CN110554102A

CN110554102A - Detection method of sugammadex sodium

Info

Publication number: CN110554102A
Application number: CN201910418301.9A
Authority: CN
Inventors: 白俊鹏; 王征; 肖宇; 孟秋月; 和潇夏; 金晶晶; 吴蕾; 吴青; 胡佰艳; 王�琦; 王利春; 王晶翼
Original assignee: Tianjin Kelun Pharmaceutical Research Co Ltd
Current assignee: Sichuan Kelun Pharmaceutical Research Institute Co Ltd
Priority date: 2018-05-30
Filing date: 2019-05-20
Publication date: 2019-12-10

Abstract

The invention relates to a detection method of sugammadex sodium, which comprises the following steps: detecting a test sample solution and the reference solution by adopting a high performance liquid chromatograph, wherein the chromatographic conditions are as follows: the chromatographic column is a C18 chromatographic column; the mobile phase A is phosphate buffer solution with the concentration of 5mmol/L to 40mmol/L, or phosphoric acid water solution with the volume concentration of 0.1 percent to 1.0 percent, and the pH value of the mobile phase A is 2.0 to 2.9; the mobile phase B is acetonitrile; the temperature of the chromatographic column is 30-45 ℃; the flow rate is 0.6ml/min to 1.2 ml/min; the wavelength of ultraviolet detection is 190nm to 210 nm. The detection method of sugammadex sodium provided by the invention can effectively separate related substances of sugammadex sodium, and has the advantages of high sensitivity, good specificity and high accuracy.

Description

Detection method of sugammadex sodium

Technical Field

the invention relates to the field of analytical chemistry, in particular to a detection method of sugammadex sodium.

background

Sugammadex Sodium (Sugammadex Sodium or Sugammadex) was first developed by oganong corporation (organo Biosciences), purchased by pionbergy company (Schering-plus) in 2007, and combined with Merck (Merck) in 2009. Sugammadex sodium is currently owned and sold by merck and has become a medium-scale product under the flag of losandong. In 2008, sugammadex sodium was first marketed in europe and subsequently in japan, the united states, etc., respectively, and is now marketed in 75 countries, and in 2016, merck corporation proposed the marketing of sugammadex sodium in china.

sugammadex sodium is a modified gamma-cyclodextrin, is the first and only selective muscle relaxation antagonist (binding agent) (SRBA) successfully developed for 20 years, can block the relaxation effect by wrapping amino steroid non-depolarizing muscle relaxants in a brand-new and only way, can quickly and predictably reverse the muscle relaxation caused by rocuronium bromide and vecuronium bromide in any strength, has small side effect, can enable the use of a muscle relaxant to be close to an ideal state, and has the effect of reversing the neuromuscular blockade more quickly and predictably than the existing drugs.

The method is used for accurately detecting the content of the sugammadex sodium and related impurities in the sugammadex sodium product or the sugammadex sodium product obtained in the production process of the sugammadex sodium, and is very important for quality evaluation of the sugammadex sodium, however, the existing High Performance Liquid Chromatography (HPLC) detection method of the sugammadex sodium cannot achieve effective separation of the sugammadex sodium related substances, and the sensitivity is low.

disclosure of Invention

based on this, there is a need for a detection method of sugammadex sodium with better resolution and detection sensitivity.

A detection method of sugammadex sodium comprises the following chromatographic conditions:

the chromatographic column is C₁₈A chromatographic column;

the mobile phase A is phosphate buffer solution with the concentration of 5mmol/L to 40mmol/L, or phosphoric acid water solution with the volume concentration of 0.1 percent to 1.0 percent, and the pH value of the mobile phase A is 2.0 to 2.9;

The mobile phase B is acetonitrile;

The temperature of the chromatographic column is 30-45 ℃;

The flow rate of the mixed liquid of the mobile phase A and the mobile phase B is 0.6ml/min to 1.2 ml/min; and

The gradient elution procedure was:

Time (minutes)	Mobile phase A (%)	mobile phase B (%)
			0	75～85	25～15
5～15	75～85	25～15
			35～45	70～82	30～18
45～55	65～78	35～22
			55～70	40～60	60～40
70～75	15～35	85～65
			75～80	15～35	85～65
80.1～85.1	75～85	25～15
			85.1～100	75～85	25～15

。

in some embodiments, a sample to be tested is dissolved in the mobile phase a to form a test solution.

In certain embodiments, the test sample solution is loaded in an amount of 5 μ l to 20 μ l.

In certain embodiments, the C₁₈the chromatographic column is an octadecylsilane chemically bonded silica gel column.

In certain embodiments, the phosphate in the phosphate buffer solution comprises at least one of potassium dihydrogen phosphate, sodium dihydrogen phosphate, ammonium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, diammonium hydrogen phosphate; potassium dihydrogen phosphate and sodium dihydrogen phosphate are preferred; more preferably potassium dihydrogen phosphate.

In certain embodiments, the eluate of the gradient elution procedure is detected using an ultraviolet-visible spectrophotometer with a detection wavelength of 190nm to 210 nm; preferably 200 nm.

In certain embodiments, the pH of mobile phase a is from 2.3 to 2.5; preferably 2.3.

In certain embodiments, the mobile phase a is a phosphate buffered solution having a concentration of 20 to 25mmol/L and a pH of 2.3 to 2.5; preferably phosphate buffer solution with the concentration of 20mmol/L and the pH value of 2.3; more preferably a potassium dihydrogen phosphate solution having a concentration of 20mmol/L and a pH of 2.3.

In certain embodiments, the column temperature of the chromatography column is 35 ℃ to 42 ℃; preferably 40 deg.c.

In certain embodiments, the flow rate of the mixture of mobile phase a and mobile phase B is 0.6ml/min to 1.2 ml/min; preferably 0.8 ml/min. In certain embodiments, the gradient elution procedure is:

preferably, the gradient elution procedure is:

Time (minutes)	Mobile phase A (%)	Mobile phase B (%)
			0	82	18
10	82	18
			40	80	20
50	75	25
			65	50	50
70	25	75
			80	25	75
80.1	82	18
			90	82	18

。

in certain embodiments, the method further comprises the steps of providing a control solution, detecting the control solution by using the high performance liquid chromatograph, and comparing the chromatogram of the test sample solution with the chromatogram of the control solution to obtain the content of the sugammadex sodium in the test sample solution, wherein the chromatographic conditions for detecting the control solution are the same as the chromatographic conditions for detecting the test sample solution.

In certain embodiments, the chromatographic conditions are:

The chromatographic column is an octadecylsilane chemically bonded silica gel column;

The mobile phase A is phosphate buffer solution with the concentration of 20mmol/L to 25mmol/L and the pH value of 2.3 to 2.5;

the mobile phase B is acetonitrile;

the temperature of the chromatographic column is 35-42 ℃;

The flow rate of the mixed liquid of the mobile phase A and the mobile phase B is 0.6ml/min to 1.2 ml/min;

The gradient elution procedure was:

In certain embodiments, the chromatographic conditions are:

The mobile phase A is a potassium dihydrogen phosphate buffer solution with the concentration of 20mmol/L and the pH value of 2.3;

The mobile phase B is acetonitrile;

The temperature of the chromatographic column is 40 ℃;

The flow rate of the mixed liquid of the mobile phase A and the mobile phase B is 0.8 ml/min;

The detection wavelength is 200 nm;

the gradient elution procedure was:

。

According to the detection method of sugammadex sodium, provided by the invention, through selecting a proper mobile phase, controlling the conditions such as the concentration and flow rate of the mobile phase, the column temperature of a chromatographic column and the like, particularly through controlling the pH value of the mobile phase and a gradient elution program, the sugammadex sodium related substances can be effectively separated, the separation degree is high, the detection sensitivity is high, the specificity is good, and the accuracy is high.

Drawings

FIGS. 1 to 6 are HPLC profiles of samples under non-destructive, acid-degradation, alkali-degradation, oxidative degradation, high temperature degradation and light degradation conditions, respectively, in the specificity test of example 13 of the present invention;

FIG. 7 is an HPLC chromatogram of a system suitability solution provided by comparative example 4 of the present invention.

Detailed Description

The chemical name of the sugammadex sodium is 6-fully deoxy-6-fully (2-carboxyethyl) thio-gamma-cyclodextrin sodium salt, and on the basis of keeping the three-dimensional structure of natural gamma-cyclodextrin, 8 side chains are added to deepen the inner cavity of the sugammadex sodium salt so that the sugammadex sodium salt can accommodate amino steroid non-depolarizing muscle relaxants with larger volume and rigidity; in turn, a negatively charged carboxyl group is added at the end of each side chain to enhance its electrostatic binding to the aminosteroid non-depolarizing muscle relaxant. The molecular formula of sugammadex sodium is shown below:

The invention provides a detection method of sugammadex sodium, which comprises the following chromatographic conditions:

the chromatographic column is C₁₈a chromatographic column;

The mobile phase B is acetonitrile;

The temperature of the chromatographic column is 30-45 ℃;

The gradient elution procedure was:

the test sample solution can be prepared from a sample to be tested. In some embodiments, the sample to be tested is a commercially available sugammadex bulk drug or a product obtained when sugammadex is prepared or a sugammadex preparation. The test sample solution can be prepared by dissolving a sample to be tested in the mobile phase A. The concentration of the test sample solution may be from 2mg/ml to 10 mg/ml. The sample amount of the test sample solution in the high performance liquid chromatograph may be 5 μ l to 20 μ l.

according to the method, the sugammadex sodium related substances can be effectively separated by selecting a proper mobile phase, controlling the concentration and flow rate of the mobile phase, the temperature of a chromatographic column and other conditions, particularly controlling the pH value of the mobile phase and a gradient elution procedure, and the method is high in detection sensitivity, good in specificity and high in accuracy.

Said C is₁₈The column refers to a column having a reversed phase stationary phase with an octadecyl carbon chain attached. Preferably, said C₁₈The chromatographic column is an ODS chromatographic column, namely an octadecylsilane chemically bonded silica gel column. Said C is₁₈the size of the column is not limited, e.g. C₁₈the particle size of the packing in the column may be 3 μm to 5 μm, the size of the column may be 150mm × 4.6mm to 250mm × 4.6mm, and so on.

The phosphate in the phosphate buffer solution may include at least one of potassium dihydrogen phosphate, sodium dihydrogen phosphate, ammonium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, and diammonium hydrogen phosphate. Preferably, the phosphate is potassium dihydrogen phosphate and sodium dihydrogen phosphate; more preferably potassium dihydrogen phosphate. The pH of the phosphate buffer solution may be adjusted by an acid, such as phosphoric acid or hydrochloric acid.

The inventor finds that when the pH value of the mobile phase A exceeds 3.0, the main peak of the sugammadex sodium in a chromatogram is delayed, so that the sugammadex sodium cannot be effectively separated from related impurities. Preferably, the pH of the mobile phase a is 2.3 to 2.5; preferably 2.3. Preferably, the concentration of the phosphate buffer solution is 20mmol/L to 25 mmol/L.

The time of segmentation and the ratio of mobile phase in the gradient elution procedure have a large influence on the time to peak and the degree of separation of the sugammadex related substances. Preferably, the gradient elution procedure is:

more preferably, the gradient elution procedure is:

。

The method also comprises the step of detecting the eluent of the gradient elution procedure by using an ultraviolet-spectroscopic visible-spectroscopic detector. The detection wavelength may be 190nm to 210nm, preferably 200 nm.

The detection method of sugammadex sodium of the invention can further comprise the steps of providing a reference substance solution and detecting the reference substance solution by using the high performance liquid chromatograph. The control solution may be made by dissolving the sugammadex sodium in the same solvent as the mobile phase a component. The chromatographic conditions for detecting the control solution may be the same as the chromatographic conditions for detecting the test sample solution.

The method can further comprise the step of comparing the chromatograms of the test sample solution and the control sample solution to obtain the content of the sugammadex sodium in the test sample solution. The content of sugammadex sodium can be calculated by either the external or internal standard method.

The sugammadex sodium and impurities used in the following examples are obtained by purifying and separating intermediate products and final products in the process of preparing the sugammadex sodium, the impurities are substances with a structure similar to that of the sugammadex sodium, and the impurities are sequentially numbered as 1-20.

Example 1

Preparation of system suitability solution:

And dissolving the sugammadex sample and the impurity 1-20 sample by using the same solvent as the mobile phase A to obtain a mixed solution, wherein the concentration of the sugammadex in the mixed solution is 5mg/ml, and the concentration of the impurity 1-20 is 5 mu g/ml respectively.

Injecting the system suitability solution into a liquid chromatograph, wherein the chromatographic conditions are as follows:

A chromatographic column: c₁₈A column (YMC ODS-AQ 150X 4.6mm, 3 μm);

Mobile phase A: preparing 20mmol/L potassium dihydrogen phosphate solution, and adjusting the pH value to 2.3 by using phosphoric acid;

Mobile phase B: acetonitrile;

Ultraviolet detection wavelength: 200 nm;

Flow rate: 0.8ml/min

Column temperature: 40 ℃;

Gradient elution procedure:

time (minutes)	Mobile phase A (%)	mobile phase B (%)
			0	83	17
10	83	17
			40	80	20
50	75	25
			65	50	50
70	25	75
			80	25	75
80.1	83	17
			90	83	17

example 2

Example 2 is essentially the same as example 1, except that the pH of mobile phase a was adjusted to 2.4 with phosphoric acid.

example 3

example 3 is essentially the same as example 1, except that the pH of mobile phase a was adjusted to 2.5 with phosphoric acid. The separation of impurities in examples 1 to 3 is shown in table 1:

TABLE 1

Example 4

Preparation of system suitability solution:

Dissolving sugammadex sodium and impurities 1-20 by using a mobile phase A to obtain a mixed solution, wherein the concentration of the sugammadex sodium in the mixed solution is 5mg/ml, and the concentration of the impurities 1-20 is 5 mu g/ml respectively.

a chromatographic column: c₁₈A column (YMC ODS-AQ 150X 4.6mm, 3 μm);

Mobile phase B: acetonitrile;

Ultraviolet detection wavelength: 200 nm;

Flow rate: 0.8 ml/min;

column temperature: 40 ℃;

Gradient elution procedure:

example 5

example 5 is essentially the same as example 4 except that mobile phase A is a 20mmol/L solution of sodium dihydrogen phosphate.

example 6

example 6 is essentially the same as example 1 except that mobile phase A is a 20mmol/L solution of ammonium dihydrogen phosphate.

the separation of each impurity in examples 4 to 6 is shown in table 2:

TABLE 2

Example 7

Example 7 is essentially the same as example 4 except that mobile phase A is a 15mmol/L solution of potassium dihydrogen phosphate.

Example 8

Example 8 is essentially the same as example 4 except that mobile phase A is a 25mmol/L solution of potassium dihydrogen phosphate.

The separation of impurities in example 4 and examples 7 to 8 is shown in table 3:

TABLE 3

example 9

Example 9 is essentially the same as example 4 except that the column temperature of the column is 35 ℃.

Example 10

Example 10 is essentially the same as example 4 except that the column temperature of the column is 42 ℃.

The separation of impurities in example 4 and examples 9 to 10 is shown in table 4:

TABLE 4

example 11:

Example 11 is essentially the same as example 4 except that the gradient elution procedure is as follows:

Time (minutes)	mobile phase A (%)	Mobile phase B (%)
			0	81	19
10	81	19
			40	80	20
50	75	25
			65	50	50
70	25	75
			80	25	75
80.1	82	18
			90	82	18

example 12:

Example 12 is essentially the same as example 4 except that the gradient elution procedure is as follows:

Time (minutes)	Mobile phase A (%)	mobile phase B (%)
			0	84	16
10	84	16
			40	80	20
50	75	25
			65	50	50
70	25	75
			80	25	75
80.1	82	18
			90	82	18

The separation of impurities in example 4 and examples 11 to 12 is shown in table 5:

TABLE 5

As can be seen from the above examples 1 to 12, the detection method provided by the present invention can effectively separate each impurity in different mobile phase systems, within different mobile phase pH ranges, under different mobile phase concentrations, under different column temperature conditions, and under different mobile phase proportions, and meet the detection requirements.

Comparative example 1

Comparative example 1 is substantially the same as example 4 except that the pH of mobile phase a was adjusted to 3.0 with phosphoric acid.

In the obtained liquid chromatogram, chromatographic peaks with retention time of 9.485min and 10.244min are overlapped, chromatographic peaks with retention time of 12.029min and 13.060min are overlapped, chromatographic peaks with retention time of 14.161min and 14.675min are overlapped, baseline separation is not achieved, and the separation degree is poor.

Comparative example 2

comparative example 2 is substantially the same as example 4 except that the mobile phase a is a 20mmol/L mixed solution (volume ratio 80:20) of potassium dihydrogen phosphate solution at pH 2.3 and acetonitrile, the mobile phase B is acetonitrile, and the gradient elution procedure is:

time (minutes)	Mobile phase A (%)	Mobile phase B (%)
			0	95	5
4	95	5
			5	83	17
25	80	20
			28	75	25
30	75	25
			40	20	80
45	20	80
			46	0	100
50	0	100
			50.1	95	5
55	95	5

In the obtained liquid chromatogram, the separation degrees of the main peak with the retention time of 15.935min, the impurity peak with the retention time of 16.731min, the impurity peaks with the retention times of 20.198min and 20.501min and the impurity peaks with the retention times of 29.556min and 29.665min are all less than 1.0.

Comparative example 3

comparative example 3 is substantially the same as example 4 except that the mobile phase a is a 25mmol/L mixed solution (volume ratio 80:20) of potassium dihydrogen phosphate solution at pH 3.0 and acetonitrile, the mobile phase B is acetonitrile, and the gradient elution procedure is:

In the obtained liquid chromatogram, the separation degrees between chromatographic peaks with retention times of 17.312min and 17.562min, chromatographic peaks with retention times of 20.334min and 20.631min and chromatographic peaks with retention times of 29.684min and 29.835min are all less than 1.0.

comparative example 4:

Comparative example 4 is substantially the same as example 4 except that the mobile phase a is a 20mmol/L mixed solution (volume ratio 80:20) of potassium dihydrogen phosphate solution at pH 4.4 and acetonitrile, the mobile phase B is acetonitrile, and the gradient elution procedure is:

Time (min)	Mobile phase A (%)	mobile phase B (%)
			0	100	0
21	100	0
			35	98	2
50	92	8
			55	75	25
60	50	50
			65	30	70
80	30	70
			81	100	0
90	100	0

The HPLC profile of comparative example 4 is shown in FIG. 7, and after the time delay of the main peak, the impurities are not effectively separated from the main peak.

example 13 specificity test

non-destroyed sample: taking 50mg of sugammadex sodium raw material medicine, precisely weighing, placing in a 10ml measuring flask, adding a diluent to dissolve and dilute to a scale, and shaking up.

acid degradation: taking 50mg of sugammadex sodium raw material medicine, precisely weighing, placing in a 10ml measuring flask, adding 1ml of 1mol/L hydrochloric acid solution, destroying in 80 ℃ water bath for 0.5h, adding 1ml of 1mol/L sodium hydroxide solution, stopping destroying, adding diluent to dilute to scale, and shaking up. And performing blank test by the same method.

Alkali degradation: taking 50mg of sugammadex sodium raw material medicine, precisely weighing, placing in a 10ml measuring flask, adding 1ml of 1mol/L sodium hydroxide solution, destroying in 80 ℃ water bath for 2h, adding 1ml of 1mol/L hydrochloric acid solution, stopping destroying, adding diluent to dilute to scale, and shaking up. And performing blank test by the same method.

oxidative degradation: taking 50mg of sugammadex sodium raw material medicine, precisely weighing, placing in a 10ml measuring flask, adding 1ml of 0.1% hydrogen peroxide solution, destroying at room temperature for 5min, adding diluent to dilute to scale, shaking uniformly, and immediately injecting sample. And performing blank test by the same method.

Solid high-temperature degradation: taking and placing the sugammadex sodium raw material medicine which is placed in a high-temperature test box at 60 ℃ for 30 days, precisely weighing, placing in a 10ml measuring flask, adding a diluent to dilute to a scale, fixing the volume to the scale, and shaking up.

solid light degradation: taking and placing the sugammadex sodium raw material medicine which is placed in a 4500 +/-500 lxs illumination test box for 30 days and is about 50mg, precisely weighing, placing in a 10ml measuring flask, adding a diluent to dilute to a scale, fixing the volume to a scale line, and shaking up.

The samples were subjected to HPLC detection under the same chromatographic conditions as in example 4, and the detection results are shown in fig. 1 to 6 and table 6:

TABLE 6

Degradation ofpathway(s)	Is not degraded	Acid destruction	alkali destruction	Oxidative destruction	High temperature	Illumination of light
							principal component	98.53	91.09	82.02	88.95	96.98	95.70
Minimum degree of separation of impurities from main peak	2.18	2.29	2.45	2.34	2.38	2.29
							main peak purity factor	1000	1000	999.99	1000	1000	1000

from table 6, it can be seen that, by using the HPLC detection method of the present invention, the degradation products of sugammadex sodium under the conditions of acid, alkali, oxidation, illumination and high temperature can be well separated from sugammadex, the minimum separation degrees are all greater than 1.5, the purity factors of the main peak of sugammadex sodium are all greater than 999, and no unknown impurity interferes with the main peak detection, which indicates that the HPLC detection method of the present invention has good specificity.

Example 14 detection and quantitation limits

Preparation of control solutions:

dissolving sugammadex sodium and impurities 1-20 by using a mobile phase A to obtain a mixed solution, wherein the concentration of the sugammadex sodium in the mixed solution is 50 mu g/ml, and the concentration of the impurities 1-20 is 5 mu g/ml respectively.

and (3) diluting the reference solution step by step until the solution with the peak height 10 times of the baseline noise is the quantitative limiting solution, and the solution with the peak height 3 times of the baseline noise is the detection limiting solution. The chromatographic conditions of HPLC were the same as in example 4, and the results are shown in Table 7:

TABLE 7

As can be seen from Table 7, the HPLC detection method of the present invention has high detection sensitivity for each impurity, and each impurity can satisfy the detection requirements.

Example 15 Linear test

Respectively taking 1-20 impurities, adding the mobile phase A to prepare a solution with the concentration of 250 mu g/ml, and preparing an impurity stock solution.

an appropriate amount of impurity stock solutions are precisely measured and diluted into limit horizontal concentrations of 10%, 50%, 100%, 120%, 150% and 200% as linear solutions. Sampling and analyzing each horizontal linear solution by adopting the chromatographic conditions of the embodiment 4, recording a chromatogram, calculating respective linear regression equations and correlation coefficients r by taking the concentrations of main components and each impurity as horizontal coordinates and the peak areas of the corresponding solutions as vertical coordinates, wherein the correlation coefficients r of each impurity are more than 99.9 percent, so that each impurity and sugammadex sodium are in the limit level of quantitative limit to 200 percent, and the linear relationship is good.

example 16 accuracy test

Taking sugammadex sodium and each impurity, adding a mobile phase A to prepare a mixed solution to obtain a reference substance solution, wherein the concentration of each impurity reference substance in the reference substance solution is 50 mu g/ml.

20% level recovery solution: taking 50mg of sugammadex sodium raw material medicine, precisely weighing, placing in a 10ml measuring flask, precisely weighing 0.2ml of each impurity stock solution, adding a diluent to scale, and shaking up to obtain the final product. 3 parts are prepared in parallel.

50% level recovery solution: taking 50mg of sugammadex sodium raw material medicine, precisely weighing, placing in a 10ml measuring flask, precisely weighing 0.5ml of each impurity stock solution, adding diluent to scale, and shaking up to obtain the final product. 3 parts are prepared in parallel.

100% level recovery solution: taking 50mg of sugammadex sodium raw material medicine, precisely weighing, placing in a 10ml measuring flask, precisely weighing 1ml of each impurity stock solution, adding a diluent to the scale, and shaking up to obtain the final product. 3 parts are prepared in parallel.

150% horizontal recovery solution: taking 50mg of sugammadex sodium raw material medicine, precisely weighing, placing in a 10ml measuring flask, precisely weighing 1.5ml of each impurity stock solution, adding a diluent to scale, and shaking up to obtain the final product. 3 parts are prepared in parallel.

The reference solution and the recovered solutions at all levels were sampled and analyzed by the chromatography conditions of example 4, and chromatograms were recorded, and the recovery rates of the respective impurities were calculated, the results of which are shown in table 8 below:

TABLE 8

impurities	Average recovery (%)	RSD value (%)
			impurity 1	100.7	0.5
Impurity 2	100.4	3.27
			Impurity 3	103.1	2.21
Impurity 4	105.4	1.21
			impurity 5	103.6	5.57
impurity 6	103.7	1.60
			Impurity 8	97.63	2.89
impurity 9	105.1	1.26
			Impurities 10	98.91	2.07
Impurity 11	104.8	1.18
			Impurities 12	100.4	1.61
Impurities 13	103.6	3.37
			impurity 14	102.9	1.18
impurity 15	97.11	3.74
			Impurity 16	97.21	2.78
Impurity 17	99.25	0.87
			Impurities 18	101.1	3.54
impurities 19	97.39	2.87
			Impurity 20	104.5	1.59

In addition, by adopting the HPLC measuring method provided by the invention, each impurity can be determined and measured within 20-150% of the limit level, and the accuracy is good.

The invention provides a detection method of sugammadex sodium, which has the advantages that the separation degrees of 20 impurities and a main peak in a system applicability solution are all larger than 1.5, the separation degrees of the impurities are all larger than 1.2, 20 impurities are detected by the same test sample solution, the detection sensitivity is high, the separation effect is good, the method has low requirements on instruments, and the detection cost is low.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. the detection method of sugammadex sodium is characterized in that the chromatographic conditions are as follows:

the chromatographic column is C₁₈A chromatographic column;

The mobile phase B is acetonitrile;

the temperature of the chromatographic column is 30-45 ℃;

The gradient elution procedure was:

。

2. the method for detecting sugammadex sodium according to claim 1, wherein a sample to be detected is dissolved in the mobile phase a to form a test solution.

3. The method for detecting sugammadex sodium according to claim 2, wherein the sample amount of the test sample solution is 5 to 20 μ l.

4. the method for detecting sugammadex sodium according to any one of claims 1 to 3, wherein C is₁₈The chromatographic column is an octadecylsilane chemically bonded silica gel column.

5. The method for detecting sugammadex sodium according to any one of claims 1 to 4, wherein the phosphate in the phosphate buffer solution comprises at least one of potassium dihydrogen phosphate, sodium dihydrogen phosphate, ammonium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, diammonium hydrogen phosphate.

6. The detection method of sugammadex sodium according to any one of claims 1 to 5, wherein the eluate of the gradient elution procedure is detected with an ultraviolet-visible light detector at a detection wavelength of 190nm to 210 nm.

7. the method for detecting sugammadex sodium according to any one of claims 1 to 6, wherein the mobile phase A has a pH of 2.3 to 2.5.

8. The method for detecting sugammadex sodium according to any one of claims 1 to 7, further comprising the steps of providing a control solution, detecting the control solution by using the high performance liquid chromatograph, and comparing the chromatogram of the test sample solution with the chromatogram of the control solution to obtain the content of sugammadex sodium in the test sample solution, wherein the chromatographic conditions for detecting the control solution are the same as the chromatographic conditions for detecting the test sample solution.

9. the method for detecting sugammadex sodium according to any one of claims 1 to 8, wherein the chromatographic conditions are as follows:

The mobile phase B is acetonitrile;

The temperature of the chromatographic column is 35-42 ℃;

the gradient elution procedure was:

Time (minutes) Mobile phase A (%) mobile phase B (％) 0 82～84 18～16 10 82～84 18～16 40 78～80 22～20 50 70～75 30～25 65 50～55 50～45 70 25～30 75～70 80 25～30 75～70 80.1 82～84 18～16 90 82～84 18～16

。

10. The method for detecting sugammadex sodium according to any one of claims 1 to 8, wherein the chromatographic conditions are as follows:

The mobile phase B is acetonitrile;

the temperature of the chromatographic column is 40 ℃;

The detection wavelength is 200 nm;

The gradient elution procedure was:

。