Vitamin C injection with low impurity content and preparation method thereof

Abstract

The invention discloses a vitamin C injection with low impurity content and a preparation method thereof, and belongs to the technical field of pharmaceutical preparation preparation. According to the vitamin C injection disclosed by the invention, D-calcium gluconate, L-cysteine and polyethylene glycol-6000 are adopted to prepare a stabilizer, and the three components have a synergistic effect, so that the stability of an injection system is remarkably improved, the oxidation chain reaction of vitamin C is effectively blocked, and the generation of impurities is reduced. In addition, a buffer system is optimized, compared with a traditional sodium bicarbonate buffer system, pH fluctuation caused by CO2 release is avoided, the content of active oxygen in the injection can be reduced through the ultra-pure deoxidation process, operation is conducted in the environment with the temperature smaller than or equal to 25 DEG C in the whole process, and light degradation is reduced by adopting an amber neutral glass ampoule. The long-acting stability of the injection is improved, and the impurity content is reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparation preparation, and more particularly to a vitamin C injection with low impurity content and a preparation method thereof. Background Art

[0002] Vitamin C, also known as ascorbic acid, is a water-soluble vitamin that cannot be synthesized in the human body and must be supplemented through food or medication. Vitamin C has strong reducing properties (antioxidant properties), can scavenge free radicals, and protect cells from oxidative damage; it promotes collagen synthesis: promotes wound healing and skin repair. Enhances immunity: promotes white blood cell function, improves anti-infection ability, promotes iron absorption and neurotransmitter synthesis, etc. Daily supplementation needs can be met by taking vitamin C tablets or effervescent tablets orally, but for critically ill patients or patients with limited gastrointestinal absorption and those who need large doses of vitamin C supplementation, the use of vitamin C injections can achieve faster and better therapeutic effects.

[0003] However, vitamin C (ascorbic acid) injection is prone to produce a variety of impurities during the preparation, storage and use process, mainly from factors such as raw materials, production process, storage conditions, etc. In terms of raw materials, vitamin C raw materials may contain residual intermediates such as 2-keto-L-gulonic acid and L-threonic acid during the chemical synthesis process. Metal ions (Fe 3+ and Cu 2+ ). In terms of production process, factors such as light, oxygen, and metal ions will catalyze the oxidation of vitamin C, accelerate degradation, and generate dehydroascorbic acid (DHAA) impurities. DHAA is further hydrolyzed into 2,3-diketo-L-gulonic acid (DKG) impurities, increasing clinical adverse reactions (such as kidney stones, phlebitis, etc.). Existing technologies usually use antioxidants (such as sodium bisulfite, EDTA) or inert gas protection to reduce oxidation, but there are the following problems: antioxidants may introduce new impurities (such as the risk of sulfite allergy); conventional nitrogen filling processes are difficult to completely deoxygenate, and long-term storage is still prone to degradation; pH adjustment relies on traditional buffer systems (such as sodium bicarbonate), which may affect stability. In addition, during high-temperature sterilization, high temperatures will promote the decomposition of vitamin C and produce harmful substances such as furfural and 5-hydroxymethylfurfural (5-HMF). In summary, the presence of the above impurities will seriously affect the safety, efficacy and stability of the drug.

[0004] Therefore, how to provide a vitamin C injection with high stability and low impurity content is a technical problem that needs to be solved urgently by those skilled in the art. Summary of the Invention

[0005] In view of this, the present invention provides a vitamin C injection with low impurity content and a preparation method thereof, which significantly improves the stability of the injection and reduces the impurity content in the injection system.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A vitamin C injection with a low impurity content contains the following components by weight per 100 mL of the vitamin C injection: 1.0-25.0 g of vitamin C, 0.01-0.05 g of L-cysteine, 0.1-0.5 g of polyethylene glycol-6000, 0.1-0.3 g of citric acid, 0.2-0.6 g of sodium citrate, 0.01-0.05 g of calcium D-gluconate, and water for injection to make up to 100 mL.

[0008] As a preferred technical solution, the vitamin C injection with low impurity content, calculated as 100 mL of vitamin C injection, contains the following components by weight: 10.0 g of vitamin C, 0.03 g of L-cysteine, 0.3 g of polyethylene glycol-6000, 0.2 g of citric acid, 0.4 g of sodium citrate, 0.03 g of calcium D-gluconate, and water for injection is added to 100 mL.

[0009] As a preferred technical solution, the water for injection is ultrapure deoxygenated water for injection.

[0010] Another object of the present invention is to provide a method for preparing the above-mentioned vitamin C injection with low impurity content, which is prepared according to the above-mentioned ratio and specifically comprises the following steps:

[0011] (1) Ultrapure deoxygenation treatment of water for injection: After vacuum degassing, the water for injection is bubbled with high-purity inert gas for 5-15 minutes to reduce the dissolved oxygen content to ≤0.5ppm. The water for injection is vacuum deoxygenated and bubbled with inert gas to reduce the dissolved oxygen content to ≤0.5ppm, which is lower than the 2-3ppm in conventional processes and avoids the oxidative degradation of vitamin C.

[0012] (2) Dissolving the stabilizer: Take the water for injection prepared in step (1), add calcium D-gluconate, L-cysteine and polyethylene glycol-6000 in sequence, and stir to dissolve; calcium D-gluconate, while avoiding the toxicity of EDTA, preferentially binds to free Fe through the coordinated coordination mechanism of polyhydroxy-carboxylic acid. 3+ / Cu 2+ Combined with, blocking its contact with vitamin C, inhibiting the oxidation chain reaction, in addition, the Ca in D-gluconate 2+ It can competitively bind to anions in the solution (such as oxalic acid) to reduce the risk of calcium oxalate precipitation.

[0013] L-cysteine, as a natural reducing agent, can directly capture free radicals in the injection solution and block the oxidation chain reaction of vitamin C. In addition, it can also reduce oxidized vitamin C (dehydroascorbic acid) and regenerate the active ingredient.

[0014] Polyethylene glycol-6000 can form a dense hydrophilic molecular film on the surface of the solution, reducing the oxygen diffusion coefficient better than traditional nitrogen protection. In addition, it can also combine with vitamin C molecules through hydrogen bonds, reducing the probability of contact with metal ions and playing a stabilizing role.

[0015] When L-cysteine and polyethylene glycol-6000 work together, a dynamic complement is formed: L-cysteine removes free radicals in the liquid phase, and PEG-6000 blocks the invasion of gaseous oxygen, covering the "source" and "propagation" links of the oxidation reaction, which has a synergistic effect.

[0016] (3) Preparation of sodium citrate-citric acid buffer solution: Take the water for injection in step (1), add citric acid and sodium citrate, stir and dissolve, and prepare sodium citrate-citric acid buffer solution; sodium citrate-citric acid buffer solution (rather than traditional sodium bicarbonate) is used to avoid pH fluctuations caused by CO2 release and maintain long-term stability.

[0017] (4) Vitamin C dissolution and pH adjustment: Add vitamin C to the stabilizer prepared in step (2), stir until completely dissolved, add the remaining water for injection, and then add sodium citrate-citric acid buffer to adjust the pH to 5.5-6.5;

[0018] (5) Filtration and filling: Sterilize by filtering through a microporous membrane, fill into amber ampoules under argon protection, and seal with amber neutral glass ampoules to reduce light degradation;

[0019] (6) Sterilization: Circulating steam sterilization.

[0020] As a preferred technical solution, the vacuum degassing pressure in step (1) is -0.08 MPa to -0.095 MPa, and the time is 20-40 min; the high-purity inert gas is high-purity argon with a purity of 99.999%.

[0021] As a preferred technical solution, the amount of water for injection in step (2) is 10-15% of the weight of the water for injection in the formula.

[0022] As a preferred technical solution, the amount of water for injection in step (3) is 20-30% by weight of the water for injection in the formula.

[0023] As a preferred technical solution, the pore size of the microporous filter membrane in step (5) is 0.22 μm.

[0024] As a preferred technical solution, the temperature of the circulating steam in step (6) is 100° C. and the sterilization time is 10-15 minutes.

[0025] As a preferred technical solution, steps (1) to (6) are all performed in an environment of ≤25°C.

[0026] It can be seen from the above technical solutions that compared with the prior art, the present invention has the following beneficial effects:

[0027] The invention provides a formula of vitamin C injection. The formula adopts D-calcium gluconate, L-cysteine and polyethylene glycol-6000 to configure a stabilizer. The three ingredients synergistically enhance the effect, significantly improve the stability of the injection system, effectively block the oxidation chain reaction of vitamin C, reduce the generation of impurities, and improve the long-term stability of the injection. In an accelerated test (40°C, RH75%, 6 months), the main component content is ≥98%, and the impurity content shows that the impurity content is significantly lower than that of a control group. DETAILED DESCRIPTION

[0028] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0029] Example 1

[0030] A vitamin C injection with low impurity content, comprising the following components by weight per 100 mL of the vitamin C injection: 1.0 g of vitamin C, 0.01 g of L-cysteine, 0.1 g of polyethylene glycol-6000, 0.1 g of citric acid, 0.2 g of sodium citrate, 0.01 g of calcium D-gluconate, and water for injection to make up to 100 mL;

[0031] The vitamin C injection is prepared by the following method:

[0032] (1) Ultrapure deoxygenation treatment of water for injection: After degassing the water for injection under vacuum (-0.08 MPa) for 20 minutes, high-purity argon (99.999%) was introduced for 5 minutes to reduce the dissolved oxygen to ≤0.5 ppm;

[0033] (2) Dissolving the stabilizer: Take 10% of the weight of the water for injection in the formula, add calcium D-gluconate, L-cysteine and polyethylene glycol-6000 in sequence, and stir to dissolve;

[0034] (3) Preparation of sodium citrate-citric acid buffer solution: Take 20% of the weight of the water for injection in the formula, add citric acid and sodium citrate, stir and dissolve, and prepare sodium citrate-citric acid buffer solution;

[0035] (4) Vitamin C dissolution and pH adjustment: Add vitamin C to the stabilizer prepared in step (2), stir until completely dissolved, add the remaining water for injection, and then add sodium citrate-citric acid buffer to adjust the pH to 5.5;

[0036] (5) Filtration and filling: Sterilize by filtering through a 0.22 μm microporous filter membrane, fill into amber ampoules under argon protection, and seal;

[0037] (6) Sterilization: 100°C circulating steam sterilization for 10 min.

[0038] Example 2

[0039] A vitamin C injection with low impurity content, comprising the following components by weight per 100 mL of the vitamin C injection: 25.0 g of vitamin C, 0.05 g of L-cysteine, 0.5 g of polyethylene glycol-6000, 0.3 g of citric acid, 0.6 g of sodium citrate, 0.05 g of calcium D-gluconate, and water for injection to make up to 100 mL;

[0040] The vitamin C injection is prepared by the following method:

[0041] (1) Ultrapure deoxygenation treatment of water for injection: After degassing the water for injection under vacuum (-0.095 MPa) for 40 minutes, high-purity argon (99.999%) was introduced for 15 minutes to reduce the dissolved oxygen to ≤0.5 ppm;

[0042] (2) Dissolving the stabilizer: Take 15% of the weight of the water for injection in the formula, add calcium D-gluconate, L-cysteine and polyethylene glycol-6000 in sequence, and stir to dissolve;

[0043] (3) Preparation of sodium citrate-citric acid buffer solution: Take 30% by weight of the water for injection in the formula, add citric acid and sodium citrate, stir and dissolve, and prepare sodium citrate-citric acid buffer solution;

[0044] (4) Vitamin C dissolution and pH adjustment: Vitamin C was added to the stabilizer prepared in step (2), stirred until completely dissolved, and the remaining water for injection was added, followed by sodium citrate-citric acid buffer, and the pH was adjusted to 6.5;

[0045] (5) Filtration and filling: Sterilize by filtering through a 0.22 μm microporous filter membrane, fill into amber ampoules under argon protection, and seal;

[0046] (6) Sterilization: 100℃ circulating steam sterilization for 15 minutes.

[0047] Example 3

[0048] A vitamin C injection with low impurity content, containing the following components by weight per 100 mL of the vitamin C injection: 10.0 g of vitamin C, 0.03 g of L-cysteine, 0.3 g of polyethylene glycol-6000, 0.2 g of citric acid, 0.4 g of sodium citrate, 0.03 g of calcium D-gluconate, and water for injection to make up to 100 mL;

[0049] The vitamin C injection is prepared by the following method:

[0050] (1) Ultrapure deoxygenation treatment of water for injection: After degassing the water for injection under vacuum (-0.09 MPa) for 30 minutes, high-purity argon (99.999%) was introduced for 10 minutes to reduce the dissolved oxygen to ≤0.5 ppm;

[0051] (2) Dissolving the stabilizer: Take 13% of the weight of the water for injection in the formula, add calcium D-gluconate, L-cysteine and polyethylene glycol-6000 in sequence, and stir to dissolve;

[0052] (3) Preparation of sodium citrate-citric acid buffer solution: Take 25% by weight of the water for injection in the formula, add citric acid and sodium citrate, stir and dissolve, and prepare sodium citrate-citric acid buffer solution;

[0053] (4) Vitamin C dissolution and pH adjustment: Vitamin C was added to the stabilizer prepared in step (2), stirred until completely dissolved, and the remaining water for injection was added, followed by sodium citrate-citric acid buffer, and the pH was adjusted to 6.0;

[0054] (5) Filtration and filling: Sterilize by filtering through a 0.22 μm microporous filter membrane, fill into amber ampoules under argon protection, and seal;

[0055] (6) Sterilization: 100°C circulating steam sterilization for 10 min.

[0056] Comparative Example 1

[0057] No ultrapure deoxygenation treatment was performed, and the remaining steps and conditions were the same as those in Example 3.

[0058] Comparative Example 2

[0059] Without adding L-cysteine, the remaining steps and conditions were the same as those in Example 3.

[0060] Comparative Example 3

[0061] Without adding polyethylene glycol-6000, the remaining steps and conditions were the same as those in Example 3.

[0062] Comparative Example 4

[0063] Calcium D-gluconate was not added, and the remaining steps and conditions were the same as those in Example 3.

[0064] Comparative Example 5

[0065] The sodium citrate-citric acid buffer system was replaced with a traditional sodium bicarbonate buffer system, and the remaining steps were the same as in Example 3.

[0066] Performance evaluation of the vitamin C injections prepared in Examples 1-3 and Comparative Examples 1-5

[0067] The injections prepared in the examples and comparative examples were subjected to accelerated tests under the following experimental conditions:

[0068] Temperature / humidity: 40°C ± 2°C, RH 75% ± 5%, time: 0, 1, 2, 3, 6 months. Sampling points: 3 batches of samples (n = 3) were taken at each time point. The specific detection methods and detection indicators are shown in Table 1.

[0069] Table 1 Detection methods and test indicators of accelerated test

[0070] Test items method Vitamin C content HPLC (UV detection, 245 nm) degradation products HPLC-MS (oxalic acid, DKG) pH Potentiometric method

[0071] The above-mentioned HPLC method was used to detect the content of vitamin C. The experimental results are shown in Table 2.

[0072] Table 2 Changes in vitamin C content (%) over time

[0073] Time (month) 0 1 2 3 6 Example 1 100.0±0.5 99.8±0.4 99.5±0.3 99.0±0.5 98.2±0.6 Example 2 100.0±0.3 99.6±0.2 99.4±0.5 99.0±0.2 98.1±0.3 Example 3 100.0±0.2 99.5±0.6 99.6±0.2 99.2±0.3 98.6±0.2 Comparative Example 1 100.0±0.5 98.1±0.4 96.5±0.4 93.8±0.5 91.2±0.2 Comparative Example 2 100.0±0.4 98.8±0.4 95.5±0.3 91.0±0.4 90.2±0.2 Comparative Example 3 100.0±0.4 97.8±0.2 95.1±0.4 90.0±0.5 88.2±0.6 Comparative Example 4 100.0±0.3 97.4±0.4 94.5±0.3 91.0±0.2 88.1±0.3 Comparative Example 5 100.0±0.5 98.1±0.4 93.3±0.2 90.2±0.6 87.1±0.2

[0074] Result analysis: As can be seen from the contents of Table 2, the vitamin C injections prepared in Examples 1-3 of the present invention have good stability. Accelerated experiments show that after 6 months, the retention rates of the vitamin C injections prepared in Examples 1-3 of the present invention are ≥98%, which is significantly higher than the retention rates of Comparative Examples 1-5.

[0075] In addition, the present invention further conducted a 24-month long-term storage experiment, and the experimental results showed that long-term storage (24 months) still met the pharmacopoeia standards.

[0076] At the same time, the present invention also measured the accumulation of key degradation products (oxalic acid, DKG, 5-HMF) after 6 months. The experimental results are shown in Table 3.

[0077] Table 3 Accumulation of key degradation products (%) in different groups

[0078] impurities oxalic acid 2,3-DKG 5-HMF Example 1 0.08±0.02 0.12±0.03 Not detected (<0.01%) Example 2 0.07±0.03 0.11±0.02 Not detected (<0.01%) Example 3 0.07±0.01 0.10±0.02 Not detected (<0.01%) Comparative Example 1 0.15±0.05 0.35±0.08 0.05±0.02 Comparative Example 2 0.16±0.04 0.38±0.06 0.06±0.01 Comparative Example 3 0.12±0.03 0.36±0.04 0.04±0.02 Comparative Example 4 0.25±0.04 0.37±0.05 0.06±0.01 Comparative Example 5 0.21±0.02 0.28±0.02 0.05±0.02

[0079] Results: As shown in Table 3, the vitamin C injection of the present invention is relatively stable. After 6 months of storage, the impurity content in the system is significantly lower than that of the comparative example group. In addition, compared with the example group, the oxalic acid content in comparative example 4 is the highest, indicating that the sodium citrate-citric acid buffer system is crucial for reducing acidic degradation pathways.

[0080] Based on the above experiments, the present invention further measured the pH changes of different groups at the initial stage of preparation and after six months of storage. The experimental results are shown in Table 4.

[0081] Table 4 Changes in pH values in different groups

[0082] Time (month) 0 6 Example 3 6.02±0.05 5.95±0.08 Comparative Example 4 5.82±0.07 5.45±0.12

[0083] Result analysis: The pH fluctuation of the citric acid buffer system was much smaller than that of the comparative example 4 group.

[0084] The present invention also conducted a safety test and found that the amount of L-cysteine used was far below its toxicity threshold (LD 50 >5g / kg, oral administration in rats), PEG-6000 is an FDA-approved pharmaceutical excipient and therefore has good safety.

[0085] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0086] The above description of the disclosed embodiments is intended to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is intended to conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A vitamin C injection with low impurity content, characterized in that: 100 mL of vitamin C injection contains the following components by weight: 1.0-25.0 g of vitamin C, 0.01-0.05 g of L-cysteine, 0.1-0.5 g of polyethylene glycol-6000, 0.1-0.3 g of citric acid, 0.2-0.6 g of sodium citrate, 0.01-0.05 g of calcium D-gluconate, and water for injection is added to make up to 100 mL.

2. A vitamin C injection with low impurity content, characterized in that: 100 mL of vitamin C injection contains the following components by weight: 10.0 g of vitamin C, 0.03 g of L-cysteine, 0.3 g of polyethylene glycol-6000, 0.2 g of citric acid, 0.4 g of sodium citrate, 0.03 g of calcium D-gluconate, and the volume is made up to 100 mL with water for injection.

3. The vitamin C injection with low impurity content according to claim 1 or 2, characterized in that: The water for injection is ultrapure deoxygenated water for injection.

4. A method for preparing the vitamin C injection with low impurity content according to any one of claims 1 to 3, characterized in that: The preparation is carried out according to the ratio described in claim 1 or 2, which specifically comprises the following steps: (1) Ultrapure deoxygenation of water for injection: After vacuum degassing, the water for injection is bubbled with high-purity inert gas for 5-15 minutes to reduce the dissolved oxygen to ≤0.5 ppm; (2) Dissolving the stabilizer: Take the water for injection prepared in step (1), add calcium D-gluconate, L-cysteine, and polyethylene glycol-6000 in sequence, and stir to dissolve; (3) Preparation of sodium citrate-citric acid buffer solution: take the water for injection prepared in step (1), add citric acid and sodium citrate, stir and dissolve, and prepare sodium citrate-citric acid buffer solution; (4) Vitamin C dissolution and pH adjustment: Add vitamin C to the stabilizer prepared in step (2), stir until completely dissolved, add the remaining water for injection, and then add sodium citrate-citric acid buffer to adjust the pH to 5.5-6.5; (5) Filtration and filling: sterilize by filtering through a microporous filter membrane, fill into amber ampoules under argon protection, and seal; (6) Sterilization: Circulating steam sterilization.

5. The method for preparing the low-impurity vitamin C injection according to claim 4, characterized in that: The vacuum degassing pressure in step (1) is -0.08 MPa to -0.095 MPa, and the time is 20-40 min; the high-purity inert gas is high-purity argon with a purity of 99.999%.

6. The method for preparing the low-impurity vitamin C injection according to claim 5, characterized in that: The amount of the water for injection in step (2) is 10-15% of the weight of the water for injection in the formula.

7. The method for preparing the low-impurity vitamin C injection according to claim 6, characterized in that: The amount of the water for injection in step (3) is 20-30% of the weight of the water for injection in the formula.

8. The method for preparing the low-impurity vitamin C injection according to claim 7, characterized in that: The pore size of the microporous filter membrane in step (5) is 0.22 μm.

9. The method for preparing the low-impurity vitamin C injection according to claim 8, characterized in that: The temperature of the circulating steam in step (6) is 100° C., and the sterilization time is 10-15 minutes.

10. The method for preparing the vitamin C injection with low impurity content according to claim 9, characterized in that: Steps (1) to (6) are all performed in an environment of ≤25°C.