Method for determining impurities in oxycodone hydrochloride sustained release tablet by high performance liquid chromatography

Abstract

The invention discloses a method for determining impurities in oxycodone hydrochloride sustained-release tablets by high performance liquid chromatography, a mixed solution of a sodium heptanesulfonate aqueous solution and methanol is used as an extracting agent to dissolve a sample, and the pretreatment process has the characteristics of simplicity and high efficiency; then, a C8 chromatographic column is adopted, only the water phase and the organic phase in different proportions are used for gradient elution, efficient separation of multiple impurities can be achieved, the separation degree meets the specification, and the method is good in durability, good in repeatability and accurate in measurement result; the method is of great significance to quality control of oxycodone hydrochloride sustained release tablets.

Description

Technical Field

[0001] The invention relates to the field of drug quality detection, and in particular to a method for determining impurities of oxycodone hydrochloride sustained-release tablets by high performance liquid chromatography. Background Art

[0002] Oxycodone hydrochloride, as a drug extracted and synthesized from the alkaloid thebaine, has a strong analgesic effect. Oxycodone hydrochloride sustained-release tablets are a new type of preparation product that has both controlled-release and sustained-release effects, and has broad application prospects in the treatment of acute pain. The impurity detection method for oxycodone hydrochloride sustained-release tablets is currently included in USP43, BP2021, and ChP2020. According to the BP standard method, impurities D and E cannot be separated, and nitrogen oxide impurities are controlled by additional methods in the import registration standards and foreign pharmacopoeias. As an impurity test, it is not simple enough in actual application. The other three methods included cannot separate the preparation excipients and impurity peaks, and the impurity detection capabilities of each method are weak. Summary of the invention

[0003] In view of this, one of the objects of the present invention is to provide a method for determining impurities in oxycodone hydrochloride sustained-release tablets by high performance liquid chromatography.

[0004] In order to achieve the above object, the present invention provides the following technical solutions:

[0005] 1. A method for determining impurities in oxycodone hydrochloride sustained-release tablets by high performance liquid chromatography, comprising the following steps:

[0006] (1) Preparation of test sample: Accurately weigh a quantity of oxycodone hydrochloride sustained-release tablets powder equivalent to 10 mg of oxycodone hydrochloride, place in a 10 mL volumetric flask, add about 8 mL of solvent, sonicate for 20 min to dissolve the oxycodone hydrochloride, cool to room temperature, dilute to scale with solvent, shake well, filter, and take the filtrate as the test sample solution;

[0007] (2) Chromatographic detection:

[0008] Specific chromatographic conditions: C8 column; column temperature: 50°C; flow rate: 1.0 mL / min; detection wavelength: 206 nm; injection volume: 10 μL, elution according to the following gradient:

[0009]

[0010] The mobile phase A is a mixed solution of sodium heptane sulfonate aqueous solution and methanol in a volume ratio of 9:1;

[0011] The mobile phase B is a mixture solution of sodium heptane sulfonate aqueous solution and methanol in a volume ratio of 1:9.

[0012] Preferably, the sodium heptane sulfonate aqueous solution is mixed with 1.01 g / L sodium heptane sulfonate solution: phosphoric acid: triethylamine in a volume ratio of 850:3:2.

[0013] Preferably, the present invention further comprises preparing a system adaptability solution and a 0.2% self-control solution;

[0014] The system adaptability solution is prepared as follows: accurately weigh impurities of noroxymorphone, 7,8-dihydro-8,14-dihydroxycodeinone, 14-hydroxycodeine, 10-hydroxyoxycodone, oxycodone hydrochloride, acrylate copolymer, and triethyl citrate into a same volumetric flask, add mobile phase A to dissolve, dilute, shake well, and filter to obtain a system adaptability solution;

[0015] The preparation method of the 0.2% self-control solution is as follows: accurately measure 1 ml of the test solution and place it in a 25 ml volumetric flask, add solvent to dilute to the scale, shake well, and then accurately measure 1 ml and place it in a 20 ml volumetric flask.

[0016] The beneficial effects of the present invention are as follows: the present invention discloses a method for determining impurities of oxycodone hydrochloride sustained-release tablets by high-performance liquid chromatography, the present invention adopts a mixed solution of sodium heptane sulfonate aqueous solution and methanol as an extractant to dissolve the sample, and the pretreatment process has the characteristics of simplicity and efficiency. Then, a C8 chromatographic column is used, and only different proportions of aqueous phase and organic phase are used for gradient elution, so that efficient separation of multiple impurities can be achieved, and the separation degree meets the requirements, the method has good durability, good repeatability, and accurate measurement results. It can better monitor the quality of oxycodone hydrochloride sustained-release tablets. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to make the purpose, technical solution and beneficial effects of the present invention clearer, the present invention provides the following drawings for illustration:

[0018] Figure 1 It is a system adaptability solution spectrum (1, impurity A; 2, noroxymorphone; 3, nitrogen oxides; 4, 10-hydroxyoxycodone (14-hydroxycodeine); 5, impurity B1; 6, 7,8-dihydro-8,14-dihydroxycodeinone; 7, impurity B2; 8, acrylate copolymer; 9, oxycodone; 10, impurity E; 11, impurity D; 12, impurity G; 13, impurity C; 14, impurity F; 15, triethyl citrate);

[0019] Figure 2 is the blank solution spectrum;

[0020] Figure 3 is the sample solution spectrum;

[0021] Figure 4 This is the spectrum of 0.2% self-control solution;

[0022] Figure 5 Detection chromatography for BP pharmacopoeia standards;

[0023] Figure 6 To detect the chromatogram using the related substance method of the raw material. DETAILED DESCRIPTION

[0024] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

[0025] Example 1

[0026] The impurity content of oxycodone hydrochloride sustained-release tablets was determined by high performance liquid chromatography. The method uses mobile phase A (a mixed solution of sodium heptane sulfonate aqueous solution and methanol) as an extractant to dissolve the sample, and the pretreatment process is simple and efficient. Then, a C8 chromatographic column is used, and only different proportions of aqueous phase and organic phase are used for gradient elution, which can achieve efficient separation of multiple impurities. The separation degree meets the requirements, the method has good durability, good repeatability, and accurate measurement results.

[0027] Specific chromatographic conditions:

[0028] Chromatographic column: C8 chromatographic column; column temperature: 50°C; flow rate: 1.0 ml / min; detection wavelength: 206 nm; injection volume: 10 μl, gradient elution ratio is shown in Table 1:

[0029] Table 1. Gradient elution ratio

[0030]

[0031] The mobile phase preparation process is as follows: the mixed solution obtained by mixing sodium heptane sulfonate buffer as the aqueous phase and methanol as the organic phase is used as the mobile phase.

[0032] Buffer solution: sodium heptane sulfonate solution (1.01 g / L sodium heptane sulfonate solution-phosphoric acid-triethylamine=850:3:2, pH adjusted to 2.40±0.02 with 50% by mass sodium hydroxide solution).

[0033] Mobile phase A: 900 ml of buffer solution, 100 ml of methanol, mix, filter and degas by ultrasonication.

[0034] Mobile phase B: 100 ml of buffer solution, 900 ml of methanol, mix, filter and degas by ultrasonication.

[0035] Testing steps:

[0036] 1) System adaptability solution: Accurately weigh impurities A, B, C, D, E, F, G, noroxymorphone, 7,8-dihydro-8,14-dihydroxycodeinone, 14-hydroxycodeine, 10-hydroxyoxycodone reference substance, oxycodone hydrochloride, acrylate copolymer, and triethyl citrate into a volumetric flask, add mobile phase A to dissolve, dilute, shake well, and filter to obtain the system adaptability solution.

[0037] 2) Preparation of test sample: Take an appropriate amount of fine powder of the product (equivalent to 10 mg of oxycodone hydrochloride), accurately weigh, place in a 10 ml volumetric flask, add about 8 ml of solvent (a mixed solution of sodium heptane sulfonate aqueous solution and methanol), ultrasonicate for 20 min to dissolve the oxycodone hydrochloride, cool to room temperature, dilute to the scale with solvent, shake well, filter, and take the filtrate as the test sample solution.

[0038] 3) 0.2% self-control solution: accurately measure 1 ml of the test solution and place it in a 25 ml volumetric flask, add solvent to dilute to the mark, shake well, and then accurately measure 1 ml and place it in a 20 ml volumetric flask, shake well.

[0039] 4) Determination: Take 10 μl of system suitability solution, blank solvent, test solution, and 0.2% self-control solution and inject them into the liquid chromatograph. Record the chromatogram. The result is as follows: Figures 1 to 4 The results show that the separation of the two can be achieved successfully.

[0040] At the same time, the method reported in the British (BP) Pharmacopoeia standard was used for testing, and the results were as follows Figure 5 The results show that impurities D and E cannot be separated.

[0041] The relevant substance method of the raw materials was used for determination according to the high performance liquid chromatography method (General Rules 0512 of Part IV of the 2020 edition of the Chinese Pharmacopoeia). The specific steps are as follows:

[0042] Test solution: Take an appropriate amount of the product, weigh accurately, dissolve it in 1% dilute acetic acid and quantitatively dilute it to make a solution containing about 1 mg per 1 ml.

[0043] Reference solution: Take appropriate amount of oxycodone impurity A, B, C, D, E, F, G reference substances, accurately weigh, dissolve and dilute with 1% dilute acetic acid to make a solution containing about 0.1 mg per 1ml, as the reference substance stock solution of each impurity. Accurately measure 2ml of the test solution, place it in a 100ml volumetric bottle, add 1% dilute acetic acid to dilute to the scale, shake well, accurately measure 5ml, place it in a 100ml volumetric bottle; accurately measure 1ml of each impurity reference substance stock solution, place it in the same 100ml volumetric bottle mentioned above, dilute to the scale with 1% dilute acetic acid, and shake well.

[0044] Chromatographic conditions: Octylsilane bonded silica gel is used as filler (Agilent poroshell 120EC-C8, 4.6×100 mm, 2.7 μm or chromatographic column with equivalent performance); 1.01 g / L sodium heptane sulfonate solution-phosphoric acid-triethylamine (850:3:2, pH adjusted to 2.40 with 50% sodium hydroxide solution) is used as mobile phase A, methanol is used as mobile phase B, and gradient elution is performed according to Table 2; column temperature is 50.0°C; detection wavelength is 206 nm, and injection volume is 10 μl.

[0045] Table 2. Gradient elution

[0046]

[0047] System suitability requirements: In the control solution chromatogram, the retention time of oxycodone should be about 10 to 12 minutes, and impurities A, B (impurity B has isomers at the 6th position, which appear as double peaks in the chromatogram, namely B1 and B2), oxycodone, E, D, G, C, and F should appear in sequence, and the separation between E, D, G, and C should not be less than 1.1.

[0048] Determination method: Accurately measure the test solution and control solution, inject them into liquid chromatograph respectively, and record the chromatogram.

[0049] Limits: If there are impurity peaks in the chromatogram of the test solution, the peak areas of impurity A, impurity B1, impurity B2, impurity C, impurity E, impurity F, and impurity G shall not be greater than the main peak area in the control solution (0.1%); the chromatographic peak area of ​​impurity D shall not be greater than twice the main peak area in the control solution (0.2%); the peak area of ​​other individual impurities shall not be greater than the main peak area of ​​the control solution (0.1%); the sum of the peak areas of all impurities shall not be greater than 10 times the peak area of ​​oxycodone in the reference solution (1.0%), see Table 3 for details.

[0050] Table 3. Impurity peak retention time

[0051]

[0052]

[0053] Note: The location of each impurity peak is based on the retention time of each impurity in the reference solution.

[0054] Chromatogram Figure 6 The results showed that although impurities A to G could be separated and nitrogen oxides in the preparation could be effectively separated from unknown impurities, the auxiliary material acrylate copolymer peak could not be effectively separated from the main peak.

[0055] The related substance method of the present invention can avoid this problem.

[0056] Example 2

[0057] (1) Repeatability experiment:

[0058] 10 μL of the system suitability solution was accurately measured and injected into the liquid chromatograph, and the injection was repeated 6 times, and the chromatogram was recorded. The average values ​​of the retention time, peak area and theoretical plate number of each impurity after 6 repeated injections are shown in Table 4.

[0059] Table 4. Repeatability experiment

[0060]

[0061]

[0062] The above results show that the separation degree and theoretical plate number between the impurities in the system applicability solution meet the requirements. After 6 injections, the relative standard deviation of the area of ​​each impurity peak and the main peak is in the range of 0.79-13.7%, both of which are not greater than 15%. The relative standard deviation of the retention time is in the range of 0.04-0.34%, both of which are not greater than 1.0%. The system applicability and injection precision of this method meet the requirements.

[0063] (2) Accuracy experiment:

[0064] The recovery tests of various impurities and oxycodone hydrochloride were carried out at 0.1% (0.25% of the concentration of the test solution for nitrogen oxides), 0.2% (0.5% of the concentration of the test solution for nitrogen oxides), and 1.0% (2.5% of the concentration of the test solution for nitrogen oxides) of the test solution of the related substances of this product; the recovery rates of 10-hydroxyoxycodone and 14-hydroxycodeine were separately examined.

[0065] Weigh three portions of each impurity reference substance accurately, place them in a 10 ml volumetric flask, add blank excipients and mixed powders according to the prescribed amount, add mobile phase A for ultrasonic dissolution, and filter to a fixed volume to obtain the test solution. Inject and test according to the chromatographic method of the relevant substances.

[0066] The average recovery rate of impurity A at three concentration levels was 99.4%; the average recovery rate of impurity B was 102.8%; the average recovery rate of impurity C was 101.5%; the average recovery rate of impurity D was 101.7%; the average recovery rate of impurity E was 100.8%; the average recovery rate of impurity F was 96.4%; the average recovery rate of impurity G was 94.5%; the average recovery rate of noroxymorphone was 98.9%; the average recovery rate of nitrogen oxides was 92.7%; the average recovery rate of 7,8-dihydro-8,14-dihydroxycodeinone was 101.8%; the average recovery rate of 14-hydroxycodeinone was 102.6%; and the average recovery rate of 10-hydroxyoxycodone was 100.2%. The 0.1% recovery rate of each impurity is within the range of 80% to 120%, and the 5.0% concentration recovery rate of nitrogen oxides and the 1.0% concentration recovery rate of each impurity are within the range of 91% to 108%, indicating that this method is accurate in determining the known impurities in the sample, and its determination results are reliable. This method can effectively and accurately determine the amount of each known impurity in the sample.

[0067] The above-described embodiments are only preferred embodiments for fully illustrating the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or changes made by those skilled in the art based on the present invention are within the protection scope of the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. A method for determining impurities in oxycodone hydrochloride sustained-release tablets by high performance liquid chromatography, characterized in that: The steps include: (1) Preparation of test sample: Accurately weigh a quantity of oxycodone hydrochloride sustained-release tablet powder equivalent to 10 mg of oxycodone hydrochloride, place in a 10 mL volumetric flask, add about 8 mL of solvent, sonicate for 20 min to dissolve the oxycodone hydrochloride, cool to room temperature, dilute to scale with solvent, shake well, filter, and take the filtrate as the test sample solution; (2) Chromatographic detection: Specific chromatographic conditions: C8 column; column temperature: 50°C; flow rate: 1.0 mL / min; detection wavelength: 206 nm; injection volume: 10 μL, elution according to the following gradient: The mobile phase A is a mixed solution of sodium heptane sulfonate aqueous solution and methanol in a volume ratio of 9:1; The mobile phase B is a mixture solution of sodium heptane sulfonate aqueous solution and methanol in a volume ratio of 1:

9.

2. The method for determining impurities of oxycodone hydrochloride sustained-release tablets by high performance liquid chromatography according to claim 1, characterized in that: The sodium heptane sulfonate aqueous solution is mixed with 1.01 g / L sodium heptane sulfonate solution, phosphoric acid and triethylamine in a volume ratio of 850:3:

2.

3. The method for determining impurities of oxycodone hydrochloride sustained-release tablets by high performance liquid chromatography according to claim 1, characterized in that: It also includes preparing a system suitability solution and a 0.2% self-control solution; The system adaptability solution is prepared as follows: accurately weigh impurities of noroxymorphone, 7,8-dihydro-8,14-dihydroxycodeinone, 14-hydroxycodeine, 10-hydroxyoxycodone, oxycodone hydrochloride, acrylate copolymer, and triethyl citrate into a same volumetric flask, add mobile phase A to dissolve, dilute, shake well, and filter to obtain a system adaptability solution; The preparation method of the 0.2% self-control solution is as follows: accurately measure 1 ml of the test solution and place it in a 25 ml volumetric flask, add solvent to dilute to the scale, shake well, and then accurately measure 1 ml and place it in a 20 ml volumetric flask.

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