A method for analyzing related substances in a compound preparation of Buluorenin tablets

Abstract

The present invention belongs to the field of analytical chemistry, and relates to a method for analyzing related substances of a compound preparation of buprenorphine tablets, wherein the method uses octadecylsilane bonded silica gel as a filler, the detection wavelength is 215nm, the column temperature is 25-35°C, the flow rate is 0.8-1.2mL / min, the injection volume is 10μL, the mobile phase A is 0.8-1.2g / L sodium octane sulfonate aqueous solution, phosphoric acid adjusts the pH to 2.80±0.02, the mobile phase B is acetonitrile aqueous solution, the volume ratio of acetonitrile to water is 80:20, and the gradient elution is performed. The method can exclude the interference of blank excipients when detecting related substances of buprenorphine tablets, and impurities PE-1, PE-2, PE-3, PE-4, PE-5, PE-6, PE-7, PE-8, PE-9, PE-10, PE-11, Ibu-1, Ibu-2, Ibu-3, Ibu-4 and Ibu-5 can all be effectively separated.

Description

Technical Field

[0001] The invention belongs to the field of analytical chemistry and relates to a method for analyzing related substances of a compound preparation of brucea renin tablets. Background Art

[0002] Ibuprofen tablets are a compound preparation containing two ingredients, ibuprofen and phenylephrine hydrochloride. Each tablet contains 200 mg of ibuprofen and 10 mg of phenylephrine hydrochloride. This product is used to temporarily relieve the following symptoms caused by the common cold or flu: headache, fever, increased sinus pressure, nasal congestion, mild body pain; reduce nasal swelling; temporarily restore nasal breathing.

[0003] Impurities are key quality attributes of drugs and can affect the safety and effectiveness of products. Impurities in drug quality standards refer to impurities introduced by the production process or raw materials, or impurities generated during storage, in drugs produced in accordance with the process and raw materials approved by the national drug supervision and administration department in accordance with the law.

[0004] At present, there is no related substance detection method for ibuprofen tablets in the pharmacopoeias of various countries. The related substance detection method in the quality standards of ibuprofen and phenylephrine hydrochloride included in the pharmacopoeias of various countries is only applicable to single-ingredient preparations, and cannot take into account the effective separation of two components and related impurities at the same time. Although the relevant literature also reports the related substance detection method of the compound preparation, the number of known impurities taken into account is small and cannot reflect the true quality of the product.

[0005] In summary, there is still no effective detection method for the related substances of Brulon tablets, and it is impossible to effectively monitor the impurities in Brulon tablets to ensure the product quality of Brulon tablets. It is urgent to develop an effective method for impurity detection suitable for this variety. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide a method for detecting related substances of a compound preparation of breccia renin tablets in view of the deficiencies in the prior art. High performance liquid chromatography (HPLC) is used. When detecting related substances of breccia renin tablets according to the method of the present invention, the interference of blank excipients can be eliminated, and each known impurity can be effectively separated.

[0007] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0008] The invention discloses a method for analyzing related substances of a compound preparation of buprenorphine tablets, wherein the related substances are impurities in the compound preparation of buprenorphine tablets, including impurity PE-1, impurity PE-2, impurity PE-3, impurity PE-4, impurity PE-5, impurity PE-6, impurity PE-7, impurity PE-8, impurity PE-9, impurity PE-10, impurity PE-11, impurity Ibu-1, impurity Ibu-2, impurity Ibu-3, impurity Ibu-4 and impurity Ibu-5;

[0009] The analytical method is high performance liquid chromatography, and the high performance liquid chromatography conditions are:

[0010] Chromatographic column: octadecylsilane bonded silica gel as filler;

[0011] Detection wavelength: 215nm;

[0012] Column temperature: 25-35°C;

[0013] Flow rate: 0.8~1.2mL / min;

[0014] Injection volume: 10 μL;

[0015] Mobile phase A: 0.8-1.2 g / L sodium octane sulfonate aqueous solution, pH adjusted to 2.80 ± 0.02 with phosphoric acid;

[0016] Mobile phase B: acetonitrile-water solution, the volume ratio of acetonitrile to water was 80:20;

[0017] Gradient elution;

[0018] The gradient elution conditions are:

[0019]

[0020]

[0021] The impurity PE-1 has the molecular formula C 11 H 15 NO4; molecular weight: 225.2440; chemical name: (R)-2-((2-hydroxy-2-(3-hydroxyphenyl)ethyl)(methyl)amino)acetic acid; chemical structure:

[0022]

[0023] The impurity PE-2 has a molecular formula of C7H6O2; a molecular weight of 122.1230; a chemical name of 3-hydroxybenzaldehyde; and a chemical structural formula of:

[0024]

[0025] The impurity PE-3 has the molecular formula C 10 H 13 NO2; molecular weight: 179.2190; chemical name: N-methyl-4,6-dihydroxy-1,2,3,4-tetrahydroisoquinoline; chemical structure:

[0026]

[0027] The impurity PE-4 has the molecular formula C 10 H 13 NO2; molecular weight: 179.2190; chemical name: N-methyl-4,8-dihydroxy-1,2,3,4-tetrahydroisoquinoline; chemical structure:

[0028]

[0029] The impurity PE-5 has the molecular formula C8H 11 NO2; molecular weight: 153.1810; chemical name: (1R)-2-amino-1-(3-hydroxyphenyl)ethanol; chemical structure:

[0030]

[0031] The impurity PE-6 has the molecular formula C9H 11 NO2; molecular weight: 165.1920; chemical name: 1-(3-hydroxyphenyl)-2-(methylamino)ethanone; chemical structure:

[0032]

[0033] The impurity PE-7 has the molecular formula C 16 H 19 NO2; molecular weight: 257.3330; chemical name: (1R)-2-(benzylmethylamino)-1-(3-hydroxyphenyl)ethanol; chemical structure:

[0034]

[0035] The impurity PE-8 has the molecular formula C 16 H 17 NO2; molecular weight: 255.3170; chemical name: 2-(benzyl(methyl)amino)-1-(3-hydroxyphenyl)ethanone; chemical structure:

[0036]

[0037] The impurity PE-9 has the molecular formula C 17 H 13NO3; molecular weight: 279.2950; chemical name: 1-(3-hydroxybenzoyl)-2-methylisoquinolin-6(2-hydrogen)-one; chemical structure:

[0038]

[0039] The impurity PE-10 has the molecular formula C 22 H 29 NO3; molecular weight: 355.4780; chemical name: (R)-1-(3-hydroxyphenyl)-2-(methylamino)ethyl-2-(4-isobutylphenyl) propionate; chemical structure:

[0040]

[0041] The impurity PE-11 has the molecular formula C 22 H 29 NO3; molecular weight: 355.4780; chemical name: N-((R)-2-hydroxy-2-(3-hydroxyphenyl)ethyl)-2-(4-isobutylphenyl)-N-methylpropionamide; chemical structure:

[0042]

[0043] The impurity Ibu-1 has a molecular formula of C 10 H 10 O3; molecular weight: 178.1870; chemical name: 2-(4-benzaldehyde) propionic acid; chemical structure:

[0044] and its enantiomers;

[0045] The impurity Ibu-2 has a molecular formula of C 13 H 16 O3; molecular weight: 220.2680; chemical name: 2-(4-isobutyrylphenyl)propionic acid; chemical structure:

[0046] and its enantiomers;

[0047] The impurity Ibu-3 has a molecular formula of C 11 H 14 O2; molecular weight: 178.2310; chemical name: 2-(4-ethylphenyl) propionic acid; chemical structure:

[0048]

[0049] The impurity Ibu-4 has a molecular formula of C 13 H 18O2; molecular weight: 206.2850; chemical name: 2-(3-isobutylphenyl) propionic acid; chemical structure:

[0050]

[0051] The impurity Ibu-5 has a molecular formula of C 12 H 16 O; molecular weight: 176.2590; chemical name: 4-isobutylacetophenone; chemical structure:

[0052]

[0053] Specifically, the chromatographic column has specifications of 4.6×250 mm and 5 μm.

[0054] Preferably, the column temperature is 30°C.

[0055] Preferably, the flow rate is 1.0 mL / min.

[0056] Preferably, the mobile phase A is a 1.0 g / L sodium octane sulfonate aqueous solution, and the pH is adjusted to 2.80±0.02 with phosphoric acid.

[0057] The application of the above method in the detection or separation of related substances in the compound preparation of Buluo Renin Tablets is also within the protection scope of the present invention.

[0058] Beneficial effects:

[0059] In the prior art, the related substance detection methods in the quality standards of ibuprofen and phenylephrine hydrochloride included in the pharmacopoeias of various countries are only applicable to single-ingredient preparations, and cannot simultaneously take into account the effective separation of two components and related impurities; although the relevant literature also reports the related substance detection methods of ibuprofen tablet compound preparations, the number of known impurities taken into account is small, and the true quality of the product cannot be reflected. Compared with the prior art, the present invention adopts a related substance analysis method to analyze various impurities in ibuprofen tablets, the number of known impurities is large, the method is simple and easy to implement, and the detection and separation of each known impurity in ibuprofen and phenylephrine hydrochloride are realized at the same time, which saves a lot of analysis and detection time compared with the use of two or more related substance analysis methods in conventional compound preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments, and the above and / or other advantages of the present invention will become more clear.

[0061] Figure 1 This is the chromatogram of the standard substance of related substances in Buluorone tablets.

[0062] Figure 2 The high performance liquid chromatogram of chromatographic condition screening 1 in Example 1.

[0063] Figure 3 The HPLC chromatogram of chromatographic condition screening 2 in Example 1.

[0064] Figure 4 The HPLC chromatogram of chromatographic condition screening 3 in Example 1.

[0065] Figure 5 This is the HPLC chromatogram of chromatographic condition screening 4 in Example 1. DETAILED DESCRIPTION

[0066] The reagents used in the examples of the present invention are all chromatographically pure, unless otherwise specified.

[0067] 1. Reagents and Materials

[0068] Sodium octane sulfonate, acetonitrile, water.

[0069] Ibuprofen (Source: China Food and Drug Inspection Institute, Batch No.: 100179-201707, Content: 99.9%);

[0070] Phenylephrine hydrochloride (Source: China Food and Drug Inspection Institute, Batch No.: 100261-202004, Content: 99.8%);

[0071] Impurity PE-1 (source: AOCS, batch number: YMP-120-20-13, purity: 98%);

[0072] Impurity PE-2 (source: sigma, batch number: WXBD3B2V, purity: 99.8%);

[0073] Impurity PE-3 (source: TRC, batch number: 1-DRR-52-2, purity: 98.8%);

[0074] Impurity PE-4 (source: TRC, batch number: 1-DRR-50-1, purity: 99.1%);

[0075] Impurity PE-5 (source: AOCS, batch number: COP-141-11, purity: 99.2%);

[0076] Impurity PE-6 (source: AOCS, batch number: YJW-001-12, purity: 97.0%);

[0077] Impurity PE-7 (source: AOCS, batch number: COP-249-30-A, purity: 99.9%);

[0078] Impurity PE-8 (source: AOCS, batch number: YJP-021-12, purity: 97.5%);

[0079] Impurity PE-9 (source: TLC, batch number: 3092-059A3, purity: 92.0%);

[0080] Impurity PE-10 (source: TLC, batch number: 3226-085A2, purity: 96.5%);

[0081] Impurity PE-11 (source: TLC, batch number: 3141-071A2, purity: 97.8%);

[0082] Impurity IBU-1 (source: China Food and Drug Inspection Institute, batch number: 101544-202101, purity: 100%);

[0083] Impurity IBU-2 (source: China Food and Drug Inspection Institute, batch number: 510108-202102, purity: 100%);

[0084] Impurity IBU-3 (source: AOCS, batch number: 0NI-140-16, purity: 98.2%);

[0085] Impurity IBU-4 (Source: China Food and Drug Inspection Institute, Batch No.: 510106-201802, Purity: 100%);

[0086] Impurity IBU-5 (source: China Food and Drug Inspection Institute, batch number: 100722-202105, purity: 100%);

[0087] Buluorenin tablets (source: self-developed, batch number: 220811);

[0088] Blank excipients (source: self-developed, batch number: 220810); wherein, the blank excipients are excipient powders mixed according to the formulation of the compound preparation of Brulonil tablets.

[0089] 2. Main instruments

[0090] Shimadzu LC-20A high performance liquid chromatograph.

[0091] 3. Structural information of impurities in Buluorenin tablets

[0092] Table 1 Structural information of impurities in Buluorenin tablets

[0093]

[0094]

[0095] Example 1: Method establishment

[0096] (1) Optimization and screening of mobile phase and gradient elution program

[0097] During the HPLC condition screening process, the common conditions are as follows:

[0098] Chromatographic column: octadecylsilane bonded silica gel as filler;

[0099] Detection wavelength: 215nm;

[0100] Column temperature: 30°C;

[0101] Flow rate: 1.0 mL / min;

[0102] Injection volume: 10 μL.

[0103] ①Chromatographic condition screening 1: Method source: EP ibuprofen quality standard

[0104] Mobile phase A: phosphoric acid-water (volume ratio of phosphoric acid to water is 0.5:1000);

[0105] Mobile phase B: acetonitrile;

[0106] The gradient schedule is as follows:

[0107] Table 2 Gradient schedule for chromatographic condition screening 1

[0108] Time, minutes Mobile phase A, volume % Mobile phase B, volume % 0 100 0 25 100 0 55 15 85 70 15 85

[0109] Test conclusion: Phenylephrine hydrochloride peaks too early, ibuprofen peak time and theoretical plate number are appropriate, but the concentration is too high, overload phenomenon occurs; ibuprofen impurity peak front; specific HPLC chromatogram see Figure 2 .

[0110] ②Chromatographic Condition Screening 2: Method Source: USP Phenylephrine Hydrochloride Standard

[0111] Mobile phase A: 0.32 wt% sodium octane sulfonate aqueous solution (pH adjusted to 2.80 ± 0.02 with phosphoric acid)-acetonitrile (90:10);

[0112] Mobile phase B: 0.32 wt% sodium octane sulfonate aqueous solution (pH adjusted to 2.80 ± 0.02 with phosphoric acid)-acetonitrile (10:90);

[0113] The gradient schedule is as follows:

[0114] Table 3 Gradient schedule for chromatographic condition screening 2

[0115]

[0116]

[0117] Test conclusion: The peak time of phenylephrine hydrochloride is appropriate, and the peak time and theoretical plate number of ibuprofen are appropriate, which can be further optimized on this basis; the specific HPLC chromatogram is shown in Figure 3 .

[0118] ③Chromatographic condition screening 3:

[0119] Mobile phase A: 0.10 wt% sodium octane sulfonate aqueous solution (pH adjusted to 2.80 ± 0.02 with phosphoric acid);

[0120] Mobile phase B: 80% acetonitrile in water;

[0121] The gradient schedule is as follows:

[0122] Table 4 Gradient schedule for chromatographic condition screening 3

[0123] Time, minutes Mobile phase A, volume % Mobile phase B, volume % 0 60 40 5 60 40 10 45 55 45 40 60 55 40 60 56 20 80 61 20 80

[0124] Detection conclusion: Impurities interfere with the main components and baseline separation cannot be achieved. Chromatographic parameters need to be further optimized. See the specific HPLC chromatogram for details. Figure 4 .

[0125] ④Chromatographic condition screening 4:

[0126] Mobile phase A: 1.0 g / L sodium octane sulfonate aqueous solution (pH adjusted to 2.80 ± 0.02 with phosphoric acid);

[0127] Mobile phase B: 80% acetonitrile in water;

[0128] The gradient schedule is as follows:

[0129] Table 5 Gradient schedule for chromatographic condition screening 4

[0130] Time, minutes Mobile phase A, volume % Mobile phase B, volume % 0 88 12 5 88 12 40 75 25 70 55 45 85 45 55 100 40 60 120 20 80 130 20 80

[0131] Test conclusion: The main peak has good symmetry, each known impurity can be completely separated from the main peak, and the blank auxiliary material does not interfere with the test; see the specific HPLC chromatogram Figure 5 .

[0132] In summary, the mobile phase and gradient elution program No. ④ were finally selected for the detection of related substances of this product.

[0133] (2) Determination of HPLC conditions

[0134] Chromatographic column: Octadecylsilane bonded silica gel as filler, specification 4.6×250mm, 5μm;

[0135] Detection wavelength: 215nm;

[0136] Column temperature: 30°C;

[0137] Flow rate: 1.0 mL / min;

[0138] Injection volume: 10 μL;

[0139] Mobile phase A: 1.0 g / L sodium octane sulfonate aqueous solution, pH adjusted to 2.80 ± 0.02 with phosphoric acid;

[0140] Mobile phase B: acetonitrile-water solution, the volume ratio of acetonitrile to water is 80:20;

[0141] Gradient elution;

[0142] The gradient elution conditions are:

[0143] Table 6 Gradient elution conditions

[0144] Time, minutes Mobile phase A, volume % Mobile phase B, volume % 0 88 12 5 88 12 40 75 25 70 55 45 85 45 55 100 40 60 120 20 80 130 20 80

[0145] Example 2: Methodological Validation

[0146] (1) Exclusivity

[0147] Ibuprofen localization solution: Take ibuprofen reference substance, weigh accurately, add diluent acetonitrile-water mixed solvent (the volume ratio of acetonitrile to water is 50:50), and dissolve and dilute to make a solution containing 1.5 μg / mL of ibuprofen per 1 mL.

[0148] Phenylephrine hydrochloride localizing solution: Take phenylephrine hydrochloride reference substance, weigh accurately, add diluent acetonitrile-water mixed solvent (the volume ratio of acetonitrile to water is 50:50), dissolve and dilute to make a solution containing 1.5μg / mL of phenylephrine hydrochloride per 1mL.

[0149] Impurity PE-1 localization solution: Take the impurity PE-1 reference substance, weigh it accurately, add the diluent acetonitrile-water mixed solvent (the volume ratio of acetonitrile to water is 50:50), and dissolve and dilute it to make a solution containing 1.5μg / mL of impurity PE-1 per 1mL.

[0150] The preparation method of the positioning solutions of impurity PE-2, impurity PE-3, impurity PE-4, impurity PE-5, impurity PE-6, impurity PE-7, impurity PE-8, impurity PE-9, impurity PE-10, impurity PE-11, impurity Ibu-1, impurity Ibu-2, impurity Ibu-3, impurity Ibu-4 and impurity Ibu-5 is the same as the preparation method of the positioning solution of impurity PE-1.

[0151] Mixed control solution: Accurately weigh each known impurity, ibuprofen and phenylephrine hydrochloride reference substance, place in the same volumetric flask, add diluent acetonitrile-water mixed solvent (the volume ratio of acetonitrile to water is 50:50), and dissolve and dilute to make a mixed control solution containing 2 mg / mL ibuprofen, 0.1 mg / mL phenylephrine hydrochloride, and 1.5 μg / mL of each known impurity per 1 mL.

[0152] Blank excipient solution: Take blank excipient, accurately weigh 220 mg, place in a 100 mL volumetric flask, add diluent acetonitrile-water mixed solvent (the volume ratio of acetonitrile to water is 50:50), shake appropriately to dissolve and dilute to the scale, shake well, and obtain.

[0153] Accurately measure 10 μL of each of the above-mentioned positioning solutions, mixed control solution and blank auxiliary material solution and inject them into the liquid chromatograph for analysis under the finally determined chromatographic conditions (serial number ④).

[0154] Table 7 Chromatographic peak information of mixed control solution

[0155]

[0156]

[0157] Test conclusion: Each component elutes in sequence and the retention time of phenylephrine hydrochloride and ibuprofen is moderate, and the separation between each component is greater than 1.5; the blank excipient does not interfere with the determination of known impurities, has good specificity, and can be used for the detection of related substances of this product; the chromatogram of the standard substance of the compound preparation ibuprofen tablets is shown in Figure 1 .

[0158] (2) Linearity and range

[0159] The linear relationship is to plot the measured response signal (peak area) as a function of the analyte concentration and perform linear regression using the least squares method. The linear regression coefficient R 2 The value should be no less than 0.990.

[0160] Take an appropriate amount of ibuprofen reference substance, add a diluent acetonitrile-water mixed solvent (the volume ratio of acetonitrile to water is 50:50) to prepare linear solutions of different concentrations, and perform sample injection analysis under the final determined chromatographic conditions (serial number ④).

[0161] Phenylephrine hydrochloride and various impurity reference substances were prepared in the same manner and analyzed.

[0162] Table 8 Ibuprofen linear determination results

[0163]

[0164] Table 9 Phenylephrine hydrochloride linear determination results

[0165]

[0166] Table 10 PE-1 linearity test results

[0167]

[0168]

[0169] Table 11 PE-2 linearity test results

[0170]

[0171] Table 12 PE-3 linearity test results

[0172]

[0173] Table 13 PE-4 linearity test results

[0174]

[0175] Table 14 PE-5 linearity test results

[0176]

[0177]

[0178] Table 15 PE-6 linearity test results

[0179]

[0180] Table 16 PE-7 linearity determination results

[0181]

[0182] Table 17 PE-8 linearity test results

[0183]

[0184] Table 18 PE-9 linearity test results

[0185]

[0186]

[0187] Table 19 PE-10 linearity test results

[0188]

[0189] Table 20PE-11 linearity test results

[0190]

[0191] Table 21 Ibu-1 linear determination results

[0192]

[0193] Table 22 Ibu-2 linearity determination results

[0194]

[0195]

[0196] Table 23 Ibu-3 linearity determination results

[0197]

[0198] Table 24Ibu-4 linearity determination results

[0199]

[0200] Table 25 Ibu-5 linearity determination results

[0201]

[0202] (3) Correction factors for known impurities

[0203] The correction factor of the known impurities of this product is determined by the standard curve method, that is, the impurity reference substance and the main component reference substance are accurately weighed, and a series of solutions (covering the reporting limit and the standard limit) are prepared and injected separately. The response factor (peak area) is linearly regressed with the test sample concentration using the least squares method to obtain two standard curves. The ratio of the slope of the main component to the known impurity is the correction factor.

[0204] Table 26 Impurity Correction Factor Table

[0205] name Slope Correction Factor Ibuprofen 0.4039 / Ibu-1 0.4642 0.87 Ibu-2 0.2994 1.35 Ibu-3 0.4576 0.88 Ibu-4 0.3905 1.03 Ibu-5 0.5146 0.78 Phenylephrine hydrochloride 0.2943 / PE-1 0.286 1.03 PE-2 1.2473 0.24 PE-3 0.3532 0.83 PE-4 0.3626 0.81 PE-5 0.3673 0.80 PE-6 0.7016 0.42 PE-7 0.4874 0.60 PE-8 0.521 0.56 PE-9 0.42 0.70 PE-10 0.355 0.83 PE-11 0.5186 0.57

[0206] (4) Detection limit and quantification limit

[0207] For known impurities, the limit of detection (LOD) and limit of quantification (LOQ) are determined based on the signal-to-noise ratio method. The impurity stock solution of known concentration is diluted to a low-concentration sample, and the measured signal is compared with the signal at the blank (baseline noise) to calculate the lowest concentration or percentage that can be reliably detected. Both the limit of detection and the limit of quantification should not be higher than the reporting limit (0.05%).

[0208] The solutions of each impurity and the main component reference substance were diluted step by step, and the LOQ was determined based on the signal-to-noise ratio S / N ≥ 10, and the LOD was determined based on S / N ≥ 3.

[0209] Table 27 Detection limit and quantification limit results

[0210]

[0211]

[0212] (5) Accuracy test

[0213] The accuracy is obtained by adding 80%, 100% and 120% of the impurity limit to the test sample and measuring the recovery rate. The accuracy of known impurities is to add a known amount of impurities and then measure the ratio (recovery rate) between the test result and the theoretical value of the known impurity in the added sample, expressed as a percentage, and the recovery rate of each sample is required to be between 95.0% and 105.0%, with RSD ≤ 5%.

[0214] Table 28 Impurity Ibu-1 Recovery

[0215]

[0216] Table 29 Impurity Ibu-2 Recovery

[0217]

[0218] Table 30 Impurity Ibu-3 Recovery

[0219]

[0220]

[0221] Table 31 Impurity Ibu-4 Recovery

[0222]

[0223] Table 32 Impurity Ibu-5 Recovery

[0224]

[0225] Table 33 Impurity PE-1 recovery rate

[0226]

[0227] Table 34 Impurity PE-2 recovery rate

[0228]

[0229]

[0230] Table 35 Impurity PE-3 recovery rate

[0231]

[0232] Table 36 Impurity PE-4 recovery rate

[0233]

[0234] Table 37 Impurity PE-5 recovery rate

[0235]

[0236]

[0237] Table 38 Impurity PE-6 Recovery Rate

[0238]

[0239] Table 39 Impurity PE-7 recovery rate

[0240]

[0241] Table 40 Impurity PE-8 Recovery Rate

[0242]

[0243] Table 41 Impurity PE-9 recovery rate

[0244]

[0245]

[0246] Table 42 Impurity PE-10 recovery rate

[0247]

[0248] Table 43 Impurity PE-11 recovery rate

[0249]

[0250] (6) Repeatability test

[0251] In the research on related substances of compound preparations, the related substances of each main component are studied and controlled separately. That is, after the attribution study, all the related substances appearing in the HPLC spectrum of the compound preparation are identified and attributed to the related substances of each main component one by one, and then the related substances are calculated for each main component respectively. In the standard, the related substances of each main component are controlled separately like single-prescription tablets, avoiding a "one-pot" type of research and quality control method.

[0252] The repeatability test is carried out by preparing 6 sample solutions and testing them, and the absolute deviation of the impurity change in the 6 data results must not exceed 0.1%.

[0253] Test solution: Take 10 tablets of Brulonin, grind into fine powder, accurately weigh 450 mg, put into a 100 mL volumetric flask, add appropriate amount of solvent (mobile phase A-acetonitrile (70:30)), shake well, ultrasonicate for 10 minutes, dilute to scale with solvent, shake well, and obtain. Prepare 6 portions in the same way.

[0254] Table 44 Related substances - Repeatability test results (area normalization method)

[0255]

[0256] (7) Stability of test solution

[0257] Test solution: Take 10 burobrene tablets, grind into fine powder, accurately weigh 450 mg, put into a 100 mL volumetric flask, add an appropriate amount of solvent (mobile phase A-acetonitrile (70:30)), shake well, ultrasonicate for 10 minutes, dilute to the scale with solvent, shake well, and obtain.

[0258] The test solution was placed at room temperature, sampled and analyzed at different time points, and the chromatogram was recorded.

[0259] Table 45 Test solution stability test results (area normalization method)

[0260]

[0261]

[0262] The present invention provides a method and method for analyzing related substances of a compound preparation of buprenorphine tablets. There are many methods and approaches to specifically implement the technical solution. The above is only a preferred embodiment of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the scope of protection of the present invention. All components not clearly defined in the present embodiment can be implemented using existing technologies.

Claims

1. A method for analyzing related substances of a compound preparation of Buluo Renin Tablets, characterized in that: The related substances are impurities in the compound preparation of Bu Luo Rensu Tablets, including impurity PE-1, impurity PE-2, impurity PE-3, impurity PE-4, impurity PE-5, impurity PE-6, impurity PE-7, impurity PE-8, impurity PE-9, impurity PE-10, impurity PE-11, impurity Ibu-1, impurity Ibu-2, impurity Ibu-3, impurity Ibu-4 and impurity Ibu-5; The analytical method is high performance liquid chromatography, and the high performance liquid chromatography conditions are: Chromatographic column: octadecylsilane bonded silica gel as filler; Detection wavelength: 215nm; Column temperature: 25-35°C; Flow rate: 0.8~1.2mL / min; Injection volume: 10 μL; Mobile phase A: 0.8-1.2 g / L sodium octane sulfonate aqueous solution, pH adjusted to 2.80 ± 0.02 with phosphoric acid; Mobile phase B: acetonitrile-water solution, the volume ratio of acetonitrile to water is 80:20; Gradient elution; The gradient elution conditions are: The impurity PE-1 has the molecular formula C 11 H 15 NO4; molecular weight: 225.2440; chemical name: (R)-2-((2-hydroxy-2-(3-hydroxyphenyl)ethyl)(methyl)amino)acetic acid; chemical structure: The impurity PE-2 has a molecular formula of C7H6O2; a molecular weight of 122.1230; a chemical name of 3-hydroxybenzaldehyde; and a chemical structural formula of: The impurity PE-3 has the molecular formula C 10 H 13 NO2; molecular weight: 179.2190; chemical name: N-methyl-4,6-dihydroxy-1,2,3,4-tetrahydroisoquinoline; chemical structure: The impurity PE-4 has the molecular formula C 10 H 13 NO2; molecular weight: 179.2190; chemical name: N-methyl-4,8-dihydroxy-1,2,3,4-tetrahydroisoquinoline; chemical structure: The impurity PE-5 has the molecular formula C8H 11 NO2; molecular weight: 153.1810; chemical name: (1R)-2-amino-1-(3-hydroxyphenyl)ethanol; chemical structure: The impurity PE-6 has the molecular formula C9H 11 NO2; molecular weight: 165.1920; chemical name: 1-(3-hydroxyphenyl)-2-(methylamino)ethanone; chemical structure: The impurity PE-7 has the molecular formula C 16 H 19 NO2; molecular weight: 257.3330; chemical name: (1R)-2-(benzylmethylamino)-1-(3-hydroxyphenyl)ethanol; chemical structure: The impurity PE-8 has the molecular formula C 16 H 17 NO2; molecular weight: 255.3170; chemical name: 2-(benzyl(methyl)amino)-1-(3-hydroxyphenyl)ethanone; chemical structure: The impurity PE-9 has the molecular formula C 17 H 13 NO3; molecular weight: 279.2950; chemical name: 1-(3-hydroxybenzoyl)-2-methylisoquinolin-6(2-hydrogen)-one; chemical structure: The impurity PE-10 has the molecular formula C 22 H 29 NO3; molecular weight: 355.4780; chemical name: (R)-1-(3-hydroxyphenyl)-2-(methylamino)ethyl-2-(4-isobutylphenyl) propionate; chemical structure: The impurity PE-11 has the molecular formula C 22 H 29 NO3; molecular weight: 355.4780; chemical name: N-((R)-2-hydroxy-2-(3-hydroxyphenyl)ethyl)-2-(4-isobutylphenyl)-N-methylpropionamide; chemical structure: The impurity Ibu-1 has a molecular formula of C 10 H 10 O3; molecular weight: 178.1870; chemical name: 2-(4-benzaldehyde) propionic acid; chemical structure: and its enantiomers; The impurity Ibu-2 has a molecular formula of C 13 H 16 O3; molecular weight: 220.2680; chemical name: 2-(4-isobutyrylphenyl)propionic acid; chemical structure: and its enantiomers; The impurity Ibu-3 has a molecular formula of C 11 H 14 O2; molecular weight: 178.2310; chemical name: 2-(4-ethylphenyl) propionic acid; chemical structure: The impurity Ibu-4 has a molecular formula of C 13 H 18 O2; molecular weight: 206.2850; chemical name: 2-(3-isobutylphenyl) propionic acid; chemical structure: The impurity Ibu-5 has a molecular formula of C 12 H 16 O; Molecular weight: 176.2590; chemical name: 4-isobutylacetophenone; chemical structure:

2. The method according to claim 1, characterized in that The chromatographic column has specifications of 4.6×250 mm and 5 μm.

3. The method according to claim 1, characterized in that The column temperature is 30°C.

4. The method according to claim 1, characterized in that The flow rate is 1.0 mL / min.

5. The method according to claim 1, characterized in that The mobile phase A is a 1.0 g / L sodium octane sulfonate aqueous solution, and the pH is adjusted to 2.80±0.02 with phosphoric acid.

6. Use of the method according to any one of claims 1 to 5 in the detection or separation of related substances in the compound preparation of Buluorenin Tablets.

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