Brimonidine tartrate eye drops photodegradation impurities and a quality detection method for brimonidine tartrate eye drops

Abstract

The present invention provides photodegradation impurities of brimonidine tartrate eye drops, as well as a quality detection method for brimonidine tartrate eye drops, belonging to the field of drug detection. The photodegradation impurities of brimonidine tartrate eye drops have the structural formulas shown in Formula I and Formula II: The present invention also provides a method for evaluating the stability of brimonidine tartrate eye drops. The detection method of the present invention can not only effectively detect other related substances, but also accurately detect photodegradation impurities, with high sensitivity, simple operation, stable method and good durability, providing a guarantee for the clinical use of brimonidine tartrate eye drops.

Description

Technical Field

[0001] The present invention relates to the photodegradation impurities of brimonidine tartrate eye drops and a quality detection method for brimonidine tartrate eye drops, belonging to the field of drug quality detection. Background Art

[0002] Brimonidine tartrate is a new type of highly efficient α2 receptor-selective intraocular pressure-lowering drug. Its affinity for the α2 receptor is 1000 times that of the α1 receptor, and its selectivity for the α2 receptor is 7 - 12 times higher than that of clonidine and 23 - 32 times higher than that of apraclonidine. In addition to having a good intraocular pressure-lowering effect, brimonidine also has potential neuroprotective effects. The domestic 0.2% brimonidine tartrate eye drops and the imported Alphagan eye drops are used to treat primary open-angle glaucoma and ocular hypertension, and their intraocular pressure-lowering effects are similar, with small systemic and local side effects.

[0003] The currently reported detection methods for brimonidine tartrate eye drops mainly focus on detecting the content of brimonidine tartrate. For example, Huang Cuiyun, etc., "Determination of the content of brimonidine tartrate eye drops by HPLC-MS / MS", Strait Pharmaceutical Journal, Vol. 32, No. 2, 2020, established an HPLC-MS / MS method to determine the content of brimonidine tartrate eye drops, aiming to improve the sensitivity of the content of brimonidine in brimonidine tartrate eye drops. Wu Hongying, etc., "Determination of the content of brimonidine tartrate eye drops by HPLC method", Qilu Pharmaceutical Affairs, 2008, Vol. 27, No. 10, established a method for determining the content of brimonidine tartrate eye drops by high-performance liquid chromatography. There are relatively few detection methods for the impurities in brimonidine tartrate eye drops. Since the impurities in commercially available brimonidine tartrate eye drops increase rapidly under light conditions, and the preservatives used in brimonidine tartrate raw materials and eye drops have certain irritation, and even contain aniline genotoxic impurities. Dai Shoufeng, etc., "Determination of aniline genotoxic impurities in brimonidine tartrate by LC-MS / MS", Chinese Journal of Modern Applied Pharmacy, Vol. 38, No. 12, June 2021, established an LC-MS / MS method to determine the content of aniline genotoxic impurities (6-aminoquinoxaline, 4-nitro-o-phenylenediamine, 6-amino-5-bromoquinoxaline) in brimonidine tartrate.

[0004] Qinghua Shui, Huai Su, Jin Li, Weiwei Lan, Lifen Wu. Influence of pH on the Stability of Brimonidine Tartrate Eye Drops [J]. Journal of Pharmaceutical Practice, 2008, (5): 364 - 366. To investigate the influence of pH on the stability of brimonidine tartrate eye drops and determine the optimal pH range and storage conditions for this product. Methods: HPLC was used to determine the content and related substances of brimonidine tartrate eye drops, and the stability of this product under strong light and high temperature conditions was investigated. The most suitable pH range for this product is slightly acidic, and the most suitable storage conditions are low temperature and light protection.

[0005] Since there are many impurities in brimonidine tartrate eye drops, it is difficult to accurately detect using existing domestic and foreign detection methods. Especially for photo - degradation impurities, there is no relevant literature report. Summary of the Invention

[0006] The present invention provides photo - degradation impurities of brimonidine tartrate eye drops. The present invention also provides a quality detection method for brimonidine tartrate eye drops, which detects photo - degradation impurities (including Photo - degradation Impurity Formula I and Formula II) in brimonidine tartrate eye drops by HPLC chromatography.

[0007] The present invention provides photo - degradation impurities of brimonidine tartrate eye drops, with the structural formulas shown as Formula I and Formula II:

[0008] 。

[0009] The present invention provides the application of the above - mentioned photo - degradation impurities of brimonidine tartrate eye drops in the quality control of brimonidine tartrate eye drops.

[0010] The present invention provides a quality detection method for brimonidine tartrate eye drops, which uses HPLC to detect the impurities shown in Formula I and Formula 2 in brimonidine tartrate eye drops;

[0011] The chromatographic conditions are as follows:

[0012] Chromatographic column: Octadecylsilyl bonded silica gel Xbridge C18, 4.6 mm × 250 mm, 5 μm;

[0013] Mobile phase: Methanol - aqueous solution containing sodium heptanesulfonate isocratic elution; Column temperature 30 °C; Flow rate 1.0 ml per minute; Detection wavelength 264 nm; Injection volume 20 μl;

[0014] The preparation method of the methanol - aqueous solution containing sodium heptanesulfonate is: Weigh 2.6 g of sodium heptanesulfonate, add 265 ml of methanol, 2.5 ml of triethylamine, 7.5 ml of glacial acetic acid, and then add water to dissolve and dilute to 1000 ml.

[0015] Specifically, it includes the following steps:

[0016] a. Preparation of the test solution:

[0017] Take the brimonidine tartrate eye drops to be tested, dilute it to the mark with water, and shake well.

[0018] b. Preparation of the control solution: Take an appropriate amount of the test solution prepared in step a, and quantitatively dilute it with water to prepare a solution containing 0.36 μg of brimonidine tartrate per 1 ml;

[0019] Sensitivity solution: Accurately measure an appropriate amount of the control solution, and quantitatively dilute it with water to prepare a solution containing approximately 0.09 μg of brimonidine tartrate per 1 ml;

[0020] c. Detect according to the described chromatographic conditions.

[0021] The retention time of the said impurity is: 36.80 - 37.10 min.

[0022] The present invention also provides a method for evaluating the stability of brimonidine tartrate eye drops, which is to detect the photodegradation impurities of the brimonidine tartrate eye drops by using the quality detection method of the brimonidine tartrate eye drops described above.

[0023] If a corresponding chromatographic peak is detected within the retention time of 36.80 - 37.10 min for the brimonidine tartrate eye drops and its content is not more than 0.1%, or no corresponding chromatographic peak of the impurity is detected within the retention time of 36.80 - 37.10 min, then the brimonidine tartrate eye drops are qualified in terms of stability.

[0024] The detection method of the present invention can not only effectively detect other related substances, but also accurately detect photodegradation impurities, with high sensitivity, simple operation, stable method and good durability, providing a guarantee for the clinical use of brimonidine tartrate eye drops. Description of the Drawings

[0025] Figure 1 It is the chromatogram of the influencing factors of related substances of brimonidine tartrate eye drops for 30 days (photodegradation);

[0026] Figure 2 It is the mass spectrum of the unknown impurity (X1) of the related substances by light degradation;

[0027] Figure 3 It is the chromatogram of the self-developed preparation after 17 days of photodegradation;

[0028] Figure 4 It is the chromatogram of the reference preparation after 17 days of photodegradation. Detailed Description of the Invention

[0029] Example 1 Discovery of Photodegradation Impurities in Brimonidine Tartrate Eye Drops of the Present Invention

[0030] In the naked sample photodegradation test of Brimonidine Tartrate Eye Drops, 1.689% of unknown photodegradation impurity (X1) was detected after 30 days under the chromatographic conditions for related substances.

[0031] Sample name: Brimonidine Tartrate Eye Drops; self-developed preparation, batch number: 221101;

[0032] Packaging form: low-density polyethylene medicinal single-dose eye drop bottle (removing the polyester / aluminum / polyethylene medicinal composite film bag);

[0033] Condition: illuminance 4500Lx ± 500Lx, ultraviolet 100μw / cm 2 , and the test results are shown in Table 1, Figure 1 :

[0034] Table 1 Test results of influencing factors (light) test for self-developed preparation (naked sample)

[0035]

[0036] The structure of impurity X1 was confirmed as follows Figure 2 shown:

[0037] The LC-MS spectrogram has M / Z peaks of 310.03 and 312.02 at RT 5.94 - 6.01 min, which are M and M+2, and the ratio of isotope peaks is 1:1, containing 1 bromine atom.

[0038] Mechanism analysis:

[0039] This impurity is the photo-oxidation impurity of brimonidine (brimonidine photodegradation impurity formula I, brimonidine photodegradation impurity formula II), and the formation mechanism is as follows:

[0040]

[0041] Among them, the prescription of the self-developed preparation is shown in Table 2:

[0042] Table 2 Prescription of self-developed preparation of Brimonidine Tartrate Eye Drops

[0043] Ingredient Dosage Function Brimonidine tartrate 0.45mg Active ingredient Sodium chloride 1.74mg Osmotic pressure regulator Calcium chloride 0.06mg Osmotic pressure regulator Magnesium chloride 0.018mg Osmotic pressure regulator Potassium chloride 0.42mg Osmotic pressure regulator Borax 0.42mg Buffer Boric acid 0.6mg Buffer Sodium carboxymethylcellulose 1.5mg Thickener Hydrochloric acid Appropriate amount pH regulator Water for injection Up to 0.3ml Solvent 0.3ml /

[0044] Example 2 Quality detection method of Brimonidine Tartrate Eye Drops of the present invention

[0045] Detection method:

[0046] Determined by high performance liquid chromatography. Operate under light protection.

[0047] For the test solution, accurately measure 3 ml of this product, place it in a 25-ml volumetric flask, dilute it to the mark with water, and shake well.

[0048] Reference solution: Accurately measure an appropriate amount of the test solution, and quantitatively dilute it with water to prepare a solution containing about 0.36 μg of brimonidine tartrate per 1 ml.

[0049] Sensitivity solution: Accurately measure an appropriate amount of the reference solution, and quantitatively dilute it with water to prepare a solution containing about 0.09 μg of brimonidine tartrate per 1 ml.

[0050] Chromatographic conditions: Use octadecylsilane chemically bonded silica gel as the filler (XBridge C18, 4.6 mm × 250 mm, 5 μm or a chromatographic column with equivalent efficiency); use a methanol aqueous solution containing sodium heptanesulfonate (weigh 2.6 g of sodium heptanesulfonate, add 265 ml of methanol, 2.5 ml of triethylamine, 7.5 ml of glacial acetic acid, and then dissolve and dilute to 1000 ml with water) as the mobile phase; the column temperature is 30 °C; the flow rate is 1.0 ml per minute; the detection wavelength is 264 nm; the injection volume is 20 μl.

[0051] System suitability requirements: In the chromatogram of the sensitivity solution, the signal-to-noise ratio of the main component peak height should be greater than 10.

[0052] Assay method: Accurately measure the test solution, reference solution, and reference substance solution, and inject them into the liquid chromatograph respectively, and record the chromatogram until 3 times the retention time of the main peak.

[0053] Among them, the test substance is brimonidine tartrate eye drops (self-developed preparation batch number: 221103; reference preparation batch number: TT5356; condition illuminance 4500 Lx ± 500 Lx, ultraviolet 100 μw / cm 2 ) stored for 17 days (reference preparation manufacturer: Allergan Sales LLC), as Total amount 、 Figure 3 Figure 4 shown.

[0054] Conclusion: Both the self-developed preparation and the reference preparation can produce an unknown photodegradation impurity X1 under strong light irradiation.

Claims

1. A quality inspection method for brimonidine tartrate eye drops, characterized in that: It uses the HPLC method to detect the photodegradation impurities in brimonidine tartrate eye drops. The structural formulas of the photodegradation impurities are shown in Formula I and Formula 2: ； The chromatographic conditions are as follows: Chromatographic column: octadecylsilane-bonded silica gel Xbridge C18, 4.6 mm × 250 mm, 5 μm; Mobile phase: methanol aqueous solution containing sodium heptanesulfonate is used as the mobile phase for isocratic elution; column temperature is 30 °C; flow rate is 1.0 ml per minute; detection wavelength is 264 nm; injection volume is 20 μl; The preparation method of the methanol aqueous solution containing sodium heptanesulfonate is as follows: weigh 2.6 g of sodium heptanesulfonate, add 265 ml of methanol, 2.5 ml of triethylamine, 7.5 ml of glacial acetic acid, and then add water to dissolve and dilute to 1000 ml; It includes the following steps: a. Prepare the test solution: Take the brimonidine tartrate eye drops to be tested, dilute to the scale with water, and shake well; b. Prepare the control solution: Take an appropriate amount of the test solution prepared in step a, and quantitatively dilute it with water to prepare a solution containing 0.36 μg of brimonidine tartrate per 1 ml; Sensitivity solution: Accurately measure an appropriate amount of the control solution, and quantitatively dilute it with water to prepare a solution containing 0.09 μg of brimonidine tartrate per 1 ml; c. Detect according to the described chromatographic conditions; The retention time of the said impurity is: 36.80 - 37.10 min.

2. A method for evaluating the stability of brimonidine tartrate eye drops, characterized in that: It uses the quality detection method of the brimonidine tartrate eye drops described in claim 1 to detect the photodegradation impurities of the brimonidine tartrate eye drops.

3. The method for evaluating the stability of brimonidine tartrate eye drops according to claim 2, wherein: If a corresponding chromatographic peak is detected within the retention time of 36.80 - 37.10 min of the brimonidine tartrate eye drops but its content is not more than 0.1%, or no corresponding chromatographic peak of the impurity is detected within the retention time of 36.80 - 37.10 min, it is a brimonidine tartrate eye drop with qualified stability.

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