CN112569270A - Lepidium meyenii glucosinolate extract and preparation method thereof - Google Patents
Lepidium meyenii glucosinolate extract and preparation method thereof Download PDFInfo
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Abstract
The invention provides a Lepidium Sativum glucosinolate extract and a preparation method thereof, belonging to the technical field of medicines. The Lepidium Sativum glucosinolate extract is prepared from Lepidium plant of Brassicaceae, wherein the total content of glucosinolate is more than 50%. The invention also provides a method for extracting the glucosinolate extract from the Lepidium Sativum, and the extraction method is simple and convenient to operate and low in cost; compared with the existing extraction method of the glucosinolate from cruciferous plants, the extraction method of the invention has the advantages that the yield of the glucosinolate extract extracted from the Lepidium meyenii Walp is high, the content of the glucosinolate in the obtained glucosinolate extract is high, the content of the glucosinolate in the glucosinolate extract is more than 50%, the problems of low yield and low content of the glucosinolate extract extracted from the cruciferous plants in the prior art are solved, and the extraction method of the glucosinolate in the Lepidium meyenii Walp has.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a Lepidium Sativum glucosinolate extract and a preparation method thereof.
Background
The Lepidium meyenii Walp is a perennial herbaceous plant of Lepidium in Lepidium of Brassicaceae, is distributed all over the world, and is mainly produced beside villages, field sides, hillsides and salinized meadows with the altitude of 1800-4250 m in inner Mongolia, Tibet, Qinghai and other places in China; the whole herbs are used as the medicine, and have the effects of clearing heat and drying dampness, and treating bacillary dysentery and enteritis.
Glucosinolates (Glucosinolates), called Glucosinolates for short, are important secondary metabolites in cruciferous vegetables and can be classified into three major classes, aliphatic, aromatic and indole, according to the difference of side chain groups. More than 120 glucosinolates have been found. About 15 species are present in cruciferous vegetables. And there are 9 types of vegetables among mustard vegetables.
Studies show that the glucosinolate has a plurality of biological activities, such as sterilization, disinsection, tumor resistance, endocrine regulation, sexual function improvement and the like. In recent years, with the increase of tumor patients in the world year by year, the demand of various novel antitumor drugs is increased rapidly, and products with glucosinolate anticancer properties on the market are mainly synthesized by chemical synthesis. Although it has a certain anticancer function, it is complicated in process, difficult to control conditions, and thus cannot be widely popularized in medicine. Therefore, the main trend of research on the glucosinolates is to develop towards pure natural extraction, safety, reliability and medicinal value. Therefore, the problem that the thioglucoside in the market is rare and expensive can be solved, various aspects of the thioglucoside are improved, the thioglucoside is easy to accept by consumers, the health problem of people can be improved, and a huge economic value can be created.
At present, glucosinolates are extracted from natural plants, such as cruciferae, papaycaceae and moringaceae, wherein cruciferae is the main family. Such as: patent CN103416733A discloses a glucosinolate extract of fresh broccoli and a preparation method thereof. It is mentioned that glucosinolates are dissolved in water and theoretically can be extracted with water, but in fact myrosinase is present in the broccoli, and glucosinolates are largely decomposed during the extraction process, and the decomposition products are unstable and cannot obtain the required functional products; the yield of the alcohol extraction is very low; therefore, the enzyme is inactivated for extraction. The content of the obtained glucosinolate accounts for 2.4 to 2.7 percent of the weight of the product, 2.9 kilograms of the product can be obtained from 100 kilograms of fresh raw materials, and the yield is 2.9 percent. Patent CN103421055A extracted glucosinolates from broccoli stem and leaf waste by using an anion exchange column, and the yield was 1.4%. Patent CN104055822B discloses a Qamgur glucosinolate extract and a preparation method thereof, which improve the average content of glucosinolate in the glucosinolate extract, and the average content of glucosinolate in the glucosinolate extract is 0.7% after the improvement through calculation.
Therefore, the prior glucosinolate extract extracted from cruciferae has the problems of low yield, low content of glucosinolate in the extract and the like. Meanwhile, the glucosinolate extract extracted from the striga asiatica is not seen at present.
Disclosure of Invention
The invention aims to provide a striga asiatica glucosinolate extract and a preparation method thereof.
The invention provides a Lepidium Sativum glucosinolate extract, which is a glucosinolate extract prepared by extracting Lepidium plants of Brassicaceae, wherein the total content of glucosinolate is more than 50%.
Further, the extract contains: sinapiside, sinigrin, 3-methylthiophenyl glucosinolate, gluconaarbolabidopathinalanin, 3-ethylsulfopropylglucosinolate.
Further, the total ion flow diagram of the extract is shown in fig. 1, and the mass spectrum conditions are as follows: UPLC-Triple-TOF 5600+ time of flight LC MS: a negative ion scanning mode; scanning range: m/z 100-1500; atomizing gas (GS 1): 55 psi; atomizing gas (GS 2): 55 psi; air curtain gas (CUR): 35 psi; ion source Temperature (TEM): 600 ℃ (positive); ion source voltage (IS): 5500V (positive); primary scanning: declustering voltage (DP): 100V; focus voltage (CE): 10V; secondary scanning: and (3) acquiring mass spectrum data by using TOF MS-Product Ion-IDA modes, wherein CID energy is-20V, -40V and-60V, and before sample injection, performing mass axis correction by using a CDS (compact disc reader) pump to ensure that the error of the mass axis is less than 2 ppm.
Further, the Lepidium plant of Brassicaceae is Lepidium apetalum or Lepidium latifolia Linnaeus.
The invention also provides a preparation method of the glucosinolate extract, which comprises the following steps:
(1) adding alcohol solution into herba Lespedezae Buergeri for extraction;
(2) centrifuging the extractive solution, and collecting supernatant;
(3) enriching the supernatant by a gel chromatographic column, washing impurities by using a buffer solution, adding sulfatase overnight, washing by using water, and drying to obtain the compound enzyme.
Further, in the step (1), the alcohol solution is a methanol solution or an ethanol solution;
and/or in the step (1), the mass-volume ratio of the cress to the alcohol solution is 1 g: 2-20 ml;
preferably, in the step (1), the alcohol solution is absolute methanol or absolute ethanol;
and/or in the step (1), the mass-volume ratio of the cress to the alcohol solution is 1 g: 10-15 ml;
more preferably, the mass-to-volume ratio of the cress to the alcohol solution is 1 g: 12.5 ml.
Further, in the step (1), the extraction is hot reflux extraction;
preferably, in the step (1), the temperature of the hot reflux extraction is 70-85 ℃; and/or in the step (1), the time for hot reflux extraction is 1-3 hours;
more preferably, in step (1), the temperature of the hot reflux extraction is 75 ℃; and/or, in the step (1), the time for hot reflux extraction is 2 hours.
Further, in the step (3), the gel chromatographic column is DEAE Sephadex A25 or DEAE Sepharose CL-6B;
and/or, in the step (3), the buffer solution is sodium acetate buffer solution;
and/or, in the step (3), the sulfatase is an aqueous sulfatase solution;
preferably, in step (3), the gel chromatography column is pre-washed with imidazolium formate;
and/or, in the step (3), the pH value of the sodium acetate buffer solution is 4;
and/or in the step (3), the concentration of the aqueous solution of the sulfatase is 10-20 mu g/mL.
Further, the air conditioner is provided with a fan,
in the step (3), the elution volume of the sodium acetate buffer solution is 2-5 times of the column volume; and/or, in step (3), the elution volume of the aqueous sulfatase solution is 1-3 times of the column volume;
preferably, in step (3), the elution volume of the sodium acetate buffer is 2 to 4 times the column volume; and/or, in step (3), the elution volume of the aqueous sulfatase solution is 2 times of the column volume.
The invention also provides a composition containing the glucosinolate extract.
The Lepidium meyenii Walp used in the invention is Lepidium apetalum or Lepidium Latifolium Linnaeus.
The invention provides a method for extracting a glucosinolate extract from Lepidium meyenii, which is simple and convenient to operate and low in cost; compared with the existing extraction method of the glucosinolate from cruciferous plants, the extraction method of the invention has the advantages that the yield of the glucosinolate extract extracted from the Lepidium meyenii Walp is high, the content of the glucosinolate in the obtained glucosinolate extract is high, the content of the glucosinolate in the glucosinolate extract is more than 50%, the problems of low yield and low content of the glucosinolate extract extracted from the cruciferous plants in the prior art are solved, and the extraction method of the glucosinolate in the Lepidium meyenii Walp has.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1 is a total ion flow diagram of the glucosinolate extract from Lepidium meyenii Walp of the present invention.
Detailed Description
The raw materials and equipment used in the embodiment of the present invention are known products and obtained by purchasing commercially available products.
Detecting the content of thioglycoside in the dried extract by high performance liquid chromatography. The instrument comprises the following steps: AcquisytTM ultra model high performance liquid chromatograph (Waters, USA); chromatographic conditions are as follows: the column was Watt's ACQUITY UPLC HSS T3(150 mm. times.2.1 mm i.d.,1.8 μm); taking 0.1% formic acid water solution as mobile phase A, taking 0.1% formic acid acetonitrile mixed solution as mobile phase B, and performing linear gradient elution for 0min 2% B; 5min 2% B; 15min 15% B; 42min 60% B; the flow rate is 0.3 ml/min; the column temperature is 50 ℃; the detection wavelength is 254 nm; sample introduction amount: 2 μ l.
Example 1 preparation of the extract of Lepidium Sativum glucoside of the invention
Taking 500g of striga asiatica, crushing, adding 5000ml of anhydrous methanol, carrying out hot reflux extraction at 75 ℃, extracting for 1 hour, centrifuging the extracting solution at high speed, taking supernatant, enriching the supernatant by using a DEAE Sephadex A25 gel chromatographic column, washing impurities by using 2 times of column volume and pH 4 sodium acetate buffer solution, adding 2 times of column volume concentration 20 mu g/ml of sulfatase aqueous solution overnight, washing by using water, and drying to obtain the glucosinolate extract. The yield of the glucosinolate extract was 5.2%, and the content of glucosinolate in the glucosinolate extract was 54%.
Example 2 preparation of the extract of Lepidium Sativum glucoside of the invention
1kg of Lepidium meyenii Walp is taken, crushed and added with 15L of absolute ethyl alcohol for hot reflux extraction at 85 ℃, the extraction time is 2 hours, the extracting solution is centrifuged at high speed, then supernatant fluid is taken, the supernatant fluid is enriched by DEAE Sepharose CL-6B gel chromatographic column, sodium acetate buffer solution with 4 times of column volume and pH value of 4 is used for washing impurities, then sulfatase aqueous solution with 3 times of column volume concentration of 20 mu g/ml is added for overnight, and then the sulfatase extract is obtained after washing and drying by water. The yield of the glucosinolate extract was 5.4%, and the content of glucosinolate in the glucosinolate extract was 55%.
Example 3 preparation of the extract of Lepidium Sativum glucoside of the invention
10kg of Lepidium meyenii Walp is taken, crushed and added with 20L of absolute ethyl alcohol for hot reflux extraction at 80 ℃, the extraction time is 3 hours, the extracting solution is centrifuged at high speed, then supernatant fluid is taken, the supernatant fluid is enriched by DEAE Sepharose CL-6B gel chromatographic column, sodium acetate buffer solution with 3 times of column volume and pH value of 4 is used for washing impurities, then sulfatase aqueous solution with 1 time of column volume concentration of 20 mu g/ml is added for overnight, and then the sulfatase extract is obtained after washing and drying by water. The yield of the glucosinolate extract was 5.5%, and the content of glucosinolate in the glucosinolate extract was 54%.
Example 4 preparation of the extract of Lepidium Sativum glucoside of the invention
Taking 2kg of striga asiatica, crushing, adding 25L of absolute ethyl alcohol, carrying out hot reflux extraction at 75 ℃, extracting for 2 hours, centrifuging the extracting solution at high speed, taking supernatant, enriching the supernatant by using a DEAE Sepharose CL-6B gel chromatographic column, washing impurities by using 2 times of column volume and pH 4 sodium acetate buffer solution, adding 2 times of column volume and 20 mu g/ml sulfatase aqueous solution overnight, washing by using water, and drying to obtain the glucosinolate extract. The yield of the glucosinolate extract was 5.4%, and the content of glucosinolate in the glucosinolate extract was 54%.
The advantageous effects of the present invention are described below by way of test examples.
Test example 1 screening of Process for producing a glycoside extract from Lepidium Sativum
1. Extraction solvent
Taking 2kg of striga asiatica, crushing, adding 25L of different extraction solvents, carrying out hot reflux extraction at 75 ℃, extracting for 2 hours, centrifuging the extracting solution at high speed, taking supernatant, enriching the supernatant by using a DEAE Sepharose CL-6B gel chromatographic column, washing impurities by using 2 times of column volume of a sodium acetate buffer solution with pH of 4, adding 2 times of column volume of a sulfatase aqueous solution with concentration of 20 mu g/ml for one night, washing by using water, and drying to obtain the glucosinolate extract.
Detecting the content of thioglycoside in the dried extract by high performance liquid chromatography. The instrument comprises the following steps: AcquisytTM ultra model high performance liquid chromatograph (Waters, USA); chromatographic conditions are as follows: the column was Watt's ACQUITY UPLC HSS T3(150 mm. times.2.1 mm i.d.,1.8 μm); taking 0.1% formic acid water solution as mobile phase A, taking 0.1% formic acid acetonitrile mixed solution as mobile phase B, and performing linear gradient elution for 0min 2% B; 5min 2% B; 15min 15% B; 42min 60% B; the flow rate is 0.3 ml/min; the column temperature is 50 ℃; the detection wavelength is 254 nm; sample introduction amount: 2 μ l.
The specific extraction solvent and the results of measuring the thioglycoside content are shown in Table 1.
TABLE 1 thioglycoside content in extracts obtained with different extraction solvents
| Extraction solvent | The content of thioglycoside in the thioglycoside extract | Yield of |
| Anhydrous methanol | 55％ | 5.2％ |
| Anhydrous ethanol | 57％ | 5.3％ |
| 70% methanol | 43％ | 4.8％ |
| 70% ethanol | 38％ | 4.5％ |
| Water (W) | 43％ | 4.8％ |
As can be seen from table 1, when absolute methanol and absolute ethanol are used as extraction solvents for reflux extraction, the content of the glucosinolates in the glucosinolate extract is greater than 55%, and the yield is higher than 5%; when 70% methanol, 70% ethanol and water are used as extraction solvents for reflux extraction, the content of the glucosinolate in the glucosinolate extract is remarkably reduced to less than 45%, and the yield is also remarkably reduced to less than 5%. Therefore, the reflux extraction effect of the eleutheroside extract is optimal by adopting absolute methanol or absolute ethanol as an extraction solvent, and the content and yield of the glucosinolate in the glucosinolate extract are obviously higher than those of the glucosinolate extract which is prepared by adopting other solvents as the extraction solvent.
2. Temperature of extraction
Taking 2kg of striga asiatica, crushing, adding 25L of anhydrous methanol or anhydrous ethanol, carrying out hot reflux extraction at different temperatures for 2 hours, centrifuging the extracting solution at a high speed, taking supernatant, enriching the supernatant by using a DEAE Sepharose CL-6B gel chromatographic column, washing impurities by using 2 times of column volume and pH 4 sodium acetate buffer solution, adding 2 times of column volume and 20 mu g/ml sulfatase aqueous solution overnight, washing by using water, and drying to obtain the glucosinolate extract.
Detecting the content of thioglycoside in the dried extract by high performance liquid chromatography. The instrument and chromatographic conditions are the same as those of the method 1 and the extraction solvent.
The specific extraction solvent and temperature and the results of measuring the thioglycoside content are shown in Table 2.
TABLE 2 thioglycoside content in extracts obtained with different extraction solvents and extraction temperatures
As can be seen from Table 2, during reflux extraction, the optimum extraction temperature is 70-85 ℃ regardless of whether absolute methanol or absolute ethanol is used for extraction, when the extraction temperature is 70-85 ℃, the yield is similar to or even superior to other extraction temperatures, but the content of glucosinolates in the glucosinolate extract is more than 50%; when the temperature is too high (90 ℃) or too low (60 ℃), the content of the glucosinolates in the glucosinolate extract is obviously reduced and is lower than 40%. Therefore, the optimum extraction temperature of the extract of the glucosinolates in the Lepidium meyenii Walp is 70-85 ℃, wherein the content is the highest when the extraction temperature is 75 ℃.
3. Extraction time
Taking 2kg of striga asiatica, crushing, adding 25L of absolute methanol or absolute ethanol, carrying out hot reflux extraction at 75 ℃ for different times, centrifuging the extracting solution at a high speed, taking supernatant, enriching the supernatant by using a DEAE Sepharose CL-6B gel chromatographic column, washing impurities by using 2 times of column volume and pH 4 sodium acetate buffer solution, adding 2 times of column volume and 20 mu g/ml sulfatase aqueous solution for one night, washing by using water, and drying to obtain the glucosinolate extract.
Detecting the content of thioglycoside in the dried extract by high performance liquid chromatography. The instrument and chromatographic conditions are the same as those of the method 1 and the extraction solvent.
The specific extraction conditions (extraction solvent and extraction time) and the extract assay results (content of thioglycoside in the extract) are shown in Table 3.
TABLE 3 thioglycoside content in extracts obtained with different extraction solvents and extraction times
As can be seen from Table 3, the optimum extraction time for 75 ℃ extraction, whether using absolute methanol or absolute ethanol, is 1-3 hours, within which the thioglycoside content of the thioglycoside extract is greater than 50%; regardless of the extraction time being too long (4h) or too short (0.5h), under the condition of similar yield, the content of the glucosinolate in the glucosinolate extract is remarkably reduced and is lower than 35 percent. Therefore, the optimum extraction time for extracting the glucosinolate extract from the Lepidium meyenii Walp is 1-3 hours, wherein the extraction time is 2 hours and the content is the highest.
Test example 2 Mass Spectrometry characterization of glycosides in the extract of Tinospora cordifolia of the present invention
Sample experiment pretreatment method
Taking 2kg of striga asiatica, crushing, adding 25L of absolute ethyl alcohol, carrying out hot reflux extraction at 75 ℃, extracting for 2 hours, centrifuging the extracting solution at high speed, taking supernatant, adding 10mmol/L of sinapioside internal standard, enriching the internal standard by using a DEAE Sepharose CL-6B gel chromatographic column after adding the internal standard, washing impurities by using 2 times of sodium acetate buffer solution with the column volume pH of 4, adding 2 times of sulfatase aqueous solution with the column volume concentration of 20 mu g/ml for one night, washing by using water, and drying to obtain the glucosinolate extract. The glucosinolate extract was dissolved and the supernatant was used for testing.
Second, instrument and equipment
UPLC-Triple-TOF/MS system: AcquisytTM ultra high performance liquid chromatograph (Waters, USA), Triple TOF 5600+ type flight time mass spectrometer, equipped with electrospray ion source (AB SCIEX, USA); eppendorf minispan centrifuge (Germany Eppendorf Co.)
Third, test conditions
Chromatographic conditions are as follows: the column was Watt's ACQUITY UPLC HSS T3(150 mm. times.2.1 mm i.d.,1.8 μm); taking 0.1% formic acid water solution as mobile phase A, taking 0.1% formic acid acetonitrile mixed solution as mobile phase B, and performing linear gradient elution for 0min to obtain 2% B; 5min 2% B; 15min 15% B; 42min 60% B; the flow rate is 0.3 ml/min; the column temperature is 50 ℃; the detection wavelength is 254 nm; sample introduction amount: 2 μ l.
Mass spectrum conditions: UPLC-Triple-TOF 5600+ time of flight LC MS: a negative ion scanning mode; scanning range: m/z 100-1500; atomizing gas (GS 1): 55 psi; atomizing gas (GS 2): 55 psi; air curtain gas (CUR): 35 psi; ion source Temperature (TEM): 600 ℃ (positive); ion source voltage (IS): 5500V (positive); primary scanning: declustering voltage (DP): 100V; focus voltage (CE): 10V; secondary scanning: and (3) acquiring mass spectrum data by using TOF MS-Product Ion-IDA modes, wherein CID energy is-20V, -40V and-60V, and before sample injection, performing mass axis correction by using a CDS (compact disc reader) pump to ensure that the error of the mass axis is less than 2 ppm.
Fourthly, analyzing the detection result and the data
The total ion flow diagram of the glucosinolate extract extracted from Lepidium Sativum is shown in FIG. 1. The glucosides in the striga asiatica contains sinapiin, sinigrin, 3-methylthiophenylglucosinolate, gluconarabidopsis thaliana, 3-ethylsulfopropylglucosinolate. And (4) measuring and calculating the content of the glucosinolate extract to be more than 50% according to the peak area (the peak-off time is 18.05min) of the internal standard.
According to the above examples and test examples: when the glucosinolate extract is extracted from the Lepidium meyenii Walp, absolute ethyl alcohol or absolute methyl alcohol is used as an extraction solvent, the extraction temperature is 70-85 ℃, the extraction time is 1-3 hours, and under the condition of high yield, the obtained glucosinolate extract has high content of glucosinolate and the extraction effect is optimal.
In conclusion, the invention provides a method for extracting the glucosinolate extract from the Lepidium meyenii, which has simple and convenient operation and low cost; compared with the existing extraction method of the glucosinolate from cruciferous plants, the extraction method of the invention has the advantages that the yield of the glucosinolate extract extracted from the Lepidium meyenii Walp is high, the content of the glucosinolate in the obtained glucosinolate extract is high, the content of the glucosinolate in the glucosinolate extract is more than 50%, the problems of low yield and low content of the glucosinolate extract extracted from the cruciferous plants in the prior art are solved, and the extraction method of the glucosinolate in the Lepidium meyenii Walp has.
Claims (10)
1. A striga asiatica glucosinolate extract is characterized in that: it is a glucosinolate extract prepared by extracting plants in the genus Lepidium of the family Brassicaceae, wherein the total content of glucosinolates is greater than 50%.
2. The glucosinolate extract of claim 1, wherein: the extract contains: sinapiside, sinigrin, 3-methylthiophenyl glucosinolate, gluconaarbolabidopathinalanin, 3-ethylsulfopropylglucosinolate.
3. The glucosinolate extract of claim 2, wherein: the total ion flow diagram of the extract is shown in figure 1, and the mass spectrum conditions are as follows: UPLC-Triple-TOF 5600+ time of flight LC MS: a negative ion scanning mode; scanning range: m/z 100-1500; atomizing gas (GS 1): 55 psi; atomizing gas (GS 2): 55 psi; air curtain gas (CUR): 35 psi; ion source Temperature (TEM): 600 ℃ (positive); ion source voltage (IS): 5500V (positive); primary scanning: declustering voltage (DP): 100V; focus voltage (CE): 10V; secondary scanning: and (3) acquiring mass spectrum data by using TOF MS-Product Ion-IDA modes, wherein CID energy is-20V, -40V and-60V, and before sample injection, performing mass axis correction by using a CDS (compact disc reader) pump to ensure that the error of the mass axis is less than 2 ppm.
4. The glucosinolate extract as claimed in any one of claims 1 to 3, characterized in that: the Lepidium plant of Brassicaceae is Lepidium apetalum or Lepidium latifolia Linnaeus.
5. The method for producing a glucosinolate extract as claimed in any one of claims 1 to 4, characterized in that: it comprises the following steps:
(1) adding alcohol solution into herba Lespedezae Buergeri for extraction;
(2) centrifuging the extractive solution, and collecting supernatant;
(3) enriching the supernatant by a gel chromatographic column, washing impurities by using a buffer solution, adding sulfatase overnight, washing by using water, and drying to obtain the compound enzyme.
6. The method of claim 5, wherein: in the step (1), the alcoholic solution is a methanol solution or an ethanol solution;
and/or in the step (1), the mass-volume ratio of the cress to the alcohol solution is 1 g: 2-20 ml;
preferably, in the step (1), the alcohol solution is absolute methanol or absolute ethanol;
and/or in the step (1), the mass-volume ratio of the cress to the alcohol solution is 1 g: 10-15 ml;
more preferably, the mass-to-volume ratio of the cress to the alcohol solution is 1 g: 12.5 ml.
7. The method of claim 5, wherein: in the step (1), the extraction is hot reflux extraction;
preferably, in the step (1), the temperature of the hot reflux extraction is 70-85 ℃; and/or in the step (1), the time for hot reflux extraction is 1-3 hours;
more preferably, in step (1), the temperature of the hot reflux extraction is 75 ℃; and/or, in the step (1), the time for hot reflux extraction is 2 hours.
8. The method of claim 5, wherein: in the step (3), the gel chromatographic column is DEAE Sephadex A25 or DEAE Sepharose CL-6B;
and/or, in the step (3), the buffer solution is sodium acetate buffer solution;
and/or, in the step (3), the sulfatase is an aqueous sulfatase solution;
preferably, in step (3), the gel chromatography column is pre-washed with imidazolium formate;
and/or, in the step (3), the pH value of the sodium acetate buffer solution is 4;
and/or in the step (3), the concentration of the aqueous solution of the sulfatase is 10-20 mu g/mL.
9. The method of claim 8, wherein:
in the step (3), the elution volume of the sodium acetate buffer solution is 2-5 times of the column volume; and/or, in step (3), the elution volume of the aqueous sulfatase solution is 1-3 times of the column volume;
preferably, in step (3), the elution volume of the sodium acetate buffer is 2 to 4 times the column volume; and/or, in step (3), the elution volume of the aqueous sulfatase solution is 2 times of the column volume.
10. A composition characterized by: it contains the glucosinolate extract as claimed in any one of claims 1 to 4.
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| CN114324723A (en) * | 2021-12-14 | 2022-04-12 | 湖北省农业科学院经济作物研究所 | Qualitative detection method for radish glucosinolates |
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| CN107367565A (en) * | 2017-07-06 | 2017-11-21 | 新时代健康产业（集团）有限公司 | Utilize the method for smell finger-print Rapid identification maca quality |
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| CN107367565A (en) * | 2017-07-06 | 2017-11-21 | 新时代健康产业（集团）有限公司 | Utilize the method for smell finger-print Rapid identification maca quality |
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| CN114324723A (en) * | 2021-12-14 | 2022-04-12 | 湖北省农业科学院经济作物研究所 | Qualitative detection method for radish glucosinolates |
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