Photodegradation impurities of brimonidine tartrate eye drops and quality detection method of brimonidine tartrate eye drops

Abstract

The invention provides brimonidine tartrate eye drop photodegradation impurities and a quality detection method of brimonidine tartrate eye drops, and belongs to the field of medicine detection. The brimonidine tartrate eye drop photodegradation impurity has a structural formula shown as a formula I and a formula II: # imgabs0. The invention also provides a method for evaluating the stability of brimonidine tartrate eye drops. According to the detection method, other related substances can be effectively detected, meanwhile, photodegradation impurities can be accurately detected, the sensitivity is high, operation is easy and convenient, the method is stable, durability is good, and a guarantee is provided for clinical use of brimonidine tartrate eye drops.

Description

Technical Field

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

[0002] Brimonidine tartrate is a new, highly effective α2 receptor selective ocular hypotensive drug. Its affinity for α2 receptors is 1000 times that of α1 receptors. Its selectivity for α2 receptors is 7-12 times higher than that of clonidine and 23-32 times higher than that of apraclonidine. In addition to its good ocular hypotensive effect, brimonidine also has potential neuroprotective effects. Domestic 0.2% brimonidine tartrate eye drops and imported aphagen eye drops are used to treat primary open-angle glaucoma and ocular hypertension. Both have similar ocular hypotensive effects and few systemic and local side effects.

[0003] The detection methods of brimonidine tartrate eye drops reported so far are mainly to detect the content of brimonidine tartrate, such as Huang Cuiyun, et al., HPLC-MS / MS determination of the content of brimonidine tartrate eye drops, Strait Pharmacy 2020 Vol. 32 No. 2, Establishment of HPLC-MS / MS method for determination of the content of brimonidine tartrate eye drops, the purpose is to improve the sensitivity of brimonidine content in brimonidine tartrate eye drops. Wu Hongying, et al., HPLC determination of the content of brimonidine tartrate eye drops, Qilu Pharmaceutical 2008 Vol.27, No.10, Establishment of a method for determination of the content of brimonidine tartrate eye drops by high performance liquid chromatography. There are few detection methods for impurities in brimonidine tartrate eye drops. The impurities of commercial brimonidine tartrate eye drops increase rapidly under light conditions, and the preservatives used in brimonidine tartrate raw materials and eye drops are irritating to a certain extent, and even contain aniline genotoxic impurities. Dai Shoufeng, et al., LC-MS / MS determination of aniline genotoxic impurities in brimonidine tartrate, Chinese Modern Applied Pharmacy, June 2021, Vol. 38, No. 12, 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] Shui Qinghua, Su Huai, Li Jin, Lan Weiwei, Wu Lifen. Effect of pH on the stability of brimonidine tartrate eye drops [J]. Journal of Pharmaceutical Practice, 2008, (5): 364-366. The effect of pH on the stability of brimonidine tartrate eye drops was investigated to determine the optimal pH range and storage conditions of this product. Methods: The content and related substances of brimonidine tartrate eye drops were determined by HPLC, 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 dark.

[0005] Since brimonidine tartrate eye drops contain a large number of impurities, it is difficult to accurately detect them using existing domestic and foreign detection methods, especially for photodegradable impurities, and there are no relevant literature reports. Summary of the invention

[0006] The present invention provides photodegradable impurities of brimonidine tartrate eye drops. The present invention also provides a quality detection method of brimonidine tartrate eye drops, which detects photodegradable impurities (including photodegradable impurities of formula I and formula II) in brimonidine tartrate eye drops by HPLC chromatography.

[0007] The present invention provides photodegradable impurities of brimonidine tartrate eye drops, the structural formulas of which are shown in Formula I and Formula II: .

[0008] The invention provides application of the photodegradable impurities of brimonidine tartrate eye drops in quality control of the brimonidine tartrate eye drops.

[0009] The invention provides a quality detection method for brimonidine tartrate eye drops, which uses an HPLC method to detect impurities represented by formulas 1 and 2 in the brimonidine tartrate eye drops; The chromatographic conditions are: Chromatographic column: octadecylsilane bonded silica Xbridge C18, 4.6 mm × 250 mm, 5 μm; Mobile phase: Isocratic elution with methanol-water solution containing sodium heptane sulfonate as mobile phase; column temperature 30°C; flow rate 1.0 ml per minute; detection wavelength 264 nm; injection volume 20 μl; The preparation method of the methanol aqueous solution containing sodium heptane sulfonate is as follows: weigh 2.6 g of sodium heptane sulfonate, add 265 ml of methanol, 2.5 ml of triethylamine, and 7.5 ml of glacial acetic acid, then add water to dissolve and dilute to 1000 ml.

[0010] Specifically, it includes the following steps: a. Prepare the test solution: Take the brimonidine tartrate eye drops to be tested, dilute it to the scale with water, and shake well.

[0011] b. Prepare the control solution: Take an appropriate amount of the test solution prepared in step a, and dilute it quantitatively 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 make a solution containing approximately 0.09 µg of brimonidine tartrate per 1 ml; c. Detect according to the chromatographic conditions described.

[0012] The retention time of the impurity is: 36.80-37.10min.

[0013] The present invention also provides a method for evaluating the stability of brimonidine tartrate eye drops, which adopts the quality detection method of brimonidine tartrate eye drops to detect the photodegradation impurities of the brimonidine tartrate eye drops.

[0014] If the corresponding chromatographic peak of brimonidine tartrate eye drops is detected within the retention time of 36.80-37.10min and its content is not greater than 0.1%, or the corresponding chromatographic peak of impurities is not detected within the retention time of 36.80-37.10min, then the brimonidine tartrate eye drops have qualified stability.

[0015] The detection method of the invention can not only effectively detect other related substances, but also accurately detect photodegradation impurities, has high sensitivity, is easy to operate, is stable, and has good durability, thus providing a guarantee for the clinical use of brimonidine tartrate eye drops. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is the 30-day (photodegradation) chromatogram of the influencing factors of the conditions of the related substances of brimonidine tartrate eye drops; Figure 2 The mass spectrum of the unknown impurity (X1) after photodegradation of the related substance; Figure 3 This is the chromatogram of the 17-day photodegradation of the self-made agent; Figure 4 This is the chromatogram of the reference preparation after 17 days of photodegradation. DETAILED DESCRIPTION

[0017] Example 1 Discovery of photodegradable impurities in the brimonidine tartrate eye drops of the present invention In the naked sample photodegradation test of brimonidine tartrate eye drops, 1.689% of unknown photodegradable impurities (X1) were detected under the chromatographic conditions of related substances within 30 days.

[0018] Sample name: Brimonidine tartrate eye drops; self-made preparation, batch number: 221101; Packaging form: Low-density polyethylene medicinal single-dose eye drop bottle (remove the polyester / aluminum / polyethylene medicinal composite film bag); Conditions: Illuminance 4500Lx±500Lx, UV 100μw / cm 2 The test results are shown in Table 1. Figure 1 : Table 1 Test results of influencing factors (light) of self-made agent (bare sample)

[0019] Confirm the structure of X1 impurity, such as Figure 2 As shown: The LC-MS spectrum has M / Z peaks of 310.03 and 312.02 at RT5.94~6.01min, which are M and M+2. The isotope peak ratio is 1:1, containing 1 bromine atom.

[0020] Mechanism analysis: This impurity is a photo-oxidation impurity of brimonidine (brimonidine photo-degradation impurity formula Ⅰ, brimonidine photo-degradation impurity formula Ⅱ), and the formation mechanism is as follows:

[0021] Among them, the self-made preparation prescription is shown in Table 2: Table 2 Prescription of self-made preparation of brimonidine tartrate eye drops Element Dosage effect Brimonidine tartrate 0.45mg Active ingredients 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 Moderate pH Adjusters Water for injection Up to 0.3ml Solvents Total 0.3ml / Example 2 Quality testing method for brimonidine tartrate eye drops of the present invention Detection method: Determine by high performance liquid chromatography. Operate in a dark environment.

[0022] Accurately measure 3 ml of the test solution, place it in a 25 ml volumetric flask, dilute to the mark with water, and shake well.

[0023] Control solution: Accurately measure an appropriate amount of the test solution and quantitatively dilute it with water to make a solution containing approximately 0.36 μg of brimonidine tartrate per 1 ml.

[0024] 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.

[0025] Chromatographic conditions: octadecylsilane bonded silica gel is used as filler (XBridge C18, 4.6mm×250mm, 5μm or chromatographic column with equivalent performance); methanol aqueous solution containing sodium heptane sulfonate (weigh 2.6g of sodium heptane sulfonate, add 265ml of methanol, 2.5ml of triethylamine, 7.5ml of glacial acetic acid, then add water to dissolve and dilute to 1000ml) is used as mobile phase; column temperature is 30℃; flow rate is 1.0ml per minute; detection wavelength is 264nm; injection volume is 20μl.

[0026] System suitability requires that the signal-to-noise ratio of the main component peak height in the sensitivity solution chromatogram should be greater than 10.

[0027] Determination method: Accurately measure the test solution, control solution and reference substance solution, inject them into the liquid chromatograph respectively, and record the chromatogram to 3 times the retention time of the main peak.

[0028] The test sample was brimonidine tartrate eye drops (self-made preparation batch number: 221103; reference preparation batch number: TT5356; condition illumination 4500Lx±500Lx, UV 100μw / cm 2 ) for 17 days (reference preparation manufacturer: Allergan Sales LLC). Figure 3 , Figure 4 shown.

[0029] Conclusion: Both the self-developed preparation and the reference preparation can produce unknown photodegradation impurity X1 under strong light irradiation.

Claims

1. Brimonidine tartrate eye drops photodegradable impurities, characterized in that: The structural formula is shown in Formula I and Formula II: 。 2. Use of the photodegradable impurities of brimonidine tartrate eye drops according to claim 1 in the quality control of brimonidine tartrate eye drops.

3. A quality detection method for brimonidine tartrate eye drops, characterized in that: It adopts HPLC method to detect impurities shown in formula I and formula 2 in brimonidine tartrate eye drops; The chromatographic conditions are: Chromatographic column: octadecylsilane bonded silica Xbridge C18, 4.6 mm × 250 mm, 5 μm; Mobile phase: Isocratic elution with methanol-water solution containing sodium heptane sulfonate as mobile phase; column temperature 30°C; flow rate 1.0 ml per minute; detection wavelength 264 nm; injection volume 20 μl; The preparation method of the methanol aqueous solution containing sodium heptane sulfonate is as follows: weigh 2.6 g of sodium heptane sulfonate, add 265 ml of methanol, 2.5 ml of triethylamine, and 7.5 ml of glacial acetic acid, then add water to dissolve and dilute to 1000 ml.

4. The quality detection method of brimonidine tartrate eye drops according to claim 3, characterized in that: It includes the following steps: a. Prepare the test solution: Take the brimonidine tartrate eye drops to be tested, dilute to the mark with water, and shake well; b. Prepare the control solution: Take an appropriate amount of the test solution prepared in step a, and dilute it quantitatively 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 make a solution containing approximately 0.09 µg of brimonidine tartrate per 1 ml; c. Detection according to the chromatographic conditions described in claim 3.

5. The quality detection method of brimonidine tartrate eye drops according to claim 3 or 4, characterized in that: The retention time of the impurity is: 36.80-37.10min.

6. A method for evaluating the stability of brimonidine tartrate eye drops, characterized in that: The method for detecting the quality of the brimonidine tartrate eye drops according to any one of claims 3 to 5 is used to detect the photodegradable impurities of the brimonidine tartrate eye drops according to claim 1.

7. The method for evaluating the stability of brimonidine tartrate eye drops according to claim 6, characterized in that: If the corresponding chromatographic peak of brimonidine tartrate eye drops is detected within the retention time of 36.80-37.10min but its content is not greater than 0.1%, or the corresponding chromatographic peak of impurities is not detected within the retention time of 36.80-37.10min, then the brimonidine tartrate eye drops have qualified stability.

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