A method for determining degradation impurities in related substances of vitamin B6 injection
By using high-performance liquid chromatography in vitamin B6 injection and using specific mobile phase and gradient elution procedures, the problem of difficult detection of degraded impurities in the prior art is solved, and the accurate detection and separation of degraded impurities is achieved, ensuring the accuracy and stability of the detection results.
Patent Information
- Application Number
- CN202311283723.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-10-07
- Publication Date
- 2025-04-08
AI Technical Summary
The prior art is difficult to effectively detect impurities degraded in vitamin B6 injection, resulting in incomplete detection, poor peak shape and low response.
High performance liquid chromatography was used, using 0.04% sodium pentanesulfonate solution and methanol as mobile phases, and the detection was carried out on the C18 column through a gradient elution procedure to ensure the separation of degraded impurities from the main peak and to achieve effective separation between the degraded peaks.
Accurate detection of degraded impurities in vitamin B6 injection has been achieved. Each peak-to-peak type is good, no solvent effect, good solution stability, accurate detection results, and comply with the pharmacopoeia standards.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the field of drug analysis, and specifically relates to a method for determining vitamin B 6 Detection method for degradation impurities in related substances of injection. Background Art
[0002] Vitamin B6 6 ) is also called pyridoxine, which includes pyridoxine, pyridoxal and pyridoxamine. It exists in the body in the form of phosphate ester. It is a water-soluble vitamin that is easily destroyed by light or alkali and is not resistant to high temperatures. Vitamin B 6 It is a component of certain coenzymes in the human body and is involved in a variety of metabolic reactions, especially closely related to amino acid metabolism. 6 The preparation is used to prevent and treat vomiting during pregnancy and vomiting due to radiation sickness.
[0003] Vitamin B 6 Not resistant to high temperatures. Under high temperature conditions, vitamin B 6 The molecules will dehydrate and dimerize to form ethers, which will then gradually decompose completely. 6 In the production process, the sterilization method is terminal sterilization. During the sterilization process, it will be exposed to high temperature conditions, so thermal degradation impurities will be produced during the sterilization process. In order to ensure the quality of vitamin B 6 The quality of the injection solution, therefore, needs to be in vitamin B 6 In the injection solution, the amount of degradation impurities must be strictly controlled.
[0004] According to the current edition of the Chinese Pharmacopoeia, USP, EP, JP and other relevant pharmacopoeia standards on vitamin B 6 and vitamin B 6 Although the quality standards for injections test relevant substances, they are basically about the control of process impurities and there is no control over degradation impurities.
[0005] The chromatographic conditions in the above-mentioned pharmacopoeia standards were used to detect heat-damaged samples, and it was found that the degradation impurities could not be completely detected, and the peak shape of some degradation peaks detected was poor and the response was not high. Summary of the invention
[0006] Purpose of the invention: The purpose of the present invention is to provide a method for determining vitamin B 6 Detection method for degradation impurities in related substances of injection.
[0007] The objective of the present invention is achieved through the following technical solutions: A method for measuring vitamin B 6 Detection method for degradation impurities in related substances of injection.
[0008] The sample was diluted to a certain concentration with a suitable solvent, and a certain volume was taken for gradient elution injection using mobile phase A and mobile phase B by high performance liquid chromatography, and detection was performed using a C18 column at a certain wavelength and column temperature.
[0009] The method is characterized in that the sample concentration is vitamin B 6 For injection, dilute with solvent to a solution of 1 mg / ml.
[0010] The method is characterized in that the diluent is a 0.04% sodium pentane sulfonate solution (the pH value is adjusted to 3.0 with acetic acid).
[0011] The method is characterized in that the mobile phase A is a 0.04% sodium pentane sulfonate solution (the pH value is adjusted to 3.0 with acetic acid).
[0012] The method is characterized in that the mobile phase B is methanol.
[0013] The method is characterized in that the gradient elution procedure is as follows: T(min) A（%） B（%） 0 90 10 20 85 15 40 50 50 45 50 50 45.1 90 10 60 90 10 The method is characterized in that the detection wavelength is 291 nm.
[0014] The method is characterized in that the C18 column is a Shimadzu VP-ODS column, 4.6×250 mm, 5 μm.
[0015] The method is characterized in that the C18 column is a Yuexu Ultimate AQ-C18 column, 4.6×250 mm, 5 μm.
[0016] The method is characterized in that the certain volume taken is 20 μl.
[0017] The method is characterized in that the column temperature is 30°C.
[0018] The method is characterized in that vitamin B 6 The related substances include at least one of impurity C, impurity D, impurity A, RRT1.35, impurity F, impurity G, and impurity E.
[0019] The present invention has the following advantages and effects over the prior art: 1. The solvent used in the present invention is 0.04% sodium pentane sulfonate solution (the pH value is adjusted to 3.0 with acetic acid). Under this solvent condition, each peak has a good shape, no solvent effect, and good solution stability.
[0020] 2. Under the chromatographic conditions of the present invention, each degradation peak can be separated from the main peak, and each degradation peak can be effectively separated from each other.
[0021] 3. Through validation tests such as specificity, quantitative limit, detection limit, linearity, stability, repeatability, accuracy and durability, all tests of this method meet the requirements and the method is good.
[0022] The present invention provides a vitamin B 6 The detection method of related substances in injection uses octadecylsilane bonded silica gel as filler, 0.04% sodium pentane sulfonate solution (pH adjusted to 3.0 with acetic acid) and methanol as mobile phase for gradient elution, which can quickly complete the detection of vitamin B under the same chromatographic adjustment. 6 Rapid separation of degradation impurities can achieve accurate test results and achieve the purpose of inspection. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Attached Figure 1 Vitamin B 6 System suitability solution chromatogram for injection.
[0024] Attached Figure 2 Vitamin B 6 Chromatogram of the test sample solution for injection.
[0025] Attached Figure 3 Vitamin B 6 Chromatogram of injection control solution. Implementation
[0026] The present invention is further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto.
[0027] The degradation impurities of the samples were detected by high performance liquid chromatography, and the chromatographic conditions were as follows: Chromatographic column: Octadecylsilane bonded silica gel is used as filler; Mobile phase A: 0.04% sodium pentane sulfonate solution (pH adjusted to 3.0 with acetic acid); Mobile phase B: methanol; Elution procedure: T(min) A（%） B（%） 0 90 10 20 85 15 40 50 50 45 50 50 45.1 90 10 60 90 10 Detection wavelength: 291nm; Column temperature: 30°C; Detection wavelength: 280nm; Injection volume: 20 μl Determination method: Accurately measure an appropriate amount of vitamin B6 injection, quantitatively dilute with solvent to a solution containing 1 mg per milliliter, shake well, and obtain the test solution; accurately measure 1 ml of the test solution and place it in a 100 ml volumetric flask, dilute with solvent to the scale, shake well, and obtain the control solution. Accurately measure 20 μl of the test solution and control solution, respectively, and inject them into the liquid chromatograph, record the chromatogram, see Figure 2-Figure 3 。
Claims
1. A HPLC method for determining degradation impurities in vitamin B6 injection, characterized in that The method comprises the following steps: diluting the sample to a certain concentration with a suitable solvent, taking a certain volume, using high performance liquid chromatography, using mobile phase A and mobile phase B for gradient elution injection, using a C18 column, and detecting at a certain wavelength and column temperature.
2. The method according to claim 1, characterized in that The sample concentration is obtained by taking an appropriate amount of vitamin B6 injection and diluting it with a solvent to obtain a solution of 1 mg / ml.
3. The method according to claim 1, characterized in that The diluent is 0.04% sodium pentane sulfonate solution (the pH value is adjusted to 3.0 with acetic acid).
4. The method according to claim 1, characterized in that The mobile phase A is 0.04% sodium pentane sulfonate solution (the pH value is adjusted to 3.0 with acetic acid).
5. The method according to claim 1, characterized in that The mobile phase B is methanol.
6. The method according to claim 1, characterized in that The gradient elution procedure is as follows:
7. The method according to claim 1, characterized in that The detection wavelength is 291 nm.
8. The method according to claim 1, characterized in that The C18 column is a Shimadzu VP-ODS column, 4.6×250 mm, 5 μm.
9. The method according to claim 1, characterized in that The C18 column is Yuexu Ultimate AQ-C18 column, 4.6×250 mm, 5 μm.
10. The method according to claim 1, characterized in that The certain volume is 20 μl.
11. The method according to claim 1, characterized in that The column temperature is 30°C.
12. The method according to claim 1, characterized in that Vitamin B6 related substances include at least one of impurity C, impurity D, impurity A, RRT1.35, impurity F, impurity G, and impurity E.