Method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride

Abstract

The invention belongs to the technical field of detection, and particularly relates to a method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride. The detection method comprises the following steps: dissolving cefcapene pivoxil hydrochloride to be detected by using N, N-dimethylformamide or N, N-dimethylacetamide to obtain a test solution; and carrying out headspace-gas chromatography analysis on the test solution to obtain a test spectrogram, comparing the test spectrogram with a predetermined reference spectrogram, determining the nature by retention time, and obtaining the content of 2-chloro-2-methylpropane in the to-be-detected cefcapene pivoxil hydrochloride according to the peak area. The detection method provided by the invention has the advantages of good separation degree of 2-chloro-2-methylpropane and other substances, high accuracy, high sensitivity, high recovery rate and high detection speed, and is suitable for detection of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride.

Description

Technical Field

[0001] The present invention belongs to the technical field of detection, and particularly relates to a method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride. Background Art

[0002] Cefcapene pivoxil hydrochloride is a monohydrate, namely (6R,7R)-7-[(2Z)-2-(2-aminothiazol-4-yl)pent-2-enamido]-3-carbamoyloxymethyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid 2,2-dimethylpropionyloxymethyl ester monohydrochloride monohydrate, and belongs to oral cephalosporin antibiotics. Cefcapene pivoxil hydrochloride is a white to slightly yellowish-white crystalline powder with a slight special odor; it is very slightly soluble in water, easily soluble in methanol and N,N-dimethylformamide, soluble in ethanol, and insoluble in ether. Its chemical structural formula is shown as follows:

[0003]

[0004] 2-chloro-2-methylpropane present in cefcapene pivoxil hydrochloride may be derived from: during the process of removing the BOC group from the protected cefcapene to generate the crude product of cefcapene pivoxil hydrochloride, the removed BOC group generates tert-butanol, and tert-butanol may further react with hydrochloric acid present in the system to produce 2-chloro-2-methylpropane. The CAS number of 2-chloro-2-methylpropane is 507-20-0, and its structural formula is as follows:

[0005]

[0006] 2-chloro-2-methylpropane belongs to haloalkanes and may have a risk of mutagenesis. According to the ICH M7 guideline: when calculating the acceptable intake based on TTC, when the daily intake of a mutagenic impurity per person is 1.5 μg, the risk is considered negligible. This value can be generally applied to most drugs as the default value of the acceptable control limit. The maximum daily dose of cefcapene pivoxil hydrochloride is 600 mg / day. According to the following formula:

[0007]

[0008] When the residual limit of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride is greater than 2.5 ppm, there will be a potential carcinogenic risk, and it is necessary to control the residual amount of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride. However, there is currently no detection method for 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride. Summary of the Invention

[0009] In view of this, the purpose of the present invention is to provide a method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride. The method for quantitatively detecting the residue of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride provided by the present invention has good resolution between 2-chloro-2-methylpropane and other substances, high accuracy, high sensitivity, high recovery rate, fast detection speed, and is applicable to the detection of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride.

[0010] In order to achieve the above purpose, the present invention provides the following technical solutions:

[0011] The present invention provides a method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride, comprising the following steps:

[0012] Dissolve the cefcapene pivoxil hydrochloride to be tested with N,N-dimethylformamide or N,N-dimethylacetamide to obtain a test solution;

[0013] Perform headspace-gas chromatography analysis on the test solution to obtain a test spectrum, compare the test spectrum with a predetermined reference spectrum, qualitatively analyze by retention time, and obtain the content of 2-chloro-2-methylpropane in the cefcapene pivoxil hydrochloride to be tested according to the peak area;

[0014] The conditions for the headspace-gas chromatography analysis include:

[0015] Headspace heating oven temperature: 90-110°C; sample equilibration time: 25-35 min;

[0016] Chromatographic column: DB-624 or equivalent chromatographic column;

[0017] Column temperature programming: maintain at 40°C for 10-20 min, increase the temperature at a rate of 20-30°C / min to 200-240°C, and maintain for 3 ～ 8 min:

[0018] Preferably, the column temperature programming is: maintain at 40°C for 20 min, increase the temperature at a rate of 30°C / min to 230°C, and maintain for 5 min.

[0019] Preferably, the specification of the chromatographic column is 60 m × 0.32 mm, 1.8 μm.

[0020] Preferably, the headspace heating oven temperature is 100°C; the sample equilibration time is 30 min.

[0021] Preferably, the concentration of cefcapene pivoxil hydrochloride in the test solution is 0.5-1 g / mL.

[0022] Preferably, the split ratio of the headspace-gas chromatography analysis is 3-10:1.

[0023] Preferably, the flow rate of the headspace-gas chromatography analysis is 1-2 mL / min.

[0024] Preferably, the detector temperature of the headspace-gas chromatography analysis is 200-250 °C.

[0025] Preferably, the inlet temperature of the headspace-gas chromatography analysis is 200-220 °C.

[0026] The present invention also provides an application of the detection method of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride described in the above technical solution in the quality control of cefcapene pivoxil hydrochloride.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The present invention provides a detection method of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride, comprising the following steps: dissolving the cefcapene pivoxil hydrochloride to be detected with N,N-dimethylformamide or N,N-dimethylacetamide to obtain a test solution; performing headspace-gas chromatography analysis on the test solution to obtain a test sample chromatogram, comparing the test sample chromatogram with a predetermined reference chromatogram, qualitatively analyzing by retention time, and obtaining the content of 2-chloro-2-methylpropane in the cefcapene pivoxil hydrochloride to be detected according to the peak area.

[0029] The present invention develops a method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride based on headspace-gas chromatography. The present invention adopts headspace sampling with low matrix effect. Combining with the column temperature rising program of the gas chromatography of the present invention, 2-chloro-2-methylpropane can be effectively separated from unknown peaks, with good separation effect, symmetric peak shape, and improved result accuracy. For the detection method of the present invention, the separation degree between 2-chloro-2-methylpropane and other substances is good, with high accuracy, high sensitivity, high recovery rate, and fast detection speed, and is applicable to the detection of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride.

[0030] Furthermore, the concentration of cefcapene pivoxil hydrochloride, the chromatographic column specification and the split ratio in the test solution of the present invention make the detection peak area of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride high, the tailing factor low, and the theoretical plate number high, further improving the detection sensitivity and accuracy.

[0031] The example data shows that the detection limit of the detection method of the present invention for the impurity of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride is 0.101 μg / mL, the quantitation limit is 0.202 μg / mL, with high sensitivity and good linear relationship in the concentration range of 0.2-3 μg / mL; the spiked recovery rate of 2-chloro-2-methylpropane is 91.03%-107.97%, with good recovery rate and high accuracy. Description of the Drawings

[0032] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0033] Figure 1 It is the chromatogram of the 2-chloro-2-methylpropane positioning solution in Example 1;

[0034] Figure 2 It is the chromatogram of the test solution in Example 1;

[0035] Figure 3 It is the chromatogram of the test solution + impurity mixed solution in Example 1;

[0036] Figure 4 It is the chromatogram of the 2-chloro-2-methylpropane detection limit solution in Example 2;

[0037] Figure 5 It is the linear curve graph of 2-chloro-2-methylpropane in Example 2;

[0038] Figure 6 It is the chromatogram of the reference substance solution at 0.6 μg / mL in Comparative Example 1;

[0039] Figure 7 It is the chromatogram of the test solution + reference substance solution at 0.6 μg / mL in Comparative Example 1;

[0040] Figure 8 It is the chromatogram of the test solution + reference substance solution at 1.2 μg / mL in Comparative Example 1;

[0041] Figure 9 It is the chromatogram of the reference substance solution in Comparative Example 2;

[0042] Figure 10 It is the chromatogram of the test solution + reference substance solution in Comparative Example 2;

[0043] Figure 11 It is the chromatogram of the test solution + reference substance solution in Comparative Example 3. Specific embodiments

[0044] The present invention provides a method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride, comprising the following steps:

[0045] Dissolve the cefcapene pivoxil hydrochloride to be tested with N,N-dimethylformamide or N,N-dimethylacetamide to obtain a test solution;

[0046] The test sample solution is subjected to headspace-gas chromatography analysis to obtain a test sample chromatogram. The test sample chromatogram is compared with a predetermined reference chromatogram for qualitative analysis based on retention time, and the content of 2-chloro-2-methylpropane in the cefcapene pivoxil hydrochloride to be measured is obtained according to the peak area.

[0047] The conditions for the headspace-gas chromatography analysis include:

[0048] Headspace heating oven temperature: 90 - 110 °C; Sample equilibration time: 25 - 35 min;

[0049] Chromatographic column: DB-624 or an equivalent chromatographic column;

[0050] Column temperature programming: Hold at 40 °C for 10 - 20 min, then increase the temperature at a rate of 20 - 30 °C / min to 200 - 240 °C, and hold for 3 ～ 8 min.

[0051] In the present invention, unless otherwise specified, the materials and equipment used are commercially available products in the art.

[0052] In the present invention, the cefcapene pivoxil hydrochloride to be measured is dissolved in N,N-dimethylformamide or N,N-dimethylacetamide to obtain a test sample solution.

[0053] In the present invention, the concentration of cefcapene pivoxil hydrochloride in the test sample solution is preferably 0.5 - 1 g / mL, more preferably 1 g / mL. The concentration of cefcapene pivoxil hydrochloride in the present invention can ensure a relatively high response value of 2-chloro-2-methylpropane during chromatographic detection, thereby improving the accuracy of detection.

[0054] In the present invention, the dissolution is preferably carried out in a headspace vial. The volume of the headspace vial is preferably 20 mL, and the volume of the test sample solution in the headspace vial is preferably 1 mL.

[0055] After obtaining the test sample solution, the test sample solution is subjected to headspace-gas chromatography analysis to obtain a test sample chromatogram. The test sample chromatogram is compared with a predetermined reference chromatogram for qualitative analysis based on retention time, and the content of 2-chloro-2-methylpropane in the cefcapene pivoxil hydrochloride to be measured is obtained according to the peak area.

[0056] The conditions for the headspace-gas chromatography analysis include:

[0057] Headspace heating oven temperature: 90 - 110 °C; Sample equilibration time: 25 - 35 min;

[0058] Chromatographic column: DB-624 or an equivalent chromatographic column;

[0059] Column temperature programming: Maintain at 40°C for 10 - 20 min, then increase the temperature to 200 - 240°C at a rate of 20 - 30°C / min, and maintain for 3 - 8 min.

[0060] In the present invention, the temperature of the headspace heating box is preferably 100°C; the sample equilibration time is preferably 30 min.

[0061] In the present invention, the specifications of the chromatographic column are preferably 60 m × 0.32 mm, 1.8 μm.

[0062] In the present invention, the column temperature programming is preferably: Maintain at 40°C for 20 min, then increase the temperature to 230°C at a rate of 30°C / min, and maintain for 5 min.

[0063] In the present invention, the detector temperature for the headspace - gas chromatography analysis is preferably 200 - 250°C, more preferably 240°C; the inlet temperature is preferably 200 - 220°C, more preferably 220°C.

[0064] In the present invention, the split ratio for the headspace - gas chromatography analysis is preferably 3 - 10:1, more preferably 5:1, and the flow rate is preferably 1 - 2 mL / min.

[0065] In the present invention, the injection volume for the headspace - gas chromatography analysis is preferably 1 mL.

[0066] In the present invention, the headspace analyzer used for the headspace - gas chromatography analysis is preferably Agilent 7697A, and the gas chromatograph is preferably Agilent 7820A.

[0067] In the present invention, the predetermined reference chromatogram is obtained by performing headspace - gas chromatography analysis on a reference solution of 2 - chloro - 2 - methylpropane. The solvent of the reference solution of 2 - chloro - 2 - methylpropane is preferably N,N - dimethylformamide (DMF) or N,N - dimethylacetamide (DMAC).

[0068] In the present invention, the concentration of the reference solution is preferably 0.202 - 3.028 μg / mL, and specifically can be 0.2 μg / mL, 0.5 μg / mL, 1.25 μg / mL, 2 μg / mL, 2.5 μg / mL, or 3 μg / mL.

[0069] In the present invention, the qualitative analysis based on the retention time is preferably: If a peak appears in the test sample chromatogram at the same position or within the error range as the peak of 2 - chloro - 2 - methylpropane in the reference chromatogram, it indicates the presence of 2 - chloro - 2 - methylpropane, otherwise it does not contain 2 - chloro - 2 - methylpropane; the error range is preferably ±1% of the retention time.

[0070] In the present invention, the content of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride to be detected based on the peak area is determined by the external standard method, and the external standard method is preferably calculated according to the following formula:

[0071] C x = A x / A R × C R ;

[0072] Wherein C x represents the concentration of 2-chloro-2-methylpropane in the test solution; A x represents the peak area of the test solution; A R represents the peak area of the reference solution; C R represents the concentration of 2-chloro-2-methylpropane in the reference solution.

[0073] The present invention also provides the application of the method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride described in the above technical solution in the quality control of cefcapene pivoxil hydrochloride.

[0074] The detection method of the present invention can realize the rapid detection and analysis of cefcapene pivoxil hydrochloride, which is of great significance for the quality control of cefcapene pivoxil hydrochloride raw materials.

[0075] In order to further illustrate the present invention, the method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride provided by the present invention will be described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the protection scope of the present invention.

[0076] Example 1

[0077] A method for detecting the potential mutagenic impurity 2-chloro-2-methylpropane includes the following steps:

[0078] (1) Instrument conditions

[0079] Instrument: Agilent 7820A gas chromatograph

[0080] Headspace instrument: Agilent 7697A

[0081] Chromatographic column: DB-624 60m×0.32mm, 1.8μm;

[0082] Detector temperature: 240 °C; Injection port temperature: 220 °C;

[0083] Column temperature: Hold at 40 °C for 20 min, then increase the temperature to 230 °C at a rate of 30 °C / min and hold for 5 min;

[0084] Split ratio: 5:1; Flow rate: 1.0 mL / min;

[0085] Heating oven temperature: 100 °C; Sample equilibration time: 30 min; Injection volume: 1 mL.

[0086] (2) Solution preparation

[0087] Reference solution (impurity localization solution): Weigh 62.5 mg of 2-chloro-2-methylpropane into a 50 mL volumetric flask, dilute with DMF to the mark, shake well, and obtain the 2-chloro-2-methylpropane stock solution. Dilute the stock solution with DMF to prepare a 2-chloro-2-methylpropane reference solution with a concentration of 2.5 μg / mL. Accurately pipette 1 mL of the 2-chloro-2-methylpropane reference solution into a 20 mL headspace vial and seal it.

[0088] Test solution: Weigh accurately 1 g of cefcapene pivoxil hydrochloride into a 20 mL headspace vial, add 1 mL of DMF, and seal it.

[0089] Test sample + reference (impurity) mixed resolution solution: Weigh accurately 1 g of cefcapene pivoxil hydrochloride into a 20 mL headspace vial, add 1 mL of the reference solution, and seal it.

[0090] Take the above solutions and perform detections respectively, record the chromatograms, and the results are shown in Figures 1 to 3 and Table 1 below:

[0091] Table 1 Specificity data table of 2-chloro-2-methylpropane

[0092] From Figures 1 to 3 and Table 1, it can be seen that the retention times of the 2-chloro-2-methylpropane peaks in the test solution and the test sample + impurity mixed resolution solution are consistent with the retention time of the corresponding 2-chloro-2-methylpropane peak in the impurity localization solution, indicating that this method can accurately locate and separate the 2-chloro-2-methylpropane peak, and has good specificity.

[0093] Example 2

[0094] Perform the following tests according to the instrument conditions in Example 1:

[0095] 1. Sensitivity

[0096] Figure 4 The chromatogram of the 2-chloro-2-methylpropane detection limit solution is shown. The quantitative limit and detection limit data table are shown in Table 2 below:

[0097] Table 2 Quantitative limit and detection limit data table of 2-chloro-2-methylpropane

[0098]

[0099] From Table 2 and Figure 4It can be seen that both the detection limit and the quantification limit of 2-chloro-2-methylpropane are below 10% of the limit concentration (2.5 ppm), indicating that this method has a high response to 2-chloro-2-methylpropane and the sensitivity of the method is good.

[0100] 2. Linear relationship

[0101] Weigh 62.5 mg of 2-chloro-2-methylpropane into a 50 mL volumetric flask, add DMF to dilute and make up to the mark, shake well, and obtain stock solution 1 of 2-chloro-2-methylpropane. Pipette 1 mL of stock solution 1 into a 50 mL volumetric flask, add DMF to dilute and make up to the mark, shake well, and obtain stock solution 2 of 2-chloro-2-methylpropane. Accurately pipette 1 mL, 2.5 mL, 4 mL, 5 mL, and 6 mL of stock solution 2 of 2-chloro-2-methylpropane into 50 mL volumetric flasks respectively, dilute with DMF to the mark, and shake well to obtain a series of linear solutions with concentrations of 20%, 50%, 80%, 100%, and 120%. Pipette 1 mL of the corresponding linear solution into a 20 mL headspace vial and seal it. Then, perform gas chromatography detection on the quantification limit solution and a series of 2-chloro-2-methylpropane solutions with different concentrations to obtain chromatograms of 2-chloro-2-methylpropane with different concentrations. The linear equation data of gas chromatography are calculated and shown in Table 3. Figure 5 It is the linear curve graph of 2-chloro-2-methylpropane.

[0102] Table 3 Linear equation data table of 2-chloro-2-methylpropane

[0103] Name Linear range Linear equation <![CDATA[Coefficient of correlation R 2 > 2-chloro-2-methylpropane solution 0.202 μg / mL to 3.028 μg / mL y = 15.534x + 1.0332 0.997

[0104] As can be seen from Table 3 and Figure 5 it can be known that for the detection method of 2-chloro-2-methylpropane impurity in cefcapene pivoxil hydrochloride, the linear relationship between the peak area and the concentration is good in the range of 0.202 μg / mL - 3.028 μg / mL.

[0105] 3. Standard addition recovery rate

[0106] Accurately weigh 1 g of cefcapene pivoxil hydrochloride into a 20 mL headspace vial, and add 1 mL of 50%, 100%, and 120% linear solutions of 2-chloro-2-methylpropane respectively to obtain 2-chloro-2-methylpropane standard addition sample solutions with different concentrations of 50%, 100%, and 120%, and prepare 3 portions for each concentration; then perform gas chromatography detection on a series of 2-chloro-2-methylpropane standard addition sample solutions with different concentrations under the same conditions to obtain chromatograms of 2-chloro-2-methylpropane standard addition sample solutions with different concentrations. The standard addition recovery rates are calculated and shown in Table 4:

[0107] Table 4 Standard addition recovery rate data table of 2-chloro-2-methylpropane standard addition sample solution

[0108] Name Spiked recovery rate RSD value of spiked recovery rate Spiked sample solution of 2-chloro-2-methylpropane 91.03％～107.97％ 5.52％

[0109] As can be seen from Table 4, the spike recovery rate of the spiked sample solution of 2-chloro-2-methylpropane was between 91.03% and 107.97%, and the RSD value of the spike recovery rate was 5.52%, indicating that the detection method for 2-chloro-2-methylpropane impurities in cefcapene pivoxil hydrochloride had high accuracy.

[0110] 4. Stability

[0111] Gas phase detection was carried out on the 2-chloro-2-methylpropane solution (2.5 μg / mL) and the mixed solution of cefcapene pivoxil hydrochloride + 2-chloro-2-methylpropane to be detected at 0 h, 12.5 h, 24 h, 53 h, 69 h, and 100 h of standing time, and the statistical data are shown in Table 5:

[0112] Table 5 Statistical data table of 2-chloro-2-methylpropane solution and mixed solution of cefcapene pivoxil hydrochloride + 2-chloro-2-methylpropane to be detected at different time points

[0113]

[0114] As can be seen from Table 5, the change in peak area was not significant at different injection times, indicating that the detection method for 2-chloro-2-methylpropane impurities in cefcapene pivoxil hydrochloride had good stability and high accuracy.

[0115] Example 3

[0116] The instrument conditions were the same as those in Example 1.

[0117] Test solution: Weigh accurately 1 g of cefcapene pivoxil hydrochloride into a 20 mL headspace vial, add 1 mL of DMF, and seal.

[0118] Reference solution: See the 100% linear solution in Example 2.

[0119] The residual 2-chloro-2-methylpropane in 3 batches of process validation batches (KP-2203001, KP-2203002, KP-2203003) and 1 batch of production batch (KP-2406001) of cefcapene pivoxil hydrochloride was detected, and the results are shown in Table 6:

[0120] Table 6 Statistical data table of the detection of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride of different batches

[0121] Sample batch number Content (ppm) KP-2406001 0.232 KP-2203001 0.240 KP-2203002 0.245 KP-2203003 0.235

[0122] As can be seen from Table 6, the detection method for 2-chloro-2-methylpropane impurity in cefcapene pivoxil hydrochloride is applicable to the determination of the residue of the genotoxic impurity 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride.

[0123] The detection method for 2-chloro-2-methylpropane impurity in cefcapene pivoxil hydrochloride of the present invention has good accuracy, high sensitivity, high recovery rate and fast detection speed.

[0124] Comparative Example 1

[0125] The differences from Example 1 are as follows:

[0126] (1) Chromatographic conditions

[0127] Instrument: Agilent 7820A gas chromatograph

[0128] Chromatographic column: DB-624 60m×0.32mm, 1.8μm;

[0129] Detector temperature: 240°C; Injection port temperature: 220°C;

[0130] Column temperature: Hold at 40°C for 25 min, increase the temperature to 230°C at a rate of 30°C / min, and hold for 2 min;

[0131] Injection volume: 5 μL; Split ratio 5:1; Flow rate 1.0 mL / min.

[0132] (2) Solution preparation

[0133] Calculated according to the limit of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride (2.5 ppm), when the concentration of the test substance is 250 mg / mL, the corresponding concentration of the 2-chloro-2-methylpropane reference substance is about 0.6 μg / mL; when the concentration of the test substance is 500 mg / mL, the corresponding concentration of the 2-chloro-2-methylpropane reference substance is about 1.2 μg / mL. (All are dissolved in DMF)

[0134] The 0.6 μg / mL reference substance solution, the test substance + 0.6 μg / mL reference substance solution, and the test substance + 1.2 μg / mL reference substance solution were investigated respectively. The test substance + reference substance solution was prepared by dissolving 1 g of the sample with the reference substance solution.

[0135] Figure 6 is the chromatogram of the 0.6 μg / mL reference substance solution; Figure 7 is the chromatogram of the test substance + 0.6 μg / mL reference substance solution; Figure 8 is the chromatogram of the test substance + 1.2 μg / mL reference substance solution.

[0136] From Figure 6 and Figure 7It can be seen that when the concentration of the reference solution is 0.6 μg / mL, the peak area of 2-chloro-2-methylpropane is about 4.6, and the response value is relatively low. Therefore, the concentration of the reference solution was continuously adjusted to 1.2 μg / mL. From Figure 8 It can be seen that at this concentration, 2-chloro-2-methylpropane and the unknown peak in the test solution could not be effectively separated, proving that the method of increasing the concentration of the test solution to improve the response value under this chromatographic condition is not feasible. Therefore, in the present invention, the headspace injection method was replaced.

[0137] Comparative Example 2

[0138] The differences from Example 1 are as follows:

[0139] (1) Chromatographic conditions

[0140] Chromatographic column: DB-624 30 m × 0.32 mm, 1.8 μm;

[0141] Column temperature: Hold at 35 °C for 25 min, then increase the temperature to 240 °C at a rate of 30 °C / min and hold for 2 min.

[0142] (2) Solution preparation

[0143] Reference solution, test article + reference solution (2.5 μg / mL): The preparation method is the same as that in Example 1.

[0144] Figure 9 is the chromatogram of the reference solution; Figure 10 is the chromatogram of the test article + reference solution, and the data statistical table is shown in Table 7.

[0145] Table 7 Statistical table of 2-chloro-2-methylpropane detection data with a 30 m chromatographic column

[0146]

[0147] From Table 7 and Figure 9 、 Figure 10 It can be seen that although when the chromatographic column is DB-624 30 m × 0.32 mm, 1.8 μm, 2-chloro-2-methylpropane also has a relatively high response value, 2-chloro-2-methylpropane in the test article + reference solution and the unknown peak were not completely separated, and the separation effect was poor.

[0148] Comparative Example 3

[0149] The differences from Example 1 are as follows:

[0150] (1) Chromatographic conditions

[0151] Column temperature: Hold at 35 °C for 20 min, then increase the temperature to 240 °C at a rate of 20 °C / min and hold for 2 min;

[0152] (2) Solution preparation

[0153] Test sample + reference solution (2.5 μg / mL): Prepared in the same manner as in Example 1.

[0154] Figure 11 is the chromatogram of the test sample + reference solution. As Figure 11 can be seen, the response value of 2-chloro-2-methylpropane detected under this condition is low, the resolution effect is poor, and the peak shape is asymmetric (tailing factor is 0.81), which is not suitable for the detection of 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride.

[0155] The gas chromatography conditions of the present invention enable the detection peak area of 2-chloro-2-methylpropane impurity in cefcapene pivoxil hydrochloride to be high, the tailing factor to be low, and the theoretical plate number to be high, realizing the rapid detection and analysis of 2-chloro-2-methylpropane residue in cefcapene pivoxil hydrochloride, facilitating the control of 2-chloro-2-methylpropane mutagenic impurity in cefcapene pivoxil hydrochloride, and providing a guarantee for the quality control of cefcapene pivoxil hydrochloride bulk drug. The detection limit of the detection method of the present invention for 2-chloro-2-methylpropane impurity in cefcapene pivoxil hydrochloride is 0.101 μg / mL (equivalent to 0.00001% of the test sample concentration), and the quantitation limit is 0.202 μg / mL (equivalent to 0.00002% of the test sample concentration), with high sensitivity; the recovery rate of 2-chloro-2-methylpropane is 91.03% - 107.97%, with high accuracy.

[0156] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all embodiments. People can also obtain other embodiments according to the embodiments of the present invention without creative labor, and these embodiments all belong to the protection scope of the present invention.

Claims

1. A method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride, characterized in that, Comprising the following steps: Dissolve cefcapene pivoxil hydrochloride to be tested with N,N-dimethylformamide or N,N-dimethylacetamide to obtain a test solution; Perform headspace-gas chromatography analysis on the test solution to obtain a test sample chromatogram, compare the test sample chromatogram with a predetermined reference chromatogram, qualitatively analyze by retention time, and obtain the content of 2-chloro-2-methylpropane in the cefcapene pivoxil hydrochloride to be tested according to the peak area; The conditions for the headspace-gas chromatography analysis include: Headspace heating oven temperature: 90-110°C; Sample equilibration time: 25-35 min; Chromatographic column: DB-624 or an equivalent chromatographic column; Column temperature rising program: maintain at 40°C for 10 - 20 min, rise the temperature to 200 - 240°C at a rate of 20 - 30°C / min, and maintain for 3 ～ 8 min.

2. The detection method according to claim 1, wherein The column temperature programming is: maintain at 40°C for 20 min, increase the temperature to 230°C at a rate of 30°C / min, and maintain for 5 min.

3. The detection method according to claim 1 or 2, characterized in that, The specification of the chromatographic column is 60 m × 0.32 mm, 1.8 μm.

4. The detection method according to claim 1, wherein The headspace heating oven temperature is 100°C; The sample equilibration time is 30 min.

5. The detection method according to claim 1 or 4, characterized in that, The concentration of cefcapene pivoxil hydrochloride in the test solution is 0.5-1 g / mL.

6. The detection method according to claim 1, wherein The split ratio of the headspace-gas chromatography analysis is 3-10:

1.

7. The detection method according to claim 1, wherein The flow rate of the headspace-gas chromatography analysis is 1-2 mL / min.

8. The detection method according to claim 1, wherein The detector temperature of the headspace-gas chromatography analysis is 200-250°C.

9. The detection method according to claim 1, wherein, The inlet temperature of the headspace-gas chromatography analysis is 200-220°C.

10. Use of the method for detecting 2-chloro-2-methylpropane in cefcapene pivoxil hydrochloride according to any one of claims 1-9 in the quality control of cefcapene pivoxil hydrochloride.

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