A method for preparing a low-bitterness cistanche extract for wine

Abstract

The present application relates to the technical field of separation and purification, and specifically discloses a preparation method of low-bitterness cistanche wine extract, which comprises the following steps: extracting base wine, adding alkali and ultrasonic dissolving, subcritical water cooking, and drying, and finally obtaining the low-bitterness cistanche wine extract. The preparation method is simple, the bitterness value of the obtained cistanche wine extract is significantly reduced, and the cistanche wine extract has good industrial production value.

Description

Technical Field

[0001] The invention relates to the technical field of separation and purification, in particular to a method for preparing a low-bitterness Cistanche deserticola wine extract. Background Art

[0002] Cistanche deserticola ( Cistanche deserticola YCMa (YCMa) is the dried, fleshy stem with scale leaves of the plant Cistanche deserticola or Cistanche tubulosa, part of the Orobanchaceae family. It is widely cultivated in Inner Mongolia, Gansu, Xinjiang, and Qinghai in my country. Cistanche deserticola contains numerous active ingredients and is often used in the preparation of health beverages and health wines. Current research indicates that the main active ingredients in Cistanche deserticola include echinacoside, isoverbascoside, verbascoside, cistancheside A, and cistancheside C. However, the application of Cistanche deserticola extracts also presents certain challenges. For example, when added to health wines, Cistanche deserticola extract imparts a certain bitterness, which can lead to low sensory pleasure for consumers. Currently, no literature has examined in detail the source of the bitterness in Cistanche deserticola.

[0003] In view of the above problems, how to reduce the bitterness of Cistanche deserticola extract has become a technical problem that needs to be solved urgently. Summary of the Invention

[0004] In view of this, the present invention proposes a preparation method of a low-bitterness Cistanche deserticola extract for wine, aiming to reduce the bitterness of the Cistanche deserticola extract so that it has better sensory properties when used as a wine additive.

[0005] The technical solution of the present invention is achieved as follows: The present invention provides a method for preparing a low-bitterness Cistanche deserticola wine extract, comprising the following steps:

[0006] Step 1: Crush the Cistanche deserticola and soak it in base wine, perform reflux extraction after soaking, and collect the extract;

[0007] Step 2: adding baking soda to the extract obtained in step 1 and dissolving it under ultrasonic conditions;

[0008] Step 3: The extract obtained after ultrasonic treatment in step 2 is subjected to subcritical water cooking, and after cooking, concentrated and dried to obtain a low-bitterness Cistanche deserticola wine extract.

[0009] In some embodiments, in step one, the base wine concentration is 30-85% vol, and the base wine solvent multiple is 3-8 times.

[0010] In some embodiments, in step 1, the soaking time is 10-18 hours.

[0011] In some embodiments, in step 1, the temperature of reflux extraction is 60-80° C., and the reflux extraction time is 1.5-3.5 h.

[0012] In some embodiments, in step 2, the ultrasonic power is 140-160 W, and the ultrasonic time is 10-20 min.

[0013] In some embodiments, in step 2, the amount of baking soda added is 0.3%-0.5% of the mass of Cistanche deserticola.

[0014] In some embodiments, in step three, the temperature of subcritical water cooking is 120-140° C., and the cooking time is 60-180 min.

[0015] In some embodiments, in step three, the concentration and drying method is vacuum drying, and the drying temperature is 50-70°C.

[0016] In a second aspect, the present invention further provides a low-bitterness Cistanche deserticola wine extract prepared by the above-mentioned preparation method.

[0017] Thirdly, the present invention also provides an application of the above-mentioned low-bitterness Cistanche deserticola extract for wine, which can be specifically applied to the preparation of alcoholic beverages containing Cistanche deserticola extract.

[0018] The present invention has the following beneficial effects compared to the prior art:

[0019] The preparation method provided by the present invention can effectively reduce the bitterness of a Cistanche deserticola extract for winemaking. Compared with a Cistanche deserticola extract obtained solely through extraction with base wine, the low-bitterness Cistanche deserticola extract for winemaking prepared by the present invention can reduce the bitterness by over 70%. The preparation process of the low-bitterness Cistanche deserticola extract for winemaking in the present invention is simple, environmentally friendly, and pollution-free, and has excellent industrial value. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0021] Figure 1 This is a liquid chromatogram of the macroporous resin eluate of the Cistanche deserticola wine extract in Example 1 of the present invention. DETAILED DESCRIPTION

[0022] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which embodiments of the present invention belong. If the definitions set forth in this section are contrary to or otherwise inconsistent with definitions set forth in the patents, patent applications, published patent applications, and other publications incorporated herein by reference, the definitions listed in this section take precedence over the definitions incorporated herein by reference.

[0024] Unless otherwise specified, the methods used in the following examples are conventional methods. The materials, reagents, and instruments used are conventional materials, reagents, and instruments in the art, unless otherwise specified, and can be obtained commercially by those skilled in the art.

[0025] When an amount, concentration or other value or parameter is expressed as a range, a preferred range or a range defined by a series of upper preferred values ​​and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any upper range limit or preferred value with any lower range limit or preferred value, regardless of whether the range is disclosed alone. For example, when a range "1 to 5" is disclosed, the described range should be interpreted as including the ranges "1 to 4", "1 to 3", "1 to 2", "1 to 2 and 4 to 5", "1 to 3 and 5", etc. When a numerical range is described herein, unless otherwise stated, the range is intended to include its endpoints and all integers and fractions within the range. In the present specification and claims, range definitions may be combined and / or interchanged, and if not otherwise stated, such ranges include all subranges contained therein.

[0026] During their early research into the bitterness of Cistanche deserticola extracts, the inventors discovered that the bitterness primarily stems from verbascoside, the active ingredient in the herb. Subsequent experiments revealed that verbascoside in Cistanche deserticola can be decomposed and converted into other substances under certain conditions, such as ultrasonic treatment under alkaline conditions and subcritical water cooking. Currently, there are no reports on the preparation of a low-bitterness Cistanche deserticola extract for winemaking, as described in the present invention.

[0027] The technical solution of the present invention is further described in detail below through specific embodiments.

[0028] Example 1

[0029] This example is about the identification of bitter substances in the Cistanche deserticola wine extract, and the specific steps are as follows:

[0030] S1. Weigh 1 kg of coarsely ground Cistanche deserticola powder and soak it in 8 L of 85% vol base wine solution for 18 h. After soaking, perform condensation reflux extraction at 80°C for 3.5 h. Collect and combine the extracts to obtain a total of 7.91 L. Divide the extract into 7 L and 0.91 L, respectively, designated as Sample 1 and Sample 2.

[0031] S2 and Sample 2 were dried at 50°C to obtain 35.2 g of the Cistanche deserticola wine extract. 30 g of the obtained Cistanche deserticola wine extract was dissolved in 900 mL of ultrapure water and passed through a macroporous resin column at a flow rate of 1 BV / h. The resin column was then eluted with 200 mL of 30% ethanol. The eluate was collected and concentrated until it was free of alcohol. The eluate concentrated to a state free of alcohol was then subjected to a high performance preparative liquid chromatography. The chromatographic conditions were as follows: column: Daisogel C18 preparative column, mobile phase: acetonitrile-0.1% phosphoric acid aqueous solution, mobile phase flow rate: 15 mL / min, detection wavelength: 238 nm, elution gradient: 0-12 min, 95%B-91.5%B; 12-15 min, 91.5%B; 15-16 min, 91.5%B-90%B; 16-25 min, 90%B-86%B; 25-50 min, 86%B; 50-62 min, 86%B-82%B; 62-70 min, 82%B-75%B; 70-100 min, 75%B. Using the above chromatographic conditions, the extract for Cistanche deserticola wine was separated and purified to obtain a chromatogram as shown below. Figure 1 As shown, the fractions within the time period corresponding to each peak were collected, and a total of 5 fractions were obtained, and the corresponding time periods were 2.132-4.418 min, 4.420-6.178 min, 45.162-47.283 min, 68.125-70.135 min, and 71.128-73.010 min, respectively.

[0032] The fractions within the time periods of 2.132-4.418 min, 4.420-6.178 min, 45.162-47.283 min, 68.125-70.135 min, and 71.128-73.010 min were collected and freeze-dried.

[0033] S3. The collected fractions were dissolved in ultrapure water to a concentration of 3 mg / mL. The bitterness value of each sample was measured using an electronic tongue. It was found that the fraction corresponding to the time period 45.162-47.283 min had a strong bitter taste, while the fractions corresponding to the other time periods had basically no bitter taste. The fraction corresponding to the time period 45.162-47.283 min was identified as verbascoside using UPLC-MS / MS technology.

[0034] The instrument models used were: Waters Xevo G2 QTof mass spectrometer, Waters ACQUITY UPLC ultra-high performance liquid chromatograph, and ACQUITY UPLC HSS T3 C18 column (2.1 mm × 100 mm, 1.8 μm), all from Waters, USA.

[0035] UPLC-MS / MS conditions: Chromatographic conditions: Chromatographic column: ACQUITY UPLC HSS T3 C18 column (2.1 mm×100 mm, 1.8 μm); flow rate: 0.4 mL / min; column temperature: 40 °C; sample injection volume: 0.5 μL, reference solution injection volume: 0.1 μL; detection wavelength: 203 nm; mobile phase: 0.1% formic acid-water solution (A), 0.1% formic acid-acetonitrile solution (B); gradient elution program: 0 min, 100%A; 5 min, 95%A; 20 min, 60%A; 28 min, 10%A; 30 min, 100%A; 32 min, 100%A. Mass spectrometry conditions: negative ion scan mode; desolvation temperature 500°C; electrospray ionization source; ion source temperature 120°C; high-purity nitrogen desolvation gas; desolvation gas flow rate 800 L / Hr; capillary voltage 3.0 kV; collision gas high-purity argon at 7 psi; collision energy 6–40 eV. The electronic tongue measurement data and UPLC-MS / MS detection data are shown in Table 1.

[0036] Table 1. Electronic tongue bitterness values ​​and UPLC-MS / MS data of each fraction in Example 1

[0037]

[0038] As can be seen from Table 1 , fraction 3 among the five fractions had a typical bitter taste and was identified as verbascoside by UPLC-MS / MS.

[0039] Example 2

[0040] This embodiment provides a method for preparing a low-bitterness Cistanche deserticola wine extract, and the specific steps are as follows:

[0041] Based on Sample 1 prepared in S1 of Example 1, 3 g of baking soda was added to the extract of Sample 1 and ultrasonically dissolved at an ultrasonic power of 140 W for 10 minutes. The ultrasonically dissolved Cistanche deserticola extract was subjected to subcritical water cooking at 120°C for 60 minutes and finally concentrated and dried to obtain 91.7 g of a low-bitterness Cistanche deserticola wine extract.

[0042] Example 3

[0043] This example is about the identification of bitter substances in the Cistanche deserticola wine extract, and the specific steps are as follows:

[0044] S1. Weigh 1 kg of coarsely ground Cistanche deserticola powder and soak it in 8 L of 85% vol base wine solution for 18 h. After soaking, perform condensation reflux extraction at 80°C for 3.5 h. Collect and combine the extracts to obtain a total of 7.91 L. Divide the extract into 7 L and 0.91 L, respectively, designated as Sample 1 and Sample 2.

[0045] S2 and Sample 2 were dried at 70°C to obtain 14.3 g of the Cistanche deserticola wine extract. 10 g of the obtained Cistanche deserticola wine extract was dissolved in 600 mL of ultrapure water and passed through a macroporous resin column at a flow rate of 2 BV / h. The resin column was then eluted with 250 mL of 60% ethanol. The eluate was collected and concentrated until it was free of alcohol. The eluate concentrated until it was free of alcohol was then subjected to high performance preparative liquid chromatography. The chromatographic conditions were as follows: column: Daisogel C18 preparative column, mobile phase: acetonitrile-0.1% phosphoric acid aqueous solution, mobile phase flow rate: 15 mL / min, detection wavelength: 238 nm, elution gradient: 0-12 min, 95%B-91.5%B; 12-15 min, 91.5%B; 15-16 min, 91.5%B-90%B; 16-25 min, 90%B-86%B; 25-50 min, 86%B; 50-62 min, 86%B-82%B; 62-70 min, 82%B-75%B; 70-100 min, 75%B. The fractions within the time periods of 2.132-4.418 min, 4.420-6.178 min, 45.162-47.283 min, 68.125-70.135 min, and 71.128-73.010 min were collected and freeze-dried.

[0046] S3. The collected fractions were dissolved in ultrapure water to a concentration of 3 mg / mL. The bitterness value of each sample was measured using an electronic tongue. It was found that the fraction corresponding to the time period 45.162-47.283 min had a strong bitter taste, while the fractions corresponding to the other time periods had basically no bitter taste. The fraction corresponding to the time period 45.162-47.283 min was identified as verbascoside using UPLC-MS / MS technology.

[0047] Example 4

[0048] This embodiment provides a method for preparing a low-bitterness Cistanche deserticola wine extract, and the specific steps are as follows:

[0049] Based on Sample 1 prepared in S1 in Example 3, 5 g of baking soda was added to the extract of Sample 1 and ultrasonically dissolved at an ultrasonic power of 160 W for 20 min. The ultrasonically dissolved Cistanche deserticola extract was subjected to subcritical water cooking at 140°C for 180 min and finally concentrated and dried to obtain 120.1 g of a low-bitterness Cistanche deserticola wine extract.

[0050] Example 5

[0051] This example is about the identification of bitter substances in the Cistanche deserticola wine extract, and the specific steps are as follows:

[0052] S1. Weigh 1 kg of coarsely ground Cistanche deserticola powder and soak it in 8 L of 65% vol base wine solution for 14 h. After soaking, perform condensation reflux extraction at 70°C for 2 h. Collect and combine the extracts to obtain a total of 4.90 L. Divide the extract into 4 L and 0.91 L, respectively, designated as Sample 1 and Sample 2.

[0053] S2 and Sample 2 were dried at 60°C to obtain 20.2 g of the Cistanche deserticola wine extract. 15 g of the obtained Cistanche deserticola wine extract was dissolved in 675 mL of ultrapure water and passed through a macroporous resin column at a flow rate of 1.5 BV / h. The resin column was then eluted with 210 mL of 45% ethanol. The eluate was collected and concentrated until it was free of alcohol. The eluate concentrated until it was free of alcohol was then subjected to high performance preparative liquid chromatography. The chromatographic conditions were as follows: column: Daisogel C18 preparative column, mobile phase: acetonitrile-0.1% phosphoric acid aqueous solution, mobile phase flow rate: 15 mL / min, detection wavelength: 238 nm, elution gradient: 0-12 min, 95%B-91.5%B; 12-15 min, 91.5%B; 15-16 min, 91.5%B-90%B; 16-25 min, 90%B-86%B; 25-50 min, 86%B; 50-62 min, 86%B-82%B; 62-70 min, 82%B-75%B; 70-100 min, 75%B. The fractions within the time periods of 2.132-4.418 min, 4.420-6.178 min, 45.162-47.283 min, 68.125-70.135 min, and 71.128-73.010 min were collected and freeze-dried.

[0054] S3. The collected fractions were dissolved in ultrapure water to a concentration of 3 mg / mL. The bitterness value of each sample was measured using an electronic tongue. It was found that the fraction corresponding to the time period 45.162-47.283 min had a strong bitter taste, while the fractions corresponding to the other time periods had basically no bitter taste. The fraction corresponding to the time period 45.162-47.283 min was identified as verbascoside using UPLC-MS / MS technology.

[0055] Example 6

[0056] This embodiment provides a method for preparing a low-bitterness Cistanche deserticola wine extract, and the specific steps are as follows:

[0057] Based on Sample 1 prepared in S1 in Example 5, 4 g of baking soda was added to the extract of Sample 1 and ultrasonically dissolved at an ultrasonic power of 150 W for 15 minutes. The ultrasonically dissolved Cistanche deserticola extract was subjected to subcritical water cooking at 130°C for 100 minutes and finally concentrated and dried to obtain 103.1 g of a low-bitterness Cistanche deserticola wine extract.

[0058] Comparative Example 1

[0059] This embodiment provides a method for preparing a low-bitterness Cistanche deserticola wine extract, and the specific steps are as follows:

[0060] Based on the sample 1 prepared by S1 in Example 1, the sample 1 was subjected to subcritical water cooking at 120° C. for 60 min, and finally concentrated and dried to obtain a low-bitterness Cistanche deserticola wine extract.

[0061] Comparative Example 2

[0062] This embodiment provides a method for preparing a low-bitterness Cistanche deserticola wine extract, and the specific steps are as follows:

[0063] Based on the sample 1 prepared in S1 of Example 1, 3 g of baking soda was added to the extract of sample 1, and ultrasonic dissolution was performed for 10 minutes at an ultrasonic power of 140 W. Finally, the extract was concentrated and dried to obtain a low-bitterness Cistanche deserticola wine extract.

[0064] Experimental Example 1

[0065] The low-bitterness Cistanche deserticola extracts for wine prepared in Examples 2, 4, 6, and Comparative Examples 1 and 2, as well as the Cistanche deserticola extracts for wine prepared in S2 of Examples 1, 3, and 5, were dissolved in alcohol to form 3 mg / ml solutions. The Cistanche deserticola extracts for wine prepared in S2 of Examples 1, 3, and 5 served as the control group. The bitterness reduction rates of the low-bitterness Cistanche deserticola extracts for wine prepared in Examples 2, 4, 6, and Comparative Examples 1 and 2 were calculated. The bitterness values ​​of the samples were measured using an electronic tongue. The specific steps were as follows: Each sample was poured into the electronic tongue sample beaker and then placed on the sample injection and analysis device. Before measuring the sample, the electronic tongue system was initialized, self-checked, and calibrated. After measurement, the system was rinsed with water for 300 seconds. The data acquisition time for the sample was 150 seconds. Each sample was tested four times, and the data from the last three times was averaged. The bitterness reduction rate was calculated using the following formula:

[0066] Bitterness reduction rate = [(AB) / A] × 100%

[0067] Wherein: A is the bitterness value of the Cistanche deserticola extract for wine; B is the bitterness value of the low-bitterness Cistanche deserticola extract for wine obtained in different embodiments and comparative examples of the present invention.

[0068] The calculated bitterness reduction rate data are as follows:

[0069]

[0070] As can be seen from the above table, the bitterness reduction rate of the low-bitter Cistanche deserticola wine extracts of Examples 2, 4, and 6 is significantly different from that of Comparative Examples 1-2. The bitterness reduction rates of Examples 2, 4, and 6 are all greater than 70%, indicating that the low-bitterness Cistanche deserticola wine extract prepared by the present invention has a significantly reduced bitterness value compared to the conventional preparation process of Cistanche deserticola wine extract. In addition, both alkali ultrasonic dissolution and subcritical water cooking in the present invention have a significant effect on the bitterness reduction rate of the Cistanche deserticola wine extract. The absence of either step cannot achieve a bitterness reduction rate of more than 70% for the Cistanche deserticola wine extract.

[0071] Comparative Example 3

[0072] This comparative example provides a method for preparing a low-bitterness Cistanche deserticola wine extract, and the specific steps are as follows:

[0073] Based on Sample 1 prepared in S1 of Example 1, 1.4 g of sodium hydroxide was added to the extract of Sample 1, and ultrasonic dissolution was performed at an ultrasonic power of 140 W for 10 minutes. The ultrasonically dissolved Cistanche deserticola extract was subjected to subcritical water cooking at 120°C for 60 minutes, and finally concentrated and dried to obtain 62.7 g of a low-bitterness Cistanche deserticola wine extract.

[0074] Comparative Example 4

[0075] This comparative example provides a method for preparing a low-bitterness Cistanche deserticola wine extract, and the specific steps are as follows:

[0076] Based on Sample 1 prepared in S1 of Example 1, 1.9 g of sodium carbonate was added to the extract of Sample 1 and ultrasonically dissolved at an ultrasonic power of 140 W for 10 minutes. The ultrasonically dissolved Cistanche deserticola extract was subjected to subcritical water cooking at 120°C for 60 minutes and finally concentrated and dried to obtain 88.6 g of a low-bitterness Cistanche deserticola wine extract.

[0077] The bitterness values ​​of the low-bitterness Cistanche deserticola wine extracts prepared in Comparative Examples 3 and 4 were measured using an electronic tongue, and the bitterness reduction rate was calculated. The results are shown in the following table:

[0078]

[0079] From the data in the above table, it can be seen that baking soda has the best effect and the highest reduction rate of bitterness, and the final yield of the low-bitterness Cistanche deserticola wine extract is higher.

[0080] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing a low-bitterness Cistanche deserticola wine extract, characterized in that: The steps include: Step 1, crushing the Cistanche deserticola and soaking it in base wine for 10-18 hours, performing reflux extraction after the soaking, collecting the extract, the base wine concentration is 30-85%vol, the volume mass ratio of the base wine to the Cistanche deserticola is 3-8, the reflux extraction temperature is 60-80°C, and the reflux extraction time is 1.5-3.5 hours; Step 2: adding baking soda to the extract obtained in step 1 and dissolving it under ultrasonic conditions, with an ultrasonic power of 140-160 W and an ultrasonic time of 10-20 min. The amount of baking soda added is 0.3%-0.5% of the mass of Cistanche deserticola; Step 3: The extract after ultrasonic treatment in step 2 is subjected to subcritical water cooking at a temperature of 120-140° C. for 60-180 min. After cooking, the extract is concentrated and dried to obtain a low-bitterness Cistanche deserticola wine extract.

2. The method for preparing the low-bitterness Cistanche deserticola wine extract according to claim 1, wherein: In step 3, the concentration and drying method is vacuum drying, and the drying temperature is 50-70°C.

3. A low bitterness Cistanche deserticola extract for wine, characterized in that: The product is prepared by the preparation method described in any one of claims 1-2.

4. The low-bitterness Cistanche deserticola extract for wine according to claim 3 is used in the preparation of alcoholic beverages containing the Cistanche deserticola extract.

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