阅读说明：本技术 一种蒙药肋柱花提取物的制备 (Preparation method of Mongolian medicine extract of costaria ) 是由 付明海 阿丽沙 于 2021-06-29 设计创作，主要内容包括：本发明提供了一种蒙药肋柱花提取物的制备；包括以下步骤：步骤1,取肋柱花,粉碎、过筛,加二氯甲烷,回流提取,滤过；步骤2,将滤液进行处理,回收二氯甲烷,滤渣中加入乙醇,回流提取,滤过,将滤液进行处理,回收乙醇至浸膏状；步骤3,将浸膏进行干燥,即可。本发明利用高灵敏的UPLC-MS技术对蒙药肋柱花提取物的化学成分进行鉴定分析,再用电子舌技术检测肋柱花提取物对碱基性苦味、酸性苦味回味、涩味回味、盐酸盐苦味、酸性苦味先味和涩味等六种味觉传感器的苦味强度,对本发明方法工艺参数进行调整,进而得到本发明方法。本发明所涉及方法制备得到的蒙药肋柱花提取物对小鼠胃排空和小肠运动研究有着指导性意义。(The invention provides a preparation method of a Mongolian medicine extract of lomatogonium carinthiacum; the method comprises the following steps: step 1, taking the lomatogonis, crushing, sieving, adding dichloromethane, performing reflux extraction, and filtering; step 2, treating the filtrate, recovering dichloromethane, adding ethanol into filter residue, performing reflux extraction, filtering, treating the filtrate, and recovering ethanol to obtain an extract; and 3, drying the extract to obtain the finished product. The invention utilizes a high-sensitivity UPLC-MS technology to identify and analyze the chemical components of the Mongolian medicine echeveria costata extract, and then uses an electronic tongue technology to detect the bitter intensity of the echeveria costata extract on six taste sensors of basic bitter taste, acidic bitter taste aftertaste, astringent aftertaste, hydrochloride bitter taste, acidic bitter taste first taste, astringent taste and the like, and adjusts the technological parameters of the method, thereby obtaining the method. The Mongolian medicine ribbonfil extract prepared by the method has guiding significance for the research of gastric emptying and small intestine movement of mice.)

1. The preparation method of the Mongolian medicine ribwort flower extract is characterized by comprising the following steps:

step 1, taking the lomatogonis, crushing, sieving, adding dichloromethane, performing reflux extraction, and filtering;

step 2, treating the filtrate, recovering dichloromethane, adding ethanol into filter residue, performing reflux extraction, filtering, treating the filtrate, and recovering ethanol to obtain an extract;

and 3, drying the extract to obtain the finished product.

2. The preparation of the Mongolian medicine extract of costellus of claim 1, wherein in step 1, the dosage ratio of costellus to dichloromethane is 1: 10.

3. The preparation of the Mongolian medicine extract of Ribes nigrum as claimed in claim 1, wherein the sieved Ribes nigrum has a particle size of 80 mesh in step 1.

4. The preparation of the extract of costaphyllum as mongolian drug according to claim 1, wherein in step 1, said reflux extraction is performed 2 times for 4 hours each.

5. The preparation of the extract of copaidia officinalis of claim 1, wherein in step 2, the recovery apparatus is a rotary evaporator RE-2010.

6. The preparation of the Mongolian medicine extract of Rizhilia as claimed in claim 1, wherein in step 2, the ratio of the amount of the residue to the amount of the ethanol is 1: 10.

7. The preparation of the extract of costaphyllum as mongolian drug according to claim 1, wherein in step 2, the reflux extraction is performed 2 times for 6 hours each time.

8. The preparation of the extract of the Mongolian medicine, ribwort, as claimed in claim 1, wherein, in step 2, the concentration of ethanol is 95 wt%.

9. The preparation of the extract of the Mongolian medicine, ribwort, as claimed in claim 1, wherein in step 3, the drying apparatus is a freeze dryer FD-1A-50.

10. The preparation of the extract of the Mongolian medicine, costaphania acuminata according to claim 1, wherein in step 3, the drying time is (3-5h) and the drying temperature is (-50 ℃).

Technical Field

The invention belongs to the field of a preparation method of a Mongolian medicine ribwort extract; in particular to a preparation method of a Mongolian medicine ribwort extract.

Background

Costal column flower is a Mongolian medicine characteristic medicinal material. Has extremely bitter taste, has the effects of calming, invigorating stomach, healing wound and clearing heat, and is commonly used for treating diseases such as heat in the morning, typhoid fever, jaundice, influenza, heatstroke and fever, plague, liver and gallbladder heat, injury of heat, stomach in the morning and evening. Costcolumn flower is also the main traditional medicine for treating liver and gall diseases by Mongolian medicine. It has been shown in previous studies to have therapeutic effects on dyspepsia, gallbladder swelling, jaundice, and inflammation of the liver and gallbladder. Later, the flavonoids of the Mongolian medicine of the costaria flowers are found to have the functions of reducing blood fat and treating obesity. In addition, researches report that swertia pseudochinensis of Gentianaceae is a traditional medicine for treating gastrointestinal diseases such as nausea, gastroparesis, gastric retardation and the like. However, the costal column flower as the Mongolian medicine has less experimental research on the gastrointestinal function and mechanism.

In recent years, with the improvement of living standard of people, the incidence rate of gastrointestinal motility disorder diseases in China is higher and higher, and the disease becomes a research hotspot of a digestion clinic. Gastrointestinal motility is an extremely complex neuromuscular activity that can ensure a person's normal digestive function, and its gastrointestinal motility disorders may lead to a variety of gastrointestinal motility disorder diseases. However, with the development of modern medicine, more and more researches show that the factors for regulating the gastrointestinal motility are diversified, mainly comprising gastrointestinal hormones, intestinal flora, neurotransmitters, some proteins and ion channels, and besides bitter taste and receptors thereof are also involved in the regulation of the gastrointestinal motility. Studies report that the bitter substance bendinylamine can inhibit gastric emptying and reduce food intake, and quinine can inhibit gastrointestinal motility and influence the release of motilin and ghrelin. In addition, many studies report that gastrointestinal motility is not only regulated by gastrin, ghrelin and nitric oxide NO [134], but is also affected by neurotransmitters such as Dopamine (DA) and 5-HT. DA is an inhibitory neurotransmitter, and has the effects of relaxing smooth muscle of stomach and delaying gastric emptying. While 5-HT is an excitatory neurotransmitter, is synthesized and secreted by EC cells, and has the effects of regulating gastrointestinal motility, contracting intestinal canals and promoting gastrointestinal motility.

In order to verify the influence of the extract of the ribbing flower on the gastrointestinal function, the ribbing flower with higher content of the swertiamarin is selected according to the detection result of the chemical components of the extract of the ribbing flower, the influence of the extract of the ribbing flower on the gastric emptying and the small intestine propulsion of a mouse is finally verified, and a gastrointestinal motility disorder model is established by DA and 5-HT, so that the influence of the Mongolian medicine ribbing flower on neurotransmitter disorder gastrointestinal motility disorder diseases is clarified.

Disclosure of Invention

The invention aims to provide a preparation method of a Mongolian medicine ribwort extract.

The invention is realized by the following technical scheme:

the invention relates to a preparation method of a Mongolian medicine extract of lomatogonium carinthiacum, which comprises the following steps:

step 1, taking the lomatogonis, crushing, sieving, adding dichloromethane, performing reflux extraction, and filtering;

step 2, treating the filtrate, recovering dichloromethane, adding ethanol into filter residue, performing reflux extraction, filtering, treating the filtrate, and recovering ethanol to obtain an extract;

and 3, drying the extract to obtain the finished product.

Preferably, in step 1, the ratio of the amount of the ribbing flower to the amount of the dichloromethane is 1: 10.

Preferably, in step 1, the sieved ribbing is 80 mesh in particle size.

Preferably, in step 1, the reflux extraction is performed for 2 times, and each time is 4 hours.

Preferably, in step 2, the recovered apparatus is a rotary evaporator RE-2010.

Preferably, in the step 2, the dosage ratio of the filter residue to the ethanol is 1: 10.

Preferably, in step 2, the reflux extraction is performed for 2 times, and each time is 6 hours.

Preferably, in step 2, the concentration of ethanol is 95 wt%.

Preferably, in step 3, the drying apparatus is a freeze dryer FD-1A-50.

Preferably, in step 3, the drying time is (3-5h), and the drying temperature is (-50 ℃).

The invention has the following advantages: the invention utilizes a high-sensitivity UPLC-MS technology to identify and analyze the chemical components of the Mongolian medicine echeveria costata extract, and then uses an electronic tongue technology to detect the bitter intensity of the echeveria costata extract on six taste sensors of basic bitter taste, acidic bitter taste aftertaste, astringent aftertaste, hydrochloride bitter taste, acidic bitter taste first taste, astringent taste and the like, and adjusts the technological parameters of the method, thereby obtaining the method.

Drawings

FIG. 1 is a radar chart showing bitterness detection of extract of lomatogonium carinthiacum according to the present invention.

Detailed Description

The present invention will be described in detail with reference to specific examples. It should be noted that the following examples are only illustrative of the present invention, but the scope of the present invention is not limited to the following examples.

Examples

The embodiment relates to a preparation method of a Mongolian medicine ribwort extract, which comprises the following steps:

step 1, taking the lomatogonis, crushing, sieving, adding dichloromethane, performing reflux extraction, and filtering;

step 2, treating the filtrate, recovering dichloromethane, adding ethanol into filter residue, performing reflux extraction, filtering, treating the filtrate, and recovering ethanol to obtain an extract;

and 3, drying the extract to obtain the finished product.

Further, in step 1, the ratio of the amount of the ribbonfil to the amount of the dichloromethane is 1: 10.

Further, in step 1, the particle size of the sieved ribbing flower is 80 meshes.

Further, in step 1, the reflux extraction is performed for 2 times, and each time is 4 hours.

Further, in step 2, the recovered apparatus is a rotary evaporator RE-2010.

Further, in the step 2, the dosage ratio of the filter residue to the ethanol is 1: 10.

Further, in step 2, the reflux extraction is performed for 2 times, and each time is 6 hours.

Further, in step 2, the concentration of ethanol is 95 wt%.

Further, in step 3, the drying apparatus is a freeze dryer FD-1A-50.

Further, in step 3, the drying time is (3-5h), and the drying temperature is (-50 ℃).

The method for analyzing the chemical components of the extract of the Mongolian medicine ribwort flower obtained by the method comprises the following specific steps:

detecting the extract of the lomatogonium costatum under a positive and negative ion mode according to UPLC-MS conditions, performing retention time correction, peak identification, peak extraction and other work by using Compound discover 3.0, performing substance identification by using a Thermo mzCloud online database and a Thermo mzValut local database according to secondary mass spectrum information, and then performing identification to obtain 127 chemical components.

A total of 80 chemical components were identified in positive ion mode, and 18 were identified with a content of more than 1%, as shown in Table 1. The content of flavonoids, organic nitrogen compounds, coumarin and derivatives thereof is high.

A total of 47 chemical compositions were identified in negative ion mode, and 12 with a content of more than 1% (Table 2). The content of organic oxygen compounds, iridoids, benzene and its derivatives, glycosides, etc. is high.

TABLE 1

TABLE 2

The bitterness of the extract of the Mongolian medicine ribwort prepared in the embodiment is detected, and the specific steps are as follows:

(1) the response results of the electrical tongue detection of the extract of the ribcage flower with different concentrations to six bitter taste sensors show that the sensitivity of the extract of the ribcage flower to the acidic bitter taste sensor (C00) is the highest, the bitterness is the highest, the acidic bitter taste is increased along with the increase of the concentration, and the bitterness degree of the extract of the ribcage flower with different concentrations is shown in Table 3.

TABLE 3

As can be seen from FIG. 1, 0.4 g/ml of extract of Crotalaria sinica^-1Each sensor response value of (1) is higher than 0.1 g.ml^-1Among them, the acidic bitter taste sensor responded the highest. The astringency of the samples at both concentrations was below the tasteless point where the hydrochloride bitterness was close to the tasteless point, and it was seen that the samples at this concentration tested failed to elicit the response of both the astringency sensor (AE1) and the hydrochloride bitterness sensor (BT 0).

According to this embodiment, the extract of Esculus costatus contains various chemical components, such as xanthones, flavonoids and their glycosides, triterpenes and their glycosides, iridoids and secoiridoids, lactones, organic acids and phenols. The iridoid has biological activities of resisting oxidation, protecting liver, promoting bile flow, relieving spasm, resisting anxiety, reducing blood lipid and blood glucose; the flavonoids have antioxidant, antitumor, antiinflammatory and gastrointestinal motility improving effects. Ribes burejense etc. is extracted with water, and then analyzed by LC-MS to detect 12 compounds including swertiamarin, quercetin, luteolin-7-0-glucoside, 6' -O- [ (E) -cinnamyl ] geniobiosylgenin, (1S, 4As, 6S, 7As) -1, 4a, 5, 6, 7, 7 a-hexahydro-6-hydroxy-7-methyl-1- (O-beta-D-glucopyranosyl) -cyclopenta [ c ] pyran-4-carbaldehyde etc., and the rest 7 are unknown compounds. Separating and purifying flos Corchorii chloroform extract to obtain 4 compounds such as flos Daturae Metelis lactone, 1-hydroxy-3, 7, 8-trimethoxyxanthone, 1-hydroxy-3, 5, 8-trimethoxyxanthone, 8-hydroxy-1, 3, 5-trimethyl xanthone, etc. Jatropha et al reported that 75% ethanol extract of echeveria costata mainly contains xanthones, flavonoids, triterpenes, iridoids and secoiridoids, and the chemical components of the 75% ethanol extract were studied to find 10 compounds, of which 6 compounds such as 1, 8-dihydroxy-3, 4, 5-trimethoxyxanthone, 1-hydroxy-3, 5, 8-trimethoxyxanthone, 1-hydroxy-3, 7, 8-trimethoxyxanthone, 8-hydroxy-1, 3, 5-trimethoxyxanthone, quercetin, 5,7,3',4',5' -pentahydroxyflavone were first isolated from echeveria costata. In addition, they analyzed the chemical components of the flower of ribbonfil in the same manner, and found 8 compounds, which were identified as 2 a-hydroxy oleanolic acid, ursolic acid 3 β -hydroxy-ursolic-11, 12-ene-28, 13 β lactone, ferulic acid, kaempferol, daucosterol, apigenin-7-0-glucoside, luteolin-7-0-glucoside, and swertiamarin, respectively. The first 5 of these are novel compounds. In this study, the method according to the invention was: firstly, performing reflux extraction for 2 times/4 h by using 10 times of dichloromethane, and performing reflux extraction for 2 times/6 h by using 10 times of 95% ethanol to obtain a costaphyllum extract; the invention adopts ultra-high performance liquid chromatography tandem mass spectrometry chromatography technology to carry out qualitative and quantitative analysis on chemical components of the extract of the ribbonfil, and finds out 127 compounds, wherein the content of 30 compounds is more than 1 percent, and the compounds mainly comprise flavonoids, iridoid, coumarin and derivatives thereof, organic oxygen compounds, carboxylic acid and derivatives thereof, lipid compounds, coumarin and derivatives thereof, cinnamic acid and derivatives thereof, benzene and substituted derivatives thereof and the like, wherein the content of bitter components such as swertiamarin, diazabenzene, gentiopicrin, amygdalin, hesperetin and coumarin accounts for 66 percent of the total proportion.

Therefore, the extraction method is the best extraction process of the bitter components of the lomatogonium carinthiacum. The electronic tongue is a novel technology which directly contacts with a sample through a sensor array, collects characteristic electric signals of the sample, and obtains the sensory characteristics of the sample by digitizing the received signals. According to the invention, the electronic tongue is used for detecting the bitterness intensity of the extract of the lomatogonium carinthiacum, and the sensitivity of the acidic bitter taste early-flavor sensor is the highest. The main bitter component source of the ribcage flower is an acidic component, and the chemical components detected by UPLC-MS mainly comprise acidity.

In combination, the extract of the lomatogonium carinthiacum mainly comprises bitter components such as swertiamarin, diazabenzene, gentiopicrin, amygdalin, hesperetin and coumarin, and mainly takes acidity as the main component. The experiment preliminarily discusses the taste characterization of Mongolian medicine ribbing flower, provides a new extraction process of ribbing flower, and provides a basis for better extracting the bitter component of ribbing flower in future.

The method utilizes a high-sensitivity UPLC-MS technology to identify and analyze the chemical components of the Mongolian medicine echeveria costata extract, and then utilizes an electronic tongue technology to detect the bitter intensity of the echeveria costata extract on six taste sensors of basic bitter taste, acidic bitter taste aftertaste, astringent aftertaste, hydrochloride bitter taste, acidic bitter taste first taste, astringent taste and the like, so as to adjust the method, thereby obtaining the method. The Mongolian medicine ribbonfil extract prepared by the method has guiding significance for the research of gastric emptying and small intestine movement of mice.

The foregoing description of specific embodiments of the present invention has been presented. It is to be understood that the present invention is not limited to the specific embodiments described above, and that various changes or modifications may be made by one skilled in the art within the scope of the appended claims without departing from the spirit of the invention.