Extraction method of dragon's blood, dragon's blood extracting solution, mouthwash composition as well as preparation method and application of mouthwash composition

Abstract

The invention provides an extraction method of dragon's blood, an extraction solution of the dragon's blood, a mouthwash composition and a preparation method and application of the mouthwash composition, and belongs to the technical field of plant pharmaceuticals, the extraction method of the dragon's blood is characterized by comprising the following steps: extracting grease-containing wood of the dragon's blood by adopting clove essential oil to obtain the dragon's blood extraction solution; the extraction conditions are as follows: the mass ratio of the resina draconis fat-containing wood to the clove essential oil is (1: 15)-(1: 60); the extraction temperature is 20-50 DEG C; the extraction time is 1-24 hours. The prepared dragon's blood mouthwash oil is simple in process and stable in quality and has a good bacteriostatic effect, and in-vitro bacteriostatic experiment tests show that the effect of inhibiting staphylococcus aureus and escherichia coli is obvious.

Description

Technical Field

[0001] The invention relates to the technical field of preparing plant medicines, and in particular to a method for extracting dragon's blood, a dragon's blood extract, a mouthwash composition, and a preparation method and application thereof. Background Art

[0002] Oral diseases mainly manifest as oral inflammation, ulcers, bad breath, etc., which will affect the patient's comfort and diet, and in severe cases will cause some systemic diseases, such as cardiovascular disease, rheumatoid arthritis, diabetes, Alzheimer's disease, etc. As people's awareness of oral care continues to increase, more people rely on using mouthwash for oral care to achieve the purpose of fresh breath and prevent periodontal diseases.

[0003] Mouthwash oil originated from oil pulling in ancient India and has been used for more than 3,000 years. It is a natural therapy that uses natural plant oils to make oral washes for daily oral hygiene to promote oral health. [3] . Studies have shown that this natural remedy can cure more than thirty systemic diseases, such as headaches, diabetes, asthma, etc. [4] ; and has a therapeutic effect on a variety of oral diseases, such as improving gum health, reducing inflammation and bleeding, whitening teeth, reducing bad breath, and improving oral hygiene.

[0004] Dragon's Blood, derived from Dracaena cochinchinensis (Lour.) SCChen of the Liliaceae family, has the effects of promoting blood circulation and dispersing blood stasis, healing sores and promoting tissue regeneration, relieving pain and stopping bleeding. It is often used to treat injuries from falls, traumatic bleeding, swelling and pain from congestion, irregular menstruation, amenorrhea, dysmenorrhea, and ecthyma that does not heal for a long time. Pharmacological studies have shown that Dragon's Blood has the effects of analgesia, anti-inflammatory, promoting skin repair and antibacterial. It also has good antibacterial activity against Staphylococcus aureus and Candida albicans. However, the poor solubility of Dragon's Blood affects its clinical application. Summary of the invention

[0005] In order to solve the problems of low solubility of dragon's blood and easy solidification of mouthwash oil at room temperature in the prior art, the present invention proposes to increase the solubility of dragon's blood in coconut oil by using clove essential oil, and add peppermint oil to prepare the mouthwash oil, and evaluate the solubility of dragon's blood, solidification point, synergistic antibacterial effect of dragon's blood and extraction solution clove essential oil, and further evaluate the antibacterial effect of dragon's blood mouthwash oil through in vitro antibacterial experiments.

[0006] In a first aspect, the present invention provides a method for extracting dragon's blood, which comprises extracting dragon's blood oil-containing wood with clove essential oil to obtain a dragon's blood extract; the extraction conditions include: a mass ratio of dragon's blood oil-containing wood to clove essential oil of 15:1 to 60:1; an extraction temperature of 20 to 50° C.; and an extraction time of 1 hour to 24 hours.

[0007] In a second aspect, the present invention also provides a dragon's blood extract, wherein the content of dragon's blood A in the dragon's blood extract is 0.5 to 10 mg·mL -1 , the content of dragon's blood B is 0.2~5mg·mL -1 .

[0008] As a preferred embodiment, the content of dragon's blood extract A is 1 to 10 mg / mL -1 , the content of dragon's blood B is 0.4~5mg·mL -1 .

[0009] In a third aspect, the present invention also provides a mouthwash composition or mouthwash oil, comprising: 1 to 10 parts of peppermint oil; 0.1 to 0.3 parts of propylparaben; 97.9 to 79.7 parts of coconut oil; 1 to 10 parts of dragon's blood extract; the dragon's blood extract is obtained by the extraction method described in the first aspect, or is from any one of the dragon's blood extracts described in the second aspect.

[0010] In a fourth aspect, the present invention also provides a method for preparing the mouthwash composition described in the third aspect, the method comprising: mixing raw materials containing peppermint oil, propylparaben, coconut oil and dragon's blood extract to prepare the mouthwash composition; the mixing conditions include: mixing temperature of 20 to 50° C. and mixing time of 1 to 60 min.

[0011] In a fifth aspect, the present invention also provides the use of the dragon's blood extract obtained by the extraction method described in the first aspect or the dragon's blood extract described in any one of the second aspects, or the mouthwash composition described in the third aspect in inhibiting Staphylococcus aureus and / or Escherichia coli.

[0012] The dragon's blood extract and dragon's blood mouthwash oil prepared by the invention have simple process, stable quality and good antibacterial effect. In vitro antibacterial experimental tests show that the antibacterial effect on Staphylococcus aureus and Escherichia coli is obvious. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 The HPLC chromatogram of extracting dragon's blood lipid-containing wood from each essential oil in the present invention (in the figure, 1 camellia oil; 2 eucalyptol; 3 olive oil; 4 peppermint essential oil; 5 clove essential oil; ① dragon's blood A; ② dragon's blood B);

[0014] Figure 2 The antibacterial graph of the normal saline solution of Comparative Example 7;

[0015] Figure 3 This is the antibacterial graph of 30% methanol in Comparative Example 8;

[0016] Figure 4This is the antibacterial graph of the 30% methanol extract of dragon's blood in comparative example 5;

[0017] Figure 5 This is the antibacterial graph of clove essential oil in Comparative Example 6;

[0018] Figure 6 This is the antibacterial graph of the clove essential oil dragon's blood extract of Example 2;

[0019] Figure 7 This is a graph showing the inhibition of Staphylococcus aureus growth by the essential oil dragon's blood extract, normal saline, and Yunnan Baiyao Jinkoujian mouthwash of the present invention;

[0020] Figure 8 The graph shows the inhibition of Escherichia coli growth by the essential oil dragon's blood extract, normal saline, and Yunnan Baiyao Jinkoujian mouthwash of the present invention. DETAILED DESCRIPTION

[0021] The present invention will be further described below in conjunction with specific embodiments, but they do not constitute any limitation to the present invention.

[0022] The instruments and materials used in the examples and comparative examples of the present invention are described as follows.

[0023] LC-2030C 3D high performance liquid chromatograph (Shimadzu);

[0024] Velocity 18R centrifuge (Dynamica);

[0025] HW-S26 electric constant temperature water bath (Shanghai Yiheng Scientific Instrument Co., Ltd.);

[0026] HZ-124 / B55 one-hundred-thousandth electronic balance (Huazhi (Fujian) Electronic Technology Co., Ltd.).

[0027] Dragon's Blood Resin-Containing Dried Wood (Batch No.: 2023110301, Xishuangbanna Jianlong Pharmaceutical Co., Ltd.);

[0028] Dragon's blood fat dried wood coarse powder: ground from dragon's blood fat dried wood of batch number 2023110301;

[0029] Dragon's blood medicinal materials were purchased from Xishuangbanna Jianlong Pharmaceutical Co., Ltd.;

[0030] Loureirin A reference substance (Batch No.: AFCK1402, Chengdu Aifa Biotechnology Co., Ltd.);

[0031] Loureirin B reference substance (Batch No.: AFCK1403, Chengdu Aifa Biotechnology Co., Ltd.);

[0032] Clove essential oil (Batch No. 20231107, Huashu Fragrance Co., Ltd.);

[0033] Camellia oil (Batch No. 20230707, Qiandao source);

[0034] Olive oil (batch number: 20231016, Yushan County Dachengcang Food Co., Ltd.);

[0035] Eucalyptol (lot number: 1J2206E010G, Gyalabs);

[0036] Peppermint oil (Batch No. 20231122, Huangshan Tianmu Mint Pharmaceutical Co., Ltd.);

[0037] Coconut oil (Batch No.: Q / WCY0001S, Wenchang Pure Coconut Food Co., Ltd.);

[0038] Propylparaben (Batch No.: P1651073, adamas);

[0039] Yunnan Baiyao Jinkoujian Mouthwash (Batch No.: NTK62, Yunnan Baiyao Group Co., Ltd.)

[0040] Acetonitrile (batch number: K54935530313, chromatographic grade, MORK);

[0041] Phosphoric acid (Batch No.: 20210315, AR, Tianjin Damao Chemical Reagent Factory);

[0042] Methanol (lot number: F23NBR203, chromatographic grade, Fisher Scientific).

[0043] Staphylococcus aureus (ATCC6538).

[0044] Antibacterial to Escherichia coli (ATCC25922).

[0045] Comparative Example 1

[0046] Weigh 200.0 mg of coarse powder of dry wood containing fat of dragon's blood, put it into 6 mL of eucalyptol, extract at 25-35° C. for 8 h, centrifuge, take 2 mL of supernatant, dilute and extract with 6 mL of methanol, and obtain dragon's blood extract 1#.

[0047] Comparative Example 2

[0048] Weigh 200.0 mg of Dragon's Blood fat-containing dry wood crude powder, place it in 6 mL of olive oil, extract at 25-35°C for 8 h, centrifuge, take 2 mL of the supernatant, dilute and extract with 6 mL of methanol to obtain Dragon's Blood extract 2#.

[0049] Comparative Example 3

[0050] Weigh 200.0 mg of Dragon's Blood fat-containing dry wood crude powder, place it in 6 mL of camellia oil, extract at 25-35°C for 8 h, centrifuge, take 2 mL of the supernatant, dilute and extract with 6 mL of methanol to obtain Dragon's Blood extract 3#.

[0051] Comparative Example 4

[0052] Weigh 200.0 mg of Dragon's Blood fat-containing dry wood crude powder, place it in 6 mL of peppermint essential oil, extract at 25-35°C for 8 h, centrifuge, take 2 mL of the supernatant, dilute and extract with 6 mL of methanol to obtain Dragon's Blood extract 4#.

[0053] Comparative Example 5

[0054] Take the dragon's blood medicinal material and grind it, pass it through a 100-mesh sieve, weigh 500 mg of the powder, put it in a 50 mL volumetric flask, dissolve it in 30% methanol and make up to 50 mL, and get 10 mg mL -1 Dragon's blood extract 5#.

[0055] Comparative Example 6

[0056] Clove essential oil, labeled as Clove Essential Oil 1#.

[0057] Comparative Example 7

[0058] Normal saline, marked as normal saline 1#.

[0059] Comparative Example 8

[0060] 30% methanol, marked as 30% methanol 1#.

[0061] Comparative Example 9

[0062] Coconut oil, labeled as coconut oil #1.

[0063] Comparative Example 10

[0064] Yunnan Baiyao Jinkoujian Mouthwash, labeled as Mouthwash 1#.

[0065] Example 1

[0066] Weigh 200.0 mg of Dragon's Blood fat-containing dry wood crude powder, place it in 6 mL of clove essential oil, extract at 25-35°C for 8 h, centrifuge, take 2 mL of the supernatant, dilute and extract with 6 mL of methanol to obtain Dragon's Blood extract 6#.

[0067] Test Example 1

[0068] Content detection of dragon's blood A and dragon's blood B in different solvents: chromatographic analysis was used to detect the content of dragon's blood A and dragon's blood B in dragon's blood extract 1#, dragon's blood extract 2#, dragon's blood extract 3#, dragon's blood extract 4#, and dragon's blood extract 6#, respectively. The chromatograms are shown in FIG. Figure 1 The test results are shown in Table 1. The data in Table 1 show that the content of dragon's blood A and dragon's blood B in dragon's blood extract 6# extracted from clove essential oil is the highest.

[0069] Chromatographic conditions: ODS-3C18 column (4.6 mm × 250 mm, 5 μm), acetonitrile (C)-0.1% phosphoric acid aqueous solution (D) as mobile phase, gradient elution: 0-3 min, 30%-30% C; 3-30 min, 30%-60% C; 30-33 min, 60%-80% C; 33-45 min, 80%-80% C; 45-50 min, 80%-70% C; 50-55 min, 70%-70% C. Column temperature 40°C; flow rate 1.0 mL / min; detection wavelength 260 nm; injection volume 10 μL.

[0070] Table 1

[0071]

[0072] “-” means not detected.

[0073] Example 2

[0074] Take 200.0 mg of dry dragon's blood fat-containing dry wood crude powder, add 6 mL of clove essential oil, extract at 25-35°C for 8 hours, and take the supernatant after high-speed centrifugation to obtain dragon's blood extract 7#.

[0075] Test Example 2

[0076] The antibacterial rate of dragon's blood extract 5#, dragon's blood extract 7#, clove essential oil 1#, normal saline 1#, and 30% methanol 1# were tested respectively:

[0077] Ask 10 testers to rinse their mouths with clean water to wash away the remaining food residues in their mouths, then rinse with 5 mL of pure water for 1 minute, collect the pure water after rinsing, take 1 mL and dilute 1000 times to make a bacterial suspension. Use a pipette to transfer 2 mL of the bacterial suspension and sample into a 5 mL centrifuge tube, mix well and set aside. Take 0.1 mL of each sample and spread it evenly on the plate. Incubate in a constant temperature incubator at 37°C for 24 hours, then take out and observe the colony growth area. The results are as follows: Figure 2-Figure 6 shown.

[0078] in, Figure 2 This is the antibacterial graph of physiological saline 1# of comparative example 7; Figure 3 This is the antibacterial graph of 30% methanol 1# in Comparative Example 8; Figure 4 This is the antibacterial graph of the 30% methanol extract of dragon's blood in comparative example 5; Figure 5 This is the antibacterial graph of clove essential oil 1# of comparative example 6; Figure 6 This is the antibacterial graph of Dragon's Blood Extract 7# in Example 2.

[0079] The results show that Figure 6 The colony growth area is the smallest, indicating that the antibacterial effect of clove essential oil dragon's blood extract (dragon's blood extract 7#) is the most significant.

[0080] Example 3

[0081] Take 200.0 mg of dry crude powder of the resinous wood of Dracaena draco, add 6 mL of clove essential oil, extract at 25-35°C for 8 h, centrifuge at high speed, and take the supernatant to obtain Dracaena draco extract 8#.

[0082] Mouthwash oil ingredients: 5 parts of dragon's blood extract 8#, 5 parts of peppermint oil, 0.1 parts of propylparaben, 89.9 parts of coconut oil, labeled as dragon's blood mouthwash oil 1#.

[0083] Test Example 3

[0084] Dragon's Blood Mouthwash Oil 1# and Coconut Oil 1# were placed in the same temperature environment to test the solidification point. The results showed that Coconut Oil 1# began to solidify at 25°C, and Dragon's Blood Mouthwash Oil 1# began to solidify at 19°C, indicating that Dragon's Blood Mouthwash Oil 1# lowered the solidification point compared to Coconut Oil 1#.

[0085] Example 4

[0086] Take 200.0 mg of dry Dragon's Blood fat-containing wood powder, add 6 mL of clove essential oil, extract at 25-35°C for 8 h, and take the supernatant after high-speed centrifugation to obtain Dragon's Blood extract 9#.

[0087] Mouthwash oil ingredients: 5 parts of dragon's blood extract 9#, 5 parts of peppermint oil, 0.1 parts of propylparaben, 89.9 parts of coconut oil, marked as dragon's blood mouthwash oil 2#.

[0088] Test Example 4

[0089] Dragon's Blood Mouthwash Oil 2#, Mouthwash 1#, and Normal Saline 1# were used to test the antibacterial rate of Staphylococcus aureus and Escherichia coli:

[0090] Use a 1 mL sterile pipette tip to dig three holes in the culture dish, transfer 50 μL of bacterial suspension and 50 μL of sample (dragon blood mouthwash oil 2#, mouthwash 1#, normal saline 1#) to each hole, place it in a 37°C incubator, take it out after 24 hours, and observe the colony growth area.

[0091] Dragon's blood mouthwash oil 2# was used as the sample group, mouthwash 1# was used as the positive group, and normal saline 1# was used as the negative group.

[0092] The results are as follows Figure 7 (Staphylococcus aureus) and Figure 8(Escherichia coli) as shown in the figure, the dragon's blood mouthwash oil 2# prepared by the present invention inhibits the growth of Escherichia coli for up to 15 hours, which is basically the same as the positive drug group, and grows colonies 6 hours later than the negative group; it inhibits Staphylococcus aureus for 14 hours, and grows colonies 5 hours later than the negative group.

[0093] Any numerical value mentioned in the present invention includes all values ​​that increase by one unit each time from the lowest value to the highest value if there is only an interval of two units between any minimum value and any maximum value. For example, if the amount of a component, or the value of a process variable such as temperature, pressure, time, etc. is declared to be 50-90, in this specification it means that 51-89, 52-88... and 69-71 and 70-71 are specifically listed. For non-integer values, 0.1, 0.01, 0.001 or 0.0001 can be appropriately considered as a unit. These are just some specially specified examples. In this application, in a similar manner, all possible combinations of numerical values ​​between the listed lowest and highest values ​​are considered to have been disclosed.

[0094] It should be noted that the embodiments described above are only used to explain the present invention and do not constitute any limitation to the present invention. The present invention has been described with reference to typical embodiments, but it should be understood that the words used therein are descriptive and explanatory words, rather than restrictive words. The present invention may be modified as specified within the scope of the claims of the present invention, and the present invention may be revised without departing from the scope and spirit of the present invention. Although the present invention described therein relates to specific methods, materials and embodiments, it does not mean that the present invention is limited to the specific examples disclosed therein, on the contrary, the present invention can be extended to all other methods and applications with the same functions.

Claims

1. A method for extracting dragon's blood, characterized in that: The lipid-containing wood of dragon's blood is extracted by using clove essential oil to obtain dragon's blood extract; The extraction conditions include: The mass ratio of dragon's blood resinous wood to clove essential oil is 15:1 to 60:1; Extraction temperature 20~50℃; Extraction time is 1h~24h.

2. Dragon's blood extract, characterized in that The content of dragon's blood extract A is 0.5-10 mg·mL -1 , the content of dragon's blood B is 0.2~5mg·mL -1 .

3. The dragon's blood extract according to claim 2, characterized in that The content of dragon's blood extract A is 1-10 mg / mL -1 , the content of dragon's blood B is 0.4~5mg·mL -1 .

4. A mouthwash composition, characterized in that contain: 1-10 parts of peppermint oil; Propylparaben 0.1-0.3 parts; Coconut oil 97.9-79.7 parts; 1-10 parts of dragon's blood extract; The dragon's blood extract is obtained by the method of claim 1, or is selected from the dragon's blood extract of claim 2 or 3.

5. The method for preparing the mouthwash composition according to claim 4, characterized in that: The method comprises: The raw materials containing peppermint oil, propylparaben, coconut oil and dragon's blood extract are mixed to obtain the mouthwash composition; the mixing conditions include: mixing temperature of 20-50° C. and mixing time of 1-60 minutes.

6. Use of the dragon's blood extract obtained by the method of claim 1, the dragon's blood extract according to claim 2 or 3, or the mouthwash composition according to claim 4 in inhibiting Staphylococcus aureus and / or Escherichia coli.

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