Methods for detecting related substances in pharmaceutical compositions containing naltrexone and risperidone
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SHENZHEN SCIENCARE PHARMACEUTICAL CO LTD
- Filing Date
- 2024-12-31
- Publication Date
- 2026-06-30
Smart Images
Figure BDA0005225847920000011 
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Abstract
Description
Technical Field
[0001] This application relates to the field of pharmaceutical testing technology, and in particular to a method for detecting related substances in a pharmaceutical composition containing naltrexone and risperidone. Background Technology
[0002] Naltrxone is a pure opioid receptor antagonist, blocking μ-, δ-, and κ-opioid receptors. It can block the relapse effect of drugs, thereby weakening both positive and negative reinforcement, playing a good role in relapse prevention. Amphetamine stimulants, in addition to increasing dopamine levels in the interproximal space, also increase serotonin levels. Risperidone has good efficacy on both positive and negative symptoms and their associated affective symptoms (such as anxiety and depression), and can alleviate affective symptoms associated with schizophrenia. Utilizing this, risperidone alleviates the mental symptoms of amphetamine stimulant addicts, reduces the frequency of drug use and cravings, and is well-tolerated.
[0003] Chinese patent CN112245434A provides a sustained-release composition of naltrexone and risperidone, which enables the simultaneous release of naltrexone and risperidone, effectively inhibiting relapse of amphetamine-type drugs while reducing the side effects of risperidone. The naltrexone, risperidone, and their main related substances in the sustained-release composition are shown in Table 1 below.
[0004] Table 1
[0005]
[0006]
[0007] Currently, there are no studies on the detection of related substances in pharmaceutical compositions containing naltrexone and risperidone; only analytical methods for related substances in naltrexone or risperidone raw materials or single-active-ingredient formulations are available. For example, the European Pharmacopoeia EP11.0 provides quality standards for naltrexone hydrochloride and risperidone hydrochloride, which include methods for the detection of related substances. However, these methods are not applicable to the detection of pharmaceutical compositions containing naltrexone and risperidone, and cannot simultaneously meet the separation requirements for related impurities of naltrexone and risperidone, nor can they effectively separate newly generated impurities in pharmaceutical compositions containing naltrexone and risperidone. Summary of the Invention
[0008] Based on this, this application provides a method for detecting related substances in pharmaceutical compositions containing naltrexone and risperidone, which can effectively separate and detect multiple related substances in such compositions.
[0009] The specific technical solution is as follows:
[0010] A method for detecting related substances in a pharmaceutical composition containing naltrexone and risperidone, comprising the following steps:
[0011] A test solution was prepared using a combination of naltrexone and risperidone.
[0012] The test solution was analyzed by high-performance liquid chromatography (HPLC), and the HPLC detection conditions included:
[0013] (1) Chromatographic column: A C18 chromatographic column and a phenyl silica bonded chromatographic column arranged in series;
[0014] (2) Mobile phase A is a buffer solution, acetonitrile and methanol in a volume percentage of (85%–95%):(4%–10%):(1%–5%); mobile phase B is a buffer solution, acetonitrile and methanol in a volume percentage of (15%–25%):(60%–70%):(10%–20%); mobile phase C is a buffer solution, acetonitrile and methanol in a volume percentage of (15%–25%):(25%–35%):(45%–55%); each of the buffer solutions is independently a sodium octanesulfonate solution with a pH of 2.25–2.35.
[0015] In one embodiment, the C18 column is selected from: ZORBAX SB-C18, 4.6mm×250mm, 5μm; Luna Omega Polar C18, 4.6mm×250mm, 5μm; XSelect HSS C18, 4.6mm×250mm, 5μm.
[0016] In one embodiment, the phenyl silica bonded column is selected from: ZORBAX Eclipse XDBPhenyl, 4.6mm×250mm, 5μm; XBridge BEH Phenyl, 4.6mm×250mm, 5μm; YMC-Pack Phenyl, 4.6mm×250mm, 5μm.
[0017] In one embodiment, the elution procedure used in the high-performance liquid chromatography includes:
[0018] From 0 min to 20 min, maintain the volume percentage of mobile phase A at 90%, mobile phase B at 10%, and mobile phase C at 0%.
[0019] From 20 min to 58 min, the volume percentage of mobile phase A changed from 90% to 73%, the volume percentage of mobile phase B changed from 10% to 27%, and the volume percentage of mobile phase C was maintained at 0%.
[0020] From 58 min to 65 min, the volume percentage of mobile phase A changed from 73% to 60%, the volume percentage of mobile phase B changed from 27% to 0%, and the volume percentage of mobile phase C changed from 0% to 40%.
[0021] From 65 min to 110 min, the volume percentage of mobile phase A changed from 60% to 45%, while the volume percentage of mobile phase B was kept at 0%, and the volume percentage of mobile phase C changed from 40% to 55%.
[0022] In one embodiment, the detection conditions of the high-performance liquid chromatography further include: a column temperature of 40°C to 50°C; optionally, a column temperature of 43°C to 50°C.
[0023] In one embodiment, the detection conditions of the high-performance liquid chromatography further include: a flow rate of 1 mL / min to 1.3 mL / min; optionally, a flow rate of 1.2 mL / min to 1.3 mL / min.
[0024] In one embodiment, the detection conditions of the high-performance liquid chromatography further include: a detection wavelength of 275 nm to 285 nm; optionally, the detection wavelength is 278 nm to 282 nm.
[0025] In one embodiment, the naltrexone and risperidone pharmaceutical composition comprises a naltrexone and risperidone combination sustained-release formulation; optionally, the naltrexone and risperidone combination sustained-release formulation comprises a lipid-soluble polymer.
[0026] In one embodiment, during the preparation of the test solution, the naltrexone and risperidone drug composition is dissolved using a dissolving agent, which includes acetonitrile.
[0027] In one embodiment, the concentration of the buffer solution is 0.8 g / L to 1.5 g / L.
[0028] The above detection method is based on high performance liquid chromatography (HPLC). By using appropriate chromatographic columns, mobile phases, and other chromatographic conditions, it can effectively separate and accurately detect a variety of related substances in drug compositions containing naltrexone and risperidone. This greatly facilitates the quality control of drug compositions containing naltrexone and risperidone and provides a basis for the safety of drug formulations.
[0029] In some embodiments, the resolution of related impurities is superior to that of the related substances method of pharmacopoeia quality standards, with the chromatographic resolution of each impurity peak and adjacent peaks ≥1.0. It can simultaneously achieve the separation of the main peaks of naltrexone and risperidone as well as the separation of each related impurity, and can also effectively separate newly discovered impurities. Attached Figure Description
[0030] Figure 1Chromatogram of the spiked solution of the test sample in Example 1.
[0031] Figure 2 The chromatogram is of the spiked solution of the test sample in Comparative Example 1.
[0032] Figure 3 The chromatogram is of the spiked solution of the test sample in Comparative Example 2.
[0033] Figure 4 The chromatogram of column conditions ① in Example 2.
[0034] Figure 5 The chromatogram of column conditions ② in Example 2.
[0035] Figure 6 The chromatogram of column condition ③ in Example 2.
[0036] Figure 7 The chromatogram of column condition ④ in Example 2.
[0037] Figure 8 The chromatogram of column condition ⑤ in Example 2.
[0038] Figure 9 The chromatogram of column conditions ⑥ in Example 2.
[0039] Figure 10 The chromatogram of column condition ⑦ in Example 2.
[0040] Figure 11 The chromatogram of sodium heptanesulfonate as the ion-pairing reagent in Example 3.
[0041] Figure 12 The chromatogram of sodium octanesulfonate as the ion-pairing reagent in Example 3.
[0042] Figure 13 The chromatogram of sodium decanesulfonate as the ion-pairing reagent in Example 3.
[0043] Figure 14 Chromatogram of the buffer solution with pH 2.50 in Example 4.
[0044] Figure 15 Chromatogram of the buffer solution with pH 2.40 in Example 4.
[0045] Figure 16 Chromatogram of the buffer solution with pH 2.35 in Example 4.
[0046] Figure 17 Chromatogram of the buffer solution with pH 2.30 in Example 4.
[0047] Figure 18 Chromatogram of the buffer solution with pH 2.25 in Example 4.
[0048] Figure 19 Chromatogram of column temperature 40°C in Example 5.
[0049] Figure 20 Chromatogram of column temperature 43°C in Example 5.
[0050] Figure 21 Chromatogram of column temperature 45°C in Example 5.
[0051] Figure 22 Chromatogram of column temperature 47°C in Example 5.
[0052] Figure 23 Chromatogram of column temperature 50°C in Example 5.
[0053] Figure 24 Chromatogram of a flow rate of 1.0 mL / min in Example 6.
[0054] Figure 25 Chromatogram of a flow rate of 1.1 mL / min in Example 6.
[0055] Figure 26 Chromatogram of a flow rate of 1.2 mL / min in Example 6.
[0056] Figure 27 Chromatogram of a flow rate of 1.3 mL / min in Example 6.
[0057] Figure 28 Chromatogram of mobile phase ratio ① in Example 7.
[0058] Figure 29 Chromatogram of mobile phase ratio ② in Example 7.
[0059] Figure 30 Chromatogram of mobile phase ratio ③ in Example 7.
[0060] Figure 31 Chromatogram of mobile phase ratio ④ in Example 7.
[0061] Figure 32 Chromatograms of the impurity reference standard stock solution, excipient blank solution, and blank solution (from bottom to top) in Example 8.
[0062] Figure 33 Chromatogram of the test sample stock solution in Example 8.
[0063] Figure 34 Overlap diagram of the solutions for locating each impurity in Example 8. Detailed Implementation
[0064] The following detailed description, in conjunction with specific embodiments, illustrates the detection method for related substances in the pharmaceutical composition containing naltrexone and risperidone of this application. This application can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and complete understanding of the disclosure of this application.
[0065] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.
[0066] As used herein, the terms “and / or,” “or / and,” and “and / or” may include any one of two or more of the related listed items, as well as any and all combinations of the related listed items, including any two related listed items, any more related listed items, or a combination of all the related listed items.
[0067] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.
[0068] In this application, numerical ranges are referred to as continuous unless otherwise specified, and include the minimum and maximum values of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.
[0069] Unless otherwise specified, the percentage content mentioned in this application refers to mass percentage for solid-liquid mixtures and solid-phase-solid mixtures, and volume percentage for liquid-phase-liquid mixtures. Unless otherwise specified, the solvent of the solution is water.
[0070] Unless otherwise specified, all percentage concentrations mentioned in this application refer to the final concentration. The final concentration refers to the proportion of the added component in the system after the addition of that component.
[0071] Unless otherwise specified, the temperature parameters in this application may be either constant temperature processing or processing within a certain temperature range. The constant temperature processing allows for temperature fluctuations within the precision range controlled by the instrument.
[0072] In this application, room temperature generally refers to 10℃~30℃, and preferably 20±5℃.
[0073] Some examples of this application provide a method for detecting related substances in a pharmaceutical composition containing naltrexone and risperidone, comprising the following steps:
[0074] A test solution was prepared using a combination of naltrexone and risperidone.
[0075] The test solution was analyzed by high-performance liquid chromatography (HPLC), and the HPLC detection conditions included:
[0076] (1) Chromatographic column: A C18 chromatographic column and a phenyl silica bonded chromatographic column arranged in series;
[0077] (2) Mobile phase A is a buffer solution, acetonitrile and methanol in a volume percentage of (85%–95%):(4%–10%):(1%–5%); mobile phase B is a buffer solution, acetonitrile and methanol in a volume percentage of (15%–25%):(60%–70%):(10%–20%); mobile phase C is a buffer solution, acetonitrile and methanol in a volume percentage of (15%–25%):(25%–35%):(45%–55%); each of the buffer solutions is independently a sodium octanesulfonate solution with a pH of 2.25–2.35.
[0078] Furthermore, mobile phase A is a buffer solution, acetonitrile, and methanol in a volume percentage of 90%:8%:2%.
[0079] Furthermore, mobile phase B is a buffer solution, acetonitrile, and methanol in a volume percentage of 20%:64%:16%.
[0080] Furthermore, mobile phase C is a buffer solution, acetonitrile, and methanol in a volume percentage ratio of 20:32%:48%.
[0081] Without limitation, the pH of each sodium octane sulfonate solution is independently 2.25, 2.3, 2.35, or any two of the foregoing. Further, each of the buffer solutions is an sodium octane sulfonate solution with a pH of 2.25 to 2.3.
[0082] In some examples, the concentration of the buffer solution is 0.8 g / L to 1.5 g / L. Without limitation, the concentration of the buffer solution includes, but is not limited to: 0.8 g / L, 0.9 g / L, 1 g / L, 1.1 g / L, 1.2 g / L, 1.3 g / L, 1.4 g / L, 1.5 g / L, or any range between the foregoing.
[0083] In some of these examples, the elution procedure used in the high-performance liquid chromatography includes:
[0084] From 0 min to 20 min, maintain the volume percentage of mobile phase A at 90%, mobile phase B at 10%, and mobile phase C at 0%.
[0085] From 20 min to 58 min, the volume percentage of mobile phase A changed from 90% to 73%, the volume percentage of mobile phase B changed from 10% to 27%, and the volume percentage of mobile phase C was maintained at 0%.
[0086] From 58 min to 65 min, the volume percentage of mobile phase A changed from 73% to 60%, the volume percentage of mobile phase B changed from 27% to 0%, and the volume percentage of mobile phase C changed from 0% to 40%.
[0087] From 65 min to 110 min, the volume percentage of mobile phase A changed from 60% to 45%, while the volume percentage of mobile phase B was kept at 0%, and the volume percentage of mobile phase C changed from 40% to 55%.
[0088] Understandably, a C18 column can be replaced by a column with equivalent performance, and a phenyl-silica bonded column can be replaced by a column with equivalent performance.
[0089] In some of these examples, the C18 column is selected from: ZORBAX SB-C18, 4.6 mm × 250 mm, 5 μm; Luna Omega Polar C18, 4.6 mm × 250 mm, 5 μm; and XSelect HSS C18, 4.6 mm × 250 mm, 5 μm.
[0090] In some of these examples, the phenyl silica bonded column is selected from: ZORBAX Eclipse XDBPhenyl, 4.6 mm × 250 mm, 5 μm; XBridge BEH Phenyl, 4.6 mm × 250 mm, 5 μm; YMC-Pack Phenyl, 4.6 mm × 250 mm, 5 μm.
[0091] Understandably, in the chromatographic column, "Amm×Bmm,Cμm" means that A represents the diameter, B represents the length, and C represents the particle size of the packing material.
[0092] In some examples, the detection conditions for the high-performance liquid chromatography (HPLC) further include a column temperature of 40°C to 50°C. Without limitation, the column temperature includes, but is not limited to, 40°C, 43°C, 45°C, 47°C, 50°C, or any range between the foregoing. Further, the column temperature is 43°C to 50°C.
[0093] In some examples, the detection conditions for the high-performance liquid chromatography (HPLC) further include a flow rate of 1 mL / min to 1.3 mL / min. Without limitation, the flow rate includes, but is not limited to, 1 mL / min, 1.1 mL / min, 1.2 mL / min, 1.3 mL / min, or any range between the foregoing. Further, the flow rate is 1.2 mL / min to 1.3 mL / min.
[0094] In some examples, the detection conditions of the high-performance liquid chromatography further include a detection wavelength of 275 nm to 285 nm. Without limitation, the detection wavelength includes, but is not limited to, 275 nm, 278 nm, 280 nm, 282 nm, 285 nm, or any range between the foregoing. Further, the detection wavelength is 278 nm to 282 nm.
[0095] In some examples, the naltrexone and risperidone pharmaceutical composition comprises a sustained-release formulation of naltrexone and risperidone. Further, the sustained-release formulation of naltrexone and risperidone comprises a lipid-soluble polymer. Without limitation, the lipid-soluble polymer may include, for example, one or more of glycolide-lactide copolymer and polylactic acid.
[0096] In some examples, during the preparation of the test solution, the naltrexone and risperidone pharmaceutical composition is dissolved using a dissolving agent, including acetonitrile. For formulations containing polymeric materials, the naltrexone and risperidone pharmaceutical composition can be dissolved first in acetonitrile and then diluted with a diluent to prepare the test solution.
[0097] In some examples, preparing the test solution includes the step of diluting the naltrexone and risperidone pharmaceutical composition using an aqueous hydrochloric acid solution as the diluent. Further, the concentration of the aqueous hydrochloric acid solution is 0.05 mol / L to 0.15 mol / L. Without limitation, the concentration of the aqueous hydrochloric acid solution includes, but is not limited to, 0.05 mol / L, 0.08 mol / L, 0.1 mol / L, 0.12 mol / L, 0.15 mol / L, or any range between the foregoing.
[0098] For experimental parameters not specified in the following specific embodiments, please refer to the guidelines given in this application document first, or refer to experimental manuals or other experimental methods known in the art, or refer to the experimental conditions recommended by the manufacturer.
[0099] The raw materials and reagents involved in the following specific embodiments can be obtained commercially or prepared by those skilled in the art using known methods.
[0100] Example 1
[0101] This embodiment describes a method for detecting related substances in a pharmaceutical composition containing naltrexone and risperidone.
[0102] (1) Solution preparation:
[0103] Stock solutions of single impurities: Take about 15 mg each of naltrexone impurities NTX-B, NTX-L, NTX-F, NTX-G, NTX-C, NTX-I, and NTX-D, and risperidone impurities RPD-C and RPD-G, and place them in 100 mL volumetric flasks. Dissolve them in an appropriate amount of methanol and dilute to the mark with 0.1 mol / L hydrochloric acid solution to prepare stock solutions of the above impurities.
[0104] Test sample stock solution: Take about 250 mg of fine powder of naltrexone and risperidone compound sustained-release preparation (composition includes naltrexone, risperidone and polylactic acid, etc., provided by Shenzhen Shankang Pharmaceutical Technology Co., Ltd.), place it in a 10 mL volumetric flask, add an appropriate amount of acetonitrile, sonicate to dissolve, dilute with acetonitrile to the mark, shake well, centrifuge, and take the supernatant to obtain the test sample.
[0105] Test solution: Accurately measure 2 mL of the test solution stock solution and place it in a 10 mL volumetric flask. Dilute to the mark with 0.1 mol / L hydrochloric acid solution, shake well, centrifuge, and take the supernatant.
[0106] Spiked solution for test sample: Accurately measure 2 mL of the test sample stock solution, 300 μL each of the stock solutions of naltrexone impurities G, C, I, and D reference standards, and 30 μL each of the stock solutions of risperidone impurities C and G reference standards, place them in a 10 mL volumetric flask, dilute to the mark with 0.1 mol / L hydrochloric acid solution, shake well, and centrifuge to obtain the solution.
[0107] Blank solvent: 0.1 mol / L hydrochloric acid solution.
[0108] (2) The chromatographic conditions are as follows:
[0109] Chromatographic columns: Two columns with different packing materials were connected in series, namely Waters Symmetry C18, 4.6×250mm, 5μm and Agilent ZORBAX SB-Phenyl, 4.6×250mm, 5μm;
[0110] Buffer solution: 1.1 g / L sodium octane sulfonate solution (adjusted to pH 2.25 with phosphoric acid);
[0111] Mobile phase A: Buffer-acetonitrile-methanol (900:80:20, v:v:v);
[0112] Mobile phase B: Buffer-acetonitrile-methanol (200:640:160, v:v:v);
[0113] Mobile phase C: buffer-acetonitrile-methanol (200:320:480, v:v:v);
[0114] The elution gradient is shown in Table 2 below:
[0115] Table 2
[0116]
[0117]
[0118] Flow rate: 1.2 mL / min;
[0119] Detection wavelength: 280nm;
[0120] Column temperature: 45℃.
[0121] Accurately measure the spiked solution of the test sample and inject it into the liquid chromatograph. Run the chromatogram according to the chromatographic conditions described above and record the chromatogram. The results are as follows: Figure 1 As shown.
[0122] Comparative Example 1
[0123] This comparative example was prepared according to the chromatographic conditions specified in the European Pharmacopoeia EP11.0 quality standard for naltrexone hydrochloride, and the spiked solution of the test sample in Example 1 was tested. The chromatographic conditions are as follows:
[0124] Chromatographic column: Symmetry Shield RP C18 4.6×150mm 5μm;
[0125] Mobile phase A: 1.1 g / L sodium octane sulfonate solution (pH 2.3);
[0126] Mobile phase B: Acetonitrile;
[0127] The elution gradient is shown in Table 3 below:
[0128] Table 3
[0129] Time (min) A(％) B(％) 0 90 10 45 55 45 47 90 10 55 90 10
[0130] Flow rate: 1.2 mL / min;
[0131] Detection wavelength: 230nm;
[0132] Column temperature: 40℃.
[0133] The results are as follows Figure 2 As shown.
[0134] Comparative Example 2
[0135] This comparative example tested the spiked solution of the test sample in Example 1 according to the chromatographic conditions specified in the quality standard of risperidone in European Pharmacopoeia EP11.0. The chromatographic conditions are as follows:
[0136] Chromatographic column: Symmetry Shield RP C18 4.6×100mm, 3μm;
[0137] Mobile phase A: 5 g / L ammonium acetate solution;
[0138] Mobile phase B: Methanol;
[0139] The elution gradient is shown in Table 4 below:
[0140] Table 4
[0141] Time (min) A(％) B(％) 0 70 30 2 70 30 17 30 70 22 30 70
[0142] Flow rate: 1.5 mL / min;
[0143] Detection wavelength: 260nm;
[0144] Column temperature: room temperature.
[0145] The results are as follows Figure 3 As shown.
[0146] comprehensive Figures 1-3 It can be seen that under the chromatographic conditions of Comparative Example 1 and Comparative Example 2, the resolution of impurities was <1.5, while under the chromatographic conditions of Example 1, the resolution of impurities was ≥1.5, which greatly improved the separation effect of related substances, especially the resolution between the naltrexone main peak and naltrexone impurity C, as well as the resolution between naltrexone impurities B and F, and realized the detection of naltrexone impurity L, which can better meet the impurity separation requirements of drug compositions containing naltrexone and risperidone.
[0147] Example 2
[0148] This example is a screening study of chromatographic columns.
[0149] The solution preparation and chromatographic conditions were the same as in Example 1, the main difference being the use of a different chromatographic column, as shown in Table 5 below:
[0150] Table 5
[0151]
[0152] Inject the spiked solution of the test sample, record the chromatogram, and the results are as follows: Figures 4-10As shown. Among the chromatographic columns under conditions ① to ⑤, only column ⑤ achieves optimal separation of impurities between NTX-B and NTX-F. However, condition ⑤ cannot effectively separate RPD and RPD-G. Column ① can effectively separate RPD and RPD-G, and effectively separate RPD-C from adjacent impurity peaks. Therefore, column ⑤ is connected in series with column ① (i.e., condition ⑥). To further optimize the separation of RPD and RPD-G, condition ⑦ is further optimized. Compared with other chromatographic conditions, this application can separate more unknown impurities, such as the unknown impurity between the newly added naltrexone main peak and naltrexone impurity C (retention time 51.044 min), and the resolution of the separated impurities all meet the requirements. Therefore, it is preferable to connect the Waters Symmetry C18 4.6×250mm, 5μm column and the Agilent ZORBAX SB-Phenyl 4.6×250mm, 5μm column in series.
[0153] Example 3
[0154] This example illustrates the screening of ion-pairing reagents.
[0155] The solution preparation and chromatographic conditions are the same as in Example 1, the main difference being the use of different ion-pairing reagents, specifically:
[0156] Buffer solution ①: 1.1 g / L sodium heptanesulfonate solution (adjusted to pH 2.25 with phosphoric acid);
[0157] Buffer solution ②: 1.1 g / L sodium octane sulfonate solution (adjusted to pH 2.25 with phosphoric acid);
[0158] Buffer solution ③: 1.1 g / L sodium decane sulfonate solution (adjusted to pH 2.25 with phosphoric acid).
[0159] Inject the spiked solution of the test sample, record the chromatogram, and the results are as follows: Figures 11-13 As shown, the separation of impurities NTX-B, NTX-L, and NTX-C from adjacent peaks is poor under heptanesulfonate sodium solution conditions. Under decanesulfonate sodium solution conditions, the separation of impurities NTX-B, NTX-L, NTX-G, NTX-D, and RPD-C from adjacent peaks is also poor. The separation of all relevant substances is best under octanesulfonate sodium solution conditions. This indicates that neither heptanesulfonate nor decanesulfonate sodium solutions are suitable; therefore, octanesulfonate sodium was chosen as the ion-pairing reagent in this application.
[0160] Example 4
[0161] This example is a screening study on the pH value of the mobile phase buffer solution.
[0162] The solution preparation and chromatographic conditions are the same as in Example 1, the main difference being the use of mobile phase buffers with different pH values, specifically:
[0163] Buffer solution ①: 1.1 g / L sodium octane sulfonate solution (adjusted to pH 2.50 with phosphoric acid);
[0164] Buffer solution ②: 1.1 g / L sodium octane sulfonate solution (adjusted to pH 2.40 with phosphoric acid);
[0165] Buffer solution ③: 1.1 g / L sodium octane sulfonate solution (adjusted to pH 2.35 with phosphoric acid);
[0166] Buffer solution ④: 1.1 g / L sodium octane sulfonate solution (adjusted to pH 2.30 with phosphoric acid);
[0167] Buffer solution ⑤: 1.1 g / L sodium octane sulfonate solution (adjusted to pH 2.25 with phosphoric acid).
[0168] Inject the spiked solution of the test sample, record the chromatogram, and the results are as follows: Figures 14-18 As shown in Table 6:
[0169] Table 6
[0170]
[0171]
[0172] It can be seen that as the pH value of the buffer solution decreases, the separation between NTX and its adjacent peaks gradually improves. The separation is worst at pH 2.50 (peak overlap), the separation is <1.0 at pH 2.40, and the impurity separation effect is best at pH 2.25. Therefore, the pH value of the buffer solution should be selected from 2.25 to 2.35, with pH 2.25 being the preferred value.
[0173] Example 5
[0174] This example is a column temperature screening study.
[0175] The solution preparation and chromatographic conditions were the same as in Example 1, the main difference being the use of different column temperatures, specifically: 40℃, 43℃, 45℃, 47℃, and 50℃.
[0176] Inject the spiked solution of the test sample, record the chromatogram, and the results are as follows: Figures 19-23 And as shown in Table 7:
[0177] Table 7
[0178]
[0179] It can be seen that at a column temperature of 40℃, NTX-C slightly overlaps with adjacent impurities. As the column temperature increases, the adjacent impurity peaks after the naltrexone main peak gradually approach the naltrexone peak, and the separation between the naltrexone impurity C and the naltrexone impurity gradually improves. At the same time, the separation between the risperidone main peak and its impurity G gradually decreases. Taking all factors into consideration, the column temperature can be selected from 43℃ to 50℃, with 45℃ being preferred.
[0180] Example 6
[0181] This example is a flow rate screening study.
[0182] The solution preparation and chromatographic conditions were the same as in Example 1, the main difference being the use of different flow rates, specifically: 1.0 mL / min, 1.1 mL / min, 1.2 mL / min, and 1.3 mL / min.
[0183] Inject the spiked solution of the test sample, record the chromatogram, and the results are as follows: Figures 24-27 As shown in Table 8:
[0184] Table 8
[0185]
[0186]
[0187] It can be seen that as the flow rate increases, the separation of the NTX or RPD main peak from its adjacent impurities gradually improves. Therefore, taking all factors into consideration, the flow rate can be selected from 1.2 mL / min to 1.3 mL / min, with 1.2 mL / min being preferred.
[0188] Example 7
[0189] This embodiment is a screening study on the proportion of mobile phase.
[0190] The solution preparation and chromatographic conditions were the same as in Example 1, the main difference being the use of different mobile phase ratios, as shown in Table 9 below:
[0191] Table 9
[0192]
[0193] Inject the spiked solution of the test sample, record the chromatogram, and the results are as follows: Figures 28-31 As shown in Table 10:
[0194] Table 10
[0195]
[0196] It is evident that reducing the methanol ratio can improve the separation of naltrexone-related impurities, but is not conducive to the separation of risperidone-related impurities. Therefore, it is necessary to increase the C phase (containing a high proportion of methanol) so that the methanol ratio is maintained at a low level in the front section of the gradient and increased in the back section. Therefore, considering all factors, the mobile phase composition is determined to be ratio ④.
[0197] Example 8
[0198] This embodiment is a methodological investigation.
[0199] (1) Exclusivity
[0200] The solution is prepared as follows:
[0201] Blank solvent: 0.1 mol / L hydrochloric acid solution.
[0202] Blank excipient solution: Weigh 125.00 mg of PLA and place it in a 10 mL volumetric flask. Dissolve and dilute to the mark with acetonitrile, shake well, accurately measure 2 mL, place it in a 10 mL volumetric flask, dilute to the mark with 0.1 mol / L hydrochloric acid solution, centrifuge, and take the supernatant.
[0203] Risperidone reference standard stock solution: Accurately weigh 20.70 mg of risperidone reference standard, place it in a 100 mL volumetric flask, add an appropriate amount of 0.1 mol / L hydrochloric acid solution to dissolve it, and dilute to the mark. Shake well to obtain the solution.
[0204] Stock solution of single impurity reference standard: Same as in Example 1.
[0205] Mixed impurity reference solution: Weigh 13.55 mg of naltrexone hydrochloride reference standard and place it in a 5 mL volumetric flask. Accurately measure 150 μL each of the stock solutions of naltrexone impurities B, F, G, C, I, and D reference standards, 2 mL of risperidone stock solution, and 380 μL of risperidone impurity stock solution, and place them in the same 5 mL volumetric flask. Dilute to the mark with 0.1 mol / L hydrochloric acid solution and shake well.
[0206] Test solution: Same as in Example 1.
[0207] The chromatographic conditions were the same as in Example 1.
[0208] Inject blank solvent, excipient blank solution, mixed impurity reference solution, and test solution, and record the chromatograms. Results are as follows: Figures 32-34 And as shown in Table 11:
[0209] Table 11
[0210]
[0211] The above results indicate that the method of this application has good specificity.
[0212] (2) Limit of Quantification and Limit of Detection
[0213] The solution is prepared as follows:
[0214] Naltrexone reference standard stock solution: Accurately weigh 24.06 mg of naltrexone hydrochloride reference standard, place it in a 100 mL volumetric flask, add an appropriate amount of 0.1 mol / L hydrochloric acid solution to dissolve it, and dilute to the mark. Shake well to obtain the solution.
[0215] Naltrexone impurity reference stock solution (24 μg / mL): Accurately measure 1.6 mL each of naltrexone impurity B, C, L, F, G, I, and D reference stock solutions (see Example 1) and 1.2 mL of naltrexone reference stock solution, place them in the same 20 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution.
[0216] Naltrexone impurity limit of quantitation solution: Accurately measure 0.95 mL of naltrexone impurity reference standard stock solution, place it in a 10 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution.
[0217] Naltrexone impurity detection limit solution: Accurately measure 3 mL of naltrexone quantitation limit solution, place it in a 10 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution.
[0218] Risperidone impurity reference stock solution (2.4 μg / ml): Accurately measure 3.2 mL each of risperidone impurity C and G reference stock solutions (see Example 1) and 2.4 mL of risperidone reference stock solution (under the specificity section of Example 8), place them in the same 20 mL volumetric flask, dilute to the mark with 0.1 mol / L hydrochloric acid solution, accurately measure 5 mL of the above solution, place it in a 50 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution to obtain the final solution.
[0219] Risperidone impurity limit of quantitation solution: Accurately measure 1.175 mL of risperidone impurity reference standard stock solution, place it in a 10 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution.
[0220] Risperidone Impurity Detection Limit Solution: Accurately measure 3 mL of risperidone Quantification Limit Solution and place it in a 10 mL volumetric flask. Dilute to the mark with 0.1 mol / L hydrochloric acid solution.
[0221] The chromatographic conditions were the same as in Example 1.
[0222] Inject naltrexone impurity limit of quantitation solution, risperidone impurity limit of quantitation solution, naltrexone impurity limit of detection solution, and risperidone limit of detection solution, and record the chromatograms. The results are shown in Tables 12 and 13:
[0223] Table 12
[0224]
[0225] Table 13
[0226]
[0227]
[0228] (3) Linear
[0229] The solution is prepared as follows:
[0230] Naltrexone impurity B linear stock solution (60 μg / mL): Accurately measure 3 mL of naltrexone reference stock solution (200 μg / mL) and 4 mL of naltrexone impurity B reference stock solution (150 μg / L), place them in the same 10 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution.
[0231] Naltrexone impurity L and C linear stock solution (60 μg / mL): Accurately measure 4 mL each of naltrexone impurity L and C reference stock solution (150 μg / mL), place them in the same 10 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution.
[0232] Naltrexone impurity F and I linear stock solution (60 μg / mL): Accurately measure 4 mL each of naltrexone impurity F and I reference stock solution (150 μg / mL), place them in the same 10 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution.
[0233] Linear stock solution of naltrexone impurities G and D (60 μg / mL): Accurately measure 4 mL each of naltrexone impurity G and D reference stock solution (150 μg / mL), place them in a 10 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution.
[0234] Risperidone linear stock solution (6 μg / mL): Accurately measure 4 mL each of risperidone impurity C and G reference stock solutions (150 μg / mL) and 3 mL of risperidone reference stock solution, place them in the same 20 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution. Accurately measure 5 mL of the above solution, place it in a 25 mL volumetric flask, and dilute to the mark with 0.1 mol / L hydrochloric acid solution.
[0235] Take linear stock solutions of naltrexone and each naltrexone impurity, and prepare linear solutions of naltrexone and each naltrexone impurity according to Table 14.
[0236] Table 14
[0237]
[0238] Take linear stock solutions of risperidone and each risperidone impurity, and prepare linear solutions of risperidone and each risperidone impurity according to Table 15.
[0239] Table 15
[0240]
[0241] The chromatographic conditions were the same as in Example 1.
[0242] Inject naltrexone and linear solutions of each naltrexone impurity, risperidone and linear solutions of each risperidone impurity, and record the chromatograms. The results are shown in Table 16.
[0243] Table 16
[0244]
[0245] (4) Repeatability
[0246] The solution is prepared as follows:
[0247] Stock solution of test sample: Take about 250 mg of the fine powder of this product, place it in a 10 mL volumetric flask, add an appropriate amount of acetonitrile, sonicate to dissolve, dilute to the mark with acetonitrile, and shake well. Prepare 6 parallel solutions.
[0248] Repeatability solution: Accurately measure 2 mL of the test sample stock solution, 300 μL of the stock solutions of naltrexone impurities G, C, I, D, and L, and 30 μL each of the stock solutions of risperidone impurities C and G, and place them in a 10 mL volumetric flask. Dilute to the mark with 0.1 mol / L hydrochloric acid solution, shake well, and centrifuge. Prepare 6 parallel aliquots.
[0249] Control solution: Accurately measure 1 mL of the repeatability solution into a 50 mL volumetric flask, dilute to the mark with 0.1 mol / L hydrochloric acid solution, and shake well. Accurately measure 5 mL of the control solution into a 50 mL volumetric flask, dilute to the mark with 0.1 mol / L hydrochloric acid solution, and shake well. Prepare 6 parallel aliquots.
[0250] The chromatographic conditions were the same as in Example 1.
[0251] Inject the repeat solution and control solution, and record the chromatograms. The results are shown in Table 17:
[0252] Table 17
[0253]
[0254]
[0255] As can be seen, the number of impurity peaks with impurity content above the reporting limit (0.1%) was consistent across the six determinations. For single and total impurities, the actual detected amounts were less than or equal to 0.1%, and the absolute values of the content changes were all no greater than 0.03%; for single and total impurities, the actual detected amounts were greater than 0.1%, and the RSD values were all less than 10%; for single and total impurities, the actual detected amounts were greater than 0.5%, and the RSD values were all less than 5%. Therefore, the method of this application exhibits good repeatability.
[0256] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.
[0257] The embodiments described above merely illustrate several implementation methods of this application to facilitate a detailed understanding of the technical solutions of this application, but should not be construed as limiting the scope of protection of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the scope of protection of this application. Technical solutions obtained by those skilled in the art based on the technical solutions provided in this application through logical analysis, reasoning, or limited experimentation are all within the scope of protection of the appended claims. Therefore, the scope of protection of this patent application should be determined by the content of the appended claims, and the specification can be used to interpret the content of the claims.
Claims
1. A method for detecting related substances in a pharmaceutical composition containing naltrexone and risperidone, characterized in that, Includes the following steps: A test solution was prepared using a combination of naltrexone and risperidone. The test solution was analyzed by high-performance liquid chromatography (HPLC), and the HPLC detection conditions included: (1) Chromatographic column: A C18 chromatographic column and a phenyl silica bonded chromatographic column arranged in series; (2) Mobile phase A is a buffer solution, acetonitrile and methanol in a volume percentage of (85%–95%):(4%–10%):(1%–5%); mobile phase B is a buffer solution, acetonitrile and methanol in a volume percentage of (15%–25%):(60%–70%):(10%–20%); mobile phase C is a buffer solution, acetonitrile and methanol in a volume percentage of (15%–25%):(25%–35%):(45%–55%); each of the buffer solutions is independently a sodium octanesulfonate solution with a pH of 2.25–2.
35.
2. The method for detecting related substances in the pharmaceutical composition containing naltrexone and risperidone according to claim 1, characterized in that, The C18 chromatographic column was selected from: ZORBAX SB-C18, 4.6mm×250mm, 5μm; Luna Omega Polar C18, 4.6mm×250mm, 5μm; and XSelect HSS C18, 4.6mm×250mm, 5μm.
3. The method for detecting related substances in the pharmaceutical composition containing naltrexone and risperidone according to claim 1, characterized in that, The phenyl-silica bonded chromatographic columns were selected from: ZORBAX Eclipse XDB Phenyl, 4.6mm×250mm, 5μm; XBridge BEH Phenyl, 4.6mm×250mm, 5μm; and YMC-Pack Phenyl, 4.6mm×250mm, 5μm.
4. The method for detecting related substances in the pharmaceutical composition containing naltrexone and risperidone according to claim 1, characterized in that, The elution procedure used in the high-performance liquid chromatography includes: From 0 min to 20 min, maintain the volume percentage of mobile phase A at 90%, mobile phase B at 10%, and mobile phase C at 0%. From 20 min to 58 min, the volume percentage of mobile phase A changed from 90% to 73%, the volume percentage of mobile phase B changed from 10% to 27%, and the volume percentage of mobile phase C was maintained at 0%. From 58 min to 65 min, the volume percentage of mobile phase A changed from 73% to 60%, the volume percentage of mobile phase B changed from 27% to 0%, and the volume percentage of mobile phase C changed from 0% to 40%. From 65 min to 110 min, the volume percentage of mobile phase A changed from 60% to 45%, while the volume percentage of mobile phase B was kept at 0%, and the volume percentage of mobile phase C changed from 40% to 55%.
5. The method for detecting related substances in a pharmaceutical composition containing naltrexone and risperidone according to any one of claims 1 to 4, characterized in that, The detection conditions for the high-performance liquid chromatography also include: a column temperature of 40℃~50℃; optionally, a column temperature of 43℃~50℃.
6. The method for detecting related substances in a pharmaceutical composition containing naltrexone and risperidone according to any one of claims 1 to 4, characterized in that, The detection conditions for the high-performance liquid chromatography also include: a flow rate of 1 mL / min to 1.3 mL / min; optionally, a flow rate of 1.2 mL / min to 1.3 mL / min.
7. The method for detecting related substances in a pharmaceutical composition containing naltrexone and risperidone according to any one of claims 1 to 4, characterized in that, The detection conditions for the high-performance liquid chromatography also include: a detection wavelength of 275 nm to 285 nm; optionally, a detection wavelength of 278 nm to 282 nm.
8. The method for detecting related substances in a pharmaceutical composition containing naltrexone and risperidone according to any one of claims 1 to 4, characterized in that, The naltrexone and risperidone pharmaceutical composition comprises a naltrexone and risperidone combination sustained-release formulation; optionally, the naltrexone and risperidone combination sustained-release formulation comprises a lipid-soluble polymer.
9. The method for detecting related substances in the pharmaceutical composition containing naltrexone and risperidone according to claim 8, characterized in that, In the preparation of the test solution, the naltrexone and risperidone drug composition is dissolved using a dissolving agent, which includes acetonitrile.
10. The method for detecting related substances in a pharmaceutical composition containing naltrexone and risperidone according to any one of claims 1 to 4, characterized in that, The concentration of the buffer solution is 0.8 g / L to 1.5 g / L.