CN110585324A - Extraction and purification method of hemerocallis fulva flower polyphenol - Google Patents
Extraction and purification method of hemerocallis fulva flower polyphenol Download PDFInfo
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- 150000008442 polyphenolic compounds Chemical class 0.000 title claims abstract description 53
- 235000013824 polyphenols Nutrition 0.000 title claims abstract description 53
- 240000009206 Hemerocallis fulva Species 0.000 title claims abstract description 44
- 235000002941 Hemerocallis fulva Nutrition 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 29
- 238000000605 extraction Methods 0.000 title claims description 30
- 238000000746 purification Methods 0.000 title claims description 6
- 239000011347 resin Substances 0.000 claims abstract description 30
- 229920005989 resin Polymers 0.000 claims abstract description 30
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 11
- 241000756137 Hemerocallis Species 0.000 claims description 30
- 239000000243 solution Substances 0.000 claims description 30
- 238000001035 drying Methods 0.000 claims description 24
- 238000001179 sorption measurement Methods 0.000 claims description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 16
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 15
- 239000012153 distilled water Substances 0.000 claims description 14
- 239000000843 powder Substances 0.000 claims description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 238000011068 loading method Methods 0.000 claims description 8
- 239000003463 adsorbent Substances 0.000 claims description 7
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 5
- 238000011049 filling Methods 0.000 claims description 5
- 238000007710 freezing Methods 0.000 claims description 5
- 230000008014 freezing Effects 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 5
- 239000002994 raw material Substances 0.000 claims description 5
- 238000007873 sieving Methods 0.000 claims description 5
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 239000003480 eluent Substances 0.000 claims description 4
- 239000012535 impurity Substances 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 abstract 1
- 238000002360 preparation method Methods 0.000 abstract 1
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 34
- 229940074391 gallic acid Drugs 0.000 description 17
- 235000004515 gallic acid Nutrition 0.000 description 17
- 238000002835 absorbance Methods 0.000 description 9
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 6
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 239000012488 sample solution Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 239000010200 folin Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 241000234280 Liliaceae Species 0.000 description 1
- 241000234269 Liliales Species 0.000 description 1
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 1
- 241001529733 Nepeta Species 0.000 description 1
- 244000184734 Pyrus japonica Species 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 150000003951 lactams Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 238000004506 ultrasonic cleaning Methods 0.000 description 1
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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
-
- 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
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
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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
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/17—Preparation or pretreatment of starting material involving drying, e.g. sun-drying or wilting
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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
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
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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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The invention relates to a method for extracting and purifying hemerocallis fulva flower polyphenol. The invention aims to provide a simple, easy and efficient method for extracting, separating and purifying hemerocallis fulva flowers. The method for extracting and purifying the polyphenol in the hemerocallis fulva flowers comprises the following steps: (1) pretreating hemerocallis fulva flowers; (2) ultrasonic-assisted extraction preparation of hemerocallis fulva polyphenol; (3) separating and purifying with macroporous resin to obtain extractive solution; (4) concentrating the extracting solution; (5) freeze drying to obtain purified hemerocallis fulva flower polyphenol.
Description
Technical Field
The invention belongs to a method for extracting and purifying hemerocallis fulva flower polyphenol, and particularly relates to a method for extracting and purifying the polyphenol from hemerocallis fulva flower.
Background
Hemerocallis fulva (known as Hemerocallis fulva) is also called Nepeta japonica, a perennial herb of Liliaceae, belonging to the order Liliales. Day lily flowers generally germinate earlier in spring, and are more in variety, and the petals are also colorful. The hemerocallis fulva, as a 'mother flower', not only has high ornamental value, but also has quite rich nutrition and medicinal value, is rich in various bioactive substances such as polyphenol, flavonoid, lactam, carotenoid, alkaloid, steroid and the like, and is a plant which can be used as both food and medicine and has great development prospect. However, at present, there is no research on the extraction, separation and purification of hemerocallis fulva polyphenol.
Compared with the traditional extraction mode, the ultrasonic-assisted extraction (UAE) has short extraction time, low energy consumption and high extraction rate. Therefore, ultrasonic-assisted extraction is used as a method for replacing the traditional extraction method, and the application range is gradually expanded. Meanwhile, compared with the traditional separation and purification method, the macroporous resin can make up for the defects of low purity and high cost, has higher stability, higher adsorption speed and desorption speed, high elution rate, mild adsorption condition, simple regeneration, reusability and no influence of inorganic matters, and brings considerable economic benefit when being applied to the industries of food, medicines and cosmetics.
Disclosure of Invention
The invention aims to provide a method for extracting, separating and purifying hemerocallis fulva flower polyphenol, which improves the quality and efficiency of separation and purification.
In order to achieve the above object, the present invention provides a method for extracting and purifying hemerocallis fulva polyphenol, which is characterized by comprising the following steps:
step 1: crushing raw materials: cleaning fresh hemerocallis fulva flower, drying in a constant-temperature forced air dryer, crushing after drying, and sieving with a 100-mesh sieve to obtain hemerocallis fulva flower powder for later use;
step 2: ultrasonic extraction: adding the daylily flower powder into distilled water by taking the distilled water as an extracting agent, wherein the mass ratio of the material liquid to the distilled water is 1 (20-30), and performing ultrasonic extraction to obtain crude extraction liquid of the daylily flower polyphenol;
and step 3: separating and purifying by macroporous resin: loading the macroporous adsorbent resin into a chromatographic column of 1.8 × 40cm by a wet method, wherein the filling height is 30cm, and obtaining the macroporous adsorbent resin chromatographic column; loading the crude extraction liquid of the hemerocallis fulva polyphenol at the flow rate of 10-20 mL/min, and performing dynamic adsorption; after adsorbing for 2-4h, eluting with 10% by volume of ethanol water solution at a flow rate of 50mL/min to remove impurities, eluting with 70-80% by volume of ethanol water solution at a flow rate of 50-60 mL/min for a second time, and collecting 70-80% by volume of ethanol water solution eluent;
and 4, step 4: and (3) vacuum concentration: concentrating the eluate in vacuum, and storing the concentrated solution for later use;
and 5: vacuum freeze drying: and (4) putting the purified and concentrated feed liquid into a vacuum freeze dryer for drying to obtain the pure hemerocallis fulva flower polyphenol product.
Preferably, in the step 1, the drying temperature is 50-80 ℃, and the drying time is 36-60 h.
Preferably, in the step 2, the ultrasonic working frequency is 40kHz, the extraction temperature is 30-60 ℃, and the extraction time is 50-80 min.
Preferably, the macroporous adsorption resin in the step 3 is one of AB-8 macroporous adsorption resin, D101 macroporous adsorption resin or HPD600 macroporous adsorption resin.
More preferably, in the step 3, the diameter-height ratio of the macroporous adsorption resin chromatographic column is 1: 10.
Preferably, in the step 3, the ethanol aqueous solution for the second elution is 75% by volume of ethanol aqueous solution.
Preferably, in the step 4, the vacuum concentration temperature is 45-55 ℃, the concentration time is 1-3 h, and the vacuum degree is: -0.8 to-0.9 Mpa.
Preferably, in the step 5, the freezing temperature is-80 ℃, the drying time is 24-48 h, and the vacuum degree is-0.8 Mpa.
Compared with the prior art, the extraction method of the hemerocallis fulva flower polyphenol has the following beneficial effects: the ultrasonic assistance is adopted to extract the polyphenol in the hemerocallis fulva flowers, so that the damage of hot processing to active ingredients is avoided, the extraction cost is reduced, and the extraction efficiency is improved.
Drawings
FIG. 1 is a standard curve of gallic acid.
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.
Description of the apparatus used in the examples of the present invention:
constant temperature forced air desiccator: shanghai Leyun laboratory instruments manufacturing Co., Ltd; an ultrasonic cleaning machine: ningbo Xinzhi Biotechnology GmbH, model SB25-12 DTDN; a vacuum freeze dryer: shanghai Bilang instruments manufacturing, Inc. model number FD-2C.
The types of macroporous adsorbent resins used in the examples of the present invention are shown in Table 1
TABLE 1 macroporous resin type
The method for measuring the content of the hemerocallis fulva flower polyphenol comprises the following steps:
drawing a gallic acid standard curve: accurately sucking 0, 1, 2, 3, 4, 5 and 6mL of gallic acid standard solution (0.5mL), pouring into a 25mL volumetric flask and fixing the volume. Sucking 5mL of the gallic acid from the volumetric flask, putting the gallic acid into a 100mL volumetric flask, adding 50mL of distilled water and 4mL of Folin reagent, shaking uniformly, standing for 5min, adding 8 mL of 10% sodium carbonate solution, fixing the volume, putting the solution into a water bath kettle (10 min), measuring the absorbance (765nm) after 10 min of color development, wherein the abscissa is the mass (mg) of the gallic acid and the ordinate is the absorbance (Abs), and drawing a standard curve of the gallic acid by using Origin8.0 drawing software. And calculating the content of the hemerocallis fulva flower polyphenol by using a standard curve equation. As shown in FIG. 1, the concentration of the daylily polyphenol is linearly related to the absorbance within the range of 1-6 ug/mL. The equation of the gallic acid standard curve is Y0.0856X +0.1475, R2 0.9998(X is absorbance value, Abs; Y is mass/mg of tawny day lily polyphenol).
Example 1
The embodiment provides a method for extracting and purifying hemerocallis fulva flower polyphenol, which comprises the following specific steps:
step 1: crushing raw materials: cleaning fresh day lily flowers, placing the cleaned fresh day lily flowers into a constant-temperature forced air dryer, drying for 36 hours at the temperature of 50 ℃, crushing after drying, and sieving by a 100-mesh sieve to obtain day lily flower powder for later use;
step 2: ultrasonic extraction: adding the daylily flower powder into distilled water by taking distilled water as an extracting agent, wherein the mass ratio of material liquid is 1:20, the ultrasonic working frequency is 40kHz, the extraction temperature is 30 ℃, the extraction time is 50min, and extracting to obtain crude daylily flower polyphenol extracting solution;
and step 3: separating and purifying by macroporous resin: putting the D101 macroporous adsorption resin into a chromatographic column of 1.8 multiplied by 40cm by a wet method, wherein the filling height is 30cm, and obtaining the macroporous adsorption resin chromatographic column; loading the crude extraction liquid of the hemerocallis fulva polyphenol at the flow rate of 10mL/min, and carrying out dynamic adsorption; after adsorbing for 2h, eluting with 10% ethanol water solution at a flow rate of 50mL/min, eluting with 70% ethanol water solution at a flow rate of 50mL/min, and collecting 70% ethanol water solution eluate;
and 4, step 4: and (3) vacuum concentration: concentrating the eluate in vacuum, and storing the concentrated solution at 45 deg.C for 1 hr under vacuum degree: -0.80 Mpa;
and 5: vacuum freeze drying: and (3) putting the purified and concentrated feed liquid into a vacuum freeze dryer for drying, wherein the quick freezing temperature is-80 ℃, the drying time is 24 hours, and the vacuum degree is as follows: and (4) under-0.8 Mpa to obtain the pure hemerocallis fulva flower polyphenol product.
Example 2
The embodiment provides a method for extracting and purifying hemerocallis fulva flower polyphenol, which comprises the following specific steps:
step 1: crushing raw materials: cleaning fresh day lily flowers, placing the cleaned fresh day lily flowers into a constant-temperature forced air dryer, drying for 48 hours at the temperature of 65 ℃, crushing after drying, and sieving by a 100-mesh sieve to obtain day lily flower powder for later use;
step 2: ultrasonic extraction: adding the daylily flower powder into distilled water by taking distilled water as an extracting agent, wherein the mass ratio of material liquid is 1:25, the ultrasonic working frequency is 40kHz, the extraction temperature is 45 ℃, the extraction time is 65min, and extracting to obtain crude daylily flower polyphenol extracting solution;
and step 3: separating and purifying by macroporous resin: loading AB-8 macroporous adsorbent resin into a chromatographic column of 1.8 × 40cm by wet method, and filling the column with a height of 30cm to obtain a macroporous adsorbent resin chromatographic column; loading the crude extraction liquid of the daylily polyphenol at the flow rate of 15mL/min, performing dynamic adsorption, eluting by using an ethanol water solution with the concentration of 10% after adsorbing for 3 hours to remove impurities, wherein the flow rate is 50mL/min, eluting by using an ethanol water solution with the volume fraction of 75%, and collecting an eluent of the ethanol water solution with the volume fraction of 75%, wherein the flow rate is 55 mL/min;
and 4, step 4: and (3) vacuum concentration: concentrating the eluate in vacuum, and storing the concentrated solution at 50 deg.C for 2 hr under vacuum degree: -0.85 Mpa;
and 5: vacuum freeze drying: and (3) putting the purified and concentrated feed liquid into a vacuum drier for drying, wherein the quick freezing temperature is-80 ℃, the drying time is 36h, and the vacuum degree is as follows: and (4) under-0.8 Mpa to obtain the pure hemerocallis fulva flower polyphenol product.
Example 3
The embodiment provides a method for extracting and purifying hemerocallis fulva flower polyphenol, which comprises the following specific steps:
step 1: crushing raw materials: cleaning fresh day lily flowers, placing the cleaned fresh day lily flowers into a constant-temperature forced air dryer, drying for 60 hours at the temperature of 80 ℃, crushing after drying, and sieving by a 100-mesh sieve to obtain day lily flower powder for later use;
step 2: ultrasonic extraction: adding the daylily flower powder into distilled water by taking distilled water as an extracting agent, wherein the mass ratio of material liquid is 1:30, the ultrasonic working frequency is 40kHz, the extraction temperature is 60 ℃, the extraction time is 80min, and extracting to obtain crude daylily flower polyphenol extracting solution;
and step 3: separating and purifying by macroporous resin: putting HPD600 macroporous adsorption resin macroporous resin into a chromatographic column of 1.8 multiplied by 40cm by a wet method, wherein the filling height is 30cm, and obtaining the macroporous adsorption resin chromatographic column; loading the crude extract of the daylily polyphenol at the flow rate of 20mL/min, performing dynamic adsorption, eluting with 10% ethanol water solution at the flow rate of 50mL/min for 4 hours to remove impurities, eluting with 80% ethanol water solution at the volume fraction of 60mL/min, and collecting the eluent of 80% ethanol water solution at the volume fraction;
and 4, step 4: and (3) vacuum concentration: concentrating the eluate in vacuum, and storing the concentrated solution at 55 deg.C for 3 hr under vacuum degree: -0.85 Mpa;
and 5: vacuum freeze drying: and (3) putting the purified and concentrated feed liquid into a vacuum drier for drying, wherein the quick freezing temperature is-80 ℃, the drying time is 48h, and the vacuum degree is as follows: and (4) under-0.8 Mpa to obtain the pure hemerocallis fulva flower polyphenol product.
Measuring the polyphenol content in the daylily flowers by adopting a Fulin-Phenol (Folin-Phenol) method, drawing a standard curve by taking gallic acid as a standard substance, and drawing the standard curve:
drawing a gallic acid standard curve: accurately sucking 0, 1, 2, 3, 4, 5 and 6mL of gallic acid standard solution, putting the gallic acid standard solution into a 25mL volumetric flask and fixing the volume. Sucking 5mL of the gallic acid from the volumetric flask, putting the gallic acid into a 100mL volumetric flask, adding 50mL of distilled water and 4mL of Folin reagent, shaking uniformly, standing for 5min, adding 8 mL of 10% sodium carbonate solution, fixing the volume, putting the solution into a water bath kettle (10 min), measuring the absorbance (765nm) after 10 min of color development, wherein the abscissa is the mass (mg) of the gallic acid and the ordinate is the absorbance (Abs), and drawing a standard curve of the gallic acid by using Origin8.0 drawing software. And calculating the content of the hemerocallis fulva flower polyphenol by using a standard curve equation. As shown in FIG. 1, the concentration of the daylily polyphenol is linearly related to the absorbance within the range of 1-6 ug/mL. The equation of the gallic acid standard curve is Y0.0856X +0.1475, R2 0.9998(X is absorbance value, Abs; Y is mass/mg of tawny day lily polyphenol).
The daylily polyphenol prepared in example 1-3 was dissolved in 0.06mg/mL solution, 1mL of sample solution was added to a 10mL colorimetric tube, 1mL of deionized water, 0.5mL of the forskolin-phenol test solution, and 1.5mL of 26.7% sodium carbonate solution were sequentially added, and then the volume was adjusted to 10mL with water, and the reaction was carried out at room temperature for 2 hours, and the absorbance was measured at 760 nm. And measuring the concentration according to a standard curve and calculating the content, wherein the polyphenol content (mg) of the sample solution is multiplied by the total volume (mL) of the extracting solution/volume (mL) of the sample solution, and the polyphenol extraction rate (%) (the polyphenol content in the sample/the weight of the day lily flower is multiplied by 100%). The results are shown in Table 1.
TABLE 1 extraction ratio of polyphenols from Hemerocallis fulva in examples 1-3
| Example 1 | Example 2 | Example 3 | |
| Extraction ratio (%) | 10.2±1.02 | 14.9±1.32 | 11.3±0.98 |
Claims (8)
1. The extraction and purification method of the hemerocallis fulva flower polyphenol is characterized by comprising the following steps:
step 1: crushing raw materials: cleaning fresh hemerocallis fulva flower, drying in a constant-temperature forced air dryer, crushing after drying, and sieving with a 100-mesh sieve to obtain hemerocallis fulva flower powder for later use;
step 2: ultrasonic extraction: adding the daylily flower powder into distilled water by taking the distilled water as an extracting agent, wherein the mass ratio of the material liquid to the distilled water is 1 (20-30), and performing ultrasonic extraction to obtain crude extraction liquid of the daylily flower polyphenol;
and step 3: separating and purifying by macroporous resin: loading the macroporous adsorbent resin into a chromatographic column of 1.8 × 40cm by a wet method, wherein the filling height is 30cm, and obtaining the macroporous adsorbent resin chromatographic column; loading the crude extraction liquid of the hemerocallis fulva polyphenol at the flow rate of 10-20 mL/min, and performing dynamic adsorption; after adsorbing for 2-4h, eluting with 10% by volume of ethanol water solution at a flow rate of 50mL/min to remove impurities, eluting with 70-80% by volume of ethanol water solution at a flow rate of 50-60 mL/min for a second time, and collecting 70-80% by volume of ethanol water solution eluent;
and 4, step 4: and (3) vacuum concentration: concentrating the eluate in vacuum, and storing the concentrated solution for later use;
and 5: vacuum freeze drying: and (4) putting the purified and concentrated feed liquid into a vacuum freeze dryer for drying to obtain the pure hemerocallis fulva flower polyphenol product.
2. The method for extracting and purifying a daylily polyphenol as claimed in claim 1, wherein in the step 1, the drying temperature is 50-80 ℃ and the drying time is 36-60 h.
3. The method for extracting and purifying the polyphenol from the flowers of hemerocallis as claimed in claim 1, wherein in the step 2, the ultrasonic working frequency is 40kHz, the extraction temperature is 30-60 ℃, and the extraction time is 50-80 min.
4. The method for extracting and purifying the hemerocallis fulva polyphenol as claimed in claim 1, wherein the macroporous adsorption resin in the step 3 is one of AB-8 macroporous adsorption resin, D101 macroporous adsorption resin or HPD600 macroporous adsorption resin.
5. The method for extracting and purifying a tawny daylily polyphenol as claimed in claim 1, wherein in the step 3, the diameter-height ratio of the macroporous adsorption resin chromatographic column is 1: 10.
6. The method for extracting and purifying a tawny daylily polyphenol as claimed in claim 1, wherein in the step 3, the ethanol aqueous solution eluted twice is 75% by volume ethanol aqueous solution.
7. The method for extracting and purifying the hemerocallis fulva polyphenol as claimed in claim 1, wherein in the step 4, the vacuum concentration temperature is 45-55 ℃, the concentration time is 1-3 h, and the vacuum degree is as follows: -0.8 to-0.9 Mpa.
8. The method for extracting and purifying the polyphenol from the flowers of hemerocallis as claimed in claim 1, wherein in the step 5, the freezing temperature is-80 ℃, the drying time is 24-48 h, and the vacuum degree is as follows: -0.8 Mpa.
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| CN111234559A (en) * | 2020-03-23 | 2020-06-05 | 运城学院 | Extraction and purification method of purple locust pigment |
| CN111773165A (en) * | 2020-07-30 | 2020-10-16 | 上海应用技术大学 | Hemerocallis fulva flower shampoo and preparation method thereof |
| CN113229309A (en) * | 2021-06-22 | 2021-08-10 | 上海应用技术大学 | Hemerocallis fulva flower flaky pastry and preparation method thereof |
| CN114081922A (en) * | 2021-11-22 | 2022-02-25 | 兰州大学 | Hemerocallis plant leaf total phenol extract and preparation method and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111234559A (en) * | 2020-03-23 | 2020-06-05 | 运城学院 | Extraction and purification method of purple locust pigment |
| CN111234559B (en) * | 2020-03-23 | 2022-05-27 | 运城学院 | Extraction and purification method of purple locust pigment |
| CN111773165A (en) * | 2020-07-30 | 2020-10-16 | 上海应用技术大学 | Hemerocallis fulva flower shampoo and preparation method thereof |
| CN111773165B (en) * | 2020-07-30 | 2022-12-09 | 上海应用技术大学 | Hemerocallis fulva flower shampoo and preparation method thereof |
| CN113229309A (en) * | 2021-06-22 | 2021-08-10 | 上海应用技术大学 | Hemerocallis fulva flower flaky pastry and preparation method thereof |
| CN114081922A (en) * | 2021-11-22 | 2022-02-25 | 兰州大学 | Hemerocallis plant leaf total phenol extract and preparation method and application thereof |
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