ultrasonic extraction is carried out once at room temperature, the ultrasonic frequency is 30 KHz, and the extraction time is 45 min / time, to obtain the extraction liquid. The extraction liquid is filtered through a double filter, and then is centrifuged in a butterfly centrifuge to obtain the clear liquid.

[0093] S2, the clear liquid is decolorized through LSA-960 resin, and 229 L of effluent is collected. The effluent is continuously filtered through an ultrafiltration membrane (with a molecular weight of 1000), and the filtrate and the retentate are collected respectively.

[0094] S3, the filtrate is concentrated through a reverse osmosis membrane at room temperature, and 18 L of concentrated liquid (with a soluble solid content concentration of 6.75%) is obtained; the concentrated liquid is continuously filtered through LX-38 macroporous resin, the loading flow rate is 1 BV / h, then 2 BV of deionized water, 2 BV of 40% (V / V) ethanol aqueous solution and 2 BV of 80% (V / V) ethanol aqueous solution are sequentially used for elution, and the elution flow rate is 1 BV / h. The eluate of the 40% ethanol aqueous solution and the eluate of the 80% ethanol aqueous solution are collected respectively, are concentrated under reduced pressure, and are microwave dried to obtain 0.108 kg of loganic acid product and 0.446 kg of gentiopicroside product respectively.

[0095] It is detected that the content of loganic acid in the loganic acid product is 91.02%, and the transfer rate is 85.48%; the content of gentiopicroside in the gentiopicroside product is 94.23%, and the transfer rate is 83.72%.

[0096] Embodiment 4

[0097] The embodiment provides a preparation method of gentiana scabra extract, and the specific process is as follows:

[0098] S1, 10 kg of dried gentiana scabra raw material is weighed and placed in an ultrasonic extraction tank, 200 L of 30% (V / V) ethanol aqueous solution is added, ultrasonic extraction is carried out once at room temperature, the ultrasonic frequency is 30 KHz, and the extraction time is 60 min / time, to obtain the extraction liquid. The extraction liquid is filtered through a double filter, and then is centrifuged in a butterfly centrifuge to obtain the clear liquid.

[0099] S2, the clear liquid is decolorized through LSA-960 resin, and 183 L of effluent is collected. The effluent is continuously filtered through an ultrafiltration membrane (with a molecular weight of 500), and the filtrate and the retentate are collected respectively.

[0100] S3, under normal temperature, the filtrate was concentrated by reverse osmosis membrane to obtain 14 L of concentrated solution (soluble solid content concentration of 6.83%); the concentrated solution was continuously passed through AB-8 macroporous resin, the loading flow rate was 1 BV / h, then 2 BV of deionized water, 2 BV of 40% (V / V) ethanol aqueous solution, and 2 BV of 70% (V / V) ethanol aqueous solution were used for elution in sequence, and the elution flow rate was 1 BV / h. The eluate of 40% ethanol aqueous solution and the eluate of 80% ethanol aqueous solution were collected respectively, and were concentrated under reduced pressure and dried by microwave to obtain 0.105 kg of loganic acid product and 0.43 kg of gentiopicroside product respectively.

[0101] It was detected that the content of loganic acid in the loganic acid product was 90.54%, and the transfer rate was 82.67%; the content of gentiopicroside in the gentiopicroside product was 95.17%, and the transfer rate was 81.52%.

[0102] Comparative Example 1

[0103] This comparative example provides a preparation method of gentian extract, which is basically the same as the preparation method of Example 1, and the only difference is that there is no resin adsorption and decolorization step. The specific process is as follows:

[0104] S1, 10 kg of dried gentian raw material was weighed and placed in an ultrasonic extraction tank, 150 L of 50% (V / V) ethanol aqueous solution was added, and ultrasonic extraction was carried out once at room temperature, the ultrasonic frequency was 30 KHz, and the extraction time was 60 min / time, to obtain the extraction liquid. After the extraction liquid was filtered by double filtration, it was transferred into a butterfly centrifuge for centrifugation to obtain the clear liquid.

[0105] S2, the clear liquid was passed through an ultrafiltration membrane (molecular weight of 1000), and the filtrate and the retentate were collected respectively.

[0106] S3, under normal temperature, the filtrate was concentrated by reverse osmosis membrane to obtain 16 L of concentrated solution (soluble solid content concentration of 7.51%); the concentrated solution was continuously passed through LSA-10 macroporous resin, the loading flow rate was 1 BV / h, then 2 BV of deionized water, 2 BV of 30% (V / V) ethanol aqueous solution, and 2 BV of 70% (V / V) ethanol aqueous solution were used for elution in sequence, and the elution flow rate was 1 BV / h. The eluate of 30% ethanol aqueous solution and the eluate of 70% ethanol aqueous solution were collected respectively, and were concentrated under reduced pressure and dried by microwave to obtain 0.121 kg of loganic acid product and 0.497 kg of gentiopicroside product respectively.

[0107] It was detected that the content of loganic acid in the loganic acid product was 80.03%, and the transfer rate was 84.21%; the content of gentiopicroside in the gentiopicroside product was 83.73%, and the transfer rate was 82.90%.

[0108] Comparative Example 2

[0109] The comparative example provides a preparation method of gentian extract, which is basically the same as the preparation method of Example 1, and the only difference is that there is no ultrafiltration step. The specific process is as follows:

[0110] S1, 10 kg of dried gentian raw material was weighed and placed in an ultrasonic extraction tank, 150 L of 50% (V / V) ethanol aqueous solution was added, ultrasonic extraction was carried out once at room temperature, the ultrasonic frequency was 30 KHz, and the extraction time was 60 min / time, to obtain the extract. The extract was filtered by double filtration, and then centrifuged in a butterfly centrifuge to obtain the clear liquid.

[0111] S2, the clear liquid was decolorized by LSA-900E resin, and 132 L of effluent was collected.

[0112] S3, under normal temperature conditions, the effluent was concentrated by reverse osmosis membrane to obtain 18 L of concentrated solution (the soluble solid content concentration was 9.02%); the concentrated solution was further passed through LSA-10 macroporous resin, the sample flow rate was 1 BV / h, then 2 BV of deionized water, 2 BV of 30% (V / V) ethanol aqueous solution and 2 BV of 70% (V / V) ethanol aqueous solution were used for elution in turn, and the elution flow rate was 1 BV / h. The eluate of 30% ethanol aqueous solution and the eluate of 70% ethanol aqueous solution were collected respectively, concentrated under reduced pressure, and microwave dried to obtain 0.202 kg of loganic acid product and 0.626 kg of gentiopicroside product respectively.

[0113] It was detected that the content of loganic acid in the loganic acid product was 50.85%, and the transfer rate was 89.32%; the content of gentiopicroside in the gentiopicroside product was 70.44%, and the transfer rate was 87.84%.

[0114] Comparative Example 3

[0115] The comparative example provides a preparation method of gentian extract, which is basically the same as the preparation method of Example 1, and the only difference is that the gradient elution solvent concentration of macroporous resin is different. The specific process is as follows:

[0116] S1, 10 kg of dried gentian raw material was weighed and placed in an ultrasonic extraction tank, 150 L of 50% (V / V) ethanol aqueous solution was added, ultrasonic extraction was carried out once at room temperature, the ultrasonic frequency was 30 KHz, and the extraction time was 60 min / time, to obtain the extract. The extract was filtered by double filtration, and then centrifuged in a butterfly centrifuge to obtain the clear liquid.

[0117] S2, the clear liquid was decolorized by LSA-900E resin, and 130 L of effluent was collected. The effluent was further passed through an ultrafiltration membrane (molecular weight 1000), and the filtrate and the retentate were collected respectively.

[0118] S3, under normal temperature, the filtrate is concentrated by reverse osmosis membrane to obtain 15 L of concentrated solution (soluble solid content concentration is 6.24%); the concentrated solution is continuously passed through LSA-10 macroporous resin, the loading flow rate is 1 BV / h, then 2 BV of deionized water, 2 BV of 50% (V / V) ethanol aqueous solution, 2 BV of 70% (V / V) ethanol aqueous solution are sequentially used for elution, the elution flow rate is 1 BV / h. The eluate of 50% ethanol aqueous solution and the eluate of 70% ethanol aqueous solution are respectively collected, concentrated under reduced pressure, and microwave dried to obtain 0.274 kg of loganic acid product and 0.266 kg of gentiopicroside product.

[0119] It is detected that the content of loganic acid in the loganic acid product is 35.12%, and the transfer rate is 83.68%; the content of gentiopicroside in the gentiopicroside product is 96.6%, and the transfer rate is 51.19%. Part of the gentiopicroside is eluted by 50% ethanol aqueous solution, so that the gentiopicroside in the eluate of 70% ethanol aqueous solution is less.

[0120] The above describes the embodiments of the present application in combination with the examples, but the present application is not limited to the above examples, and various changes can be made within the knowledge range possessed by those skilled in the art without departing from the purpose of the present application.

Claims

1. A method for preparing a gentian extract, characterized by, The method comprises the following steps: S1, ultrasonic extraction of gentian with a first ethanol aqueous solution, solid-liquid separation, to obtain a clear liquid; in the first ethanol aqueous solution, the volume percentage of ethanol is 30%-50%; the ultrasonic extraction temperature is 25±5℃; S2, the clear liquid is sequentially subjected to decolorization treatment and ultrafiltration to obtain a filtrate; the resin used for the decolorization treatment is selected from at least one of LSA-900E and LSA-960; the ultrafiltration has a molecular weight cut-off of 500-1000; S3, column chromatography of the filtrate is performed using a macroporous resin, and the column is sequentially eluted with water, a second ethanol aqueous solution and a third ethanol aqueous solution; the second ethanol aqueous solution eluate is collected to obtain a loganic acid product; and the third ethanol aqueous solution eluate is collected to obtain a gentiopicroside product; in the second ethanol aqueous solution, the volume percentage of ethanol is 20%-40%; in the third ethanol aqueous solution, the volume percentage of ethanol is 60%-80%; in step S3, the type of the macroporous resin is selected from at least one of LSA-10, LX-12, LX-38 and AB-8.

2. The production method according to claim 1, characterized by, The solid-liquid ratio of the gentian and the first ethanol aqueous solution is 1 kg: 15 L-25 L.

3. The preparation method according to claim 1, characterized in that, in step S3, the amount of the second ethanol aqueous solution is 1 BV-3 BV.

4. The production method according to claim 1, characterized by, in step S3, the amount of the third ethanol aqueous solution is 1 BV-3 BV.

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