CN108164541B - Method for extracting dendrobine from dendrobium officinale - Google Patents
Method for extracting dendrobine from dendrobium officinale Download PDFInfo
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- C07—ORGANIC CHEMISTRY
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- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
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Abstract
The invention discloses a method for extracting dendrobine from dendrobium officinale, which comprises the following steps: firstly, taking dendrobium officinale, crushing, and adding water into the dendrobium officinale according to the material-liquid ratio of 1:10-14 to obtain a product A; regulating the pH of the product A to be 9 by using saturated lime water; thirdly, microwave extracting the product B for 3 times under the power of 300-320W, extracting for 15-20min each time, and merging the extracting solutions to obtain a product C; fourthly, filtering the product C, and concentrating the filtrate in vacuum to obtain a product D; fifthly, adding 3-4 times of ethanol into the product D, and standing at 1-4 ℃ for 10-12h to obtain a product E; sixthly, filtering the product E, washing the precipitate for 3 to 4 times by using ethanol, and drying the precipitate to constant weight to obtain the product. When the process disclosed by the invention is used for extracting the dendrobium officinale, the extraction rate of the dendrobine is high, and the development and utilization of the dendrobine in the dendrobium officinale are promoted.
Description
Technical Field
The invention relates to a method for extracting dendrobium officinale, in particular to a method for extracting dendrobine from dendrobium officinale.
Background
The dendrobium officinale is named as dendrobium officinale, chlorophytum comosum, Yunnan iron sheet and chlorophytum comosum, is a perennial herb of the dendrobium genus of the orchidaceae family, is the top-grade of dendrobium medicinal materials, has the element of the Chinese mesona, namely the gold in the medicine, is used as a medicine by stems, contains dendrobine, alkaloid and other effective components, has the effects of nourishing yin and clearing heat, promoting the production of body fluid and benefiting the stomach, moistening throat and improving eyesight and the like, and has good curative effects in the aspects of resisting tumors, improving human immunity, treating gastrointestinal diseases, resisting aging, resisting oxidation, resisting platelet aggregation, reducing blood sugar, treating cataract and the like.
Modern pharmacological studies show that the dendrobium officinale has an enhancement effect on nonspecific and specific immunity, and has a remarkable pharmacological effect in the aspects of resisting tumors, resisting oxidation, reducing blood sugar, treating cataract and the like. The main reason for the efficacy of Dendrobium officinale is that it contains many active ingredients, such as alkaloid, dendrobine, amino acid, etc., but the efficacy of each active ingredient is different. In clinical application, the dendrobium officinale mainly takes effect by decocting dendrobium officinale medicinal materials into decoction for taking, and the efficacy of each active ingredient is not applied specifically, so that the utilization of the dendrobium officinale mainly stays at the low-end primary stage, and the medicinal value of the dendrobium officinale is further developed. Researches show that dendrobine in dendrobium officinale has the effects of immunoregulation, anti-inflammation, blood vessel dilation and the like, and has good potential development value. If the dendrobine is extracted and then is utilized, the method has important significance for further development of the dendrobium officinale.
At present, the extraction of dendrobine has certain limitation, so that the extraction rate of dendrobine is low, and the development and utilization of active ingredients of dendrobium candidum dendrobine are limited.
Disclosure of Invention
The invention aims to provide a method for extracting dendrobine from dendrobium officinale.
The invention is realized by the following steps: the method for extracting dendrobine from dendrobium officinale comprises the following steps:
firstly, taking dendrobium officinale, crushing, and adding water into the dendrobium officinale according to the material-liquid ratio of 1:10-14 to obtain a product A;
regulating the pH of the product A to be 9 by using saturated lime water to obtain a product B;
thirdly, microwave extracting the product B for 3 times under the power of 300-320W, extracting for 15-20min each time, and merging the extracting solutions to obtain a product C;
fourthly, filtering the product C, and concentrating the filtrate in vacuum to obtain a product D;
fifthly, adding 3-4 times of ethanol into the product D, and standing at 1-4 ℃ for 10-12h to obtain a product E;
sixthly, filtering the product E, washing the precipitate for 3 to 4 times by using ethanol, and drying the precipitate to constant weight to obtain the product.
In the method for extracting dendrobine from dendrobium officinale, the step III is as follows: the power is 310W.
In the method for extracting dendrobine from dendrobium officinale, in the step III, the extraction time is 18min each time.
In the method for extracting dendrobine from dendrobium officinale, in the fourth step, the filtrate is subjected to vacuum concentration; concentrating twice, controlling the vacuum degree at 0.07-0.08MPa for the first time, controlling the vacuum degree at 0.08-0.09MPa for the second time, and controlling the vacuum concentration time at 5-10min for the second time.
In the method for extracting dendrobine from dendrobium officinale, the filtrate is subjected to vacuum concentration; concentrating twice, controlling the vacuum degree at 0.07-0.08MPa for the first time, controlling the vacuum degree at 0.08-0.09MPa for the second time, and controlling the vacuum concentration time at two times to be 8 min.
In the method for extracting dendrobine from dendrobium officinale, the fifth step is; adding 3 times of ethanol into product D, and standing at 1-4 deg.C for 11 hr to obtain product E.
The applicant has carried out a large number of experimental studies on the present invention, some of which are as follows:
experimental example 1 dendrobine extraction Rate Observation
1 Material
Taking 3 parts of dendrobium officinale with the same weight, and preparing an extract 1, an extract 2 and an extract 3 respectively according to the following methods.
Extract 1: extraction was performed as in example 1.
And (2) extraction: the extraction method differs from example 1 in that:
(1) in the step I, the ratio of the material to the liquid is 1: 20;
(2) and fourthly, during concentration in the fourth step, the first vacuum degree is 0.06MPa, and the second vacuum degree is 0.07 MPa.
2 methods and results.
Weighing the obtained extract 1 and extract 2, and calculating the extraction rate according to the result, wherein the extraction rate calculation formula is that the extraction rate (%) = (fresh weight of dendrobium officinale-extract weight) × 100%/fresh weight of dendrobium officinale. The final result is:
the extraction rate of extract 1 =21.38%
The extraction rate of extract 2 =15.01%
The extraction rate of extract 3 = 16.02%.
Therefore, the extraction process has the highest extraction rate.
Experimental example 2 examination of purity
1 materials and methods
1.1 materials
1.1.1 Dendrobium officinale 1: extraction was performed as in example 1.
1.1.2 Dendrobium officinale 2: the extraction method is the same as that of extract 2.
1.2 instruments and reagents
Dendrobine standard substance: wodsmant Biotechnology Ltd, MUST-12080608. A E240 electronic balance: MEITLER, switzerland; HH-s numerical control constant temperature water bath: experimental facilities for the Wen Hua Kong education in Jinhua; BM 510 rotary evaporator: YAM ATO, Japan; 6890N gas chromatograph: american national security
Jieren corporation; UV-2450 ultraviolet spectrophotometer: shimadzu, Japan; methanol (chromatographically pure): tianjin, Kemiou chemical reagents, Inc.; the other reagents are analytically pure; phenol: shanghai Lingfeng Chemicals, Inc.; sulfuric acid: shanghai Lingfeng Chemicals, Inc.
2 method
2.1 preparation of test solutions
2.1.1 preparation of test article solution 1:
taking 3.34g of dendrobium officinale 1 powder, adding 200mL of 90% ethanol, refluxing for 1 hour, filtering, pouring out supernatant, adding 100mL of 90% ethanol, refluxing for 1 hour, pouring out supernatant, combining the supernatant for 2 times, concentrating in a rotary evaporator, evaporating to dryness at 50 ℃ in a water bath kettle, adding methanol to dissolve a dried product, and fixing the volume to 25m L to obtain a test solution 1.
2.1.2 preparation of test solution 2
Taking 3.34g of dendrobium officinale 2 powder, adding 200mL of 90% ethanol, refluxing for 1 hour, filtering, pouring out supernatant, adding 100mL of 90% ethanol, refluxing for 1 hour, pouring out supernatant, combining the supernatant for 2 times, concentrating in a rotary evaporator, evaporating to dryness at 50 ℃ in a water bath kettle, adding methanol to dissolve a dried product, and fixing the volume to 25m L to obtain a test solution 2.
2.2 preparation of control solutions
Precisely weighing 50.01lmg to 100mL of dendrobine reference substance in a measuring flask, adding methanol to a constant volume to a scale, and shaking up to obtain 500.1m g/L of dendrobine reference substance mother liquor. 50.01m g/L solution as control solution for use.
2.3 chromatographic conditions
A chromatographic column: d B-1 capillary column (30 mm. times.0.25 mm, 0.25 um); temperature programming: the initial temperature is 80 ℃, the temperature is increased to 250 ℃ at the speed of 10 ℃/min, and the temperature is kept for 5 minutes; the injection inlet temperature is 250 ℃, the detector temperature is 250 ℃, the injection amount is 10uL, and the theoretical plate number is not less than 10000 calculated according to dendrobine.
2.4 Standard Curve preparation
Precisely measuring control solutions 0.2, 0.5, 0.8, 1.0, 2.0, and 4.0m L, respectively, placing in 10m L measuring flask, adding methanol to the scale, and shaking. Measured according to the chromatographic conditions described above. Taking the peak area integral as a vertical coordinate, taking the sample amount of a reference substance as a horizontal coordinate to perform linear regression, wherein the regression equation is A = 0.7522C-0.0252, and r = 0.9997, which shows that the dendrobine has a good linear relation in the range of 0.10-2.00 ug.
2.5 precision test
Precisely absorbing l0ul of the dendrobine reference substance solution, continuously injecting samples for 6 times, recording the peak area, wherein the RSD of the dendrobine peak area is 1.58%, which indicates that the instrument precision is good.
2.6 repeatability test
6 parts of test solution 1 and test solution 2 were prepared as described above and measured under the above chromatographic conditions. RSD of the dendrobine peak area is 2.05% and 2.06%, respectively.
2.7 stability test
Taking the same test sample solution 1, standing at room temperature, injecting samples for 0, 2, 4, 6, 8 and l2 hours respectively, recording dendrobine peak area, and RSD is 2.88%. The results show that the test article solution 1 is stable within 12 hours.
Taking the same sample solution 2, standing at room temperature, injecting samples for 0, 2, 4, 6, 8 and l2 hours respectively, recording dendrobine peak area, and RSD is 2.89%. The results show that the test solution 2 is stable within 12 hours.
2.8 recovery test
Taking 6 parts of dendrobium officinale powder with known content of 0.46 mg/g), each part is about 1.65g, and precisely weighing. Accurately adding 2 mL of dendrobine reference product mother liquor, preparing a test solution 1 and a test solution 2 according to the method, measuring under the chromatographic conditions, and calculating the recovery rate. As a result, the average recovery rates were 97.1% and 96.8%, and the RSD was 2.5% and 2.6%, respectively.
2.9 measurement of sample content
The preparation method and the determination conditions of the test solution 1, the test solution 2 and the test solution 3 are characterized in that the content of dendrobine in 6 parts of the extract 1 sample and 6 parts of the dendrobium officinale 2 sample is determined, the results are recorded, the average value of the results is obtained, and the results are shown in table 1.
As can be seen from the table, when the process provided by the invention is used for extracting dendrobium officinale, each gram of the extract contains 910.23g of dendrobine, the purity of dendrobine is 91.0%, and the purity of dendrobine in the extract 2 is 71.0%.
Compared with the prior art, when the process is used for extracting the dendrobium officinale, the extraction rate is high, and the purity of the dendrobine is up to 91.0%.
Detailed Description
Example 1.
The method for extracting dendrobine from dendrobium officinale comprises the following steps:
firstly, taking the dendrobium officinale, crushing, and adding water into the dendrobium officinale according to the material-liquid ratio of 1:12 to obtain a product A;
regulating the pH of the product A to be 9 by using saturated lime water;
③ microwave extracting the product B for 3 times under the power of 310W, each time for 18min, and mixing the extractive solutions to obtain product C;
fourthly, filtering the product C, concentrating the filtrate in two times, controlling the vacuum degree to be 0.07-0.08MPa for the first time, controlling the vacuum degree to be 0.08-0.09MPa for the second time, and performing vacuum concentration for 8min for the two times to obtain a product D;
fifthly, adding 3 times of ethanol into the product D, and standing for 11 hours at the temperature of 1-4 ℃ to obtain a product E;
sixthly, filtering the product E, washing the precipitate for 3 to 4 times by using ethanol, and drying the precipitate to constant weight to obtain the product.
Example 2.
The method for extracting dendrobine from dendrobium officinale comprises the following steps:
firstly, taking the dendrobium officinale, crushing, and adding water into the dendrobium officinale according to the material-liquid ratio of 1:10 to obtain a product A;
regulating the pH of the product A to be 9 by using saturated lime water;
③ microwave extracting product B for 3 times at power of 320W for 20min, and mixing extractive solutions to obtain product C;
fourthly, filtering the product C, concentrating the filtrate in two times, controlling the vacuum degree to be 0.07-0.08MPa for the first time, controlling the vacuum degree to be 0.08-0.09MPa for the second time, and obtaining a product D after 10min of vacuum concentration for the two times;
fifthly, adding 4 times of ethanol into the product D, and standing at 1-4 ℃ for 12h to obtain a product E;
sixthly, filtering the product E, washing the precipitate with ethanol for 4 times, and drying to constant weight to obtain the product.
Example 3.
The method for extracting dendrobine from dendrobium officinale comprises the following steps:
firstly, taking the dendrobium officinale, crushing, and adding water into the dendrobium officinale according to the material-liquid ratio of 1:14 to obtain a product A;
regulating the pH of the product A to be 9 by using saturated lime water;
③ microwave extracting the product B for 3 times at 300W for 15min, and mixing the extractive solutions to obtain product C;
fourthly, filtering the product C, concentrating the filtrate in two times, controlling the vacuum degree to be 0.07-0.08MPa for the first time, controlling the vacuum degree to be 0.08-0.09MPa for the second time, and obtaining a product D after 5min of vacuum concentration for the two times;
fifthly, adding 3 times of ethanol into the product D, and standing for 10 hours at the temperature of 1-4 ℃ to obtain a product E;
sixthly, filtering the product E, washing the precipitate with ethanol for 3 times, and drying to constant weight to obtain the product.
Claims (5)
1. The method for extracting dendrobine from dendrobium officinale is characterized by comprising the following steps: the method comprises the following steps:
firstly, taking dendrobium officinale, crushing, and adding water into the dendrobium officinale according to the material-liquid ratio of 1:10-14 to obtain a product A;
regulating the pH of the product A to be 9 by using saturated lime water to obtain a product B;
thirdly, microwave extracting the product B for 3 times under the power of 300-320W, extracting for 15-20min each time, and merging the extracting solutions to obtain a product C;
fourthly, filtering the product C, and concentrating the filtrate in vacuum to obtain a product D;
fifthly, adding 3-4 times of ethanol into the product D, and standing at 1-4 ℃ for 10-12h to obtain a product E;
sixthly, filtering the product E, washing the precipitate for 3 to 4 times by using ethanol, and drying the precipitate to constant weight to obtain the product;
the filtrate is concentrated in vacuum; concentrating twice, controlling the vacuum degree at 0.07-0.08MPa for the first time, controlling the vacuum degree at 0.08-0.09MPa for the second time, and controlling the vacuum concentration time at 5-10min for the second time.
2. The method for extracting dendrobine from dendrobium officinale of claim 1, wherein the method comprises the following steps: step three: the power is 310W.
3. The method for extracting dendrobine from dendrobium officinale of claim 1, wherein the method comprises the following steps: in the step III, the extraction time is 18min each time.
4. The method for extracting dendrobine from dendrobium officinale of claim 1, wherein the method comprises the following steps: the filtrate is concentrated in vacuum; concentrating twice, controlling the vacuum degree at 0.07-0.08MPa for the first time, controlling the vacuum degree at 0.08-0.09MPa for the second time, and controlling the vacuum concentration time at two times to be 8 min.
5. The method for extracting dendrobine from dendrobium officinale of claim 1, wherein the method comprises the following steps: the fifth step is; adding 3 times of ethanol into product D, and standing at 1-4 deg.C for 11 hr to obtain product E.
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| CN109705128A (en) * | 2018-12-21 | 2019-05-03 | 云南金九地生物科技有限公司 | A method of extracting high-purity dendrobine from dendrobium nobile |
| CN113332383A (en) * | 2021-03-05 | 2021-09-03 | 杭州蚕宝生物技术有限公司 | Method for efficiently extracting alkaloid from dendrobium officinale |
| CN113425694A (en) * | 2021-07-02 | 2021-09-24 | 杭州蚕宝生物技术有限公司 | Dendrobium alkaloid orally disintegrating tablet and preparation method thereof |
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