Method for improving quality control of cefditoren pivoxil granules

Abstract

A method for improving quality control of cefditoren pivoxil granules, comprising: injecting a sample solution of cefditoren pivoxil granules into a high performance liquid chromatograph for analysis to test impurities in the cefditoren pivoxil granules, wherein: a mixed solution having a volume ratio of an ammonium formate solution, acetonitrile, and methanol of (43-47):(23-27):(23-27) is used as a mobile phase A; a mixed solution having a volume ratio of an ammonium formate solution, acetonitrile, and methanol of (43-47):(53-57):(53-57) is used as a mobile phase B; gradient elution is employed; and the sample solution of cefditoren pivoxil granules is prepared using the mobile phase A as a diluent. The method ameliorates peak fronting of a cefditoren pivoxil ring-opened impurity peak and improves the resolution of unknown impurity peaks in the cefditoren pivoxil granules, and exhibits excellent reproducibility and high accuracy, thereby improving the quality control level of the cefditoren pivoxil granules.

Description

Methods to improve the quality control of ceftriaxone granules Technical Field

[0001] This invention belongs to the field of pharmaceutical analysis. Specifically, this invention relates to a method for improving the quality control of ceftoranil granules, which includes injecting a ceftoranil granule sample solution into a high-performance liquid chromatograph for analysis to detect impurities in the ceftoranil granules. Background Technology

[0002] The chemical name of cefotaxime is 2,2-dimethylpropionyloxymethyl(6R,7R)-7-[(Z)-2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-[(Z)-2-(4-methyl-1,3-thiazolyl-5-yl)vinyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, and it has the following structural formula:

[0003] After being absorbed by the human body, ceftoranil is metabolized in the intestinal wall into ceftoran, which exerts its antibacterial activity. It exhibits strong antibacterial activity against Staphylococcus spp., Streptococcus spp., Peptostreptococcus spp., Moraxella catarrhalis, Propionibacterium acnes, Escherichia coli, Citrobacter spp., Klebsiella spp., Enterobacter spp., Serratia spp., Proteus spp. (Proteus mirabilis, Proteus vulgaris), Morganella spp., Providencia spp., Haemophilus influenzae, and Bacteroides spp., and has a good therapeutic effect on infections caused by these bacteria. However, ceftoranil granules also have many adverse reactions. For example, allergic reactions include rash, itching, urticaria, and fever; gastrointestinal reactions include nausea, vomiting, and diarrhea, and, as with other broad-spectrum antibiotics, rare pseudomembranous colitis may occur; hematologic reactions include eosinophilia and leukopenia; there have been reports of positive Coombs reactions when treated with cephalosporins; renal function changes include occasional increases in blood urea nitrogen (BUN) and serum creatinine; and liver function changes include occasional increases in aspartate aminotransferase (AST), alanine aminotransferase (ALT), alanine aminotransferase (ALT), and alkaline phosphatase (Al-P).

[0004] Adverse drug reactions are sometimes caused not only by the drug itself, but also by impurities in the drug. Therefore, controlling impurities in drugs is very important and has a significant impact on drug quality.

[0005] Obtaining high-quality pharmaceuticals has always been a human pursuit. High quality means high purity of the active ingredient, while minimizing the types and amounts of impurities to ensure drug safety and efficacy, and to avoid unforeseen adverse effects. However, existing methods cannot completely identify all unknown impurities or detect related substances in ceftorepimex granules with high resolution, which is detrimental to drug quality control. Therefore, further improvements are needed in the analytical methods for related substances in ceftorepimex granules to enhance the quality control level of ceftorepimex granules. Summary of the Invention

[0006] To address the above problems, this invention provides a method for improving the quality control of ceftriaxone granules, the method comprising:

[0007] The ceftriaxone particle sample solution was injected into a high-performance liquid chromatograph for analysis to detect impurities in the ceftriaxone particles. Gradient elution was performed using mobile phase A and mobile phase B as eluents. Mobile phase A was a mixture of ammonium formate solution, acetonitrile, and methanol in a volume ratio of (43-47):(23-27):(23-27), and mobile phase B was a mixture of ammonium formate solution, acetonitrile, and methanol in a volume ratio of (43-47):(53-57):(53-57).

[0008] Gradient elution was employed. In the gradient elution program, from 0 to 5 minutes, the volume of mobile phase A was 100% and the volume of mobile phase B was 0%. From 5 to 25 minutes, the volume of mobile phase A decreased from 100% to 0%, while the volume of mobile phase B decreased from 0% to 100%. Then, the volume of mobile phase A was maintained at 0% and the volume of mobile phase B at 100% for 17 to 19 minutes.

[0009] The ceftoranil granules were dissolved in mobile phase A as a diluent and prepared into a solution, which was then injected into a high-performance liquid chromatograph for analysis.

[0010] Preferably, in the method of injecting the ceftoranil granule sample solution into a high-performance liquid chromatograph for analysis to detect impurities in the ceftoranil granules, the elution gradient shown in Table 1 below is used.

[0011] Table 1

[0012] Unexpectedly, it was found that by changing the diluent used to dilute and prepare ceftorpilofibrate particle samples and by improving the elution gradient in the high-performance liquid chromatography (HPLC) method for detecting related substances in ceftorpilofibrate particle samples, a superior method for detecting related substances compared to existing technologies can be provided, thereby improving the quality control of ceftorpilofibrate particles. The advantages of this method include improved peak precession of ceftorpilofibrate ring-opening impurities in ceftorpilofibrate particles, identification and accurate detection of unknown impurity peaks, good reproducibility, and high accuracy. In particular, the improvement in ceftorpilofibrate ring-opening impurity peaks allows for more precise control of impurities in ceftorpilofibrate particles, improving the quality level of ceftorpilofibrate particles, as shown in the embodiments of this application. Attached Figure Description

[0013] Figure 1 is a chromatogram of the impurity mixture solution of ceftoranil granules prepared in Example 1 using a 75% acetonitrile-water mixture as a diluent.

[0014] Figure 2 is a chromatogram of the impurity mixture solution of ceftoranil granules prepared in Example 2 using mobile phase A as a diluent.

[0015] Figure 3 is a chromatogram of the impurity mixture of ceftorepnitin particles obtained by analyzing the method of the present invention (chromatographic condition 1) in Example 3.

[0016] Figure 4 is a chromatogram of the impurity mixture of ceftriaxone particles obtained by analyzing the standard method (chromatographic condition 2) in Example 4. Detailed Implementation

[0017] The specific embodiments of the present invention are described in detail below. This description is intended to better illustrate the invention, and not to limit it.

[0018] In this application, all numerical values ​​used to express quantities, percentages, or proportions should be understood to be modified by the term "about" in all cases. As used herein, the term "about" may include a range of ±5%, ±4%, ±3%, ±2%, ±1%, or ±0.5% of the stated numerical value.

[0019] The quality of a drug not only determines its effectiveness in preventing and treating diseases but also directly affects patient safety. Therefore, a clinical drug should possess qualified quality to ensure safe and effective use. According to the 2020 edition of the Pharmacopoeia of the People's Republic of China, Part III, General Provisions 19, quality control of pharmaceutical products should include safety, efficacy, and controllability. Impurities are a key quality attribute of pharmaceuticals and can affect the safety and efficacy of the product. Impurities in pharmaceutical quality standards refer to impurities introduced into pharmaceuticals produced according to processes and raw materials approved by the National Medical Products Administration, either through the manufacturing process or by raw materials, or generated during storage. This excludes new impurities resulting from changes in the manufacturing process or raw materials, as well as foreign substances that are added or contaminated. Pharmaceutical impurities are generally classified into: organic impurities, inorganic impurities, and residual solvents. Organic impurities can be introduced during the production or storage of pharmaceuticals, or generated by the interaction between the drug and excipients or packaging structures. These impurities may be identified or unidentified, volatile or non-volatile. Organic impurities with chemical structures similar to or related to the active ingredient are usually referred to as related substances. The impurities in ceftorepnitin granules mentioned in this article are organic impurities with chemical structures similar to or related to the active ingredient, i.e., related substances.

[0020] According to the "Registration Standard for Imported Drugs" issued by the State Food and Drug Administration (standard number JX20090123), the quality standards for ceftriaxone granules should include the following:

[0021] A. Properties;

[0022] B. Identification: The maximum absorption was determined by ultraviolet-visible spectrophotometry (Appendix IVA of the 2010 edition of the Chinese Pharmacopoeia) at a wavelength of 231±2 nm.

[0023] C. Inspection: (1) Loss on drying: The loss on drying shall not exceed 3.0%; (2) Dissolution; (3) Related substances; (4) Microbial limits; (5) Particle size;

[0024] D. Content determination.

[0025] In the quality control of ceftoranil granules, the limits and control of related substances are a crucial aspect of the inspection. Improving the detection methods for related substances, resulting in more accurate detection of their contents, helps to better control the individual and total contents of related substances in ceftoranil granules, thereby improving the quality control of ceftoranil granules.

[0026] As described above, the present invention provides a method for improving the quality control of ceftriaxone granules, and the various method parameters involved in the method are described in more detail below.

[0027] In some preferred embodiments, mobile phase A may be a mixture of ammonium formate solution, acetonitrile, and methanol in a volume ratio of (44-46):(24-26):(24-26), and mobile phase B may be a mixture of ammonium formate solution, acetonitrile, and methanol in a volume ratio of (44-46):(54-56):(54-56). More commonly, mobile phase A is a mixture of ammonium formate solution, acetonitrile, and methanol in a volume ratio of 45:25:25, and mobile phase B is a mixture of ammonium formate solution, acetonitrile, and methanol in a volume ratio of 45:55:55.

[0028] The ammonium formate solution is an aqueous solution with an ammonium formate concentration of 1.56-1.60 g / L. This aqueous solution is adjusted to a pH of 4.3-4.7, preferably to 4.4-4.6, and more preferably to 4.5, using diluted formic acid. The diluted formic acid solution can be an aqueous solution of formic acid diluted 80-120 times, preferably 100 times, with water. Specifically, the ammonium formate solution can be prepared as follows: dissolve 1.58 g of ammonium formate in 900 ml of water, adjust the pH to 4.5 with diluted formic acid (1→100), and then dilute with water to 1000 ml.

[0029] In some implementations, elution can be performed using the elution gradients shown in Table 2 below:

[0030] Table 2

[0031] In the process of researching and developing a method for detecting impurities in ceftorepimidine granules, the inventors discovered a peak-leading phenomenon in the ring-opening impurity peak of ceftorepimidine. Through various experiments and trials, the inventors ultimately found that this problem could be solved by changing the diluent used to dilute the ceftorepimidine granule sample. Therefore, the method provided by this invention solves the peak-leading phenomenon of the ring-opening impurity peak of ceftorepimidine, improves the peak shape of the impurity, and makes the determination of the impurity content more accurate. Furthermore, the method provided by this invention has excellent reproducibility. In addition, by improving the elution gradient, the method of this invention can more accurately detect more unknown impurities. Since these unknown impurities are also included in the calculation when processing the relevant impurity peaks in the chromatographic software, the improvement of the elution gradient ultimately makes the final result of the impurity content more accurate, thereby improving the quality control level of ceftorepimidine granules.

[0032] In the method described according to this application, octadecylsilane-bonded silica gel is used as the stationary phase.

[0033] In the method described in this application, an ultraviolet detector is used for detection, and the detection wavelength is 254 nm.

[0034] In the method according to this application, the injection volume can be 8-15 μl, preferably 9-12 μl, more preferably 10 μl, and the measurement sample of ceftoranil particles can be a solution diluted with mobile phase A to a concentration of 0.5-1.1 mg / ml, preferably 0.7-0.9 mg / ml, more preferably 0.8 mg / ml; the mobile phase flow rate can be 0.8-1.2 ml / min, preferably 0.9-1.1 ml / min, most preferably 1 ml / min; the column temperature can be 20-50℃, preferably 25-45℃, more preferably 35-42℃; the elution time is more than 44 min, preferably not more than 70 min, and more commonly 50-65 min.

[0035] In the method described according to this application, an impurity / ghost peak trapping column is preferably used. The trapping column, also known as a trapping column, can be installed between the gradient mixer and the injector of a high-performance liquid chromatograph to adsorb and remove weakly polar impurities in the system, such as impurities in the mobile phase and impurities in the tubing and mixer, thereby preventing impurity peaks in the system from interfering with the target peak. The injected sample does not flow through the trapping column. Preferably, a model [model number missing] purchased from [company name missing] Scientific Instruments (Suzhou) Co., Ltd. is used. Ghost-Sniper Column, a 4.6mm x 50mm capture column or a capture column with equivalent performance.

[0036] When using the method of this invention to detect impurities in ceftoranil granules, if the content of cefotaxime and its Δ3 isomer is above 0.30%, the related substance II test should be performed. The related substance II test can refer to the method for related substance ② in standard number JX20090123 included in the "Imported Drug Registration Methods". Specifically, as follows:

[0037] Chromatographic conditions and system suitability test:

[0038] The column was packed with octadecylsilane-bonded silica gel; the mobile phase was ammonium acetate solution (1.0 g of ammonium acetate dissolved in water to make 1000 ml) - methanol (66:34); the flow rate was 1.0 ml / min; the detection wavelength was 230 nm; and the column temperature was 25 °C. 10 μL of the reference solution was injected into the liquid chromatograph, and the chromatogram was recorded. Peaks were elute in the order of ceftriaxone and aminopyrine, and the resolution between the two peaks should not be less than 4.0. For repeated injections, the relative standard deviation (RSD) of the ratio of the ceftriaxone peak area to the aminopyrine peak area should not exceed 1.5%.

[0039] Related substances determination method:

[0040] Accurately weigh approximately 40 mg of ceftoranol sample and place it in a 50 ml volumetric flask. Vigorously shake with a small amount of 80% methanol aqueous solution, then accurately add 10 ml of internal standard solution (0.1 ml of aminopyrine diluted with water to 1000 ml, concentration approximately 0.01%). Dilute to the mark with 80% methanol aqueous solution, shake well, filter, and use the filtrate as the sample solution. Separately, accurately weigh approximately 10 mg of ceftoranol reference standard and place it in a 50 ml volumetric flask. Dissolve and dilute to the mark with water, shake well, and accurately measure 2.5 ml of the solution. Place it in a 50 ml volumetric flask, accurately add 10 ml of internal standard solution, and dilute to the mark with 80% methanol aqueous solution. Shake well to obtain the reference solution. Accurately measure 10 μL each of the sample solution and reference solution and inject them separately into the liquid chromatograph. Record the chromatograms and calculate the content of P7 and ceftoranol using the following formula: P7 content % = (Q T ×W S ×sample dilution factor × 1000 × 1.1 × 100%) / (Q S ×W T × Dilution factor of reference standard) Ceftriaxone content % = (Q T ×W S ×sample dilution factor × 1000 × 100%) / (Q S ×W T (×dilution factor of reference standard)

[0041] Q T Q is the ratio of the peak area of ​​the impurity to the peak area of ​​aminopyrine in the sample solution. S W is the ratio of the area of ​​the main peak in the reference solution to the area of ​​the aminopyrine peak. T W S These represent the sample and reference weights, respectively, with 1.1 being a correction factor for ceftriaxone. The content of both P7 and ceftriaxone must not exceed 0.3%.

[0042] The solutions for each substance in the above experiment were prepared as follows:

[0043] Internal standard solution: Take an appropriate amount of aminopyrine, dissolve and dilute it with water to prepare a solution containing approximately 0.5 mg of aminopyrine per 1 ml.

[0044] Reference stock solution: Take an appropriate amount of ceftoranic acid, dissolve it in water and prepare a solution containing approximately 0.20 mg of ceftoranic acid per 1 ml.

[0045] Reference solution: Accurately measure 1 ml of the reference stock solution and 0.4 ml of the internal standard solution into a 20 ml volumetric flask, dilute to the mark with 80% methanol-water, and shake well to obtain the reference solution.

[0046] In the method for improving the quality control of impurities in ceftoranil particles according to the present invention, the detectable substances in ceftoranil particles are as follows:

[0047] (1) Ceftoranic acid

[0048] CAS No.: 104145-95-1

[0049] Chemical structural formula:

[0050] (2) Ceftoranic acid Δ3 isomer

[0051] Molecular formula: C 19 H 18 N6O5S3

[0052] Molecular weight: 506.60

[0053] Chemical name: (6R,7R)-7-[(Z)-2-(2-amino-1,3-thiazolyl-4-yl)-2-(methoxyimino)acetamido]-3-[(Z)-2-(4-methyl-1,3-thiazolyl-5-yl)vinyl]-8-oxo-5-thiazolyl[4.2.0]oct-3-ene-2-carboxylic acid

[0054] Chemical structural formula:

[0055] (3) Ceftorepimidine ring-opening impurities

[0056] Molecular formula: C 25 H 30 N6O8S3

[0057] Molecular weight: 638.76

[0058] Chemical name: 2-((Z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetamido)-2-(5-((Z)-2-(4-methylthiazol-5-yl)vinyl)-4-((valeroyloxy)methoxy)carbonyl)-3,6-dihydro-2H-1,3-thiazin-2-yl)acetic acid

[0059] Chemical structural formula:

[0060] (4) Cefotaxime hydrochloride

[0061] CAS No.: 111696-23-2

[0062] Chemical structural formula:

[0063] (5) Ceftoreprin Δ3 isomer

[0064] CAS No.: 148774-47-4

[0065] Chemical structural formula:

[0066] (6) Methoxymethylcefotaxime

[0067] Molecular formula: C 27 H 32 N6O8S3

[0068] Molecular weight: 664.78

[0069] Chemical name: (Valanoyloxy)methyl(6R,7R)-7-((Z)-2-(methoxyimino)-2-(2-((methoxymethyl)amino)thiazolyl-4-yl)acetamido)-3-((Z)-2-(4-methylthiazolyl-5-yl)vinyl)-8-oxo-5-thio-1-azabicyclo[4.2.0]oct-2-en-2-carboxylic acid ester

[0070] Chemical structural formula:

[0071] (7) Ceftoreprin 3E isomer

[0072] CAS No.: 138514-32-6

[0073] Chemical structural formula:

[0074] (8) Ceftoranil impurity P6

[0075] Molecular formula: C 28 H 33 N7O7S3

[0076] Molecular weight: 675.79

[0077] Chemical name: (valeroyloxy)methyl(6R,7R)-7-((2Z)-2-(2-((dimethylamino)methylene)amino)thiazolyl-4-yl)-2-(methoxyimino)acetamido)-3-((Z)-2-(4-methylthiazolyl-5-yl)vinyl)-8-oxo-5-thio-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid ester

[0078] Chemical structural formula:

[0079] (9) Ceftoranil dipropionate

[0080] Molecular formula: C 31 H 38 N6O9S3

[0081] Molecular weight: 734.86

[0082] Chemical name: (Valanoyloxy)methyl(6R,7R)-7-((Z)-2-(methoxyimino)-2-(2-((Valanoyloxy)methyl)amino)thiazol-4-yl)acetamido)-3-((Z)-2-(4-methylthiazol-5-yl)vinyl)-8-oxo-5-thio-1-azabicyclo[4.2.0]oct-2-en-2-carboxylic acid ester

[0083] Chemical structural formula:

[0084] (10) Ceftoranol pyridamole pentylamide

[0085] Molecular formula: C 30 H 36 N6O8S3

[0086] Molecular weight: 704.86

[0087] Chemical name: (valeroyloxy)methyl(6R,7R)-7-((Z)-2-(methoxyimino)-2-(2-thiazol-4-yl)acetamido)-3-((Z)-2-(4-methylthiazol-5-yl)vinyl)-8-oxo-5-thio-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid ester

[0088] Chemical structural formula:

[0089] (11) N-methoxybenzylcefotaxime

[0090] Molecular formula: C 33 H 36 N6O8S3

[0091] Molecular weight: 740.89

[0092] Chemical name: (Valanoyloxy)methyl(6R,7R)-7-((Z)-2-(2-((4-methoxybenzyl)amino)thiazolyl-4-yl)-2-(methoxyimino)acetamido)-3-((Z)-2-(4-methylthiazolyl-5-yl)vinyl)-8-oxo-5-thio-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid ester

[0093] Chemical structural formula:

[0094] (12) Ceftoran diphenoxylate

[0095] Molecular formula: C 31 H 39 N6O 10 S3

[0096] Molecular weight: 751.89

[0097] Chemical name: (Valanoyloxy)methyl 2-((Z)-4-(2-aminothiazol-4-yl)-13,13-dimethyl-5,8,12-trioxo-2,9,11-trioxa-3,6-diazatetradec-3-en-7-yl)-5-((Z)-2-(4-methylthiazol-5-yl)vinyl)-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid ester

[0098] Chemical structural formula:

[0099] (13) Ceftoranil dimer

[0100] Molecular formula: C 51 H 56 N 12 O 14 S6

[0101] Molecular weight: 1253.50

[0102] Chemical name: bis((valeroyloxy)methyl)7,7'-(((2Z,2'Z)-2,2'-((methylene bis(azadiyl))bis(thiazol-2,4-diyl))bis(2-(methoxyimino)acetyl))bis(azadiyl))(6R,6'R,7R,7'R)-bis(3-((Z)-2-(4-methylthiazol-5-yl)vinyl)-8-oxo-5-thio-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate)

[0103] Chemical structural formula:

[0104] (14) Ceftoran open-ring dimer

[0105] Molecular formula: C 50 H 55 N 12 O 14 S6

[0106] Molecular weight: 1240.48

[0107] Chemical name: (Valtoyloxy)methyl(6R,7R)-7-((Z)-2-(2-((R)-2-((Z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetamido)-2-((R)-5-((Z)-2-(4-methylthiazol-5-yl)vinyl)-4-((Valtoyloxy)methoxy)carbonyl)-3,6-dihydro-2H-1,3-thiazin-2-yl)acetamido)thiazol-4-yl)-2-(methoxyimino)acetamido 3-((Z)-2-(4-methylthiazol-5-yl)vinyl)-8-oxo-5-thio-1-azabicyclo[4.2.0]oct-2-en-2-carboxylate-3-6-dihydro-2H-1,3-thiazin-2-yl)acetamido)thiazol-4-yl)-2-(methoxyimino)acetamido)-3-((Z)-2-(4-methylthiazol-5-yl)vinyl)-8-oxo-5-thio-1-azabicyclo[4.2.0]oct-2-en-2-carboxylate

[0108] Chemical structural formula:

[0109] The present invention is further illustrated below with reference to specific embodiments. In the following embodiments, various processes and methods not described in detail are conventional methods known in the art, and all reagents used, unless otherwise specified in their source and specifications, are commercially available analytical grade or chromatographic grade.

[0110] In this embodiment, the method under the "Related Substances" section of the ceftoranil granule method (standard number JX20090123) in the "Registration Standard for Imported Drugs of the State Food and Drug Administration" was used as a comparison. For the sake of simplicity, this method is referred to as the standard method in this article.

[0111] Example

[0112] I. General Description

[0113] A. Material Information

[0114] The material information used in this embodiment is as follows:

[0115] The ceftoranil sample was purchased from Qilu Antipharm Pharmaceutical Co., Ltd.

[0116] Ceftoranil reference standard, from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0117] Propylparaben, analytical grade, purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0118] Ammonium formate, analytical grade, purchased from Guangzhou Chemical Reagent Factory;

[0119] Ceftoranic acid: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0120] Ceftoranic acid Δ3 isomer: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0121] Ceftoranil ring-opening impurities: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0122] Ceftoranil Δ3 isomer: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0123] Ceftoranil 3E isomer: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0124] Ceftoranol tebufenamide: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0125] N-Methoxybenzylcefotospirate: From Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0126] Ceftoran diclofenac: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0127] Ceftoram Dimer: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0128] Ceftoran open-ring dimer: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0129] Cefotaxime hydrochloride: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0130] Ceftoranil impurity P6: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0131] Methoxymethylcefotaxime: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0132] Ceftoran dipropionate: from Guangzhou Aiqixi Pharmaceutical Technology Co., Ltd.;

[0133] Formic acid, analytical grade, purchased from Tianjin Concord Technology Co., Ltd.

[0134] Acetonitrile, chromatographic grade, purchased from Tianjin Concord Technology Co., Ltd.;

[0135] Methanol, chromatographic grade, was purchased from Tianjin Concord Technology Co., Ltd.

[0136] B. Solution preparation

[0137] The solutions used in this embodiment were prepared as follows.

[0138] Ammonium formate solution: Weigh 1.58g of ammonium formate and dissolve it in 900ml of water. Adjust the pH to 4.5 with diluted formic acid solution (dilute formic acid 100 times with water), and dilute with water to 1000ml to obtain an ammonium formate solution of 1.58g / L.

[0139] Mobile phase A solution: Measure 450 ml of ammonium formate solution, 250 ml of acetonitrile, and 250 ml of methanol using a graduated cylinder, mix them thoroughly, and obtain mobile phase A solution with a volume ratio of ammonium formate solution, acetonitrile, and methanol of 45:25:25.

[0140] Mobile phase B solution: Measure 450 ml of ammonium formate solution, 550 ml of acetonitrile, and 550 ml of methanol using a graduated cylinder, mix them thoroughly, and obtain mobile phase B solution with a volume ratio of ammonium formate solution, acetonitrile, and methanol of 45:55:55.

[0141] Sample solution: Accurately weigh 40 mg of ceftoranil sample and place it in a 50 ml brown volumetric flask. Add 10 ml of mobile phase A, shake vigorously to dissolve the ceftoranil sample, dilute to the mark with mobile phase A, shake well, filter, and collect the filtrate. A sample solution with a ceftoranil concentration of 0.8 mg / ml is obtained.

[0142] Reference solution: Accurately weigh 1 mg of ceftoreprin reference standard, place it in a 100 ml brown volumetric flask, add 20 ml of mobile phase A to dissolve it, and dilute to the mark with mobile phase A to obtain a reference solution with a ceftoreprin concentration of 0.01 mg / ml (i.e., 10 μg / ml).

[0143] System suitability solution: Accurately weigh 10 mg of ceftoranil reference standard and 2.5 mg of propylparaben, place them in a 10 ml volumetric flask, dissolve and dilute to the mark with acetonitrile, and shake well. This yields a system suitability solution with a ceftoranil concentration of 1 mg / ml and a propylparaben concentration of 0.25 mg / ml.

[0144] All solutions mentioned above should be freshly prepared before use. Sample solutions, reference solutions, and system suitability solutions should be stored and used in the dark.

[0145] C. Chromatographic conditions

[0146] The high-performance liquid chromatograph used in this embodiment is an Ultimate 3000, purchased from Thermo Fisher Scientific.

[0147] The chromatographic conditions used in the method of this invention are as follows:

[0148] Detector: Ultraviolet detector;

[0149] Chromatographic column: Octadecylsilane-bonded silica gel packed column, Titank C18 4.6×250mm3μm, purchased from Analytical Scientific Instruments (Suzhou) Co., Ltd.

[0150] Impurities / Ghost Peak Capture Columns: Ghost-Sniper Column, 4.6mm×50mm, purchased from M&S Scientific Instruments (Suzhou) Co., Ltd.

[0151] Mobile phase A: The mobile phase A solution prepared above;

[0152] Mobile phase B: The mobile phase B solution prepared above;

[0153] Flow rate: 1.0 ml / min;

[0154] Detection wavelength: 254nm;

[0155] Injection volume: 10 μl;

[0156] Column temperature: 40℃;

[0157] Elution method: gradient elution, the elution program is shown in Table 3 below:

[0158] Table 3

[0159] The chromatographic conditions for the standard method are as follows:

[0160] Detector, column, impurity / ghost peak trapping column, mobile phase A, mobile phase B, flow rate, detection wavelength, injection volume, column temperature: same as chromatographic conditions 1;

[0161] The elution method is gradient elution, and the elution procedure is shown in Table 4 below:

[0162] Table 4

[0163] D. System suitability test

[0164] According to the system suitability requirements in the standard method, the resolution between the propylparaben peak and the ceftriaxone peak in the system suitability solution chromatogram should be no less than 5.0.

[0165] Test method:

[0166] The prepared system suitability solution was injected into the liquid chromatograph for analysis, and the chromatographic conditions were as described above: chromatographic condition 1 and chromatographic condition 2.

[0167] The resolution results obtained under chromatographic condition 1 are shown in Table 5:

[0168] Table 5

[0169] The resolution results obtained under chromatographic condition 2 are shown in Table 6:

[0170] Table 6

[0171] The formula for calculating the resolution is as follows:

[0172] In the formula The retention time of the preceding peak in two adjacent chromatographic peaks

[0173] The retention time of the preceding peak in two adjacent chromatographic peaks

[0174] W1 and W2 are the peak widths of the two next adjacent chromatographic peaks, respectively.

[0175] As shown in Table 5-6 above, the resolutions of the propylparaben peak and the ceftriaxone peak obtained under chromatographic conditions 1 and 2 are 8.60 and 8.53, respectively, both not less than 5.0, indicating that the system has met the specified requirements.

[0176] II. Peak Shape Improvement Test

[0177] When reproducing the standard method, the inventors discovered that the ring-opening impurities of ceftorepimidine exhibited significant peak pre-delay, resulting in poor peak shape and affecting the accuracy of the detection results. To address this problem, the inventors conducted various studies and unexpectedly discovered that changing the diluent from 75% acetonitrile-water in the standard method to mobile phase A resolved the peak pre-delay issue of the ceftorepimidine ring-opening impurities. This experiment provides chromatograms before and after the diluent change. Specific experimental information is as follows.

[0178] The impurity mixture solutions used in this experiment were prepared using a 75% acetonitrile-water mixture and mobile phase A, respectively. The concentrations of each impurity in the prepared mixture solutions are as follows:

[0179] Ceftolanic acid approximately 2.4 μg / ml; cefotaxime Δ3 isomer approximately 2.4 μg / ml; ceftolanpic acid ring-opening impurity approximately 8 μg / ml; ceftolanpic acid Δ3 isomer approximately 17.6 μg / ml; ceftolanpic acid 3E isomer approximately 4 μg / ml; ceftolanpic acid pentylamide approximately 3.2 μg / ml; N-methoxybenzylcefotaxime approximately 2.4 μg / ml; ceftolanpic acid ring-opening diphenoxylate approximately 2.4 μg / ml; ceftolanpic acid dimer approximately 13.6 μg / ml; ceftolanpic acid ring-opening dimer approximately 12.8 μg / ml; cefetamet hydrochloride approximately 1.6 μg / ml; ceftolanpic acid impurity P6 approximately 1.6 μg / ml.

[0180] Example 1 (Comparative Example)

[0181] The impurity mixture diluted with the 75% acetonitrile-water mixture specified in the standard method was injected into the high-performance liquid chromatograph for analysis under chromatographic conditions 2 (the conditions in the standard method). The resulting chromatogram is shown in Figure 1.

[0182] Figure 1 shows the chromatogram of the impurity mixture diluted with 75% acetonitrile-water mixture in the standard method. As can be seen from the figure, the ring-opening impurity peak of ceftriaxone with a retention time of 7.930 has a significant forward extension.

[0183] Example 2 (Invention Example)

[0184] The impurity mixture solution diluted with mobile phase A was injected into the high performance liquid chromatograph for analysis, using the same chromatographic conditions 2, to obtain chromatogram 2, see Figure 2.

[0185] As can be seen from Figure 2, the peak shape of the ceftorepnitin ring-opening impurity peak at a retention time of 7.943 is very good, with no peak prolongation.

[0186] It should be noted that although chromatographic condition 2 was used in all experiments, the chromatographic conditions had virtually no effect on the improvement of peak shape.

[0187] This experiment, especially Figures 1 and 2, demonstrates that changing the diluent can improve the peak shape of the ceftorepimidine ring-opening impurity peak.

[0188] III. Separation Test

[0189] Example 3 (Invention Example)

[0190] The impurity mixture solution was prepared using mobile phase A, as used in Example 2. The prepared impurity mixture solution was injected into a high-performance liquid chromatograph for analysis under chromatographic conditions 1 as described above. Chromatogram 3 was recorded (see Figure 3).

[0191] The obtained chromatogram data were analyzed, and the resolution results are shown in Table 7 below.

[0192] Table 7 (Chromatographic Conditions 1)

[0193] As can be seen from Table 7 and Figure 3 above, the peak shapes of the ceftoreprin ring-opening impurity peak with a retention time of 7.010 and the unknown impurity 1 peak with a retention time of 6.080 are very good, and the two are well separated. This indicates that using mobile phase A as a diluent to prepare the ceftoreprin granule sample solution and using the elution gradient in chromatographic condition 1 can accurately separate and identify unknown impurity 1 and the ceftoreprin ring-opening impurity, and can accurately detect these two impurities. This allows technicians to better control the quality of ceftoreprin granules.

[0194] Example 4 (Comparative Example)

[0195] The impurity mixture solution was prepared using the 75% acetonitrile-water mixture used in Example 1 as a diluent. The prepared impurity mixture solution was injected into a high-performance liquid chromatograph for analysis under chromatographic conditions 2 as described above. The chromatogram was recorded (see Figure 4).

[0196] The obtained chromatogram data were analyzed, and the resolution results are shown in Table 8 below.

[0197] Table 8 (Chromatographic Conditions 2)

[0198] Note: In Table 7-8, cefotaxime / cefotaxime Δ3 isomer represents two impurities, cefotaxime and cefotaxime Δ3 isomer. Since these two impurities cannot be effectively separated in either method, they are listed together.

[0199] As can be seen from Table 8 and Figure 4 above, the ceftoreprin ring-opening impurity peak at retention time 6.813 shows a forward extension, and unknown impurity 1 could not be identified. It is speculated that unknown impurity 1 and the ceftoreprin ring-opening impurity peak have merged and could not be separated. This indicates that using the 75% acetonitrile-water mixture as a diluent to prepare the ceftoreprin granule sample solution, as specified in the standard method, and employing the elution gradient in chromatographic condition 2, cannot identify unknown impurity 1, nor can it accurately detect the content of ceftoreprin ring-opening impurities and unknown impurity 1, thus failing to achieve good quality control of ceftoreprin granules.

[0200] IV. Reproducibility Test

[0201] This experiment was conducted by two separate researchers. Each researcher performed a system suitability test before each experiment. The reference solution and sample solution prepared as described above were used. Mobile phase A was used as the diluent in all solution preparations. Chromatographic conditions 1 were employed for analysis.

[0202] In the system suitability test, the reference solution was analyzed 5 times repeatedly. In the reproducibility test, the sample solution was analyzed 6 times repeatedly.

[0203] The experimental results of experimenter 1 are as follows.

[0204] The system applicability results are shown in Table 9.

[0205] Table 9

[0206] Note: In Table 9 and Table 11 below, the product coefficient is the ratio of concentration to peak area.

[0207] The repetitive data are shown in Table 10.

[0208] Table 10

[0209] The experimental results of experimenter 2 are as follows.

[0210] The system applicability results are shown in Table 11.

[0211] Table 11

[0212] The repeatability results are shown in Table 12.

[0213] Table 12

[0214] Acceptance criteria: The test results of the six samples should meet the requirements shown in Table 13 below, based on the compound concentration level and total impurity level.

[0215] Table 13

[0216] The test results of the 12 samples should meet the requirements shown in Table 14 below.

[0217] Table 14

[0218] The repeatability test results of Experimenter 1 are as follows: The average detection value of cefotaxime / cefotaxime Δ3 isomer in the 6 sample solutions was 0.20%, with an RSD of 3.16%, less than 20%; the average detection value of ring-opening impurities in cefotaxime was 0.55%, with an RSD of 0.00%, less than 5.0%; the average detection value of impurities with RRT=0.838 was 0.14%, with an RSD of 0.00%, less than 20%; the average detection value of cefotaxime hydrochloride was 0.02%, with an RSD of 0.00%, less than 25%; the average detection value of impurities with RRT=0.953 was 0.07%, with an RSD of 0.00%, less than 25%; the average detection value of cefotaxime Δ3 isomer was 1.33%, with an RSD of 0.57%, less than 5.0%; cefotaxime... The average detection value of ester 3E isomer was 0.12%, with an RSD of 3.45%, less than 20%; the average detection value of RRT=1.348 was 0.07%, with an RSD of 0.00%, less than 25%; the average detection value of ceftoranol diammonium ester was 0.002%, with an RSD of 22.13%, less than 25%; the average detection value of ceftoranol diammonium ester pentamamide was 0.09%, with an RSD of 0.00%, less than 5.0%; the average detection value of ceftoranol diammonium ester was 0.61%, with an RSD of 0.00%, less than 25%; the average detection value of ceftoranol diammonium ester was 0.41%, with an RSD of 0.00%, less than 10%; the average detection value of total impurities was 2.87%, with an RSD of 0.29%, less than 5.0%. All the above results met the requirements. Analytical repeatability was passed.

[0219] The repeatability test results of Experimenter 2 are as follows: The average detection value of cefotaxime / cefotaxime Δ3 isomer in 6 sample solutions by a single person was 0.20%, with an RSD of 0.00%, which is less than 20%. The RSD for 12 samples (including the 6 samples from Experimenter 1, the same below) was 2.13%. The average detection value of ring-opening impurities of cefoperazone was 0.51%, with an RSD of 0.00%, which is less than 5.0%. The RSD for 12 samples was 3.94%. The average detection value of RRT=0.837 was 0.14%, with an RSD of 0. 0.00%, less than 20%, 12-part RSD 0.00%; cefotaxime hydrochloride average detection value 0.02%, RSD 0.00%, less than 25%, 12-part RSD 0.00%; RRT = 0.954 average detection value 0.07%, RSD 0.00%, less than 25%, 12-part RSD 0.00%; ceftoranol Δ3 isomer average detection value 1.31%, RSD 0.31%, less than 5.0%, 12-part RSD 0.90%; ceftoranol The average detection value of ceftriaxone 3E isomer was 0.19%, with an RSD of 7.44%, less than 20%, and a total RSD of 9.94% for 12 samples; the average detection value of ceftriaxone with an RRT of 1.341 was 0.07%, with an RSD of 0.00%, less than 25%, and a total RSD of 0.00% for 12 samples; the average detection value of ceftriaxone diphenoxylate was 0.005%, with an RSD of 33.70%, less than 25%, and a total RSD of 24.76% for 12 samples; the average detection value of ceftriaxone diphenoxylate pentoxylate was 0.08%, with an RSD of 0.0%. The average detection value of ceftriaxone dimer was 0%, less than 25%, with an RSD of 4.77% for 12 samples; the average detection value of ceftriaxone ring-opening dimer was 0.65%, with an RSD of 0.00%, less than 5.0%, and an RSD of 1.58% for 12 samples; the average detection value of ceftriaxone ring-opening dimer was 0.43%, with an RSD of 0.95%, less than 10%, and an RSD of 8.06% for 12 samples; the average detection value of total impurities was 3.68%, with an RSD of 0.49%, less than 5.0%, and an RSD of 0.41% for 12 samples. All the above results met the requirements. The intermediate precision was good.

[0220] As can be seen from the above results, the method of the present invention has excellent reproducibility.

[0221] Overall, Figures 1 and 2 in the peak shape improvement examples of this application demonstrate that simply changing the diluent can effectively improve the peak shape of the ceftoreprin ring-opening impurity peak, thereby making the detection of the ceftoreprin ring-opening impurity content more accurate.

[0222] In the separation examples, Tables 7-8 and Figures 3-4 demonstrate that using mobile phase A as a diluent to prepare the ceftorpirum granule sample solution and employing the elution gradient in chromatographic condition 1 of this invention can accurately separate and identify unknown impurity 1 and ceftorpirum granule ring-opening impurities, and can accurately detect both impurities. This allows technicians to better control the quality of ceftorpirum granules. However, using the 75% acetonitrile-water mixture specified in the standard method as a diluent to prepare the ceftorpirum granule sample solution and employing the elution gradient in chromatographic condition 2 cannot identify unknown impurity 1, nor can it accurately detect the content of ceftorpirum granule ring-opening impurities and unknown impurity 1, thus failing to achieve good quality control of ceftorpirum granules.

Claims

1. A method for improving the quality control of ceftriaxone granules, the method comprising: The ceftriaxone particle sample solution was injected into a high-performance liquid chromatograph for analysis to detect impurities in the ceftriaxone particles. Gradient elution was performed using mobile phase A and mobile phase B as eluents. Mobile phase A was a mixture of ammonium formate solution, acetonitrile, and methanol in a volume ratio of (43-47):(23-27):(23-27), and mobile phase B was a mixture of ammonium formate solution, acetonitrile, and methanol in a volume ratio of (43-47):(53-57):(53-57). Gradient elution was employed. In the gradient elution program, from 0 to 5 minutes, the volume of mobile phase A was 100% and the volume of mobile phase B was 0%. From 5 to 25 minutes, the volume of mobile phase A decreased from 100% to 0%, while the volume of mobile phase B decreased from 0% to 100%. Then, the volume of mobile phase A was maintained at 0% and the volume of mobile phase B at 100% for 17 to 19 minutes. The ceftoranil granules were dissolved in mobile phase A as a diluent and prepared into a solution, which was then injected into a high-performance liquid chromatograph for analysis.

2. The method for improving the quality control of ceftriaxone particles according to claim 1, wherein the elution gradient is as follows:

3. The method for improving the quality control of ceftoranil granules according to claim 1, wherein the mobile phase A is a mixture of ammonium formate solution, acetonitrile, and methanol in a volume ratio of (44-46):(24-26):(24-26), and the mobile phase B is a mixture of ammonium formate solution, acetonitrile, and methanol in a volume ratio of (44-46):(54-56):(54-56).

4. The method for improving the quality control of ceftorepnitin granules according to claim 1, wherein the ammonium formate solution is an aqueous solution with an ammonium formate concentration of 1.56-1.60 g / L, and the aqueous solution is adjusted to 4.4-4.6 with formic acid.

5. The method for improving the quality control of ceftorepnidazole particles according to claim 1, wherein the injection volume is 8-15 μl, and wherein the sample solution of ceftorepnidazole particles is a solution diluted with mobile phase A to a concentration of 0.5-1.1 mg / ml.

6. The method for improving the quality control of ceftriaxone particles according to claim 1, wherein the mobile phase flow rate is 0.9-1.1 ml / min.

7. The method for improving the quality control of ceftriaxone particles according to claim 1, wherein octadecylsilane-bonded silica gel is used as the stationary phase.

8. The method for improving the quality control of ceftriaxone particles according to claim 1, wherein an ultraviolet detector is used for detection at a wavelength of 254 nm.

9. The method for improving the quality control of ceftriaxone particles according to claim 1, wherein the column temperature is 35-42℃.

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