Extraction method and application of rose active substances

Abstract

The invention discloses an extraction method and application of active substances of roses, and the extraction method comprises the following steps: (1) picking fresh roses, cleaning, drying and crushing to obtain rose powder; (2) dispersing the rose powder into a choline chloride type eutectic solvent, and carrying out ultrasonic extraction or microwave extraction; and (3) centrifuging the extracted solution, taking supernate, adding a freeze-drying protective agent into the supernate, and performing freeze-drying to obtain the rose active substance. The method can realize comprehensive extraction of the active substances of the roses, maximally retains the active substances of the roses, improves the utilization rate of the roses, and is simple, reliable, green and environment-friendly.

Description

Technical Field

[0001] This invention belongs to the field of biochemical technology, specifically relating to a method for extracting active substances from roses and their application. Background Technology

[0002] As people's living standards improve, the demand for health care and the efficacy of cosmetics are increasing. Roses are an important application in the fields of cosmetics, food, and health care. They are rich in a variety of active substances and have high public recognition. Therefore, the extraction of active substances from roses has promising applications.

[0003] The active substances in roses (polyphenols, flavonoids, volatile oils, and vitamin C, etc.) in cosmetics delay skin aging through powerful antioxidant effects, strengthen the skin barrier, moisturize and repair, and inhibit melanin to brighten skin tone; at the same time, their soothing ingredients can reduce redness by 20%. In the health supplement field, oral extracts of polyphenols scavenge free radicals and enhance immunity; aromatic volatile oils regulate mood and improve sleep, and work synergistically with dietary fiber to protect the liver and promote digestion, forming a dual pathway of "external protection and internal nourishment"—from skin rejuvenation to physical and mental balance, achieving holistic health value.

[0004] Common methods for extracting active substances from roses include water extraction and organic solvent extraction. However, water extraction has poor selectivity, while organic solvent extraction has problems such as organic solvent residue and toxicity. Summary of the Invention

[0005] To solve the above-mentioned technical problems, the present invention provides a method for extracting active substances from rose petals, using a choline chloride-type eutectic solvent as the extractant to assist in ultrasonic or microwave extraction. The extracted active substances from rose petals have a high extraction rate, ranging from 12.2% to 15.2%, and the extracted active substances from rose petals have high antioxidant activity.

[0006] The technical solution adopted in this invention is as follows:

[0007] A method for extracting active substances from rose petals, the extraction method comprising the following steps:

[0008] (1) Fresh roses are picked, washed, dried and crushed to obtain rose pollen;

[0009] (2) Disperse rose pollen in a choline chloride-type eutectic solvent and perform ultrasonic or microwave extraction;

[0010] (3) Centrifuge the extracted solution, take the supernatant, add a freeze-drying protectant to it, and freeze-dry to obtain the active substance of rose.

[0011] The choline chloride-type eutectic solvent is composed of a hydrogen bond acceptor and a hydrogen bond donor; wherein the hydrogen bond acceptor is choline chloride, and the hydrogen bond donor is at least one of lactic acid, ethanol, ethylene glycol, propylene glycol, glycerol, or butanediol; the molar ratio of the hydrogen bond donor compound to the hydrogen bond acceptor compound is 1 to 5.

[0012] The water content of the choline chloride-type eutectic solvent is 10% to 90%, preferably 20%.

[0013] In step (2), the solid-liquid ratio of rose pollen to choline chloride-type eutectic solvent is 1:5 to 20.

[0014] In step (2), the conditions for ultrasonic extraction are: temperature 30-50℃, power 300-1000W, and ultrasonic time 30-60min.

[0015] In step (2), the conditions for microwave extraction are: the total extraction time is 30 to 60 minutes, and microwave extraction is stopped for 1 minute after each 9-minute extraction, and then microwave extraction is continued for at least three times.

[0016] In step (3), the freeze-drying protectant is polyethylene glycol; the freeze-drying conditions are: first cool to 0-3℃, then freeze to below -25℃, and dry in a vacuum freeze dryer at a drying temperature of -50 to -60℃ for more than 24 hours.

[0017] The polyethylene glycol is polyethylene glycol 6000, and the amount added is 0.5-5% of the mass of the supernatant in the lower layer, preferably 1%.

[0018] The present invention also provides active substances from roses extracted using the extraction method described in the present invention.

[0019] The present invention also provides the application of the aforementioned active substances from roses in the preparation of antioxidant drugs, food, cosmetics, and health products.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] This invention utilizes a low-eutectic solvent-assisted ultrasonic or microwave extraction process to efficiently extract active substances from roses. This extraction method is a green extraction method that does not require the use of large amounts of organic reagents, overcoming the problems of poor selectivity, organic solvent residues, and toxicity associated with traditional solvents such as water. Furthermore, it is non-toxic and environmentally friendly.

[0022] The extraction method provided by this invention is simple and reliable, achieving comprehensive extraction of active substances from roses with a high extraction rate, maximizing the retention of active ingredients in roses, and improving the utilization rate of roses.

[0023] The active substances from roses prepared by the method of this invention can be used in the production and formulation of foods, cosmetics and health foods with functions such as anti-oxidation, anti-sensitivity, analgesia, wound healing promotion, anti-aging, and beauty enhancement, and have important economic value.

[0024] The active substances from roses prepared by the method of this invention can be used with food additives to prepare functional foods or health foods, or they can be combined with other nutrients for the processing and preparation of functional foods or health foods to exert dietary therapeutic functions such as anti-oxidation, anti-sensitivity, analgesia, wound healing promotion, anti-aging, and beauty enhancement. Detailed Implementation

[0025] The present invention will now be described in detail with reference to the embodiments.

[0026] The eutectic solvent (DES) used in the following examples is choline chloride type DES, and its composition is shown in Table 1.

[0027] Table 1 Different types of DES

[0028]

[0029]

[0030] The preparation methods for each DES in Table 1 are as follows:

[0031] (1) Preparation of DES-1

[0032] Accurately weigh 11.170 g (0.08 mol) of hydrogen bond acceptor (HBA) and 14.413 g (0.16 mol) of lactic acid (HBD) using an analytical balance. Add the mixture to a round-bottom flask and place it in a heated magnetic stirrer at 80°C for 2 hours. Once a homogeneous and stable liquid has formed, stop the reaction and remove the flask. Cool the flask to room temperature. After cooling, the liquid will still be colorless, transparent, and homogeneous. Store the flask in a sealed desiccator. When using the flask, mix DES solvent with deionized water to achieve a water content of 20%.

[0033] (2) Preparation of DES-2

[0034] The preparation of DES-1 was the same as in (1), except that the hydrogen bond donor (HBD) was replaced with 7.371 g (0.16 mol) of ethanol.

[0035] (3) Preparation of DES-3

[0036] The preparation of DES-1 was the same as in (1), except that the hydrogen bond donor (HBD) was replaced with 9.931 g (0.16 mol) of ethylene glycol.

[0037] (4) Preparation of DES-4

[0038] The preparation of DES-1 was the same as in (1), except that the hydrogen bond donor (HBD) was replaced with 12.174 g (0.16 mol) of propylene glycol.

[0039] (5) Preparation of DES-5

[0040] The preparation of DES-1 was the same as in (1), except that the hydrogen bond donor (HBD) was replaced with 14.734 g (0.16 mol) of glycerol.

[0041] (6) Preparation of DES-6

[0042] The preparation of DES-1 was the same as in (1), except that the hydrogen bond donor (HBD) was replaced with 14.419 g (0.16 mol) of butanediol.

[0043] Example 1

[0044] The extraction solvent was a choline chloride-lactic acid eutectic solvent (DES-1).

[0045] A method for extracting active substances from roses with high antioxidant activity includes the following steps:

[0046] (1) Fresh roses were dried to constant weight at 40°C using a vacuum dryer and then pulverized through a 100-mesh sieve to obtain rose powder.

[0047] (2) Accurately weigh 500mg of rose pollen, add it to a eutectic solvent of choline chloride-lactic acid (DES-1) as the extraction solvent, and perform ultrasonic extraction at a solid-liquid ratio of 1:10, a temperature of 50℃ and a power of 600W for 60min.

[0048] (3) Centrifuge the extract at 8000 rpm / min for 10 min. The resulting supernatant is the clear extract. Add 1% of the extract mass of the protective agent polyethylene glycol 6000 to the extract and cool it to 0℃. Then freeze it to -25℃ and place it in a vacuum freeze dryer with a vacuum degree of 10 Pa and a freeze drying temperature of -60℃. Maintain the vacuum degree for drying for more than 24 hours to obtain freeze-dried powder of active rose substances.

[0049] Example 2

[0050] The extraction solvent was a choline chloride-ethanol eutectic solvent (DES-2).

[0051] A method for extracting active substances from roses with high antioxidant activity includes the following steps:

[0052] (1) Fresh roses were dried to constant weight at 40°C using a vacuum dryer and then pulverized through a 100-mesh sieve to obtain rose powder.

[0053] (2) Accurately weigh 500mg of rose pollen, add it to a eutectic solvent of choline chloride-ethanol type (DES-2) as the extraction solvent, and perform ultrasonic extraction at a solid-liquid ratio of 1:10, a temperature of 50℃ and a power of 600W for 60min.

[0054] (3) Centrifuge the extract at 8000 rpm / min for 10 min. The resulting supernatant is the clear extract. Add 1% of the extract mass of the protective agent polyethylene glycol 6000 to the extract and cool it to 0℃. Then freeze it to -25℃ and place it in a vacuum freeze dryer with a vacuum degree of 10 Pa and a freeze drying temperature of -60℃. Maintain the vacuum degree for drying for more than 24 hours to obtain freeze-dried powder of active rose substances.

[0055] Example 3

[0056] The extraction solvent was a choline chloride-ethylene glycol eutectic solvent (DES-3).

[0057] A method for extracting active substances from roses with high antioxidant activity includes the following steps:

[0058] (1) Fresh roses were dried to constant weight at 40°C using a vacuum dryer and then pulverized through a 100-mesh sieve to obtain rose powder.

[0059] (2) Accurately weigh 500 mg of rose pollen, add it to a eutectic solvent of choline chloride-ethylene glycol (DES-3) as the extraction solvent, and the water content of the extraction solvent is 20%. Under the conditions of solid-liquid ratio of 1:10, temperature of 50℃ and power of 600W, perform ultrasonic extraction for 60 min.

[0060] (3) Centrifuge the extract at 8000 rpm / min for 10 min. The resulting supernatant is the clear extract. Add 1% of the extract mass of the protective agent polyethylene glycol 6000 to the extract and cool it to 0℃. Then freeze it to -25℃ and place it in a vacuum freeze dryer with a vacuum degree of 10 Pa and a freeze drying temperature of -60℃. Maintain the vacuum degree for drying for more than 24 hours to obtain freeze-dried powder of active rose substances.

[0061] Example 4

[0062] The extraction solvent was a choline chloride-propylene glycol eutectic solvent (DES-4).

[0063] A method for extracting active substances from roses with high antioxidant activity includes the following steps:

[0064] (1) Fresh roses were dried to constant weight at 40°C using a vacuum dryer and then pulverized through a 100-mesh sieve to obtain rose powder.

[0065] (2) Accurately weigh 500mg of rose pollen, add it to a eutectic solvent of choline chloride-propylene glycol (DES-4) as the extraction solvent, and perform ultrasonic extraction at a solid-liquid ratio of 1:10, a temperature of 50℃ and a power of 600W for 60min.

[0066] (3) Centrifuge the extract at 8000 rpm / min for 10 min. The resulting supernatant is the clear extract. Add 1% of the extract mass of the protective agent polyethylene glycol 6000 to the extract and cool it to 0℃. Then freeze it to -25℃ and place it in a vacuum freeze dryer with a vacuum degree of 10 Pa and a freeze drying temperature of -60℃. Maintain the vacuum degree for drying for more than 24 hours to obtain freeze-dried powder of active rose substances.

[0067] Example 5

[0068] The extraction solvent was a choline chloride-glycerol eutectic solvent (DES-5).

[0069] A method for extracting active substances from roses with high antioxidant activity includes the following steps:

[0070] (1) Fresh roses were dried to constant weight at 40°C using a vacuum dryer and then pulverized through a 100-mesh sieve to obtain rose powder.

[0071] (2) Accurately weigh 500mg of rose pollen, add it to a eutectic solvent of choline chloride-glycerol type (DES-5) as the extraction solvent, and perform ultrasonic extraction at a solid-liquid ratio of 1:10, a temperature of 50℃ and a power of 600W for 60min.

[0072] (3) Centrifuge the extract at 8000 rpm / min for 10 min. The resulting supernatant is the clear extract. Add 1% of the extract mass of the protective agent polyethylene glycol 6000 to the extract and cool it to 0℃. Then freeze it to -25℃ and place it in a vacuum freeze dryer with a vacuum degree of 10 Pa and a freeze drying temperature of -60℃. Maintain the vacuum degree for drying for more than 24 hours to obtain freeze-dried powder of active rose substances.

[0073] Example 6

[0074] The extraction solvent was a choline chloride-butanediol eutectic solvent (DES-6).

[0075] A method for extracting active substances from roses with high antioxidant activity includes the following steps:

[0076] (1) Fresh roses were dried to constant weight at 40°C using a vacuum dryer and then pulverized through a 100-mesh sieve to obtain rose powder.

[0077] (2) Accurately weigh 500mg of rose pollen, add it to a eutectic solvent of choline chloride-butanediol type (DES-6) as the extraction solvent, and perform ultrasonic extraction at a solid-liquid ratio of 1:10, a temperature of 50℃ and a power of 600W for 60min.

[0078] (3) Centrifuge the extract at 8000 rpm / min for 10 min. The resulting supernatant is the clear extract. Add 1% of the extract mass of the protective agent polyethylene glycol 6000 to the extract and cool it to 0℃. Then freeze it to -25℃ and place it in a vacuum freeze dryer with a vacuum degree of 10 Pa and a freeze drying temperature of -60℃. Maintain the vacuum degree for drying for more than 24 hours to obtain freeze-dried powder of active rose substances.

[0079] Comparative Example 1

[0080] Deionized water was used as the extraction solvent.

[0081] A method for extracting active substances from roses with high antioxidant activity includes the following steps:

[0082] (1) Fresh roses were dried to constant weight at 40°C using a vacuum dryer and then pulverized through a 100-mesh sieve to obtain rose powder.

[0083] (2) Accurately weigh 500mg of rose pollen, use deionized water as the extraction solvent, and perform ultrasonic extraction on it under the conditions of solid-liquid ratio of 1:10, temperature of 50℃ and power of 600W for 60min.

[0084] (3) Centrifuge the extract at 8000 rpm / min for 10 min. The resulting supernatant is the clear extract. Add 1% of the extract mass of the protective agent polyethylene glycol 6000 to the extract and cool it to 0℃. Then freeze it to -25℃ and place it in a vacuum freeze dryer with a vacuum degree of 10 Pa and a freeze drying temperature of -60℃. Maintain the vacuum degree for drying for more than 24 hours to obtain freeze-dried powder of active rose substances.

[0085] Comparative Example 2

[0086] Using 70% ethanol as the extraction solvent

[0087] A method for extracting active substances from roses with high antioxidant activity includes the following steps:

[0088] (1) Fresh roses were dried to constant weight at 40°C using a vacuum dryer and then pulverized through a 100-mesh sieve to obtain rose powder.

[0089] (2) Accurately weigh 500mg of rose pollen, use 70% ethanol as the extraction solvent, and perform ultrasonic extraction on it under the conditions of solid-liquid ratio of 1:10, temperature of 50℃ and power of 600W for 60min.

[0090] (3) Centrifuge the extract at 8000 rpm / min for 10 min. The resulting supernatant is a clear extract. Add 1% of the extract mass of the protective agent polyethylene glycol 6000 to the extract and cool it to 0℃. Then freeze it to -25℃ and place it in a vacuum freeze dryer with a vacuum degree of 10 Pa and a freeze drying temperature of -60℃. Maintain the vacuum degree for drying for more than 24 hours to obtain freeze-dried powder of active rose substances.

[0091] Test case

[0092] Weigh the freeze-dried rose active substance powder from each of the above embodiments and comparative examples, and calculate the extraction rate of the rose active substance:

[0093]

[0094] In the formula, the mass of the lyophilized active substance = the mass of the lyophilized rose active substance powder - the mass of the blank group. The blank group is defined as follows: 1% of the mass of the blank eutectic solvent (polyethylene glycol 6000) is added to 5g of blank eutectic solvent and cooled to 0℃, then frozen to -25℃, placed in a vacuum freeze dryer with a vacuum degree of 10Pa and a freeze-drying temperature of -60℃. The vacuum degree is maintained for drying for more than 24 hours, and the weight is then measured. This weight is the mass of the blank group.

[0095] The lyophilized products from each example and comparative example were prepared into rose active substance dilutions with concentrations of 1 mg / mL, 0.5 mg / mL, 0.25 mg / mL, 0.1 mg / mL, 0.05 mg / mL, and 0.01 mg / mL, respectively, using ethanol as the solvent. 1 mL of the rose active substance dilution was taken and 1 mL of 80 μg / mL DPPH ethanol solution was added. After mixing, the mixture was reacted at room temperature in the dark for 30 min, and this was used as the sample group. Control and blank groups were also set up, as shown in Table 2.

[0096] Table 2

[0097] Group composition Blank group 1 mL DPPH ethanol solution + 1 mL ethanol control group 1 mL rose active ingredient dilution + 1 mL ethanol Sample group 1 mL DPPH ethanol solution + 1 mL rose active ingredient dilution

[0098] Then, the absorbance of each group was measured at 517 nm, and the clearance rate was calculated according to the following formula:

[0099]

[0100] In the formula: A1 represents the absorbance of the sample group; A2 represents the absorbance of the control group; A0 represents the absorbance of the blank group.

[0101] A concentration-scavenging rate curve was plotted with the diluted rose active ingredient solution as the x-axis and the DPPH free radical scavenging rate as the y-axis. The concentration at which the scavenging rate reached 50%, i.e., the half-inhibitory concentration (IC50), was determined by fitting the curve using nonlinear regression. 50 With half-inhibitory concentration (IC50) 50 () indicates the antioxidant activity of active substances in roses.

[0102] The test results are shown in Table 3.

[0103] Table 3

[0104]

[0105]

[0106] As can be seen from Table 3, the extraction rates of active substances from rose obtained in the examples ranged from 12.2% to 15.2%, and the IC50 concentration was [missing information]. 50 The values ​​ranged from 0.126 to 0.154 mg / mL. Compared with the comparative example, the extraction method in this example significantly improved the extraction rate and antioxidant activity of the active substances extracted from roses, indicating that the extraction process yielded a high extraction rate of active substances from roses and exhibited high antioxidant activity.

[0107] The above detailed description of a method for extracting and applying active substances from roses, with reference to the embodiments, is illustrative rather than limiting. Several embodiments may be listed within the defined scope. Therefore, variations and modifications that do not depart from the overall concept of the present invention should be within the protection scope of the present invention.

Claims

1. A method for extracting active substances from rose petals, characterized in that, The extraction method includes the following steps: (1) Fresh roses are picked, washed, dried and crushed to obtain rose pollen; (2) Disperse rose pollen in a choline chloride-type eutectic solvent and perform ultrasonic or microwave extraction; (3) Centrifuge the extracted solution, take the supernatant, add a freeze-drying protectant to it, and freeze-dry to obtain the active substance of rose.

2. The method for extracting active substances from roses according to claim 1, characterized in that, The choline chloride-type eutectic solvent is composed of a hydrogen bond acceptor and a hydrogen bond donor; wherein the hydrogen bond acceptor is choline chloride, and the hydrogen bond donor is at least one of lactic acid, ethanol, ethylene glycol, propylene glycol, glycerol, or butanediol; the molar ratio of the hydrogen bond donor compound to the hydrogen bond acceptor compound is 1 to 5.

3. The method for extracting active substances from roses according to claim 1 or 2, characterized in that, The water content of choline chloride-type eutectic solvents is 10%–90%.

4. The method for extracting active substances from roses according to claim 1 or 2, characterized in that, In step (2), the solid-liquid ratio of rose pollen to choline chloride-type eutectic solvent is 1:5 to 20.

5. The method for extracting active substances from roses according to claim 1 or 2, characterized in that, In step (2), the conditions for ultrasonic extraction are: temperature 30-50℃, power 300-1000W, and ultrasonic time 30-60min.

6. The method for extracting active substances from roses according to claim 1 or 2, characterized in that, In step (2), the conditions for microwave extraction are: the total extraction time is 30 to 60 minutes, and microwave extraction is stopped for 1 minute after each 9-minute extraction, and then microwave extraction is continued for at least three times.

7. The method for extracting active substances from roses according to claim 1 or 2, characterized in that, In step (3), the freeze-drying protectant is polyethylene glycol; the freeze-drying conditions are: first cool to 0-3℃, then freeze to below -25℃, and dry in a vacuum freeze dryer at a drying temperature of -50 to -60℃ for more than 24 hours.

8. The method for extracting active substances from roses according to claim 7, characterized in that, The polyethylene glycol is polyethylene glycol 6000, and the amount added is 0.5-5% of the mass of the supernatant.

9. The active substances of rose obtained by the extraction method according to any one of claims 1-8.

10. The application of the active rose substance as described in claim 9 in the preparation of antioxidant drugs, food, cosmetics, and health products.

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