CN102520096A - Separating identification and application of Farges meehania root volatile oil component - Google Patents

Abstract

The invention belongs to the technical field of research and development of medical products and relates to separating identification and application of a Farges meehania root volatile oil component. A steam distillation method is adopted for extracting Farges meehania root volatile oil, the obtained Farges meehania root volatile oil is analyzed by using a 6890/5973N gas chromatograph-mass spectrometer and comprises the main chemical components as follows: 19.922% of carypohyllene oxide, 8.018% of cubeb oil elaeoptene, 6.179% of alpha-cadinol, 6.173% of globulol and 5.191% of carypohyllene. The Farges meehania root volatile oil component disclosed by the invention has different degrees of effects on inhibiting escherichia coli, streptococcus, salmonella, pasteurella and staphylococcus aureus, and can be developed into a natural antibacterial agent to be applied to food and medicine industries to prepare various forms of products with bacteriostasis effects.

Description

A kind of isolation identification and purposes of U.S. Chinese grass volatile oil component

Technical field

Medical product research and development technical field under the present invention.The present invention relates to a kind of isolation identification and purposes of U.S. Chinese grass volatile oil component.

Background technology

U.S. Chinese grass is the dry aerial parts that Labiatae (Labiatae) tap grass belongs to (Meehania) plant sesame flower Meehania urticifolia (Miq.) Makino; Another name folium urticae tap grass; Sesame flower (Liaoning), U.S. Chinese flower, different name water rattletop (Sichuan) is per nnial herb; Be distributed in ground such as Tonghua, Huijiang, Liuhe, Jian, Fusong, Jingyu, Jiaohe, Shulan, Antu, Dunhua, autonomous region of long white Korean nationality, Tonghua County, also there are distribution in Korea, Japan.Hot, bitter, the cold nature of U.S. Chinese grass flavor, all herbal medicine can be used as medicine among the people, has relieving superficies by cooling, effects such as inducing diuresis for removing edema.Be used for cold, fever, rush down dysentery stomachache, hepatitis, cholecystitis, difficult urination, diseases such as snake bite.In the Chinese herbal medicine usefulness of helping among the people, herb can be used as medicine, and as cold medicine, has the heat-clearing of delivering, effects such as eliminating damp, detoxifying, and its natural resources are very abundant, and the amount of containing is big.

At present, about the rarely seen report of research of U.S. Chinese grass volatile oil, U.S. Chinese grass volatile oil was not done antibacterial research.Therefore, the present invention adopts steam distillation that the U.S. Chinese grass of medicinal plant volatile oil is extracted, and utilizes the 6890/5973N gas chromatography to separate with the mass spectrometry instrument and identifies, and the careless hair oil of the U.S. Chinese has been carried out bacteriostatic test.

U.S. Chinese grass volatile oil of the present invention is inhibited to gram-positive bacteria, Gram-negative bacteria, and this bacteriostasis has positive using value.

Summary of the invention

1, a kind of isolation and identification method of U.S. Chinese grass volatile oil component is characterized in that may further comprise the steps:

Step (1): take by weighing dry U.S. Chinese grass meal 100g, put after crushed in the glass container, add ultrasonic 1h behind the 500ml-600ml distilled water immersion 6h-8h, temperature is 30 ℃, and power is 80W;

Step (2): (1) Sino-U.S. Chinese grass meal and soak solution are together poured in the volatile oil extractor, adopted steam distillation to extract 4-5h, the floating WS that volatile oil is arranged above obtaining;

Step (3): have the WS of volatile oil to emit water layer top floating in (2); Wash volatile oil extractor repeatedly with ether; U.S. Chinese grass volatile oil fully is dissolved in the ether, the diethyl ether solution that is dissolved with volatile oil is poured in the beaker, in beaker, add the anhydrous sodium sulfate suck dry moisture again; Promptly obtain pure volatile oil after reclaiming ether, it is subsequent use to put into 4 ℃ of preservations of refrigerator;

Step (4): with 6890/5973N gas chromatography and GC-MS analysis, its main chemical compositions is the carypohyllene of 19.922% carypohyllene, 8.018% cubebene, α-cadinol of 6.179%, 6.173% basket eucalyptus pure and mild 5.191% with the U.S. Chinese grass of gained in the step (3) volatile oil.

2, U.S. Chinese grass volatile oil suppresses Escherichia coli, streptococcus, salmonella, Pasteur bacterium, staphylococcus aureus.

According to the present invention, " % " among the present invention is percentage by weight.

Embodiment

The isolation identification and the purposes of a kind of U.S. Chinese grass volatile oil component of the present invention comprise following examples, and following embodiment can further specify the present invention, but does not limit the present invention in any way.

Embodiment 1:

1, a kind of isolation and identification method of U.S. Chinese grass volatile oil component is characterized in that may further comprise the steps:

Step (1): take by weighing dry U.S. Chinese grass meal 100g, put after crushed in the glass container, add ultrasonic 1h behind the 500ml distilled water immersion 6h, temperature is 30 ℃, and power is 80W;

Step (2): (1) Sino-U.S. Chinese grass meal and soak solution are together poured in the volatile oil extractor, adopted steam distillation to extract 4-5h, the floating WS that volatile oil is arranged above obtaining;

Step (3): have the WS of volatile oil to emit water layer top floating in (2); Wash volatile oil extractor repeatedly with ether; U.S. Chinese grass volatile oil fully is dissolved in the ether, the diethyl ether solution that is dissolved with volatile oil is poured in the beaker, in beaker, add the anhydrous sodium sulfate suck dry moisture again; Promptly obtain pure volatile oil after reclaiming ether, it is subsequent use to put into 4 ℃ of preservations of refrigerator;

Step (4): with 6890/5973N gas chromatography and GC-MS analysis, its main chemical compositions is the carypohyllene of 19.922% carypohyllene, 8.018% cubebene, α-cadinol of 6.179%, 6.173% basket eucalyptus pure and mild 5.191% with the U.S. Chinese grass of gained in the step (3) volatile oil.

Step (5): (3) Sino-U.S. Chinese grass volatile oil is carried out bacteriostatic test, and U.S. Chinese grass volatile oil all has inhibiting effect in various degree to Escherichia coli, streptococcus, salmonella, Pasteur bacterium, staphylococcus aureus.

Embodiment 2:

1, a kind of isolation and identification method of U.S. Chinese grass volatile oil component is characterized in that may further comprise the steps:

Step (1): take by weighing dry U.S. Chinese grass meal 100g, put after crushed in the glass container, add ultrasonic 1h behind the 600ml distilled water immersion 8h, temperature is 30 ℃, and power is 80W;

Step (2): (1) Sino-U.S. Chinese grass meal and soak solution are together poured in the volatile oil extractor, adopted steam distillation to extract 4-5h, the floating WS that volatile oil is arranged above obtaining;

Step (3): have the WS of volatile oil to emit water layer top floating in (2); Wash volatile oil extractor repeatedly with ether; U.S. Chinese grass volatile oil fully is dissolved in the ether, the diethyl ether solution that is dissolved with volatile oil is poured in the beaker, in beaker, add the anhydrous sodium sulfate suck dry moisture again; Promptly obtain pure volatile oil after reclaiming ether, it is subsequent use to put into 4 ℃ of preservations of refrigerator;

Step (4): with 6890/5973N gas chromatography and GC-MS analysis, its main chemical compositions is the carypohyllene of 19.922% carypohyllene, 8.018% cubebene, α-cadinol of 6.179%, 6.173% basket eucalyptus pure and mild 5.191% with the U.S. Chinese grass of gained in the step (3) volatile oil.

Step (5): (3) Sino-U.S. Chinese grass volatile oil is carried out bacteriostatic test, and U.S. Chinese grass volatile oil all has inhibiting effect in various degree to Escherichia coli, streptococcus, salmonella, Pasteur bacterium, staphylococcus aureus.

The bacteriostasis of the U.S. Chinese grass of pharmacological testing volatile oil

1 experiment material

Escherichia coli, streptococcus, salmonella, Pasteur bacterium and staphylococcus aureus all come from American Type Culture Collecti (ATCC), in the peptone steamed beef soup, cultivate 12h for 37 ℃.

Nutrient culture media is a beef-protein medium.Use fluid nutrient medium when activation of bacterium and cultivation, consist of: beef extract: peptone: sodium chloride: distilled water (3: 10: 5: 1000).Use solid medium when preservation of bacterium and bacteriostatic experiment, consist of: beef extract: peptone: sodium chloride: agar: distilled water (3: 10: 5: 22: 1000).

2 experimental techniques

2.1 culture medium preparation

Get beef extract 0.6g, peptone 2g, sodium chloride 1g, distilled water 200ml, mix and stir that it is dissolved fully, be made into fluid nutrient medium.Measure fluid nutrient medium with graduated cylinder, its average mark is contained in the triangular flask of 10 50ml every bottle of 20ml.Get beef extract 3g, peptone 10g, sodium chloride 5g, agar 22g, distilled water 1000ml, be placed on the Electric stove after the mixing and heat, agar is dissolved fully, be made into solid medium.Measure solid medium with graduated cylinder, its average mark is contained in the triangular flask of 5 150ml every bottle of 100ml.The triangular flask bottleneck is all wrapped up with the tampon jam-pack and with kraft.

2.2 sterilization

The solid medium for preparing, fluid nutrient medium are all put into high-pressure sterilizing pot sterilize, when temperature rose to 121 ℃, 20min picked up counting.Experiment equipments such as double dish, graduated cylinder, tweezers, scissors, 6mm filter paper (being placed in the small beaker), dropper, ammonia bottle are all put into 170 ℃ of sterilizations of baking oven 2h.Experiment is preceding with 75% ethanol wiping superclean bench and ultraviolet sterilization 20min.37 ℃ of constant incubators need be used 75% ethanol wiping sterilization before use.

2.3 the cultivation of bacterium with look into bacterium

5 bacterial classifications are poured in the triangular flask that fluid nutrient medium is housed on superclean bench, under the spirit lamp environment, and every kind of bacterium is cultivated two parts.These 10 triangular flasks are put into shaking table shake bacterium 6h for 37 ℃.Microscope begins to look into bacterium after doing microscopy, and record data also calculate bacteria concentration, if 5 medium square total bacteria counts are A, bacterium liquid extension rate is B, then total bacteria count=A/5 * 25 * 10000 * 13=50000AB in the 1ml bacterium liquid

2.4 the preparation of sample liquid and filter paper

U.S. Chinese grass volatile oil is mixed with the solution (can add 0～4 μ L dimethyl sulfoxide (DMSO) and help appearances) of 5mg/ml, pressed 1: 2 ⁿBe diluted to the solution of 2.5mg/ml, 1.25mg/ml, 0.625mg/ml, 0.3125mg/ml.Use card punch to break into the disk of diameter filter paper, at superclean bench filter paper is put in each gradient sample and soaked 2h as 6mm.

2.5 the dull and stereotyped preparation of bacterio-agar is arranged

The bacterium liquid that dilution is good mixes with solid medium and rocks evenly, and this moment, the concentration of bacterium should reach 3.6 * 10 ⁸CFU/ml.To there be bacterium culture medium to pour into successively in each double dish, and rock double dish and make nutrient culture media be layered on the double dish bottom equably, and be cooled to and solidify.

2.6 bacteriostatic experiment method

, in that label on the bacterio-agar flat board is arranged soaked filter paper is pressed from both sides out with tweezers with marking pen, puts into the double dish of writing corresponding contrast numbering, each sample do three parallel, put into 37 ℃ of constant incubators and cultivate.

3 experimental results

Fungistatic effect is represented (mm of unit) with antibacterial circle diameter DD.

12h after bacteriostatic experiment finishes, 24h, 36h, 48h observe the growing state of bacterium, measure antibacterial circle diameter and record.

The bacteriostatic activity of the U.S. Chinese grass of table 1 volatile oil

The result shows: U.S. Chinese grass volatile oil all has inhibiting effect in various degree to Escherichia coli, streptococcus, salmonella, Pasteur bacterium, staphylococcus aureus.

Explain that through the practical implementation instance U.S. Chinese grass volatile oil can be used as natural antibacterial agent and is used for food, medicine trade, replace the synthetic antibacterial agents of toxic side effect.

The analysis result of the U.S. Chinese grass of table 2 chemical composition of volatile oil

Claims (2)

1. the isolation and identification method of a U.S. Chinese grass volatile oil component is characterized in that may further comprise the steps:

Step (1): take by weighing dry U.S. Chinese grass meal 100g, put after crushed in the glass container, add ultrasonic 1h behind the 500ml-600ml distilled water immersion 6h-8h, temperature is 30 ℃, and power is 80W;

Step (2): (1) Sino-U.S. Chinese grass meal and soak solution are together poured in the volatile oil extractor, adopted steam distillation to extract 4-5h, the floating WS that volatile oil is arranged above obtaining;

Step (3): have the WS of volatile oil to emit water layer top floating in (2); Wash volatile oil extractor repeatedly with ether; U.S. Chinese grass volatile oil fully is dissolved in the ether, the diethyl ether solution that is dissolved with volatile oil is poured in the beaker, in beaker, add the anhydrous sodium sulfate suck dry moisture again; Promptly obtain pure volatile oil after reclaiming ether, it is subsequent use to put into 4 ℃ of preservations of refrigerator;

Step (4): with 6890/5973N gas chromatography and GC-MS analysis, its main chemical compositions is the carypohyllene of 19.922% carypohyllene, 8.018% cubebene, α-cadinol of 6.179%, 6.173% basket eucalyptus pure and mild 5.191% with the U.S. Chinese grass of gained in the step (3) volatile oil.

2. the purposes of a kind of U.S. Chinese grass volatile oil as claimed in claim 1 in the suppressant of preparation inhibition Escherichia coli, streptococcus, salmonella, Pasteur bacterium, staphylococcus aureus.