CN107307262A - A kind of calamondin essential oil with bacteriostatic activity and preparation method and application - Google Patents
A kind of calamondin essential oil with bacteriostatic activity and preparation method and application Download PDFInfo
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- CN107307262A CN107307262A CN201710692083.9A CN201710692083A CN107307262A CN 107307262 A CN107307262 A CN 107307262A CN 201710692083 A CN201710692083 A CN 201710692083A CN 107307262 A CN107307262 A CN 107307262A
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- calamondin
- essential oil
- bacteriostatic activity
- rascal
- leaf
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- 235000004332 Citrus madurensis Nutrition 0.000 title claims abstract description 79
- 230000003385 bacteriostatic effect Effects 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
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- 241001156382 Lansium Species 0.000 claims abstract description 30
- 239000007788 liquid Substances 0.000 claims abstract description 10
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- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 206010016952 Food poisoning Diseases 0.000 description 2
- 240000009088 Fragaria x ananassa Species 0.000 description 2
- 241001093501 Rutaceae Species 0.000 description 2
- 241000295644 Staphylococcaceae Species 0.000 description 2
- 235000009754 Vitis X bourquina Nutrition 0.000 description 2
- 235000012333 Vitis X labruscana Nutrition 0.000 description 2
- 240000006365 Vitis vinifera Species 0.000 description 2
- 235000014787 Vitis vinifera Nutrition 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- 235000021393 food security Nutrition 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- XMGQYMWWDOXHJM-UHFFFAOYSA-N limonene Chemical compound CC(=C)C1CCC(C)=CC1 XMGQYMWWDOXHJM-UHFFFAOYSA-N 0.000 description 2
- CDOSHBSSFJOMGT-UHFFFAOYSA-N linalool Chemical compound CC(C)=CCCC(C)(O)C=C CDOSHBSSFJOMGT-UHFFFAOYSA-N 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- NDVASEGYNIMXJL-UHFFFAOYSA-N sabinene Chemical compound C=C1CCC2(C(C)C)C1C2 NDVASEGYNIMXJL-UHFFFAOYSA-N 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- NDVASEGYNIMXJL-NXEZZACHSA-N (+)-sabinene Natural products C=C1CC[C@@]2(C(C)C)[C@@H]1C2 NDVASEGYNIMXJL-NXEZZACHSA-N 0.000 description 1
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- CDOSHBSSFJOMGT-JTQLQIEISA-N (R)-linalool Natural products CC(C)=CCC[C@@](C)(O)C=C CDOSHBSSFJOMGT-JTQLQIEISA-N 0.000 description 1
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- 244000007645 Citrus mitis Species 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 244000089759 Fortunella margarita Species 0.000 description 1
- 235000019123 Fortunella margarita Nutrition 0.000 description 1
- 244000307700 Fragaria vesca Species 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 206010019851 Hepatotoxicity Diseases 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 206010023126 Jaundice Diseases 0.000 description 1
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- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 235000019082 Osmanthus Nutrition 0.000 description 1
- 241000333181 Osmanthus Species 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- KQAZVFVOEIRWHN-UHFFFAOYSA-N alpha-thujene Natural products CC1=CCC2(C(C)C)C1C2 KQAZVFVOEIRWHN-UHFFFAOYSA-N 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000001857 anti-mycotic effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000002543 antimycotic Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229930006722 beta-pinene Natural products 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 239000005452 food preservative Substances 0.000 description 1
- 235000019249 food preservative Nutrition 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 231100000304 hepatotoxicity Toxicity 0.000 description 1
- 230000007686 hepatotoxicity Effects 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229940087305 limonene Drugs 0.000 description 1
- 235000001510 limonene Nutrition 0.000 description 1
- 229930007744 linalool Natural products 0.000 description 1
- 235000013622 meat product Nutrition 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000417 nephrotoxicity Toxicity 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 235000020995 raw meat Nutrition 0.000 description 1
- 235000021067 refined food Nutrition 0.000 description 1
- 229930006696 sabinene Natural products 0.000 description 1
- 208000013223 septicemia Diseases 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
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- 239000002023 wood Substances 0.000 description 1
- WTARULDDTDQWMU-UHFFFAOYSA-N β-pinene Chemical compound C1C2C(C)(C)C1CCC2=C WTARULDDTDQWMU-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D2011/005—Co-current extraction
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Botany (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a kind of calamondin essential oil with bacteriostatic activity, including the essential oil prepared respectively with the leaf, rascal or Calusena lansium of calamondin.Comprise the following steps during preparation：Step 1: leaf, rascal or the Calusena lansium of collection calamondin, it is crushed respectively；Step 2: with steam distillation solid-liquid ratio be 1/5 ~ 1/3, distillation time be 50 ~ 70min under conditions of extract calamondin tangerine leaf essential oil, calamondin rascal essential oil or calamondin Calusena lansium essential oil respectively；The calamondin essential oil of the present invention can as a kind of natural plants bacteriostatic agent development and application, while the extraction process is simple, without other additives or composition, have the advantages that safely, effectively, nontoxic, environmental protection.
Description
Technical field
The invention belongs to essential oil application field, it is related to calamondin essential oil and preparation method thereof and the suppression to food-borne pathogens
Make and use, more particularly to calamondin essential oil is in the application in food antiseptic, preserving fruit and vegetable utilizing field.
Background technology
Food security is global public health problem, and annual about 420,000 people of WHO data displays in 2016 are because of food-borne
Disease and it is dead, research show food-borne pathogens be cause morbidity even main causes of death.As Listeria monocytogenes can
Cause serious food poisoning, then trigger septicemia, meningitis even dead.Chemical synthesis preservative abuse is brought simultaneously
The negative effect such as residual and drug resistance also directly threatens the health of the mankind.Therefore safely and effectively food preservative is found and fresh-keeping
Agent is extremely urgent.Antibacterial Constituents are screened from natural plants as study hotspot in recent years.
Calamondin (Citrus mitis Blanco) is Rutaceae (Rutaceae) Citrus (Citrus L.) evergreen filling
Wood or dungarunga, are the cenospecies of cumquat (Fortunella margarita) and citrus (Citrus reticulata).Distribution
In subtropical and tropical zones, such as Philippine, Central America, Japan etc..There is plantation all over China, south cultivation is more normal
See, such as Guangdong, Guangxi etc..Medical value substantially, have resolving sputum, cough-relieving, qi-regulating, the effects such as help digestion.At present, on calamondin essence
Seldom, only research also focuses mostly in terms of chemical composition, very limited to the research in terms of bioactivity for the research of oil.But
It is to be found from the research of other plant essential oil, main chemical compositions linalool, limonene, sabinene, the moon in calamondin essential oil
Osmanthus alkene, nopinene have suppression microbial activity, therefore speculate that calamondin essential oil has bacteriostatic activity.And the other plant of Citrus
Essential oil has proven to antibacterial, antimycotic, anti-oxidant, desinsection, improves the multiple biological activities such as hepatotoxicity wind agitation and renal toxicity, extensive
In fields such as health care, food antiseptic, preserving fruit and vegetable utilizing, control of insect, eco-tour and daily chemical products.
To sum up, calamondin plants essential oil has good development prospect as plant-based bacteriostat, needs further
Research.
The content of the invention
The present situation do not studied further calamondin essential oil for prior art, is developed a kind of with bacteriostatic activity
Calamondin essential oil.
The technical scheme is that：A kind of calamondin essential oil with bacteriostatic activity, including respectively with the leaf of calamondin,
Essential oil prepared by rascal or Calusena lansium.The part of use is different, and the essential oil fungistatic effect of preparation is also different.
The preparation method of the calamondin essential oil with bacteriostatic activity in this programme, comprises the steps：
Step 1: leaf, rascal or the Calusena lansium of collection calamondin, it is crushed respectively；
Step 2: with steam distillation solid-liquid ratio be 1/5~1/3, distillation time be 50~70min under conditions of point
Indescribably take calamondin tangerine leaf essential oil, calamondin rascal essential oil or calamondin Calusena lansium essential oil.
It is preferred that, the particle diameter crushed in the step 1 is 10-20 mesh.It both can guarantee that raw material were crushed under this particle diameter
Degree can fully be contacted with water, reduce distance of the essential oil from diffusion inside to surface, and the powder that particle diameter is too small and occurs is avoided that again
The phenomenon of end caking.
It is preferred that, raw material need to be divided into 5 parts, steam by several times by steam distillation in concrete operations in the step 2
Evaporate, the raw material that every 50~70min more renews.In order to avoid the heat time is long, derived essential oil is scattered and disappeared, steamed by several times so taking
The mode evaporated, and also allow for mixing essential oil, in operation, it is 95~105 DEG C that need to control temperature, makes to protect always in still-process
Hold slight boiling condition, it is to avoid hot-spot makes the chemical composition of essential oil change.
Calamondin tangerine leaf, rascal, Calusena lansium essential oil yield are respectively 0.28%, 1.06%, 0.80% obtained by this method, with document
Middle citrus plant essential oil yield is basically identical：Leaf essential oil yield 0.23~0.29%, differing maturity pericarp essential oil yield
0.75%~1.75%.
The extraction process is simple, without other additives or composition, on the premise of recovery rate is ensured, shortens add as far as possible
The hot time, reduce heating-up temperature, it is to avoid derived essential oil scatters and disappears and changed, with safely, effectively, nontoxic, environmental protection etc. it is excellent
Point.
Further, described bacterium is food-borne pathogens, and food origin disease caused by microorganism is the main of food security
Problem, the bacteriostatic agent found effectively, safe can solve problem from source.
It is preferred that, the food-borne pathogens be Staphylococcus aureus, salmonella typhimurium, Listeria monocytogenes or
Escherichia coli, they are most common foodborne bacterial pathogenses in China's food, because food poisoning occupies main positions caused by it.
Application of the calamondin essential oil with bacteriostatic activity in food antiseptic, preserving fruit and vegetable utilizing field.Above-mentioned pathogenic bacteria are main
Be present in the food such as veterinary antibiotics, raw meat, cold cuts, can by essential oil champignon, immersion, sprinkling or directly as food additive
Plus agent processed food etc. method, carry out effectively it is antibacterial, prevent food pollution.
The operation principle and beneficial effect of the present invention：The present invention discloses calamondin essential oil as the application of plant-based bacteriostat
Potentiality and advantage.Tangerine leaf essential oil, rascal essential oil or the Calusena lansium essential oil of calamondin are extracted by steam distillation, using filter paper
Piece agar diffusion method and micro-broth dilution method they to Staphylococcus aureus, salmonella typhimurium, single increase Li Si
Special bacterium, the bacteriostatic activity of Escherichia coli.Calamondin different parts essential oil is shown by contrast experiment, to gram-positive bacteria (gold
Staphylococcus aureus, Listeria monocytogenes) and Gram-negative bacteria (salmonella typhimurium, Escherichia coli) have suppression make
With, and bacteriostatic activity order is tangerine leaf essential oil ＞ rascal essential oil ＞ Calusena lansium essential oils.Suppression of the calamondin tangerine leaf essential oil to this 4 kinds of bacterium
Effect is all extremely quick (26.76-32.20mm)；To golden yellow grape bacillus, salmonella typhimurium, Escherichia coli minimum suppression
Bacteria concentration (MIC) is all 1.88 μ l/ml, and the MIC to Listeria monocytogenes is 30 μ l/ml.Rascal essential oil is pole to Escherichia coli
Quick (22.76mm), (12.60-13.70mm) quick in being to gold-coloured staphylococci, salmonella typhimurium, Listeria monocytogenes,
Be 60 μ l/ml to the MIC of Escherichia coli, to golden yellow grape bacillus, salmonella typhimurium, Listeria monocytogenes MIC all
For 240 μ l/ml.Calusena lansium essential oil is quick in being to Listeria monocytogenes (10.03mm), is muting sensitive (7.53- to other 3 kinds of bacterium
9.13mm)；240 μ l/ml are both greater than to the MIC of four kinds of bacterium.It these results suggest that calamondin essential oil can be as a kind of natural plants
Bacteriostatic agent development and application, while the extraction process is simple, without other additives or composition, with safely, effectively, it is nontoxic, green
The advantages of colour circle is protected.
Brief description of the drawings
Fig. 1 is fungistatic effect figure of the calamondin essential oil to gold-coloured staphylococci；
Fig. 2 is fungistatic effect figure of the calamondin essential oil to salmonella typhimurium；
Fig. 3 is fungistatic effect figure of the calamondin essential oil to Listeria monocytogenes；
Fig. 4 is fungistatic effect figure of the calamondin essential oil to Escherichia coli.
Embodiment
The technical method of the present invention is described in detail with reference to specific embodiment：
Referring to Fig. 1~Fig. 4, tangerine leaf, rascal, Calusena lansium essential oil are sequentially followed successively by from left to right.
1. the preparation of calamondin bacteriostatic activity essential oil
Embodiment 1：A kind of preparation method of the calamondin essential oil with bacteriostatic activity, comprises the following steps：
Step one：Fresh calamondin leaf is collected, is cleaned, distilled water is crossed, dries at room temperature, size-reduced machine obtains powder
900.110g。
Step 2：Tangerine leaf powder is equally divided into 5 parts, point 5 distillations, with steam distillation in solid-liquid ratio 1:3, temperature
100 DEG C, prepare under conditions of time 60min, obtain pale yellow transparent essential oil 2.535g, ρLeaf=.0.881g/ml, recovery rate is
0.28%.Calamondin extraction of essential oil rate (%)=(quality of quality/raw material of essential oil) × 100%.
Step 3：Using filter paper agar diffusion method, 10 are drawn with sterile liquid-transfering gun6The cfu/ml μ l of outstanding bacterium solution 200,
Nutrient broth solid medium plate even spread.10 μ l calamondin tangerine leaf essential oils are taken gently to adsorb at sterilizing filter paper piece (6mm)
On, filter paper is put down gently after plate containing bacterium, 37 DEG C of culture 24h are observed and with vernier caliper measurement antibacterial circle diameter size.With
Sterile distilled water is blank control, and benzyl penicillin susceptibility piece does positive control.Every group of sample does three groups of parallel controls, averages.
Experimental result refers to table 1.Essential oil is to the criterion of the fungistatic effect of strain：Antibacterial circle diameter size ＜ 6mm are without sensitivity
Bacteriostasis, the effect of 6-9mm less sensitives, 10-14mm medium sensitivity inhibitory action, the extremely sensitive inhibitory action of 15-20mm, ＞
20mm extreme sensitivity inhibitory action.
Step 4：Using micro broth dilution method, a series of calamondin tangerine folic acid oil solution of gradient concentrations is prepared first
(1.88-480μl/ml).The outstanding bacterium solution prepared and each 50 μ l of essential oil solution are added to 96 orifice plates, is well mixed, bacterium is most
Final concentration of 5 × 105cfu/ml.Not to be inoculated with bacterium solution, the fluid nutrient medium for being not added with essential oil solution does blank control, to be inoculated with phase
With quantity bacterium solution, the fluid nutrient medium for being not added with essential oil solution is compareed as culture.Each sample sets three repetitions.96 orifice plates 37
DEG C culture 20h after observe, record MIC.Experimental result refers to table 2.
Embodiment 2：A kind of preparation method of the calamondin essential oil with bacteriostatic activity, comprises the following steps：
Step one：Fresh calamondin rascal is collected, is cleaned, distilled water is crossed, dries at room temperature, powder is obtained after size-reduced machine
900.007g。
Step 2：Green Tangerine Peel is equally divided into 5 parts, point 5 distillations, with steam distillation in solid-liquid ratio 1:3, temperature
100 DEG C, prepare under conditions of time 60min, obtain water white transparency essential oil 9.546g, ρRascal=0.817g/ml, recovery rate is
1.06%.
Step 3：Using filter paper agar diffusion method, 10 are drawn with sterile liquid-transfering gun6The cfu/ml μ l of outstanding bacterium solution 200,
Nutrient broth solid medium plate even spread.10 μ l calamondin rascal essential oils are taken gently to adsorb at sterilizing filter paper piece (6mm)
On, filter paper is put down gently after plate containing bacterium, 37 DEG C of culture 24h are observed and with vernier caliper measurement antibacterial circle diameter size.With
Sterile distilled water is blank control, and benzyl penicillin susceptibility piece does positive control.Every group of sample does three groups of parallel controls, averages.
Experimental result refers to table 1.
Step 4：Using micro broth dilution method, a series of calamondin rascal essential oil solution of gradient concentrations is prepared first
(1.88-480μl/ml).The outstanding bacterium solution prepared and each 50 μ l of essential oil solution are added to 96 orifice plates, is well mixed, bacterium is most
Final concentration of 5 × 105cfu/ml.Not to be inoculated with bacterium solution, the fluid nutrient medium for being not added with essential oil solution does blank control, to be inoculated with phase
With quantity bacterium solution, the fluid nutrient medium for being not added with essential oil solution is compareed as culture.Each sample sets three repetitions.96 orifice plates 37
DEG C culture 20h after observe, record MIC.Experimental result refers to table 2.
Embodiment 3：A kind of preparation method of the calamondin essential oil with bacteriostatic activity, comprises the following steps：
Step one：Fresh calamondin Calusena lansium is collected, is cleaned, distilled water is crossed, dries at room temperature, size-reduced machine obtains powder
900.108g。
Step 2：Calusena lansium powder is equally divided into 5 parts, point 5 distillations, with steam distillation in solid-liquid ratio 1:3, temperature
100 DEG C, prepare under conditions of time 60min, obtain water white transparency essential oil 7.203g, ρCalusena lansium=0.816g/ml, recovery rate is
0.80%.
Step 3：Using filter paper agar diffusion method, 10 are drawn with sterile liquid-transfering gun6The cfu/ml μ l of outstanding bacterium solution 200,
Nutrient broth solid medium plate even spread.10 μ l calamondin Calusena lansium essential oils are taken gently to adsorb at sterilizing filter paper piece (6mm)
On, filter paper is put down gently after plate containing bacterium, 37 DEG C of culture 24h are observed and with vernier caliper measurement antibacterial circle diameter size.With
Sterile distilled water is blank control, and benzyl penicillin susceptibility piece does positive control.Every group of sample does three groups of parallel controls, averages.
Experimental result refers to table 1.
Step 4：Using micro broth dilution method, a series of calamondin Calusena lansium essential oil solution of gradient concentrations is prepared first
(1.88-480μl/ml).The outstanding bacterium solution prepared and each 50 μ l of essential oil solution are added to 96 orifice plates, is well mixed, bacterium is most
Final concentration of 5 × 105cfu/ml.Not to be inoculated with bacterium solution, the fluid nutrient medium for being not added with essential oil solution does blank control, to be inoculated with phase
With quantity bacterium solution, the fluid nutrient medium for being not added with essential oil solution is compareed as culture.Each sample sets three repetitions.96 orifice plates 37
DEG C culture 20h after observe, record MIC.Experimental result refers to table 2.
Embodiment 4：A kind of preparation method of the calamondin essential oil with bacteriostatic activity, comprises the following steps：
Step one：Collect fresh calamondin Calusena lansium to clean, cross distilled water, dry at room temperature, powder is obtained after size-reduced machine
900.211g。
Step 2：Calusena lansium powder is equally divided into 5 parts, point 5 distillations, with steam distillation in solid-liquid ratio 1:5, temperature
100 DEG C, primary material is changed per 70min, water white transparency essential oil 7.301g, ρ are obtained after the completion of 5 distillationsCalusena lansium=0.818g/ml, is carried
It is 0.81% to take rate.
The calamondin essential oil inhibition zone size of table 1
Note：Difference not significantly (P is represented with column data shoulder mark same letter>0.05), different letters represent significant difference (P<
0.05)；The standard of sensitiveness：≤ 6mm is unwise, 6-9mm muting sensitives, quick in 10-14mm, and 15-20mm Gao Min, ＞ 20mm are extremely quick.
The calamondin essential oil minimum inhibitory concentration (μ l/ml) of table 2
Tables 1 and 2 be calamondin essential oil to staphylococcus aureus, salmonella typhimurium, Listeria monocytogenes and
The related data of Escherichia coli fungistatic effect and Mlc is enumerated.
2. the application of calamondin essential oil
Application examples 1：Calamondin essential oil comprises the following steps to the fresh-keeping effect of fruit：
Step one：Select well-balanced, the fresh strawberry of size as fresh-keeping experiment material, clear water is cleaned, dried, random point
Group, is divided into one group per 300g, totally four groups.
Step 2：By calamondin tangerine leaf, rascal, Calusena lansium essential oil, dissolved with a small amount of ethanol, add appropriate Tween 80 abundant
Emulsification, distilled water constant volume to 20ml is configured to essential oil fresh-keeping liquid, respective concentration is diluted to water again when using.First three component
It is not put into 3.0% calamondin tangerine leaf, rascal, Calusena lansium essential oil dilution and soaks 5min.4th group is simply placed in leaching in blank solution
Steep 5min.Strawberry takes out nature and drained, and it is storage under conditions of 85%~90% to be placed on 25 ± 2 DEG C of room temperatures, air humidity.Through
Rotten the postponed number of days of than the 4th group appearance of strawberry crossed after calamondin tangerine leaf, rascal, the processing of Calusena lansium essential oil is respectively 1.5
My god, 1 day, 0.5 day.
Application examples 2：Calamondin essential oil comprises the following steps to the fresh-keeping effect of vegetables：
Step one：Select well-balanced, the fresh green vegetables of size as fresh-keeping experiment material, go yellow leaf, clear water is cleaned, dried,
Random packet, is divided into one group per 200g, totally four groups.
Step 2：By calamondin tangerine leaf, rascal, Calusena lansium essential oil, dissolved with a small amount of ethanol, add appropriate Tween 80 abundant
Emulsification, distilled water constant volume to 20ml is configured to essential oil fresh-keeping liquid, respective concentration is diluted to water again when using.First three component
It is not put into 3.0% calamondin tangerine leaf, rascal, Calusena lansium essential oil dilution and soaks 5min.4th group is simply placed in leaching in blank solution
Steep 5min.Green vegetables take out nature and drained, and it is storage under conditions of 85%~90% to be placed on 25 ± 2 DEG C of room temperatures, air humidity.Through
The number of days point that the green vegetables crossed after calamondin tangerine leaf, rascal, the processing of Calusena lansium essential oil occur blade face jaundice than the 4th group, rot postponed
Wei not be 4 days, 3 days, 1 day.
Application examples 3：Calamondin essential oil comprises the following steps to the antiseptic effect of meat：
Step one：Chopping board, cutter are wiped with 75% alcohol, fresh halogen pork is cut into about 50g or so cube meat, at random
Packet, every 6 pieces are divided into one group, totally four groups.
Step 2：By calamondin tangerine leaf, rascal, Calusena lansium essential oil, dissolved with a small amount of ethanol, add appropriate Tween 80 abundant
Emulsification, distilled water constant volume to 20ml is configured to essential oil fresh-keeping liquid, respective concentration is diluted to water again when using.First three component
It is not put into 3.0% calamondin tangerine leaf, rascal, Calusena lansium essential oil dilution and soaks 5min.4th group is simply placed in leaching in blank solution
Steep 5min.Halogen meat taking-up nature is drained, in the food bag for putting into sterilizing, places and is stored at 4 ± 1 DEG C.According to GB47892-
2016 methods, determine total number of bacteria.According to《GB2726-2016 national food safety standard cooked meat products》, test with total plate count
≥1×105Cfu/g is used as the putrid and deteriorated reference index of halogen meat.After calamondin tangerine leaf, rascal, the processing of Calusena lansium essential oil
Than the 4th group exceeded postponed number of days of halogen meat total plate count of halogen meat is respectively 5 days, 3 days, 1 day.
The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention, for the skill of this area
For art personnel, it can still modify to the technical scheme described in foregoing embodiments, or to which part skill
Art feature carries out equivalent substitution.Within the spirit and principles of the invention, any modification, equivalent substitution and improvements done etc.,
It should be included in the scope of the protection.
Claims (7)
1. a kind of calamondin essential oil with bacteriostatic activity, it is characterised in that：Leaf, rascal or Huang including using calamondin respectively
The standby essential oil of leather.
2. the preparation method of the calamondin essential oil according to claim 1 with bacteriostatic activity, it is characterised in that including with
Lower step：Step 1: leaf, rascal or the Calusena lansium of collection calamondin, it is crushed respectively；
Step 2: with steam distillation solid-liquid ratio be 1/5 ~ 1/3, distillation time be 50 ~ 70min under conditions of extract respectively
Calamondin tangerine leaf essential oil, calamondin rascal essential oil or calamondin Calusena lansium essential oil.
3. the preparation method of the calamondin essential oil according to claim 2 with bacteriostatic activity, it is characterised in that the step
The particle diameter crushed in rapid 1 is 10-20 mesh.
4. the preparation method of the calamondin essential oil according to claim 2 with bacteriostatic activity, it is characterised in that the step
Raw material need to be divided into 5 parts, distill by several times, the original that every 50 ~ 70min more renews by steam distillation in concrete operations in rapid 2
Material.
5. the application of the calamondin essential oil according to claim 1 with bacteriostatic activity, it is characterised in that described bacterium is
Food-borne pathogens.
6. the application of the calamondin essential oil according to claim 5 with bacteriostatic activity, it is characterised in that the food-borne
Pathogenic bacteria are Staphylococcus aureus, salmonella typhimurium, Listeria monocytogenes or Escherichia coli.
7. the calamondin essential oil according to claim 1 with bacteriostatic activity food antiseptic, preserving fruit and vegetable utilizing field should
With.
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| CN111418648A (en) * | 2020-02-25 | 2020-07-17 | 浙江工商大学 | Essential oil antiseptic insect repellent for waxberries |
| CN113416608A (en) * | 2021-06-29 | 2021-09-21 | 中国热带农业科学院热带作物品种资源研究所 | Extraction method of intelligence-developing essential oil for inhibiting listeria monocytogenes and intelligence-developing essential oil |
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