CN113521183B - Method for efficiently improving content of phenolic acid and flavone in lily functional components - Google Patents
Method for efficiently improving content of phenolic acid and flavone in lily functional components Download PDFInfo
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- CN113521183B CN113521183B CN202110801297.1A CN202110801297A CN113521183B CN 113521183 B CN113521183 B CN 113521183B CN 202110801297 A CN202110801297 A CN 202110801297A CN 113521183 B CN113521183 B CN 113521183B
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- methyl jasmonate
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- 241000234435 Lilium Species 0.000 title claims abstract description 107
- 238000000034 method Methods 0.000 title claims abstract description 21
- 229930003944 flavone Natural products 0.000 title claims abstract description 20
- 235000011949 flavones Nutrition 0.000 title claims abstract description 20
- 150000007965 phenolic acids Chemical class 0.000 title claims abstract description 20
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 title claims abstract description 19
- 150000002212 flavone derivatives Chemical class 0.000 title claims abstract description 19
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- 238000011282 treatment Methods 0.000 claims abstract description 98
- GEWDNTWNSAZUDX-WQMVXFAESA-N (-)-methyl jasmonate Chemical compound CC\C=C/C[C@@H]1[C@@H](CC(=O)OC)CCC1=O GEWDNTWNSAZUDX-WQMVXFAESA-N 0.000 claims abstract description 81
- GEWDNTWNSAZUDX-UHFFFAOYSA-N methyl 7-epi-jasmonate Natural products CCC=CCC1C(CC(=O)OC)CCC1=O GEWDNTWNSAZUDX-UHFFFAOYSA-N 0.000 claims abstract description 81
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- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical group C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 claims description 6
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 claims description 6
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- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 229930013930 alkaloid Natural products 0.000 description 2
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 description 2
- 230000003796 beauty Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 2
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- 239000006228 supernatant Substances 0.000 description 2
- KMVWNDHKTPHDMT-UHFFFAOYSA-N 2,4,6-tripyridin-2-yl-1,3,5-triazine Chemical compound N1=CC=CC=C1C1=NC(C=2N=CC=CC=2)=NC(C=2N=CC=CC=2)=N1 KMVWNDHKTPHDMT-UHFFFAOYSA-N 0.000 description 1
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 1
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- 241000234280 Liliaceae Species 0.000 description 1
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- YMGFTDKNIWPMGF-QHCPKHFHSA-N Salvianolic acid A Natural products OC(=O)[C@H](Cc1ccc(O)c(O)c1)OC(=O)C=Cc2ccc(O)c(O)c2C=Cc3ccc(O)c(O)c3 YMGFTDKNIWPMGF-QHCPKHFHSA-N 0.000 description 1
- YMGFTDKNIWPMGF-UCPJVGPRSA-N Salvianolic acid A Chemical compound C([C@H](C(=O)O)OC(=O)\C=C\C=1C(=C(O)C(O)=CC=1)\C=C\C=1C=C(O)C(O)=CC=1)C1=CC=C(O)C(O)=C1 YMGFTDKNIWPMGF-UCPJVGPRSA-N 0.000 description 1
- 208000013738 Sleep Initiation and Maintenance disease Diseases 0.000 description 1
- 208000031971 Yin Deficiency Diseases 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 230000003178 anti-diabetic effect Effects 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
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- 239000003472 antidiabetic agent Substances 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
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- 239000000872 buffer Substances 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 229910001431 copper ion Inorganic materials 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
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- 239000000706 filtrate Substances 0.000 description 1
- 150000002213 flavones Chemical class 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
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- 206010022437 insomnia Diseases 0.000 description 1
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- -1 iron ion Chemical class 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
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- 235000005412 red sage Nutrition 0.000 description 1
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- 229930183842 salvianolic acid Natural products 0.000 description 1
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- 238000007873 sieving Methods 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
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Images
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8967—Lilium, e.g. tiger lily or Easter lily
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
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- 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
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/13—Preparation or pretreatment of starting material involving cleaning, e.g. washing or peeling
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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Abstract
The invention discloses a method for efficiently improving the content of phenolic acid and flavone which are functional components of lily, wherein lily scales are peeled off and cleaned, and the redundant water on the surface is dried to obtain a processing material; fully mixing methyl jasmonate with water to obtain a treatment solution, wherein the concentration of the treatment solution is 50-200 mu M; adding the treatment material into the treatment solution, and soaking for 30 minutes; and (3) moisturizing the soaked treatment material in the atmosphere of methyl jasmonate with corresponding concentration, and storing the treatment material at room temperature for a certain time. The invention provides a simple and efficient method for improving the effective components and biological activity of lily, wherein the effective component content in lily bulbs can be improved by treating lily scales with methyl jasmonate, the antioxidant activity of a lily bulb extract can be improved, the antibacterial activity of the lily bulb extract is enhanced, and a simple and efficient method is provided for improving the effective component content of lily and enhancing the health-care and medicinal effects of lily by treating lily bulb scales with methyl jasmonate.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a method for efficiently improving the contents of phenolic acid and flavone which are functional components of lily.
Background
The secondary metabolism species of the plant are rich and are important sources of natural medicines. Lily is a general term of plants in the genus of lily of the family liliaceae, contains more than one hundred wild species and tens of thousands of lily hybrid varieties, and is a world-known plant integrating ornamentation, edibility and medicinal functions. Lily contains abundant effective components such as phenolic acid, flavone, saponin, alkaloid, terpenes, etc., and has important functions of nutrition, health promotion and disease treatment. The medicinal lily specified in Chinese pharmacopoeia is dried meat-like scales of 3 kinds of lily, including red sage root, lily bulb and lilium tenuifolia, and has the functions of nourishing yin, moistening lung, clearing away heart fire and tranquilizing mind, and may be used in treating dry cough due to yin deficiency, over-strained cough with blood, dysphoria, insomnia, dreaminess and absentmindedness. Lilium plants have been the focus of chemical research on natural products for over two decades and have potential anti-inflammatory, antifungal, platelet aggregation inhibiting, anti-diabetic and anti-cancer activities. Phenolic acid and flavone are important nutritional ingredients of lily, belong to important components of secondary metabolites, and have strong antioxidant, anticancer and anti-inflammatory activities; researches in recent years show that some alkaloids and saponins in lily have better antibacterial and anticancer effects. Meanwhile, the saponin component in the lily also has a certain effect of promoting wound healing, and can be used for beauty treatment, health care and skin repair.
In conclusion, the effective components in the lily have important medicinal and nutritional health-care values, but at present, researches on the effective components of the lily mainly focus on component identification and function researches, and the researches on improving the content and the biological activity of the effective components in the lily are rarely reported. Methyl jasmonate is a commonly used plant regulator, which can be synthesized by the plant itself and participates in many defense and regulation reactions of the plant. At present, researches on improvement of stress resistance and influence on secondary metabolism by treatment of methyl jasmonate in the growth period of plants are related, but researches on improvement of effective components of plants by treatment of methyl jasmonate after harvesting are fresh, and reports on improvement of functional components in lily by methyl jasmonate are not available. Therefore, a method for efficiently improving the contents of phenolic acid and flavone which are functional components of lily is provided.
Disclosure of Invention
Based on the technical problems in the background art, the invention provides a method for efficiently improving the content of phenolic acid and flavone which are functional components of lily, so as to solve the problems in the background art.
The invention provides the following technical scheme: a method for efficiently improving the contents of phenolic acid and flavone which are functional components of lily comprises the following steps:
A. preparing materials: peeling off and cleaning the harvested lily bulb scales, and airing the excessive moisture on the surface to obtain a treatment material;
B. preparing a solution: fully mixing methyl jasmonate with water to obtain a treatment solution, wherein the concentration of the treatment solution is 50-200 mu M;
C. soaking: adding the treatment material into the treatment solution, and soaking for 30 minutes;
D. culturing: and (3) moisturizing the soaked treatment material in the atmosphere of methyl jasmonate with corresponding concentration, and storing the treatment material at room temperature for a certain time.
Preferably, the lily in the step A is red lead.
Preferably, the concentration range of the methyl jasmonate in the step B is 50-200 mu M; preferably, the methyl jasmonate concentration is 50. mu.M, 100. mu.M or 200. mu.M, more preferably 100. mu.M or 200. mu.M.
Preferably, the culture time in the step D is 3 days.
The invention provides a method for efficiently improving the contents of phenolic acid and flavone which are functional components of lily, wherein the content of the phenolic acid and the flavone which are the functional components in lily bulbs can be improved by treating lily scales with methyl jasmonate, the antioxidant activity of a lily bulb extract can be improved, the antibacterial activity of the lily bulb extract is enhanced, and a simple and effective method is provided for improving the content of effective components of lily and enhancing the health-care and medicinal effects of lily by treating with methyl jasmonate.
Drawings
FIG. 1 is a graph showing the effect of methyl jasmonate treatment at different concentrations on the content of salvianolic acid in the present invention;
FIG. 2 is a graph showing the effect of methyl jasmonate treatment at different concentrations on the content of lily flavones in accordance with the present invention;
FIG. 3 is a graph comparing the effect of methyl jasmonate treatment on lily DPPH free radical scavenging activity according to the present invention;
FIG. 4 is a graph comparing the effect of methyl jasmonate treatment on lily FRAP of the present invention;
FIG. 5 is a graph comparing the effect of methyl jasmonate treatment on lily CUPRAC of the present invention;
FIG. 6 is a graph showing the effect of methyl jasmonate treatment at different concentrations on lily bacteriostatic activity.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1-6, the present invention provides a technical solution: a method for efficiently improving the contents of phenolic acid and flavone which are functional components of lily comprises the following steps:
A. preparing materials: peeling and cleaning the lily scales, and airing the redundant water on the surface to obtain a treatment material;
B. preparing a solution: fully mixing methyl jasmonate with water to obtain a treatment solution, wherein the concentration of the treatment solution is 50-200 mu M;
C. soaking: adding the treatment material into the treatment solution, and soaking for 30 minutes;
D. culturing: and (3) moisturizing the soaked treatment material in the atmosphere of methyl jasmonate with corresponding concentration, and storing the treatment material at room temperature for a certain time.
Example one
Peeling and cleaning the lily scales, and airing the redundant water on the surface to obtain a treatment material; fully mixing methyl jasmonate with water to obtain a treatment solution, wherein the concentration of the treatment solution is 50-200 mu M; adding the treatment material into the treatment solution, and soaking for 30 minutes; and (3) moisturizing the soaked treatment material in the atmosphere of methyl jasmonate with corresponding concentration, and storing the treatment material at room temperature for a certain time.
Example two
Peeling and cleaning the lily scales, airing the redundant water on the surface to obtain a treatment material, wherein the lily is selected from the lilium brownie; fully mixing methyl jasmonate with water to obtain a treatment solution, wherein the concentration of the treatment solution is 50-200 mu M; adding the treatment material into the treatment solution, and soaking for 30 minutes; and (3) moisturizing the soaked treatment material in the atmosphere of methyl jasmonate with corresponding concentration, and storing the treatment material at room temperature for a certain time.
EXAMPLE III
Peeling and cleaning the lily scales, airing the redundant water on the surface to obtain a treatment material, wherein the lily is selected from the lilium brownie; fully mixing methyl jasmonate with water to obtain a treatment solution, wherein the concentration of the treatment solution is 50-200 mu M; preferably the methyl jasmonate concentration is 50. mu.M, 100. mu.M or 200. mu.M, more preferably 100. mu.M or 200. mu.M; adding the treatment material into the treatment solution, and soaking for 30 minutes; and (3) moisturizing the soaked treatment material in the atmosphere of methyl jasmonate with corresponding concentration, and storing the treatment material at room temperature for a certain time.
Example four
Peeling and cleaning the lily scales, airing the redundant water on the surface to obtain a treatment material, wherein the lily is selected from the lilium brownie; fully mixing methyl jasmonate with water to obtain a treatment solution, wherein the concentration of the treatment solution is 50-200 mu M, and the concentration range of the methyl jasmonate is 50-200 mu M; preferably the methyl jasmonate concentration is 50. mu.M, 100. mu.M or 200. mu.M, more preferably 100. mu.M or 200. mu.M; adding the treatment material into the treatment solution, and soaking for 30 minutes; and (3) moisturizing the soaked treatment material in the methyl jasmonate atmosphere with the corresponding concentration, and storing the treatment material at room temperature for a certain period of time, wherein the culture time is 3 days.
Preparing 200 mu M methyl jasmonate solution: 458.5 μ L of methyl jasmonate (from sigma aldrich trade ltd) was added to 1L of distilled water and stirred with a magnetic suspension stirrer until completely dissolved;
preparing a 100 mu M methyl jasmonate solution: 229.2 μ L of methyl jasmonate (from sigma aldrich trade ltd) was added to 1L of distilled water and stirred with a magnetic suspension stirrer until completely dissolved;
preparing 50 mu M methyl jasmonate solution: mu.L of methyl jasmonate (from Sigma Aldrich, Inc.) was added to 1L of distilled water and stirred with a magnetic suspension stirrer until completely dissolved.
C, taking out the scales treated in the step C, spreading the scales in a tray, covering the scales with gauze stained with methyl jasmonate with corresponding concentration for moisture preservation, and storing the scales under the conditions of room temperature and darkness for culture;
after the treatment for 12, 24, 48 and 72 hours respectively, collecting the treated scales, and carrying out freeze drying treatment;
pulverizing the freeze-dried material with a wall breaking machine, sieving with a 40-mesh sieve, and collecting sieved Bulbus Lilii powder.
Extracting effective components:
(1) weighing 0.1g of crushed lily powder in a 10mL centrifuge tube;
(2) adding 5mL of methanol, fully shaking and mixing uniformly, performing ultrasonic extraction at 400W and 30 ℃ for 30min, and shaking and mixing uniformly once at 10 th and 20 th min respectively;
(3) centrifuging the extracted solution at 8000rpm and 25 deg.C for 5 min;
(4) sucking the supernatant liquid into a new centrifuge tube;
(5) repeating the steps (2) to (4), and merging the two supernatants;
(6) and (4) passing the combined extracting solution through a 0.22 mu m organic filter, and collecting the filtrate in a new centrifugal tube to obtain the lily active ingredient extracting solution.
And (3) determining the content of phenolic acid:
mu.L of the obtained extract was put in a 5mL centrifuge tube, 3.95mL of distilled water was added, 0.25mL of 50% Folin Ciocalteu reagent (Folin Ciocalteu was diluted 1: 1 with distilled water) was added, the mixture was allowed to stand at room temperature for 2min, 750. mu.L of 15% Na2CO3 was added, the mixture was mixed well and allowed to stand in the dark at room temperature for 2h, and 200. mu.L of the reaction solution was measured for absorbance at 765 nm. And drawing a standard curve by taking gallic acid as a standard substance, wherein the content of the total phenol is expressed as the gallic acid content (mg) equivalent (mgGAE/g DW) in each gram of the dried lily powder.
And (3) measuring the flavone content:
mixing the obtained extract 100 μ L with ultrapure water 400 μ L and 5% NaNO2 40 μ L, and standing in dark at room temperature for 5 min; adding 60 μ L10% AlCl3, and standing in the dark at room temperature for 6 min; 400. mu.L of 0.5mol/L NaOH was added. The absorbance of the reaction solution was measured at 510 nm. Taking quercetin with different concentrations as standard curve, and expressing the content of flavone as the content (mg) equivalent (mg QE/gDW) of quercetin in each gram of lily dry powder.
Influence of methyl jasmonate treatment on content of effective components of phenolic acids of lily
Three different methyl jasmonate concentration gradients were set: 50. at 100. mu.M and 200. mu.M, lily material was treated with methyl jasmonate at various concentrations as described above, and the phenolic acid content was extracted and determined, and the best treatment was 100. mu.M treatment for 3 days (as shown in FIG. 1).
As can be seen from FIG. 1, the exogenous methyl jasmonate significantly promoted the accumulation of the effective components of the phenolic acids of lily, and the phenolic acid content was increased to 1.82, 2.59 and 2.27 times of the control at 3d after the treatment of 50, 100 and 200 μ M methyl jasmonate. Therefore, a simple and effective method is provided for improving the content of the effective components of the lily and enhancing the health-care and medicinal effects of the lily by using the methyl jasmonate.
Influence of methyl jasmonate treatment on content of effective components of lily flavonoids
Three different methyl jasmonate concentration gradients were set: 50. at 100, 200. mu.M, lily material was treated with methyl jasmonate at various concentrations as described above, and the flavone content was extracted and determined, and the best treatment was 100. mu.M for 3 days (as shown in FIG. 2).
As can be seen from figure 2, the exogenous methyl jasmonate remarkably promotes the accumulation of effective flavonoids in lily, and after 50, 100 and 200 mu M methyl jasmonate is treated for 3 days, the flavone content is respectively increased to 1.65, 2.59 and 2.73 times of that of a control, so that a simple and effective method is provided for improving the effective component content of lily and enhancing the health-care and medicinal effects of lily by using methyl jasmonate for treatment.
Function of exogenous methyl jasmonate in improving antioxidant activity of lily extract
The method for measuring the antioxidant capacity of the effective components of the lily extracted after treatment of different concentrations of methyl jasmonate in the embodiment 1 comprises the following steps:
determination of 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH) radical scavenging Activity
Adding 50 μ L of the extract obtained in step 6 into 950 μ L of DPPH solution (final concentration in methanol is 0.1mM), mixing, standing in the dark at room temperature for 30min, measuring the absorbance of the reaction product at 517nm, and calculating DPPH free radical scavenging activity of the lily extract according to a standard curve drawn by a standard Trolox, wherein the result is expressed as Trolox content (μ g) equivalent (μ gTE/mL) per mL of the extract.
Determination of iron ion reducing ability (FRAP)
Adding 900 μ LFe into 100 μ L extractive solution of the extract obtained in step 63+And (3) uniformly mixing the components in a TPTZ working solution, standing for 5min at room temperature, measuring the absorbance at 593nm, calculating the FRAP of the lily extract according to a standard curve drawn by a standard substance Trolox, and expressing the result as the Trolox content (mu g) equivalent (mu g TE/mL) in each milliliter of the extract.
Determination of copper ion reducing Capacity (CUPRAC)
To a 2mL sterile centrifuge tube, 100. mu.L of the extract obtained in step 6 was added 0.5mL ammonium acetate buffer (1M, pH 7), 0.5mL copper sulfate solution (10mM), and cuprous reagent (7.5mM), respectively, followed by mixing well, standing for 30min, and absorbance was measured at 450 nm. The CUPRAC of the lily extract was calculated from a standard curve drawn with the standard substance Trolox, and the result was expressed as Trolox content (. mu.g) equivalent (. mu.g TE/mL) per mL of the extract
Effect of methyl jasmonate treatment on Lily antioxidant Capacity
Effect of methyl jasmonate treatment on Lily DPPH radical scavenging Activity
As can be seen from FIG. 3, the DPPH radical scavenging ability of the lily extract was improved after 50, 100, and 200. mu.M treatments with methyl jasmonate, compared to the control (0. mu.M). Wherein the DPPH free radical scavenging capacity is increased to 3.50 times, 5.94 times and 6.05 times of that of a control at 3d after 50, 100 and 200 mu M methyl jasmonate treatment. It is shown that the oxidation resistance of lily is improved after methyl jasmonate treatment, wherein the enhancement effect of 100 and 200 μ M methyl jasmonate treatment on the oxidation resistance of lily is more obvious (as shown in figure 3).
Effect of methyl jasmonate treatment on Lily FRAP
As can be seen from FIG. 4, FRAP of the lily extract was improved in the cases of 50, 100 and 200. mu.M methyl jasmonate treatment compared with the control (0. mu.M). Wherein, the FRAP is respectively improved to 3.18 times, 5.54 times and 5.57 times of the contrast at 3d after the treatment of 50, 100 and 200 mu M methyl jasmonate, and the enhancement effect of the treatment of 100 and 200 mu M methyl jasmonate on the oxidation resistance of lily is more obvious (as shown in figure 4).
Effect of methyl jasmonate treatment on Lily CuPRAC
As can be seen in FIG. 5, the CuPRAC of the lily extract was improved in the cases of 50, 100 and 200. mu.M methyl jasmonate treatment compared with the control (0. mu.M). Wherein at 3d after 50, 100 and 200 μ M methyl jasmonate treatment, the CUPRAC was increased to 2.92, 4.67 and 4.68 times of the control respectively, wherein the enhancing effect of 100 and 200 μ M methyl jasmonate treatment on the oxidation resistance of lily is more obvious (as shown in FIG. 5).
Therefore, the exogenous methyl jasmonate treatment can simply and effectively enhance the antioxidation of the lily and improve the functions of the lily in the aspects of beauty treatment, health care and disease treatment.
Influence of exogenous methyl jasmonate on lily bacteriostasis
Carrying out bacteriostatic ability determination on lily effective components extracted 72h after treatment of different concentrations of methyl jasmonate, wherein the method comprises the following steps:
adjusting the activated staphylococcus aureus solution to 1 × 106CFU/mL, then, using a pipette 100 u L of the bacterial solution evenly spread on 90X 90mm TSA containing Petri dish. Soaking a filter paper disc with the diameter of 6mm in the lily extract treated by 50, 100 and 200 mu M methyl jasmonate for 3 days respectively for 2 hours, taking out the lily extract, placing the lily extract on the surface of a TSA culture medium coated with staphylococcus aureus liquid, and taking methanol as a negative control. The plates were incubated at 37 ℃ for 24 hours in an inverted state. The diameters of the inhibition zones of the lily extract treated by different concentrations of methyl jasmonate (as shown in fig. 6) were photographed and compared.
Influence of methyl jasmonate treatment on lily bacteriostatic ability
As can be seen from FIG. 6, the different treated lily extracts all had a certain bacteriostatic ability compared to the negative control (FIG. 6 a); meanwhile, the inhibition zones (fig. 6c-e) of the lily extract after 3 days of treatment with 50, 100 and 200 mu M methyl jasmonate are all larger than that of the material without treatment with methyl jasmonate (fig. 6b), which shows that the inhibition capacity of lily after treatment with methyl jasmonate is improved, wherein the enhancement effect of the treatment with 100 and 200 mu M methyl jasmonate on the inhibition capacity of lily is more obvious.
Therefore, a simple and effective method is provided for enhancing the bacteriostatic and anti-inflammatory effects of the lily by using methyl jasmonate.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (2)
1. A method for efficiently improving the contents of phenolic acid and flavone which are functional components of lily is characterized in that: the method comprises the following steps:
A. preparing materials: peeling off and cleaning the scales of the collected lily bulbs, and airing the redundant water on the surface to obtain a treatment material;
B. preparing a solution: fully mixing methyl jasmonate with water to obtain a treatment solution, wherein the concentration of the treatment solution is 100 mu M;
C. soaking: adding the treatment material into the treatment solution, and soaking for 30 minutes;
D. culturing: preserving moisture of the soaked treatment material in the atmosphere of methyl jasmonate with corresponding concentration, and storing the treatment material at room temperature for 3 days;
wherein the phenolic acid is gallic acid, and the flavone is quercetin.
2. The method for efficiently increasing the contents of phenolic acid and flavone in lily functional ingredients according to claim 1, wherein the method comprises the following steps: and in the step A, the lily is selected from the lilium brownie.
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