Freeze-dried powder of natural vitamin c

Abstract

The present invention provides a novel freeze-dried powder of vitamin C from Rosa Roxbunghii and a method of preparation thereof. The freeze-dried powder of the present invention surprisingly provides light-color, uniform and stable cake appearance in penicillin vial and efficiently prevents degradation of vitamin C comprised in the powder.

Description

[0001] FREEZE-DRIED POWDER OF NATURAL VITAMIN C

[0002] Technical field

[0003]

[0001] The present invention is related to a new formulation of natural vitamin C. Particularly it is related to a new freeze-dried powder of vitamin C from natural resource such as Rosa Roxbunghii.

[0004] Background of the invention

[0005]

[0002] Vitamin C, also named ascorbic acid, is one of the water-soluble vitamins. Vitamin C is an essential element for life and has been widely used in health care, beverages and beauty.

[0006]

[0003] Vegetable and fruits are the primary sources of vitamin C for human being. However, daily intake of vitamin C from vegetable and fruits is not sufficient and normally it is necessary to supplement additional vitamin C. Nowadays, most of vitamin C in the market is synthetic, and natural vitamin C, which is obtained from natural plant such as vegetable and fruits, is becoming more and more popular.

[0007]

[0004] One good resource of natural vitamin C is Rosa Roxbunghii, Fresh fruits of Rosa Roxbunghii are rich of vitamin C, upto 35411 mg / kg (see Chinese patent publication CN 101926496 B). At present, Rosa Roxbunghii is often made to fruit juice. However, Rosa Roxbunghii juice cannot be applied widely in industry as it cannot be stored for a long time and has also acidity and bitter flavor.

[0008]

[0005] Chinese patent publication CN 101073433 B discloses a process for processing Rosa Roxbunghii juice to solid products, comprising adding gelatin and chitosan into Rosa Roxbunghii juice, followed by adding auxiliary material for granulating. Described possible auxiliary materials include a long list of starch, dextrin, polypeptide, protein, dietary fiber, monose, disaccharides, compound sugar, polysaccharide, maltitol, xylitol, stevioside glycosides, albumen sugar and sucralose.

[0009]

[0006] Chinese patent publication CN 101156670 B discloses a process for producing Rosa Roxbunghii powders, comprising treating Rosa Roxbunghii juice by pectinase, adding carriers for drying after ultrafiltration and concentration to provide Rosa Roxbunghii powder. However, these solid products comprise the natural vitamin C in a low content (less than 15wt%) and are unstable.

[0007] Therefore, there is demand in solid products of Rosa Roxbunghii with a high natural vitamin C content and which exhibit an improved stability.

[0010] Summary of the invention

[0011]

[0008] The present invention provides a novel freeze-dried powder of vitamin C from Rosa Roxbunghii and a method of preparation thereof.

[0012]

[0009] Accordingly, in a first aspect, the invention provides a freeze-dried powder of vitamin C from Rosa Roxbunghii, comprising mannitol as excipient.

[0010] In a second aspect, the invention provides a process for the preparation of such freeze- dried powder, comprising the step of freeze drying a solution comprising Rosa Roxbunghii juice and mannitol.

[0013]

[0011] The freeze-dried powder of the present invention surprisingly provides light-color, uniform and stable cake appearance in penicillin vials and efficiently prevents degradation of vitamin C contained in the powder. Such is illustrated by the examples below.

[0014] Description of the Figures

[0015]

[0012] The present invention is illustrated by the following figures:

[0016]

[0013] Figure 1 shows appearance of the freeze-dried powders with different excipients: a) 4wt% mannitol; b) 5wt% mannitol; c) 6wt% mannitol; d) 7wt% mannitol; e) 8wt% mannitol; f) 5wt% mannitol+ 1wt% trehalose; and g) from left to right: with 6wt% mannitol, 6wt% lactose, 6wt% NaCI, 6wt% glycine, 6wt% citric acid, 6wt% surcose, 6wt% mycose and 6wt% malto-dextrin; and

[0017]

[0014] Figure 2 shows appearance of the freeze-dried powders with different adjuvants and with N2 bubbling and ultrasonic treatment, from left to right: Group B1 , Group C1 , Group C3, Group C4, Group D1 , Group D2, Group D3, Group E5 and Group E6.

[0018] Detailed description of the invention

[0019]

[0015] The present invention provides a novel freeze-dried powder of vitamin C from Rosa Roxbunghii, comprising mannitol as excipient.

[0020]

[0016] Preferably, the freeze-dried powder of the present invention comprises 20-60wt%, more preferably 25-55wt%, the most preferably 30-50wt% of mannitol, based on the total weight of the powder.

[0021]

[0017] Preferably, the freeze-dried powder of the present invention comprises an additional adjuvant. The adjuvant may be selected from the group consisting of cysteine or salts thereof, citric acid or salts thereof, Na2SO3, NaCI and CaCh, and mixture thereof, preferably cysteine or salts thereof and citric acid or salts thereof, and more preferably cysteine hydrochloride.

[0022]

[0018] In all embodiments of the present invention, the freeze-dried powder preferably comprises 1-10wt%, more preferably 2-8wt%, most preferably 3-7wt% such as 3.5wt%, 4wt%, 4.5wt% and 5wt% of an adjuvant, based on the total weight of the powder such as in particular cysteine hydrochloride.

[0023]

[0019] In one particular embodiment, the freeze-dried powder of the present invention comprises mannitol as excipient and cysteine hydrochloride as adjuvant. In a preferable embodiment, the freeze-dried powder of the present invention comprises 25-55wt% mannitol and 1-10wt% cysteine hydrochloride, based on the total weight of the powder.

[0024]

[0020] In all embodiments of the present invention, the freeze-dried powder preferably comprises 15-30wt%, more preferably 17-25wt% such as 18wt%, 19wt%, 20wt% and 21wt% vitamin C, based on the total weight of the powder.

[0021] Preferably, the freeze-dried powder of the present invention further comprises 5-15wt%, preferably 8-10wt% flavonoids, and / or 3-12wt%, preferably 5-10wt% polyphenols, based on the total weight of the powder. Without wishing to be bound by any kind of theory it is believed that the presence of such flavonoids and / or polyphenols may result in advantageous anti-oxidant properties.

[0025]

[0022] Surprisingly, by using mannitol as excipient, the freeze-dried powder of the present invention provides lighter, more uniform and stabler cake appearance in penicillin vials compared to the powders comprising other excipients. In addition, by adding the adjuvant as described above, the freeze-dried powder of the present invention efficiently prevents degradation of vitamin C contained therein.

[0026]

[0023] Particularly, under a stability test at 40 °C for 7 days, for example, as described in Example 3 of the present application, the freeze-dried powder of the present invention has less than 10wt%, preferably less than 5wt%, more preferably less than 3wt%, the most preferably less than 2wt% of degradation of vitamin C, measured by HPLC according to standard methods in the art.

[0027]

[0024] More particularly, under a hygroscopicity test at 22 °C and 61 .7% humidity, for example, as described in Example 4 of the present application, the freeze-dried powder of the present invention exhibits less than 5wt%, preferably less than 3wt%, more preferably less than 2wt% of weight increase in 60 mins, illustrating a low hygroscopicity of the powder.

[0028]

[0025] More particularly, under a solubility test, for example, as described in Example 4 of the present application, 350 mg of the freeze-dried powder of the present invention is completely dissolved in 1 mL water at room temperature in 60 seconds, preferably 30 seconds, more preferably 20 seconds.

[0029]

[0026] In all embodiments of the present invention preferably mannitol is used as main excipient. The term main excipient refers to an amount of at least 80wt%, preferably at least 83wt% of mannitol, based on the total amount of excipients. Most preferably mannitol is the sole excipient.

[0030]

[0027] In all embodiments of the present invention preferably the freeze-dried powder consists essentially of vitamin C, flavonoid(s) and polyphenol(s) from Rosa Roxbunghii, and cysteine or a salt thereof such as preferably cysteine hydrochloride, and mannitol, with all the concentration ranges and preferences as given herein.

[0031]

[0028] The present invention also provides a process for the preparation of the freeze-dried powder of the present invention, which comprises the step of freeze drying a solution comprising Rosa Roxbunghii juice and mannitol to obtain the freeze-dried powder.

[0032]

[0029] Surprisingly, inventors found mannitol to behave quite different in the preparation process from many other excipients. Mannitol has an advantageous low solubility in water (only 20g of mannitol can be soluble in 100 g water at 20 °C), it advantageously allows for a high eutectic temperature (-1 .5 °C), and it exhibits a low hygroscopicity. Without wishing to be bound by any kind of theory it is believed that within the process of the invention, the mannitol may advantageously increase the tendency towards crystallization during freezing and / or may advantageously reduce or avoid melting and / or may advantageously increase sublimation rate.

[0030] In the process of the present invention, the Rosa Roxbunghii juice is any juice from Rosa Roxbunghii fruits which may be obtained commercially or by crushing, squeezing and / or extracting Rosa Roxbunghii fruits or pomace according to known processes in the art.

[0033]

[0031] Preferably, the Rosa Roxbunghii juice used in the process of the present invention is subjected to one, two or three treatments selected from fermentation with yeast, concentration and / or activated carbon treatment. More preferably, the Rosa Roxbunghii juice used in the process of the present invention is subjected to fermentation with yeast and concentration or activated carbon treatment. Most preferably, the Rosa Roxbunghii juice used in the process of the present invention is subjected to fermentation with yeast and activated carbon treatment.

[0034]

[0032] As anticipated by any person known in the art, the treatments of fermentation with yeast, concentration and / or activated carbon treatment can be included in the process of the present invention in any sequence. For example, the Rosa Roxbunghii juice used in the process of the present invention is subjected to fermentation with yeast, followed by concentration and then activated carbon treatment, or followed by activated carbon treatment and then concentration.

[0035]

[0033] In one embodiment, the Rosa Roxbunghii juice used in the process of the present invention is subjected to fermentation with yeast followed by activated carbon treatment.

[0036]

[0034] In the process of the present invention, the fermentation with yeast and the concentration may be carried out as disclosed in patent publication WO 2016146029 A1 .

[0037]

[0035] In the process of the present invention, activated carbon treatment may be carried out at 20-60 °C. The activated carbon may be used in an amount of 1-5wt% based on the weight of the juice to be treated.

[0038]

[0036] Preferably, in the process of the present invention, the Rosa Roxbunghii juice used comprises 12-20wt%, preferably 14-18wt% of solid content.

[0039]

[0037] Preferably, an adjuvant is added into the Rosa Roxbunghii juice in the process of the present invention. In one embodiment, the adjuvant is added before any of the above treatments. More preferably, the adjuvant is added after the fermentation with yeast and before the activated carbon treatment.

[0040]

[0038] The adjuvant suitable for use in the process of the present invention may be selected from the group consisting of cysteine or salts thereof, citric acid or salts thereof, Na2SOs, NaCI and CaCh, and mixture thereof, preferably from cysteine or salts thereof and citric acid or salts thereof. Most preferably, the adjuvant is cysteine hydrochloride. The adjuvant may be added in an amount of 0.1-1wt% based on the weight of the juice.

[0041]

[0039] In the process of the present invention, the Rosa Roxbunghii juice and mannitol are admixed to provide the solution. The mannitol may be added in an amount of 4-8wt% based on the weight of the juice.

[0042]

[0040] Optionally, the solution may be subjected to further treatments including but not limited to nitrogen bubbling treatment and / or ultrasonic treatment. Without wishing to be bound by any theory, it is believed that these treatments would be helpful for good cake appearance of the freeze-dried powder of the present invention in penicillin vials.

[0041] In a particular advantageous embodiment the process for the preparation of a freeze-dried powder according to the present invention is a process comprising the consecutive steps of a) Fermentation of Rosa Roxbunghii (RR) juice with yeast to obtain a fermented RR mixture, b) Filtration of the fermented RR mixture to obtain a fermented RR filtrate, c) Optionally adding to the fermented RR filtrate an adjuvant with all the definitions and preferences as given herein, d) Optionally treating the fermented RR filtrate obtained in b) or c) with active charcoal, followed by e) Addition of mannitol, followed by f) Freeze drying.

[0043]

[0042] It is well understood that all the definitions and preferences, including the concentrations as given herein, apply to the process individually as well as in combination.

[0044]

[0043] In a preferred embodiment, the yeast is incubated before addition to the Rosa Roxbunghii Juice.

[0045]

[0044] In the process of the present invention, the freeze-drying is run according to the manufacture manual of freeze-dryer.

[0046]

[0045] According to the process of the present invention, the freeze-dried powder is provided in a light-colored, uniform and stable appearance with high content of vitamin C (up to 30wt%) and has reduced degradation of vitamin C.

[0047]

[0046] As anticipated by any person skilled in the art, the freeze-dried powder of the present invention can be used in various applications such as food, nutritional products, medicines and / or cosmetic products. Accordingly, the present invention also provides a food, a nutritional product, a medicine and / or cosmetic product comprising freeze-dried powder of the present invention.

[0048]

[0047] The present invention further provides use of the freeze-dried powder of the present invention in the preparation of a food, a nutritional product, a medicine and / or a cosmetic product.

[0049]

[0048] The present invention will be further illustrated in the examples below.

[0050] Examples

[0051] Example 1 : Appearance of the freeze-dried powders with different excipients

[0052]

[0049] 18 g yeast was added into 90 g water at 40°C. The mixture was put into a shaking incubator and incubated with 120 rpm / min at 40 °C for 30 mins. The obtained mixture was then added into 1 kg commercially purchased Rosa Roxbunghii juice (Hengliyuan group, China) which was preheated to 40 °C. The juice was fermented in a shaking incubator with 120 rpm / min at 40 °C for 6 h. After fermentation, the juice was filtered. To the obtained filtrate was added various excipients (4wt% mannitol, 5wt% mannitol, 6wt% mannitol, 7wt% mannitol, 8wt% mannitol, 5wt% mannitol+ 1wt% trehalose, 6wt% mycose, 6wt% lactose, 6wt% sucrose, 6wt% glycine, 6wt% NaCI, 6wt% citric acid and 6wt% malto-dextrin, respectively) and injected into penicillin vials. All the vials were put in a freeze-drier for freeze drying. Only the powders with mannitol as excipient had uniform and stable cake appearance in the vials as shown in Figure 1 .

[0053] Example 2: Appearance of the freeze-dried powders with different adjuvants and treatments

[0054]

[0050] 18 g yeast was added into 90 g water at 40 °C. The mixture was put into a shaking incubator and incubated with 120 rpm / min at 40 °C for 30 mins. The obtained mixture was then added into 1 kg commercially purchased Rosa Roxbunghii (Hengliyuan group, China) which was pre-heated to 40 °C. The juice was fermented in a shaking incubator with 120 rpm / min at 40 °C for 6 h. After fermentation, the juice was filtered. The obtained filtrate was treated in different ways as follow:

[0055] • Group B1 : pH of filtrate was adjusted to 3 with citric acid

[0056] • Group C1 : filtrate was added 0.5 wt% NaCI

[0057] • Group C3: filtrate was added 0.5 wt% CaCh

[0058] • Group C4: filtrate was added 0.5 wt% cysteine hydrochloride

[0059] • Group D2: filtrate was bubbled with nitrogen gas for 20 mins

[0060] • Group D3: filtrated was treated with ultrasound for 20 mins

[0061] • Group E5: filtrate was added 0.4 wt% Na2SO3

[0062] • Group D1 , Group E6: no extra treatment.

[0063]

[0051] After respective treatment, each group was added 5wt% activated charcoal and heated at 40 °C for an hour. The activated charcoal was filtered. For each group except Group D1 , obtained filtrate was added 6wt% mannitol and Group D1 was added 6 wt% modified starch as excipient. All groups were injected into penicillin vials. All the vials were put in a freeze-drier for freeze drying. As shown in Figure 2, the treatments Group B1 , Group C4, Group D2, Group D3, Group E5 and Group E6 provided acceptable appearance in the vials, while Group C1 and Group C3 provided cake with higher granularity and rougher surface, and Group D1 provided a shrunken cake. Example 3: Tests of vitamin C degradation in the freeze-dried powders

[0064]

[0052] The freeze-dried powder produced according to Example 2 were put in a thermostat at 40°C for 7 days for stability test. Before and after the test, vitamin C content (wt%) of each group was measure by HPLC. The results are shown in Table 1 .

[0065] Table 1

[0066] Example 4: Solubility and hygroscopicity tests of the freeze-dried powders.

[0067]

[0053] 18 g yeast was added into 90 g water at 40 °C. The mixture was put into a shaking incubator and incubated with 120 rpm / min at 40 °C for 30 mins. The obtained mixture was then added into 1 kg commercially purchased Rosa Roxbunghii juice (Hengliyuan group, China) which was preheated to 40 °C. The juice was fermented in a shaking incubator with 120 rpm / min at 40 °C for 6 h. After fermentation, the juice was filtered. The obtained filtrate was added 0.5wt% L-cysteine hydrochloride, 2wt% activated charcoal and heated at 40 °C for an hour. The activated charcoal was filtered. The obtained filtrate was added 6wt% mannitol and injected into penicillin vials. All the vials were put in a freeze-drier for freeze drying to obtain a freeze-dried powder with light-colored, uniform, and stable appearance with 19.12wt% vitamin C content, 8.8wt% flavonoid and 5.6wt% polyphenol measured via HPLC.

[0068] Solubility test

[0069]

[0054] 350 mg of the obtained freeze-dried powder was put into 1 mL water at room temperature. It was observed that the freeze-dried powder was fully dissolved and turned into a clear solution in 20 seconds.

[0070] Hygroscopicity test

[0055] Two samples (300 mg for each) were taken and kept open in air at 22 °C with humidity 61.7%. The weight increase was measured at 5, 10, 15, 20, 20, 30, 60 mins and the results are shown in Table 1 . As shown in Table 2, the freeze-dried powder of the present invention has less than 1wt% weight increase in 30 mins and less than 2wt% weight increase in 60 mins.

[0071] Table 2

Claims

CLAIMS1 . A freeze-dried powder of vitamin C from Rosa Roxbunghii, comprising mannitol as excipient.

2. The freeze-dried powder of claim 1 , which comprises 20-60wt%, more preferably 25-55wt%, most preferably 30-50wt% of mannitol, based on the total weight of the powder.

3. The freeze-dried powder of claim 1 and / or 2, which comprises an additional adjuvant selected from the group consisting of cysteine or salts thereof, citric acid or salts thereof, Na2SOs, NaCI and CaCh, and mixture thereof, preferably from cysteine or salts thereof and citric acid or salts thereof, most preferably the adjuvant is cysteine hydrochloride.

4. The freeze-dried powder of any one or more of claims 1 to 3, which comprises 25-55wt% mannitol and 1-10wt% cysteine hydrochloride, based on the total weight of the powder.

5. The freeze-dried powder of any one or more of claims 1-4, which comprises 15-30wt%, preferably 17-25wt% vitamin C, based on the total weight of the powder6. The freeze-dried powder of any one or more of claims 1-5, which comprises 5-15wt%, preferably 8-10wt% flavonoid, and / or 3-12wt%, preferably 5-10wt% polyphenol, based on the total weight of the powder.

7. The freeze-dried powder of any one or more of claims 1-6, which upon storage at 40°C for 7 days exhibits less than 10wt%, preferably less than 5wt%, more preferably less than 3wt%, most preferably less than 2wt% of degradation of vitamin C.

8. The freeze-dried powder of any one or more of claims 1-7, which under a hygroscopicity test exhibits less than 5wt%, preferably less than 3wt%, more preferably less than 2wt% of weight increase in 60 mins.

9. The freeze-dried powder of any one or more of claims 1-8, wherein 350 mg of the powder is completely dissolved in 1 mL water at room temperature in 60 seconds, preferably 30 seconds, more preferably 20 seconds.

10. A process for the preparation of the freeze-dried powder of any one of claims 1-9, comprising the step of freeze drying a solution comprising Rosa Roxbunghii juice and mannitol.11 . The process of claim 10, wherein the Rosa Roxbunghii juice is subjected to one, two or three treatments selected from the group consisting of fermentation with yeast, concentration and / or activated carbon treatment.

12. The process of claim 10, wherein the Rosa Roxbunghii juice is subjected to fermentation with yeast followed by activated carbon treatment.

13. The process of any one of claims 10-12, wherein an adjuvant is added into the Rosa Roxbunghii juice.

14. The process of claim 13, wherein the adjuvant is selected from the group consisting of cysteine or salts thereof, citric acid or salts thereof, Na2SOs, NaCI and CaCh, and mixture thereof, preferably cysteine or salts thereof and citric acid or salts thereof, and more preferably cysteine hydrochloride.

15. The process of claim 13, wherein the adjuvant is added in an amount of 0.1-1wt%, based on the weight of the juice.

16. The process of any one of claims 10-15, wherein mannitol is added in an amount of 4-8wt%, based on the weight of the juice.

17. The process of any one of claims 10-16, wherein the solution is subjected to nitrogen bubbling treatment and / or ultrasonic treatment.

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