CN116139055A - Preparation method and application of vitamin C-enriched roxburgh rose extract - Google Patents
Preparation method and application of vitamin C-enriched roxburgh rose extract Download PDFInfo
- Publication number
- CN116139055A CN116139055A CN202211602768.7A CN202211602768A CN116139055A CN 116139055 A CN116139055 A CN 116139055A CN 202211602768 A CN202211602768 A CN 202211602768A CN 116139055 A CN116139055 A CN 116139055A
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- CN
- China
- Prior art keywords
- vitamin
- extract
- juice
- rosa roxburghii
- roxburgh rose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
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Classifications
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Landscapes
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Abstract
The invention discloses a preparation method of a roxburgh rose extract rich in vitamin C, which comprises the following steps: (1) Cleaning fructus Rosae Normalis, squeezing juice or freezing, respectively, and squeezing juice; (2) taking the roxburgh rose juice, diluting the roxburgh rose juice with an ultrafiltration membrane to obtain filtrate; (3) Concentrating the filtrate by reverse osmosis membrane to obtain fructus Rosae Normalis concentrated juice; (4) Collecting fructus Rosae Normalis concentrated juice, adding ethanol, centrifuging to obtain supernatant, concentrating, and recovering ethanol to obtain ethanol precipitation concentrate; (5) Eluting with macroporous resin, collecting eluate, and concentrating to obtain extract; (6) Drying the extract to obtain fructus Rosae Normalis extract rich in vitamin C, and its application are also disclosed. The preparation method can fully utilize fresh Rosa roxburghii fruits, enriches the vitamin C through the combination of the processes of the steps, is a natural vitamin C extract, remarkably enhances the antioxidant and whitening functions of the extract, and provides a theoretical basis and an application basis for improving the development of the Rosa roxburghii industrial chain.
Description
Technical Field
The invention belongs to the technical field of organic compound extraction, and particularly relates to a preparation method and application of a vitamin C-enriched roxburgh rose extract.
Background
Rosa roxburghii is a single-petal silk reeling flower and a fruit of the silk reeling flower of Rosaceae plants, is a noble and minority characteristic medicinal material, is listed in the quality standard of Chinese medicinal materials and national medicinal materials in Guizhou province (2003 edition), and is listed as common food. Fructus Rosae Normalis has effects of invigorating stomach, resolving food stagnation, strengthening body constitution, astringing essence, relieving diarrhea, etc., contains multiple chemical components such as polyphenols, flavone, triterpene, etc., especially contains vitamin C and SOD (superoxide dismutase), and the content of vitamin C in pulp is in crown of various fruits. The modern pharmacological research results show that the roxburgh rose has various effects of resisting oxidation, resisting tumor, delaying aging, whitening skin, removing spots and the like, and has potential health-care application prospect.
At present, the research on the Rosa roxburghii extract is more, vitamin C is used as one of the key efficacy components of the Rosa roxburghii, has the effects of resisting oxidation, delaying aging, resisting scurvy, treating anemia and the like, is widely applied to medicines, health products, foods and cosmetic products, and the Rosa roxburghii extract with high vitamin C content is directly related to the development of the Rosa roxburghii industry, but the Rosa roxburghii extract obtained by the preparation method disclosed by the prior art has lower vitamin C content, complex process and higher production cost, and cannot be suitable for large-scale industrial production. Such as: patent application CN106036270A, CN111700247B, CN109232487a, etc.
Therefore, there is an urgent need to establish a method for preparing a Rosa roxburghii extract with high content of vitamin C, so as to meet the application requirements of the Rosa roxburghii extract with high content of vitamin C in industrial application.
Disclosure of Invention
The invention provides a preparation method and application of a roxburgh rose extract rich in vitamin C, wherein the extracts obtained through the steps of freezing and juicing fresh roxburgh rose fruits, membrane filtration and impurity removal, reverse osmosis concentration, alcohol precipitation and impurity removal, chromatography enrichment and the like all contain higher vitamin C, and the antioxidation and whitening effects of the extract are obviously enhanced.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a preparation method of a fructus Rosae Normalis extract rich in vitamin C comprises the following steps:
(1) Cleaning fructus Rosae Normalis, freezing, and squeezing to obtain fructus Rosae Normalis juice;
(2) Diluting fructus Rosae Normalis juice with purified water, and filtering with ultrafiltration membrane to obtain filtrate;
(3) Concentrating the filtrate with reverse osmosis membrane to obtain fructus Rosae Normalis concentrated juice;
(4) Collecting fructus Rosae Normalis concentrated juice, adding ethanol, stirring, standing at low temperature, centrifuging to obtain supernatant, and recovering ethanol at vacuum degree and concentration temperature to obtain ethanol precipitation concentrate;
(5) Purifying the ethanol precipitation concentrate with macroporous resin, eluting with purified water and ethanol-containing water solution, collecting eluate, and concentrating to obtain extract;
(6) Drying the extract to obtain fructus Rosae Normalis extract rich in vitamin C.
The preparation method of the vitamin C-enriched roxburgh rose extract comprises the following steps:
(1) Cleaning fresh Rosa roxburghii fruit, freezing for more than 24 hours at the temperature of-4 to-20 ℃, taking out the frozen fresh Rosa roxburghii fruit, thawing, squeezing juice at the pressure of 3-7 kg for 0.5-1.5 hours, and filtering to obtain Rosa roxburghii juice; thawing and squeezing after freezing is more beneficial to the dissolution of vitamin C, and the content of vitamin C in the finished product is improved;
(2) Diluting the roxburgh rose juice with purified water with the volume of 1-7 times of that of the roxburgh rose juice, and filtering with a 10-100KD ultrafiltration membrane to obtain filtrate, wherein the ultrafiltration membrane can effectively remove impurities, especially macromolecular substances, in the roxburgh rose juice;
(3) Concentrating the filtrate with reverse osmosis membrane under a controlled pressure of 1-6 kg to obtain concentrated juice of fructus Rosae Normalis, and removing most of water in the juice by reverse osmosis membrane filtration to facilitate alcohol precipitation;
(4) Taking concentrated juice of roxburgh rose, adding 80-95% ethanol with the mass of 8-12 times, fully stirring, standing for 10-16 hours at the temperature of 2-8 ℃, centrifuging, taking supernatant, concentrating the supernatant under reduced pressure and vacuum degree of 0.08-0.09 mpa, concentrating at the temperature: 40-55 ℃ to obtain an alcohol precipitation concentrated solution; alcohol precipitation can effectively remove substances insoluble in alcohol in the roxburgh rose juice, and further remove impurities;
(5) Taking macroporous resins such as AB-8, HPD-100, D101 and the like on an alcohol precipitation concentrated solution, adopting purified water and an aqueous solution containing 5-30% of ethanol as eluent, concentrating the eluent to an extract with the relative density of 1.10-1.20, and adopting macroporous resin chromatography to enrich vitamin C;
(6) The extract is dried in vacuum: vacuum degree is 0.08-0.09 mpa, and drying temperature is 40-55 ℃; or freeze drying: prefreezing temperature: -10-50 ℃, the vacuum degree of 10-30 Pa, the temperature of the water catcher of 35-45 ℃ and the temperature of the baffle of-10 ℃ to obtain the roxburgh rose extract rich in vitamin C.
In the step (1), the freezing temperature is-12 ℃, the juice squeezing pressure is 5kg, and the juice squeezing time is 1 hour.
In the step (2), the extract is diluted by 4 times of the volume of the Rosa roxburghii juice purified water and filtered by a 50KD membrane;
the preparation method of the vitamin C-enriched Rosa roxburghii extract comprises the step (3), wherein the membrane concentration pressure is controlled to be 3kg.
In the preparation method of the vitamin C-enriched Rosa roxburghii extract, in the step (4), 10 times of 85% ethanol is adopted, the ethanol is precipitated for 13 hours at the temperature of 6 ℃, the ethanol recovery vacuum degree is 0.085mpa, and the concentration temperature is 43 ℃.
In the step (5), macroporous resin is HPD-100, purified water is used as eluent for eluting, and the purified water eluent is concentrated to extract with a relative density of 1.15.
In the step (6), the extract is dried in vacuum, the vacuum degree is 0.085mpa, and the drying temperature is 47 ℃.
The extract of fructus Rosae Normalis rich in vitamin C prepared by any one of the above methods has vitamin C content of more than or equal to 50%.
The application of vitamin C-enriched fructus Rosae Normalis extract in resisting oxidation and whitening skin is provided.
The beneficial technical effects of the invention are as follows:
1. the invention provides a preparation method of a roxburgh rose extract rich in vitamin C, which comprises the steps of freezing and juicing fresh roxburgh rose fruits, filtering and removing impurities by an ultrafiltration membrane, concentrating by a reverse osmosis membrane, removing impurities by alcohol precipitation, enriching by macroporous resin chromatography, drying and the like, so that the roxburgh rose extract with high content of natural vitamin C is obtained, wherein the vitamin C content in the roxburgh rose extract is higher than 50 percent.
2. According to the invention, the fresh rosa roxburghii fruits are frozen, thawed and juiced, so that the dissolution of vitamin C is facilitated, macromolecular impurities in the rosa roxburghii juice are filtered out by the ultrafiltration membrane, substances such as dissolved salts in the rosa roxburghii juice are removed by the reverse osmosis membrane, substances insoluble in alcohol in the rosa roxburghii juice are further removed by the alcohol precipitation step, the vitamin C is further enriched by the macroporous resin, and the physical and chemical properties of the substances are removed according to the requirement, and specific technological parameter combinations and technological step sequences are screened through experiments, so that the rosa roxburghii extract with high vitamin C content is obtained, and the process steps are simple, easy and convenient to operate and suitable for large-scale industrial production.
3. The in vitro antioxidation function test and whitening efficacy test research result shows that the antioxidation and whitening effects of the finished product of the roxburgh rose extract rich in vitamin C are obviously enhanced, the invention provides technical support for applying the vitamin C in the roxburgh rose to medicines, health care products, foods and cosmetic products, and provides theory and application foundation for the development of the roxburgh rose industry and the big health industry.
Detailed Description
Example 1:
a method for preparing fructus Rosae Normalis extract rich in vitamin C comprises:
(1) Cleaning fructus Rosae Normalis, freezing at-4deg.C for more than 24 hr, taking out frozen fructus Rosae Normalis, thawing, squeezing juice under pressure of 3kg for 0.5 hr, and filtering to obtain fructus Rosae Normalis juice;
(2) Diluting the roxburgh rose juice obtained in the step (1) with purified water with the volume of 1 time of the roxburgh rose juice, and filtering with a 10KD ultrafiltration membrane to obtain a filtrate;
(3) Taking the filtrate obtained in the step (2), controlling the pressure to be 1kg, and concentrating by using a reverse osmosis membrane to obtain a roxburgh rose concentrated juice;
(4) Taking the roxburgh rose concentrated juice obtained in the step (3), adding 80% ethanol with the mass of 8 times, fully stirring, standing at the temperature of 2 ℃ for 10 hours, centrifuging to obtain supernatant, and concentrating the supernatant under reduced pressure to recover ethanol: vacuum degree is 0.08mpa, concentration temperature is 55 ℃, and alcohol precipitation concentrate is obtained;
(5) Taking AB-8 macroporous resin on the alcohol precipitation concentrated solution obtained in the step (4), eluting with purified water as an eluent, and concentrating the eluent to obtain an extract with the relative density of 1.10;
(6) And (5) taking the extract obtained in the step (5), and adopting vacuum drying: vacuum degree is 0.08mpa, drying temperature is 55deg.C, and fructus Rosae Normalis extract rich in vitamin C is obtained.
The obtained fructus Rosae Normalis extract rich in vitamin C has vitamin C content of 51.52%.
Example 2:
a method for preparing fructus Rosae Normalis extract rich in vitamin C comprises:
(1) Cleaning fructus Rosae Normalis, freezing at-20deg.C for more than 24 hr, taking out frozen fructus Rosae Normalis, thawing, squeezing juice under pressure of 7kg for 1.5 hr, and filtering to obtain fructus Rosae Normalis juice;
(2) Diluting the roxburgh rose juice obtained in the step (1) with 7 times of purified water with the volume of the roxburgh rose juice, and filtering with a 100KD ultrafiltration membrane to obtain a filtrate;
(3) Taking the filtrate obtained in the step (2), controlling the pressure to be 6kg, and concentrating by using a reverse osmosis membrane to obtain a roxburgh rose concentrated juice;
(4) Taking the roxburgh rose concentrated juice obtained in the step (3), adding 95% ethanol with the mass of 10 times, fully stirring, standing at 8 ℃ for 16 hours, centrifuging to obtain supernatant, and concentrating the supernatant under reduced pressure to recover ethanol: vacuum degree is 0.09mpa, concentration temperature is 40 ℃, and alcohol precipitation concentrated solution is obtained;
(5) Loading the ethanol precipitation concentrated solution obtained in the step (4) on macroporous resin such as D101, eluting with an ethanol water solution with the ethanol content of 5% as an eluent, and concentrating the eluent to obtain an extract with the relative density of 1.20;
(6) And (5) taking the extract obtained in the step (5), and adopting vacuum drying: vacuum degree is 0.09mpa, drying temperature is 40 ℃, and fructus Rosae Normalis extract rich in vitamin C is obtained.
The obtained fructus Rosae Normalis extract rich in vitamin C has a vitamin C content of 53.78%.
Example 3:
a method for preparing fructus Rosae Normalis extract rich in vitamin C comprises:
(1) Cleaning fructus Rosae Normalis, freezing at-12deg.C for more than 24 hr, taking out frozen fructus Rosae Normalis, thawing, squeezing juice under 5kg pressure for 1.0 hr, and filtering to obtain fructus Rosae Normalis juice;
(2) Diluting the roxburgh rose juice obtained in the step (1) with 4 times of purified water with the volume of the roxburgh rose juice, and filtering with a 50KD ultrafiltration membrane to obtain a filtrate;
(3) Taking the filtrate obtained in the step (2), controlling the pressure to be 3kg, and concentrating by using a reverse osmosis membrane to obtain a roxburgh rose concentrated juice;
(4) Taking the roxburgh rose juice obtained in the step (3), adding 87% ethanol with the mass of 12 times, fully stirring, standing at the temperature of 5 ℃ for 13 hours, centrifuging to obtain supernatant, and concentrating the supernatant under reduced pressure to recover ethanol: vacuum degree is 0.085mpa, concentration temperature is 47 ℃, and alcohol precipitation concentrated solution is obtained;
(5) Loading HPD-100 macroporous resin on the alcohol precipitation concentrated solution obtained in the step (4), eluting with 30% alcohol aqueous solution as eluent, and concentrating the eluent to obtain extract with relative density of 1.15;
(6) And (5) taking the extract obtained in the step (5), and adopting vacuum drying: vacuum degree is 0.085mpa, drying temperature is 47 ℃, and fructus Rosae Normalis extract rich in vitamin C is obtained.
The obtained fructus Rosae Normalis extract rich in vitamin C has vitamin C content of 53.46%.
Example 4:
a method for preparing fructus Rosae Normalis extract rich in vitamin C comprises:
(1) Cleaning fructus Rosae Normalis, freezing at-15deg.C for more than 24 hr, taking out frozen fructus Rosae Normalis, thawing, squeezing juice under pressure of 6kg for 1.2 hr, and filtering to obtain fructus Rosae Normalis juice;
(2) Diluting the roxburgh rose juice obtained in the step (1) with purified water with the volume of 5 times of the roxburgh rose juice, and filtering with a 90KD ultrafiltration membrane to obtain a filtrate;
(3) Taking the filtrate obtained in the step (2), controlling the pressure to be 5kg, and concentrating by using a reverse osmosis membrane to obtain a roxburgh rose concentrated juice;
(4) Taking the roxburgh rose concentrated juice obtained in the step (3), adding 83% ethanol with the mass of 10 times, fully stirring, standing at the temperature of 6 ℃ for 12 hours, centrifuging to obtain supernatant, concentrating the supernatant under reduced pressure, and recovering ethanol: vacuum degree is 0.09mpa, concentration temperature is 47 ℃, and alcohol precipitation concentrated solution is obtained;
(5) Loading HPD-100 macroporous resin on the alcohol precipitation concentrated solution obtained in the step (4), eluting with 17% alcohol aqueous solution as eluent, and concentrating the eluent to obtain extract with relative density of 1.15;
(6) And (5) freeze-drying the extract obtained in the step (5): the pre-freezing temperature is-10deg.C, the vacuum degree is 10Pa, the temperature of water catcher is 35deg.C, and the temperature of baffle plate is-10deg.C to obtain fructus Rosae Normalis extract rich in vitamin C.
The obtained fructus Rosae Normalis extract rich in vitamin C has a vitamin C content of 54.17%.
Example 5:
a method for preparing fructus Rosae Normalis extract rich in vitamin C comprises:
(1) Cleaning fructus Rosae Normalis, freezing at-10deg.C for more than 24 hr, taking out frozen fructus Rosae Normalis, thawing, squeezing juice under pressure of 4kg for 1.0 hr, and filtering to obtain fructus Rosae Normalis juice;
(2) Diluting the roxburgh rose juice obtained in the step (1) with 3 times of purified water with the volume of the roxburgh rose juice, and filtering with a 60KD ultrafiltration membrane to obtain a filtrate;
(3) Taking the filtrate obtained in the step (2), controlling the pressure to be 6kg, and concentrating by using a reverse osmosis membrane to obtain a roxburgh rose concentrated juice;
(4) Taking the roxburgh rose concentrated juice obtained in the step (3), adding 80% ethanol with the mass of 8 times, fully stirring, standing at the temperature of 2 ℃ for 10 hours, centrifuging to obtain supernatant, and concentrating the supernatant under reduced pressure to recover ethanol: vacuum degree is 0.08mpa, concentration temperature is 40 ℃, and alcohol precipitation concentrated solution is obtained;
(5) Loading the alcohol precipitation concentrated solution obtained in the step (4) on macroporous resin such as D101 and eluting with purified water as an eluent, and concentrating the eluent to obtain extract with the relative density of 1.10;
(6) And (5) freeze-drying the extract obtained in the step (5): the pre-freezing temperature is 50 ℃, the vacuum degree is 30Pa, the temperature of the water catcher is 40 ℃, and the temperature of the baffle plate is 0 ℃, so that the Rosa roxburghii extract rich in vitamin C is obtained.
The obtained fructus Rosae Normalis extract rich in vitamin C has a vitamin C content of 52.17%.
Example 6:
a method for preparing fructus Rosae Normalis extract rich in vitamin C comprises:
(1) Cleaning fructus Rosae Normalis, freezing at-8deg.C for more than 24 hr, taking out frozen fructus Rosae Normalis, thawing, squeezing juice under pressure of 3kg for 0.5 hr, and filtering to obtain fructus Rosae Normalis juice;
(2) Diluting the roxburgh rose juice obtained in the step (1) with purified water with the volume of 5 times of the roxburgh rose juice, and filtering with a 100KD ultrafiltration membrane to obtain a filtrate;
(3) Taking the filtrate obtained in the step (2), controlling the pressure to be 3kg, and concentrating by using a reverse osmosis membrane to obtain a roxburgh rose concentrated juice;
(4) Taking the roxburgh rose concentrated juice obtained in the step (3), adding 95% ethanol with the mass of 10 times, fully stirring, standing at 8 ℃ for 16 hours, centrifuging to obtain supernatant, and concentrating the supernatant under reduced pressure to recover ethanol: vacuum degree is 0.09mpa, concentration temperature is 55 ℃, and alcohol precipitation concentrated solution is obtained;
(5) Taking AB-8 macroporous resin on the alcohol precipitation concentrated solution obtained in the step (4), eluting by using an ethanol water solution with the alcohol content of 5% as an eluent, and concentrating the eluent to obtain an extract with the relative density of 1.20;
(6) And (5) freeze-drying the extract obtained in the step (5): the pre-freezing temperature is 20 ℃, the vacuum degree is 20Pa, the temperature of the water catcher is 45 ℃, the temperature of the baffle plate is 10 ℃, and the Rosa roxburghii extract rich in vitamin C is obtained.
The obtained fructus Rosae Normalis extract rich in vitamin C has a vitamin C content of 50.85%.
Example 7:
a method for preparing fructus Rosae Normalis extract rich in vitamin C comprises:
(1) Cleaning fructus Rosae Normalis, freezing at-14deg.C for more than 24 hr, taking out frozen fructus Rosae Normalis, thawing, squeezing juice under 5kg pressure for 1.1 hr, and filtering to obtain fructus Rosae Normalis juice;
(2) Diluting the roxburgh rose juice obtained in the step (1) with purified water with the volume of 5 times of the roxburgh rose juice, and filtering with a 40KD ultrafiltration membrane to obtain a filtrate;
(3) Taking the filtrate obtained in the step (2), controlling the pressure to be 4kg, and concentrating by using a reverse osmosis membrane to obtain a roxburgh rose concentrated juice;
(4) Taking the roxburgh rose juice obtained in the step (3), adding 87% ethanol with the mass of 12 times, fully stirring, standing at the temperature of 5 ℃ for 13 hours, centrifuging to obtain supernatant, and concentrating the supernatant under reduced pressure to recover ethanol: vacuum degree is 0.085mpa, concentration temperature is 47 ℃, and alcohol precipitation concentrated solution is obtained;
(5) Loading HPD-100 macroporous resin on the alcohol precipitation concentrated solution obtained in the step (4), eluting with 30% alcohol aqueous solution as eluent, and concentrating the eluent to obtain extract with relative density of 1.15;
(6) And (5) taking the extract obtained in the step (5), and adopting vacuum drying: vacuum degree is 0.085mpa, drying temperature is 47 ℃, and fructus Rosae Normalis extract rich in vitamin C is obtained.
The obtained fructus Rosae Normalis extract rich in vitamin C has a vitamin C content of 53.92%.
Example 8:
a method for preparing fructus Rosae Normalis extract rich in vitamin C comprises:
(1) Cleaning fructus Rosae Normalis, freezing at-20deg.C for more than 24 hr, taking out frozen fructus Rosae Normalis, thawing, squeezing juice under 5kg pressure for 0.8 hr, and filtering to obtain fructus Rosae Normalis juice;
(2) Diluting the roxburgh rose juice obtained in the step (1) with purified water with the volume of 6 times of the roxburgh rose juice, and filtering with a 70KD ultrafiltration membrane to obtain a filtrate;
(3) Taking the filtrate obtained in the step (2), controlling the pressure to be 4kg, and concentrating by using a reverse osmosis membrane to obtain a roxburgh rose concentrated juice;
(4) Taking the roxburgh rose concentrated juice obtained in the step (3), adding 95% ethanol with the mass of 10 times, fully stirring, standing at 8 ℃ for 16 hours, centrifuging to obtain supernatant, and concentrating the supernatant under reduced pressure to recover ethanol: vacuum degree is 0.084mpa, concentration temperature is 43 ℃, and alcohol precipitation concentrated solution is obtained;
(5) Taking AB-8 macroporous resin on the alcohol precipitation concentrated solution obtained in the step (4), eluting by using an ethanol water solution with the alcohol content of 17% as an eluent, and concentrating the eluent to obtain an extract with the relative density of 1.10;
(6) And (5) freeze-drying the extract obtained in the step (5): the pre-freezing temperature is 25 ℃, the vacuum degree is 23Pa, the temperature of the water catcher is 42 ℃, and the temperature of the baffle plate is 5 ℃, so that the Rosa roxburghii extract rich in vitamin C is obtained.
The obtained fructus Rosae Normalis extract rich in vitamin C has a vitamin C content of 55.35%.
Example 9:
a method for preparing fructus Rosae Normalis extract rich in vitamin C comprises:
(1) Cleaning fructus Rosae Normalis, freezing at-12deg.C for more than 24 hr, taking out frozen fructus Rosae Normalis, thawing, squeezing juice under 5kg pressure for 1.0 hr, and filtering to obtain fructus Rosae Normalis juice;
(2) Diluting the roxburgh rose juice obtained in the step (1) with 4 times of purified water with the volume of the roxburgh rose juice, and filtering with a 50KD ultrafiltration membrane to obtain a filtrate;
(3) Taking the filtrate obtained in the step (2), controlling the pressure to be 3kg, and concentrating by using a reverse osmosis membrane to obtain a roxburgh rose concentrated juice;
(4) Taking the roxburgh rose concentrated juice obtained in the step (3), adding 85% ethanol with the mass of 10 times, fully stirring, standing at the temperature of 6 ℃ for 13 hours, centrifuging to obtain supernatant, and concentrating the supernatant under reduced pressure to recover ethanol: vacuum degree is 0.085mpa, concentration temperature is 43 ℃, and alcohol precipitation concentrated solution is obtained;
(5) Loading HPD-100 macroporous resin on the alcohol precipitation concentrated solution obtained in the step (4), eluting with purified water as an eluent, and concentrating the eluent to obtain extract with a relative density of 1.15;
(6) And (5) taking the extract obtained in the step (5), and adopting vacuum drying: vacuum degree is 0.085mpa, drying temperature is 47 ℃, and fructus Rosae Normalis extract rich in vitamin C is obtained.
The obtained fructus Rosae Normalis extract rich in vitamin C has a vitamin C content of 57.46%.
Example 10:
application of fructus Rosae Normalis extract rich in vitamin C:
the vitamin C-enriched Rosa roxburghii extract obtained in examples 1-9 can be directly used as raw material of antioxidant and whitening functional products.
The method for measuring the content of the ascorbic acid (vitamin C) in the Rosa roxburghii extract rich in the vitamin C (refer to the measurement of the ascorbic acid in food safety national standard food (standard number: GB 5009.86-2016)) comprises the following steps:
1. vitamin C content determination
1.1 chromatographic conditions:
chromatographic column: c18 column, column length 250mm, inner diameter 4.6mm, particle size 5 μm, or chromatographic column of equivalent performance; a detector: a diode array detector or an ultraviolet detector; mobile phase: 6.8g of monopotassium phosphate and 0.91g of cetyltrimethylammonium bromide, dissolved with water and fixed to a volume of 1L (pH was adjusted to 2.5-2.8 with phosphoric acid); and B, 100% of methanol. Mixing according to A:B=98:2, filtering with a 0.45 μm filter membrane, and ultrasonic degassing; flow rate: 0.7mL/min; detection wavelength: 245nm; column temperature: 25 ℃; sample injection amount: 20. Mu.L.
1.2 preparation of control vitamin C solution:
1.2.1L (+) -ascorbic acid standard stock solution (1.000 mg/mL): 0.01g (accurate to 0.01 mg) of L (+) -ascorbic acid standard substance is accurately weighed, and the volume is fixed to 10mL by using 20g/L metaphosphoric acid solution.
1.2.2D (-) -ascorbic acid standard stock solution (1.000 mg/mL): 0.01g (accurate to 0.01 mg) of D (-) -ascorbic acid standard is accurately weighed, and the volume is fixed to 10mL by using 20g/L metaphosphoric acid solution.
1.2.3 ascorbic acid Mixed Standard series working fluids: l (+) -ascorbic acid and D (-) -ascorbic acid standard stock solutions 0mL,0.05mL,0.50mL,1.0mL,2.5mL,5.0mL were pipetted, respectively, and the volumes were fixed to 100mL with 20g/L metaphosphoric acid solution. The concentrations of L (+) -ascorbic acid and D (-) -ascorbic acid in the standard series of working fluids were 0. Mu.g/mL, 0.5. Mu.g/mL, 5.0. Mu.g/mL, 10.0. Mu.g/mL, 25.0. Mu.g/mL, 50.0. Mu.g/mL, respectively. Is prepared in the clinic.
1.3 sample solution preparation: about 0.5g-2g (accurate to 0.001 g) of the homogenized sample relative to the sample was weighed, or 2mL-10mL of the liquid sample was aspirated [ about 0.03mg-6mg of L (+) -ascorbic acid was contained in the sample taken in a 50mL beaker ], and the sample was transferred to a 100mL volumetric flask with a 20g/L metaphosphoric acid solution, dissolved by shaking, and fixed in volume. Shaking, centrifuging at 4000r/min for 5min, sucking 1mL of supernatant, placing in a 10mL volumetric flask, diluting to scale with 20g/L metaphosphoric acid solution, shaking, filtering with 0.45 μm aqueous phase filter membrane, and measuring the filtrate.
1.4 assay: sucking 10 μl of each of the control solution and the sample solution, injecting into a liquid chromatograph, and calculating.
1.5 vitamin C-enriched Rosa roxburghii extract contains vitamin C for comparison, see Table 1.
Table 1 comparison table of the juice of Rosa roxburghii and vitamin C contained in the extract of Rosa roxburghii rich in vitamin C
Compared with the dry powder of the roxburgh rose juice prepared by directly squeezing and drying, the dry powder of the roxburgh rose extract rich in vitamin C has the advantages that the vitamin C content is greatly improved, the upper limit of the vitamin C content of the natural roxburgh rose juice is broken through, and the vitamin C content of the dry powder of the roxburgh rose extract rich in vitamin C is 257.50 percent higher than that of the natural roxburgh rose juice.
Conclusion compared with the prior art for the preparation of vitamin C:
comparative example 1:
the method comprises the following steps: the method of adopting the comparison document 1 ' a method for concentrating natural vitamin C of roxburgh rose ' of CN1060362709A ' comprises the following steps:
(1) And (3) cleaning at low temperature: fresh roxburgh rose fruit is put into a 2-section type cleaning tank. Removing sand, floating leaves and rotten fruits to obtain clean fructus Rosae Normalis with temperature of 15-20deg.C. The purpose of 2-section cleaning is that the first section of water is primary cleaning and needs to be discharged; the second stage of water is fine washing, filtering and circulating are needed, and the temperature of the center of the rosa roxburghii tratt fruit is controlled.
(2) Crushing and juicing at low temperature with oxygen isolation: the paddle beating machine is arranged above the feeding hole of the squeezer, and the whole body is sealed and insulated. Introducing carbon dioxide into the bottom of the equipment, removing oxygen in the equipment by using a feed inlet of the beating machine as an exhaust port, and keeping the whole equipment at micro positive pressure (10-100 Pa). The whole equipment has a heat preservation layer, the treatment time is 5 minutes, and the temperature of the materials is ensured to be 20-25 ℃.
(3) Removing tannin in a vacuum tank: spraying the squeezed juice into a vacuum tank for temporary storage (with the relative vacuum degree of-90 to-20 KPa), adding 0.1-1% of deacetylated chitin (meeting the use standard of GB2760 food additives) into the vacuum tank, mixing, standing for 5-30 minutes, and combining with tannin for precipitation. Filtering with plate-frame filter with diatomite as filter aid to remove tannin. The vacuum tank is arranged to remove air contained in fructus Rosae Normalis pulp, so as to avoid oxygen from damaging vitamin C. At this point the temperature is about 20-25 degrees.
(4) Microfiltration: micro-filtering with 0.1-0.5 μm ceramic membrane to obtain clear juice. In the process, the microfiltration also plays a role in removing microorganisms such as bacteria, prevents the membrane concentration system from being infected with bacteria, and ensures the product sanitation index. At this point the temperature is about 20-30 degrees.
(5) Concentrating by a reverse osmosis membrane: the clarified juice enters a membrane concentration system. The membrane material is RO reverse osmosis membrane, and adopts polyamide composite membrane with high low temperature permeability, the operation temperature is 25-35 ℃ and the operation pressure is 5-25MPa. A concentrated juice (corresponding to 8-fold concentration) with 64% solids was obtained. In view of the preservation property of vitamin C, the concentrated juice is packaged and sealed and stored in a-18-DEG refrigerator.
Conclusion: the cold processing and oxygen isolation technology in the preparation process has higher requirements on equipment and higher cost, is not suitable for being applied to large-scale industrial production, and the content of vitamin C in the concentrated juice of the roxburgh rose prepared by the method is lower than 18 percent, so that the method is not suitable for enriching the vitamin C.
Comparative example 2:
the method comprises the following steps: the method of adopting a vitamin C-enriched roxburgh rose powder and a preparation method thereof in a comparison document 2 of CN111700247B comprises the following steps:
(1) Squeezing fresh fructus Rosae Normalis to obtain fructus Rosae Normalis juice;
(2) Transferring fructus Rosae Normalis juice into a fermentation tank, adding 0.3% activated Saccharomyces cerevisiae, setting the temperature to 25-40deg.C, and stirring and fermenting for 4 hr to obtain fermented fructus Rosae Normalis juice;
(3) Filtering the fermentation liquor with 250 mesh filter cloth, filtering the filtrate with ceramic membrane with built-in 80 nanometer filter core, and collecting filtrate for use;
(4) Concentrating the filtered fructus Rosae Normalis juice under vacuum with vacuum concentration degree of-0.04 MPa and temperature of 45-75deg.C until solid content is 20-40% to obtain extract;
(5) Mixing the above extract with 25% modified starch, and spray drying at 160-205 deg.C and 70-100 deg.C to obtain dry powder; sieving to obtain fructus Rosae Normalis powder.
Conclusion: the preparation process comprises adding 25% modified starch, and spray drying, which is unfavorable for enriching vitamin C, so that the content of vitamin C in the fructus Rosae Normalis powder prepared by the method is low and is more than 17%, and the method is not suitable for enriching vitamin C.
Comparative example 3:
the method comprises the following steps: the method for extracting vitamin C from Rosa roxburghii by adopting comparative document 3, namely a method for extracting vitamin C from Rosa roxburghii by using the following steps:
(1) Grinding and enzymolysis: firstly, cleaning and sterilizing air-dried fructus Rosae Normalis, pulverizing into 90-120 meshes, adding water with volume 1 times of fructus Rosae Normalis fruit, stirring for 5min, and adding ethanol vapor and CO 2 Grinding for 6min in a grinding tank as protective gas, adding compound enzyme with mass of 0.05% of that of fructus Rosae Normalis fruit, wherein the compound enzyme is A, B, wherein A is cellulase, amylase and protease, and B is superoxide dismutase, adding compound enzyme A and compound enzyme B, continuously grinding for 20-40min, and sterilizing for 5-8min to obtain fructus Rosae Normalis mixed solution;
(2) Extracting: adding tea saponin and chitosan into sterilized fructus Rosae Normalis mixed solution, continuously oscillating for 10min, adding 24% sodium chloride, centrifuging, and filtering to obtain clear fructus Rosae Normalis permeate;
(3) Separating and concentrating: adding mixed acid into the clear permeate of the roxburgh rose obtained in the step 2, separating by using separation membrane equipment to obtain a separation membrane permeate, concentrating the separation membrane permeate by using ultrafiltration membrane equipment again, adding alkali, neutralizing the mixed acid in the concentrate to obtain vitamin C concentrate, freeze-drying at-15 ℃, and crystallizing to obtain the vitamin C concentrate.
Conclusion: the method has the advantages that the variety of auxiliary materials used is large, the production cost is increased, the grinding treatment step is suitable for dry Rosa roxburghii fruit, the preparation process is complex, the vitamin C content in the product obtained by the method is 398mg/g, the content is low, the technological process is complex, and the method is not suitable for industrial production application.
Experimental example:
the inventor of the invention carries out in-vitro antioxidant function test and whitening efficacy test research on the vitamin C-enriched Rosa roxburghii extract prepared by the invention, and part of test contents are extracted as follows:
1. in vitro antioxidant function test:
DPPH radical scavenging Rate measurement experiment
DPPH solution preparation
Precisely weighing DPPH reagent, adding absolute ethyl alcohol to prepare 0.1mmol/L DPPH solution, and refrigerating in dark place; sample liquid preparation: respectively weighing 0.5g, 0.4g, 0.3g, 0.2g, 0.1g and 0.05g of the Rosa roxburghii extract rich in vitamin C, rosa roxburghii juice and positive control solution (containing 1mg/mL of vitamin C), placing the Rosa roxburghii extract, the Rosa roxburghii juice and the positive control solution into a 10mL volumetric flask, adding distilled water to prepare solutions with different concentration gradients, shaking uniformly, and measuring.
2. Determination of DPPH removal Rate of raw Material
Taking 2mL of samples with different concentration gradients, adding 2.0mL of DPPH solution into each test tube, reacting for 30min under the condition of avoiding light in a greenhouse, fully shaking uniformly, measuring the absorbance value, and recording the absorbance value at 517nm as A j The method comprises the steps of carrying out a first treatment on the surface of the 2mL of each of the samples of various gradients was taken, 2.0mL of distilled water was added to each tube, and the absorbance at 517nm was recorded as A i 。
The control group was 2mL DPPH solution with 2mL distilled water, and the absorbance value was recorded as A c 。
As positive control, vitamin C solution at equal concentration was used.
3. DPPH radical scavenging (K) was calculated as follows:
the DPPH radical scavenging ability test results of the vitamin C-enriched Rosa roxburghii extract and Rosa roxburghii juice are shown in Table 2.
TABLE 2 determination of DPPH radical scavenger of vitamin C enriched Rosa roxburghii extract
Note that: p <0.05, P <0.01 compared to vitamin C enriched rosa roxburghii extract.
4. Conclusion(s)
The extract of Rosa roxburghii containing rich vitamin C has obviously enhanced DPPH free radical scavenging ability compared with Rosa roxburghii juice and positive control vitamin C.
(II) OH radical scavenger measurement experiment
1. Preparation of stock solutions
Precisely weighing salicylic acid, adding distilled water to prepare 10mmol/mL solution, and marking as stock solution A; precisely weighing ferrous sulfate, adding distilled water to prepare 10mmol/mL solution, and marking as stock solution B; precisely measuring hydrogen peroxide, adding distilled water to prepare 100mmol/mL solution, and marking the solution as stock solution C for preparation at present; preparing a sample to be tested: respectively taking 0.5g, 0.4g, 0.3g, 0.2g, 0.1g and 0.05g of the Rosa roxburghii extract rich in vitamin C, rosa roxburghii juice and positive control solution (containing 1mg/mL of vitamin C), placing the Rosa roxburghii extract rich in vitamin C, rosa roxburghii juice and positive control solution into a 10mL volumetric flask, adding distilled water to prepare solutions with different concentration gradients, shaking uniformly, and measuring.
2. Determination of OH radical scavenging Rate of Rosa roxburghii extract rich in vitamin C
Sequentially taking 0.5mL of each concentration gradient sample, adding 0.5mL of stock solution A,0.5mL of stock solution B and 3.5mL of distilled water into a test tube, adding 5mL of stock solution C to start the reaction, shaking uniformly, and measuring the absorbance value A at 510nm m The method comprises the steps of carrying out a first treatment on the surface of the Taking 0.5mL of each concentration gradient sample, adding only stock solution A and 4mL of distilled water, adding 5mL of stock solution C to start reaction, shaking, and measuring absorbance value A at 510nm n 。
The control group had no sample solution, only 0.5mL of each of the stock solutions A, B and 4mL of distilled water were added, and 5mL of the stock solution C was added, and the absorbance value thereof was recorded as A c 。
As a positive control, a vitamin C (hereinafter referred to as vitamin C) solution of equal concentration was used.
3. The OH radical scavenging rate (K) was calculated as follows:
the test results of the OH free radical scavenging ability of the vitamin C-enriched Rosa roxburghii extract and Rosa roxburghii juice are shown in Table 3.
TABLE 3 determination of OH free radical scavenger of vitamin C enriched Rosa roxburghii extract
Note that: p <0.05, P <0.01 compared to vitamin C enriched rosa roxburghii extract.
4. Conclusion(s)
The extract of Rosa roxburghii rich in vitamin C has obviously enhanced OH free radical scavenging ability compared with positive control vitamin C and Rosa roxburghii juice.
2. Whitening experiment (tyrosinase inhibition experiment)
1. The tyrosinase-catalyzed L-dopa is detected to generate the dopa pigment, the tyrosinase-catalyzed L-dopa pigment has a characteristic absorption peak at 475nm, the generation of the dopa pigment is inhibited in the presence of an antioxidant, and the tyrosinase inhibition capacity of a sample can be measured and calculated by measuring the absorbance of the dopa pigment at 475 nm.
2. Extracting tyrosinase, namely adding 0.1g of potatoes into 1mL of extracting solution for homogenate grinding, centrifuging, and taking supernatant to obtain tyrosinase extracting solution.
3. And (3) preparing the Rosa roxburghii juice and the Rosa roxburghii extract rich in vitamin C from the sample solution, and respectively diluting the Rosa roxburghii juice and the Rosa roxburghii extract rich in vitamin C into sample solutions with different concentration gradients, wherein the total reaction system is 200mL. Taking 20mL of samples to be tested with different concentration gradients, respectively and sequentially adding 140mL of a first reagent and 20mL of tyrosinase extract, reacting for 10min at 37 ℃, measuring the absorbance at 475nm, replacing a blank with purified water, measuring 3 times in parallel for each sample, and taking a vitamin C control solution as a positive control. Calculating tyrosinase inhibition rate according to the formula, drawing a correlation curve of concentration gradient of the standard extract and the tyrosinase inhibition rate, and calculating IC of the sample 50 The concentration (the concentration of the sample required for 50% tyrosinase inhibition) was compared to the tyrosinase inhibitory capacity of the Rosa roxburghii juice and the vitamin C enriched Rosa roxburghii extract.
4. Calculation formula
Tyrosinase inhibition ratio% = [ (a-B) - (C-D) ]/(a-B) ×100
Wherein A is the absorbance value of a reaction system containing tyrosinase but no sample; b is the absorbance value of the reaction system without tyrosinase and without sample; c is the absorbance value of the reaction system containing tyrosinase and containing samples; d is the absorbance value of the reaction system containing no tyrosinase but containing samples.
The tyrosinase inhibition test results of the vitamin C-enriched Rosa roxburghii extract and Rosa roxburghii juice are shown in Table 4.
TABLE 4 vitamin C enriched Rosa roxburghii extract tyrosinase inhibition rate assay table
Note that: compared with the extract of Rosa roxburghii rich in vitamin C, P<0.05，**P<0.01， Δ P>0.05。
5. Conclusion(s)
The extract of Rosa roxburghii containing rich vitamin C has remarkably enhanced tyrosinase inhibition rate compared with Rosa roxburghii juice, and has enhanced tendency compared with positive control vitamin C.
Although the present invention has been described in detail by combining preferred embodiments with experimental examples, the present invention is not limited thereto. Various equivalent modifications and substitutions for embodiments of the invention may be made by those skilled in the art without departing from the spirit and scope of the invention, and these modifications and substitutions are intended to be within the scope of the invention. Any person skilled in the art can easily think of changes or substitutions within the technical scope of the present disclosure, and the present disclosure is intended to be covered by the present disclosure. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (10)
1. A preparation method of a roxburgh rose extract rich in vitamin C is characterized by comprising the following steps:
(1) Cleaning fructus Rosae Normalis, freezing, and squeezing to obtain fructus Rosae Normalis juice;
(2) Diluting fructus Rosae Normalis juice with purified water, and filtering with ultrafiltration membrane to obtain filtrate;
(3) Concentrating the filtrate obtained in the step (2) by using a reverse osmosis membrane to obtain a roxburgh rose concentrated juice;
(4) Collecting fructus Rosae Normalis concentrated juice, adding ethanol, stirring, standing at low temperature, centrifuging to obtain supernatant, and recovering ethanol at vacuum degree and concentration temperature to obtain ethanol precipitation concentrate;
(5) Purifying the ethanol precipitation concentrate with macroporous resin, eluting with purified water and ethanol-containing water solution, collecting eluate, and concentrating to obtain extract;
(6) Drying the extract to obtain fructus Rosae Normalis extract rich in vitamin C.
2. The method for preparing a vitamin C enriched rosa roxburghii extract according to claim 1, comprising the steps of:
(1) Cleaning fresh Rosa roxburghii fruit, freezing for more than 24 hours at the temperature of-4 to-20 ℃, taking out the frozen fresh Rosa roxburghii fruit, thawing, squeezing juice at the pressure of 3-7 kg for 0.5-1.5 hours, and filtering to obtain Rosa roxburghii juice;
(2) Diluting the roxburgh rose juice with purified water with the volume of 1-7 times of that of the roxburgh rose juice, and filtering with a 10-100KD ultrafiltration membrane to obtain a filtrate;
(3) Collecting filtrate, controlling the pressure to be 1-6 kg, and concentrating by using a reverse osmosis membrane to obtain concentrated juice of roxburgh rose;
(4) Taking concentrated juice of roxburgh rose, adding 80-95% ethanol with the mass of 8-12 times, fully stirring, standing for 10-16 hours at the temperature of 2-8 ℃, centrifuging, taking supernatant, concentrating the supernatant under reduced pressure and vacuum degree of 0.08-0.09 mpa, concentrating at the temperature: 40-55 ℃ to obtain an alcohol precipitation concentrated solution;
(5) Taking AB-8 or HPD-100 or D101 macroporous resin on the alcohol precipitation concentrated solution, adopting purified water or 5-30% aqueous solution containing ethanol as eluent, concentrating the eluent to extract with relative density of 1.10-1.20;
(6) The extract is dried in vacuum: vacuum degree is 0.08-0.09 mpa, and drying temperature is 40-55 ℃; or freeze drying: prefreezing temperature: -10-50 ℃, the vacuum degree of 10-30 Pa, the temperature of the water catcher of 35-45 ℃ and the temperature of the baffle of-10 ℃ to obtain the roxburgh rose extract rich in vitamin C.
3. The method for preparing the vitamin C enriched rosa roxburghii tratt extract according to claim 2, characterized in that: in the step (1), the freezing temperature is-12 ℃, the juice squeezing pressure is 5kg, and the juice squeezing time is 1 hour.
4. A process for the preparation of vitamin C enriched rosa roxburghii tratt extract according to claim 3, characterized in that: in the step (2), 4 times of the volume of the rosa roxburghii tratt juice is used for purifying water for dilution, and a 50KD ultrafiltration membrane is used for filtering.
5. The method for preparing the vitamin C-enriched Rosa roxburghii extract according to claim 4, wherein: in the step (3), the control pressure is 3kg.
6. The method for preparing the vitamin C-enriched Rosa roxburghii extract according to claim 5, wherein: in the step (4), 10 times of 85% ethanol is adopted, the ethanol is subjected to ethanol precipitation at 6 ℃ for 13 hours, the ethanol recovery vacuum degree is 0.085mpa, and the concentration temperature is 43 ℃.
7. The method for preparing the vitamin C enriched rosa roxburghii tratt extract according to claim 6, wherein the steps of: in the step (5), macroporous resin is HPD-100, purified water is used as eluent for eluting, and eluent is concentrated to extract with the relative density of 1.15.
8. The method for preparing the vitamin C enriched rosa roxburghii tratt extract according to claim 7, wherein the method comprises the following steps: in the step (6), the extract is dried in vacuum, the vacuum degree is 0.085mpa, and the drying temperature is 47 ℃.
9. The vitamin C-enriched Rosa roxburghii extract prepared by the preparation method according to any one of claims 1 to 8, wherein the vitamin C content of the extract is more than or equal to 50%.
10. The use of the vitamin C enriched extract of rosa roxburghii as claimed in claim 8 for anti-oxidation and whitening.
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| CN117503661B (en) * | 2024-01-04 | 2024-05-07 | 珠海远大美业生物科技有限公司 | Preparation method of Rosa roxburghii extract and application of freeze-dried powder thereof in cosmetics |
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