CN115856125A

CN115856125A - Method for detecting 16 impurities in risperidone capsule

Info

Publication number: CN115856125A
Application number: CN202211515595.5A
Authority: CN
Inventors: 沈幼红; 肖志勇; 张蝶雯; 房纯纯; 宋慧慧
Original assignee: Ningbo Dahongying Medicine Co ltd
Current assignee: Ningbo Dahongying Medicine Co ltd
Priority date: 2022-11-29
Filing date: 2022-11-29
Publication date: 2023-03-28

Abstract

The invention discloses a method for detecting 16 impurities in a risperidone capsule, which comprises the following steps: (1) preparing a test solution: precisely weighing 16 impurities of the risperidone capsule and a reference substance of the risperidone; (2) chromatographic conditions: performing gradient elution by using a high performance liquid chromatograph and an ultraviolet detector, wherein a chromatographic column filler is octadecylsilane chemically bonded silica, a water-acetonitrile-trifluoroacetic acid mixed solution is used as a mobile phase A, and an organic phase acetonitrile is used as a mobile phase B; precisely measuring a sample solution, injecting the sample solution into a liquid chromatograph, injecting a sample, recording a chromatogram, and calculating the separation degree between each impurity peak and the impurity peak and between each impurity peak and a main peak. The method can effectively detect the impurities of the risperidone capsule; the method has strong specificity, good reproducibility and high accuracy, can ensure the controllability of risperidone impurities, and can be applied to the quality control of risperidone capsules.

Description

Method for detecting 16 impurities in risperidone capsule

Technical Field

The invention relates to the technical field of pharmaceutical analysis, in particular to a method for detecting 16 impurities in a risperidone capsule.

Background

The safety of medicines is closely related to the life health of people, and with the increasing strictness of the approval system of medicines in various countries, impurities closely related to the quality, safety and curative effect of medicines are used as important evaluation items in approval. The impurities of the medicine generally have no therapeutic effect, and some impurities also have certain toxicity, which is a main factor influencing the safety of the medicine. Adverse reactions generated by the drugs in clinical use are not only related to the pharmacological activities of the drugs, but also are sometimes greatly related to impurities in the drugs. Therefore, pharmaceutical manufacturers are urgently required to develop a proper analysis method for controlling impurities and quantitatively detecting the impurities, so that the high-quality standard of the medicines is ensured, and the safety and the effectiveness of the medicines are ensured.

Risperidone is used for treating acute and chronic schizophrenia, especially has good curative effect on positive and negative symptoms and accompanying emotional symptoms (such as anxiety, depression and the like), and can also relieve the emotional symptoms related to schizophrenia.

Risperidone chemical name: 3- [2- [4- (6-fluoro-1, 2-benzisoxazol-3-yl) -1-piperidinyl ] ethyl ] -6,7,8, 9-tetrahydro-2-methyl-4H-pyrido [1, 2-. Alpha. ] pyrimidin-4-one.

The risperidone capsule only collects the detection of related substances of the risperidone capsule in Chinese pharmacopoeia, but does not collect the related substances in other national pharmacopoeias such as USP, JP and BP. The USP only contains related substance detection methods of risperidone tablets, orally disintegrating tablets and oral solution dosage forms.

The analysis of impurities in the synthesis process of the raw material medicaments and the stability investigation and analysis of finished products show that the risperidone contains 13 process impurities such as impurities A, B, C, D, E, F, G, H, I, J, K, L, M and the like and 3 degradation substances such as bicyclic risperidone, trans-nitrogen oxide, cis-nitrogen oxide and the like. The chemical formula of the 16 risperidone impurities is shown in table 1.

TABLE 1 chemical formulas of 16 Risperidone impurities

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The method comprises the steps of detecting risperidone and 16 mixed solutions of risperidone impurities by adopting an impurity reference substance positioning mode and a related substance detection method of risperidone capsules in 'Chinese pharmacopoeia' 2015 edition, a related substance detection method of USP42 risperidone raw material medicine, risperidone tablets, orally disintegrating tablets, oral solution and EP9.0 risperidone, wherein all the impurities or the impurities and the risperidone cannot be completely separated.

Fig. 1 to 6 are high performance liquid chromatograms obtained by detecting a mixed solution of risperidone and 16 risperidone impurities by using a method for detecting related substances of risperidone capsules, a method for detecting related substances of USP42 risperidone bulk drug, a method for detecting related substances of USP42 risperidone tablets, a method for detecting related substances of USP42 risperidone orally disintegrating tablets, a method for detecting related substances of USP42 risperidone oral solution, and a method for detecting related substances of european pharmacopoeia EP9.0 risperidone bulk drug, respectively, and the measurement results of the methods of the pharmacopoeia of each country are shown in table 2.

Table 2 results of determining 16 risperidone impurities using pharmacopoeia methods of various countries

Wherein, the chromatographic conditions of the related substance detection method of risperidone capsules in the 'Chinese pharmacopoeia' 2015 edition in table 2 are as follows:

and (3) chromatographic column: octadecylsilane chemically bonded silica;

mobile phase:

methanol-0.05 mol/1 ammonium acetate solution (pH 7.0) (60: 40);

detection wavelength: 234nm

Sample introduction volume: 20 mu l of the mixture;

control solution: each 1ml of mobile phase solution contains 5 mu g of risperidone;

test solution: each 1ml of mobile phase solution contains 0.5mg of risperidone.

The chromatographic conditions of the USP42 risperidone bulk drug related substance detection method are as follows:

a chromatographic column: octane silane bonded silica gel (4.6 mm × 25cm;

a mobile phase A:5g/l ammonium acetate solution (pH 6.0);

mobile phase B: acetonitrile-methanol (4;

gradient elution:

time (min)	A(％)	B(％)
			0	70	30
12	65	35
			18	65	35
25	35	65
			35	30	70
40	30	70
			42	70	30
50	70	30

Detection wavelength: 260nm;

sample introduction volume: 10 mu l of the mixture;

flow rate: 2ml/min;

column temperature: 50 ℃;

control solution: each 1ml of methanol solution contains 20 mu g of risperidone;

test solution: each 1ml of methanol solution contained 10mg of risperidone.

The chromatographic conditions of the related substance detection method of USP42 risperidone tablet are as follows:

a chromatographic column: octadecylsilane bonded silica (4.6 mm × 15cm;

diluent agent: 80% methanol;

mobile phase:

mobile phase A: water, acetonitrile and trifluoroacetic acid (80: 19.5: 0.1), mixing uniformly, and adjusting the pH value to 3.0 by ammonia water;

mobile phase B: water, methanol and trifluoroacetic acid (61: 39: 0.1), mixing, adjusting pH to 3.0 with ammonia water;

gradient elution:

time (min)	A(％)	B(％)
			0	100	0
8	100	0
			16	0	100
20	0	100
			21	100	0
30	100	0

Detection wavelength: 275nm;

sample introduction volume: 20 mu l of the mixture;

flow rate: 2.5ml/min;

control solution: every 1ml 80% methanol solution contains 100 mug risperidone;

test solution: each 1ml of 80% methanol solution contained 100. Mu.g of risperidone.

The chromatographic conditions of the related substance detection method of the USP42 risperidone orally disintegrating tablet are as follows:

and (3) chromatographic column: octadecylsilane chemically bonded silica (3.0 mm × 15cm;

diluent agent: methanol-0.1 NHCl (40: 60);

mobile phase:

mobile phase: water, acetonitrile, trifluoroacetic acid (800: 200: 1.5);

detection wavelength: 275nm

Sample introduction volume: 10 mu l of the mixture;

flow rate: 0.8ml/min;

control solution: each 1ml of diluent solution contains 100 mu g of risperidone;

test solution: each 1ml of the diluent solution contained 100. Mu.g of risperidone.

The chromatographic conditions of the substance detection method related to the USP42 risperidone oral solution are as follows:

a chromatographic column: octadecylsilane bonded silica (4.6 mm. Times.10cm;

diluent agent: 5.0g/1 ammonium acetate solution, methanol;

mobile phase:

mobile phase: acetonitrile: 5.0g/1 ammonium acetate solution (11: 39);

detection wavelength: 275nm;

sample introduction volume: 10 mul;

flow rate: 1.5ml/min;

control solution: 1 mu g risperidone is contained in each 1ml of diluent solution;

test solution: each 1ml of the diluent solution contained 200. Mu.g of risperidone.

The chromatographic conditions of the related substance detection method of the raw material drug of the risperidone in the European pharmacopoeia EP9.0 are as follows:

a chromatographic column: octadecylsilane bonded silica (4.6 mm. Times.10cm;

mobile phase:

mobile phase A:5g/l ammonium acetate solution;

and (3) mobile phase B: methanol;

gradient elution:

time (min)	A(％)	B(％)
			0	70	30
2	70	30
			17	30	70
22	30	70

Detection wavelength: 260nm;

sample introduction volume: 10 mu l of the mixture;

flow rate: 1.5ml/min;

test solution: each 1ml of the methanol solution contained 10mg of risperidone.

Therefore, it is necessary to develop a better detection and analysis method for risperidone capsule-related substances for controlling the content of the risperidone capsule-related substances, ensuring the quality of the product and the safety of the medicine.

Disclosure of Invention

The invention aims to solve the technical problems that the existing detection method cannot effectively respond and separate 16 impurities of risperidone, and has the defects of low impurity control and the like, and provides a method for detecting 16 impurities of risperidone capsules. The method can effectively detect the impurities of the risperidone capsule; the method has strong specificity, good reproducibility and high accuracy, can ensure the controllability of risperidone impurities, and can be applied to the quality control of risperidone capsules; the detection method can achieve the aim of effectively ensuring the quality of the risperidone product.

The technical scheme adopted by the invention for solving the technical problems is as follows:

a method for detecting 16 impurities in a risperidone capsule comprises the following steps:

(1) Preparing a test solution: precisely weighing 16 impurities of a risperidone capsule and a reference substance of the risperidone, wherein the 16 impurities of the risperidone capsule are impurities A, B, C, D, E, F, G, H, I, J, K, L, M, bicyclic risperidone, trans-nitric oxide and cis-nitric oxide, and preparing a mixed solution by using 80% methanol, wherein each 1ml of the mixed solution contains 0.5mg of risperidone, 1 mug of each of the impurities A-M, 2.5 mug of bicyclic risperidone, 2.5 mug of cis-nitric oxide and 1 mug of trans-nitric oxide; the chemical formulas of 16 impurities of the peimine capsule are shown in table 1;

(2) Chromatographic conditions are as follows: performing gradient elution by using a high performance liquid chromatograph and an ultraviolet detector, wherein a chromatographic column filler is octadecylsilane chemically bonded silica, a water-acetonitrile-trifluoroacetic acid mixed solution is used as a mobile phase A, and an organic phase acetonitrile is used as a mobile phase B; gradient elution parameters were set as:

0-30 min, the volume ratio of acetonitrile is 0%;

30-40 min, wherein the volume ratio of acetonitrile is changed from 0% to 5%;

the volume proportion of acetonitrile is changed from 5 percent to 40 percent within 40 to 60 min;

60-65 min, the volume of acetonitrile is 40%;

65-66 min, the volume proportion of acetonitrile is changed from 40% to 0%;

66-80 min, the volume proportion of acetonitrile is 0%;

precisely measuring a sample solution, injecting the sample solution into a liquid chromatograph, injecting a sample, recording a chromatogram, and calculating the separation degree between each impurity peak and the impurity peak and between each impurity peak and a main peak.

Preferably, in the step (1), the risperidone concentration in the test solution is 0.5mg/ml.

Preferably, in the step (2), the detection wavelength of the ultraviolet detector is 275nm.

Preferably, in the step (2), the column temperature of the chromatographic column is 43 to 47 ℃, and more preferably 45 ℃.

Preferably, in the step (2), the ratio of the water, the acetonitrile and the trifluoroacetic acid in the mobile phase A is 79: 21: 0.1-81: 19: 0.1, and more preferably 80:20:0.1.

Preferably, in the step (2), the pH of the mobile phase a is 2.6 to 3.0, and more preferably 2.8.

Preferably, in the step (2), the flow rate of the gradient elution is 0.9 to 1.1ml/min, and more preferably 1.0ml/min.

Preferably, in the step (2), 10 μ l of the test solution is precisely measured.

Aiming at risperidone raw material drug synthesis process impurities and finished product stability degradation impurities, 16 impurities are identified, namely impurities A, B, C, D, E, F, G, H, I, J, K, L, M, bicyclic risperidone, trans-nitrogen oxide and cis-nitrogen oxide.

According to the invention, conditions such as the proportion composition of the mobile phase A, the pH value, the flow rate, the column temperature, the gradient elution program and the like are inspected and screened, the separation requirements of 16 impurities and 17 components in total of risperidone are considered, and finally, a proper related substance analysis method is established, so that the main component and each related substance can be separated to the limit, and the purpose of the invention is achieved.

Compared with the prior art, the invention has the beneficial effects that:

(1) The invention collects the risperidone impurity detection methods of different dosage forms of Chinese pharmacopoeia, united states pharmacopoeia and European pharmacopoeia about risperidone raw material medicines, risperidone tablets, risperidone orally disintegrating tablets, risperidone oral solutions, risperidone capsules and the like, the methods in the pharmacopoeia can not detect all impurities or overlap part of impurities or can not completely separate the impurities to different degrees, but the method can well detect 16 impurities in risperidone, the separation degree of risperidone from adjacent peaks is not less than 1.5, the separation between the impurity peaks is not less than 1.2, and good separation is achieved; in the actual inspection process, the actual impurity content of the product can be effectively detected, the impurity control is more effective, and the product quality is ensured;

(2) The method adopts high performance liquid chromatography, uses octadecyl bonded silica gel as chromatographic column filler, adopts an ultraviolet detector, selects a proper organic phase and a proper buffer solution as a mobile phase, adopts gradient elution to effectively complete the separation of risperidone and related impurities, and simultaneously remarkably improves the detection capability and ensures the effective detection of the impurities compared with the related detection methods of international pharmacopoeia at home and abroad; meanwhile, in the method, water and an organic phase are adopted as a mobile phase, and the buffer solution does not contain inorganic salt, so that the loss of the chromatographic column is reduced, and the service life of the chromatographic column is prolonged; the method has strong specificity and good repeatability, and can ensure the controllability of related substances of risperidone; the method can effectively separate the stability degradation impurities and the synthesis process impurities which may remain from the main component, and can be applied to the quality control of the risperidone capsule.

Drawings

FIG. 1 is a high performance liquid chromatogram for detecting a mixed solution of risperidone and 16 risperidone impurities by using a method for detecting related substances of risperidone capsules in the 'Chinese pharmacopoeia' 2015 edition;

FIG. 2 is a high performance liquid chromatogram for detecting a mixed solution of risperidone and 16 risperidone impurities by using a USP42 risperidone bulk drug related substance detection method;

FIG. 3 is a high performance liquid chromatogram for detecting a mixed solution of risperidone and 16 risperidone impurities by a USP42 risperidone tablet related substance detection method;

FIG. 4 is a high performance liquid chromatogram for detecting a mixed solution of risperidone and 16 risperidone impurities by a USP42 risperidone orally disintegrating tablet related substance detection method;

FIG. 5 is a high performance liquid chromatogram for detecting a mixed solution of risperidone and 16 risperidone impurities by USP42 risperidone oral solution related substance detection method;

FIG. 6 is a high performance liquid chromatogram for detecting a mixed solution of risperidone and 16 risperidone impurities by using the method for detecting related substances in the raw material drug of the European pharmacopoeia EP9.0 risperidone;

FIG. 7 is a chromatogram for locating risperidone related substances in example 1.

Detailed Description

For a better understanding of the present invention, reference is made to the following detailed description and accompanying drawings. It is to be understood that these examples are for further illustration of the invention and are not intended to limit the scope of the invention. In addition, it should be understood that the invention is not limited to the above-described embodiments, but is capable of various modifications and changes within the scope of the invention.

The chemical formulas of 16 impurities of the risperidone capsule are detailed in table 1.

Methodological validation of chromatographic methods

Example 1 (test for substance localization)

(1) Preparing a test solution: precisely weighing 16 impurities of a risperidone capsule and a reference substance of the risperidone, wherein the 16 impurities of the risperidone capsule are impurities A, B, C, D, E, F, G, H, I, J, K, L, M, bicyclic risperidone, trans-nitric oxide and cis-nitric oxide, and preparing a mixed solution by using 80% methanol, wherein each 1ml of the mixed solution contains 0.5mg of risperidone, 1 mug of each of the impurities A-M, 2.5 mug of bicyclic risperidone, 2.5 mug of cis-nitric oxide and 1 mug of trans-nitric oxide;

0-30 min, the volume ratio of acetonitrile is 0%;

30-40 min, wherein the volume ratio of acetonitrile is changed from 0% to 5%;

60-65 min, the volume of acetonitrile is 40%;

65-66 min, the volume proportion of acetonitrile is changed from 40% to 0%;

66-80 min, the volume proportion of acetonitrile is 0%;

precisely measuring 10 mu l of test solution, injecting into a liquid chromatograph, injecting and recording a chromatogram, and calculating the separation degree between each impurity peak and between each impurity peak and a main peak.

In the step (1), a test solution containing risperidone with a concentration of 0.5mg/ml is prepared.

In the step (2), the detection wavelength of the ultraviolet detector is 275nm.

In the step (2), the column temperature of the chromatographic column is 45 ℃.

In the step (2), the ratio of water, acetonitrile and trifluoroacetic acid in the mobile phase A is 80:20:0.1.

In the step (2), the pH of the mobile phase A is 2.8.

In the step (2), the flow rate of the gradient elution is 1.0ml/min.

In the embodiment, 16 impurities, namely impurities a, B, C, D, E, F, G, H, I, J, K, L, M, bicyclic risperidone, trans-nitroxide, and cis-nitroxide, are identified for risperidone bulk drug synthesis process impurities and finished product stability degradation impurities.

In the embodiment, conditions such as the proportion composition of the mobile phase A, the pH value, the flow rate, the column temperature, the gradient elution program and the like are considered and screened, the separation requirements of 16 impurities and 17 components in total of risperidone are considered, and finally, a proper related substance analysis method is established, so that the main component and each related substance can be separated to the limit, and the purpose of the embodiment is achieved.

The results of the separation parameters of risperidone and impurities in the mixed solution described in this example are shown in table 3, and the positioning chromatogram of risperidone-related substances is shown in fig. 7.

Table 3 risperidone separation parameter results from each impurity in the mixed solution described in example 1

	Retention time (min)	Relative retention time	Degree of separation
				Impurity L	8.078	0.27	/
Impurity M	12.327	0.41	13.58
				Impurity A	14.010	0.46	3.99
Impurity B	18.191	0.60	4.10
				Bicyclic risperidone	20.299	0.67	3.53
Impurity K	22.003	0.73	2.79
				Impurity C	23.775	0.79	2.74
Impurity H	24.736	0.82	1.35
				Impurity D	27.837	0.92	4.17
Risperidone	30.201	1.00	1.78
				Impurity G	33.838	1.12	2.50
Trans-nitrogen oxides	40.710	1.35	7.93
				Cis-nitroxides	42.208	1.40	2.25
Impurity E	44.508	1.47	4.04
				Impurity F	55.543	1.84	30.54
Impurity J	60.148	1.99	22.06
				Impurity I	61.695	2.04	7.63

The results show that: the separation degree between risperidone and adjacent impurities is not less than 1.5, and the separation degree between the impurities is not less than 1.2, so that good separation is achieved.

Example 2 (quantitative limit test)

And (3) determining the quantitative limit of the risperidone and each related substance thereof by a signal-to-noise ratio method. Respectively preparing stock solutions of risperidone and related substances thereof. Diluting to a certain concentration, injecting, calculating peak area and noise ratio (signal to noise ratio), wherein the detected amount of the sample with the signal to noise ratio (S/N) of about 10 is taken as a quantitative limit, and the result is shown in Table 4.

Table 4 example 2 quantitative limit test results

The results show that: the method has high response to various related substances and can accurately determine the content of the various related substances.

Example 3 (Linear Curve)

Risperidone and 16 impurities thereof are precisely weighed, diluted by 80% methanol and prepared into 7 parts of solutions with different concentrations. And (3) respectively injecting samples and recording chromatograms, and taking the concentration as a horizontal coordinate and the peak area as a vertical coordinate to obtain a linear regression equation of risperidone and each related substance thereof, wherein the result is shown in table 5.

Table 5 example 3 linear curve test results

The results show that: the risperidone and 16 impurities in the method show good linearity in a certain concentration range.

Example 4 (accuracy)

Precisely weighing blank auxiliary materials of the risperidone capsule, preparing 12 parts, taking three parts as one group, and respectively adding a proper amount of impurities and risperidone into each group to ensure that the concentrations of the impurities and the risperidone in the three groups of solutions are respectively 50% of the quantitative limit concentration, 100% of the relative limit and 150% of the relative limit. And (5) injecting and recording a chromatogram, and calculating the recovery rate, wherein the result is shown in a table 6.

Table 6 example 4 accuracy experimental results

Compound (I)	Quantitative limiting concentration	Limit concentration of 50%	Limit concentration of 100%	Limit concentration of 150%
					Risperidone	110.6％	102.8％	101.9％	98.3％
Impurity A	108.5％	103.0％	101.6％	100.8％
					Impurity B	102.9％	100.9％	99.9％	100.0％
Impurity C	111.9％	106.5％	99.9％	96.3％
					Impurity D	102.2％	100.1	102.2	99.1
Impurity E	110.0％	101.0	99.8	98.4
					Impurity F	111.3％	105.2	103.2	102.8
Impurity G	/	100.9	98.8	95.2
					Impurity H	84.4％	99.4	99.2	99.3
Impurity I	102.4％	103.3	102.1	102.6
					Impurity J	107.9％	103.8	102.4	101.8
Impurity K	103.4％	102.5	101.5	100.9
					Impurity L	105.0％	101.2	100.3	99.2
Impurity M	101.3％	99.2	98.7	97.0
					Bicyclic risperidone	100.8％	98.6	97.7	97.2
Cis-nitroxides	97.8％	101.4	103.7	103.8
					Trans-nitrogen oxides	105.1％	98.7	98.5	97.7

The results show that: the recovery rate of the quantitative limit concentration of the method is between 80% and 115%, and the recovery rate of 50% of the relative limit, 100% of the relative limit and 150% of the relative limit is between 85% and 110%, which shows that the method has good accuracy.

Example 5 (forced degradation test)

The contents of the risperidone capsules were subjected to a forced degradation test under acid, alkali, oxidation, high temperature, and light conditions, and the results are shown in table 7.

Table 7 example 5 forced degradation test

The results show that: the method can well detect the degradation product generated by the risperidone destruction test, and the material balance rate is between 95% and 105%; the separation degree of the degraded impurities and the main peak is not less than 1.5, which meets the requirement; the purity of the main peak is less than the threshold value of the purity of the main peak, which meets the requirement. Thus, the above chromatographic system can be used to determine impurities and their degradation products in risperidone capsules.

Example 6 (durability test)

Precisely weighing the content of the risperidone capsule, preparing 2 parts, respectively adding appropriate cis-nitrogen oxide, trans-nitrogen oxide and dicyclic risperidone reference substances, and preparing a sample solution by using 80% methanol, wherein each 1ml of the sample solution contains 0.5mg of risperidone, 2.5 mu g of dicyclic risperidone, 2.5 mu g of cis-nitrogen oxide and 1 mu g of trans-nitrogen oxide.

The ability of the method to be unaffected by variations in the parameters was examined by deliberately varying the parameters of the method. The following parameters will be considered independently in this study: mobile phase composition (. + -. 1%), mobile phase pH (. + -. 2%), flow rate (. + -. 0.1 ml/min), column temperature (. + -. 2 ℃). The results are shown in Table 8.

And (4) sampling the sample solution, injecting the sample solution into a liquid chromatograph according to the changed chromatographic conditions, recording the chromatogram, and calculating the content of the impurities. The durability test results are shown in Table 8.

Table 8 example 6 durability test

The results show that: the content of each impurity and the total content of the impurities after the conditions are changed are not more than 0.1 percent of the difference value compared with the content under the normal conditions. The results show that the method is durable to small changes of mobile phase proportion, mobile phase pH value, flow velocity and column temperature.

Example 7 measurement of related substances in 4 lots of Risperidone tablets original research drug (manufactured by Qiangsheng, USA)

Precisely weighing 5 risperidone tablets in 4 batches, respectively placing into 10ml measuring bottles, and dissolving and diluting with 80% methanol to obtain a test solution containing 0.5mg risperidone per 1 ml. The test solution was diluted with 80% methanol to prepare a control solution containing 2.5. Mu.g risperidone per 1 ml.

Chromatographic conditions are as follows: the high performance liquid chromatograph, the ultraviolet detector and the chromatographic column filler are octadecylsilane chemically bonded silica; adjusting pH of the mixed solution of water, acetonitrile and trifluoroacetic acid (80: 20: 0.1) to 2.8 with ammonia water (mobile phase A) and an organic phase acetonitrile (mobile phase B), and carrying out gradient elution; gradient elution parameters were set as:

0-30 min, wherein the volume ratio of acetonitrile is 0%;

30-40 min, wherein the volume ratio of acetonitrile is changed from 0% to 5%;

the volume proportion of the acetonitrile is changed from 5 percent to 40 percent within 40-60 min;

60-65 min, the volume of acetonitrile is 40%;

65-66 min, the volume proportion of acetonitrile is changed from 40% to 0%;

66-80 min, the volume proportion of acetonitrile is 0%;

the detection wavelength is 275nm; the flow rate is 1.0ml/min; the column temperature was 45 ℃; precisely measuring 10 μ l of each of the reference solution and the sample solution, respectively injecting into a liquid chromatograph, injecting and recording chromatogram, and calculating the content of each impurity by using a self-comparison method, wherein the results are shown in Table 9.

TABLE 9 EXAMPLE 7 comparison of the results of the four Risperidone tablet test in the original drug research

Impurities/batch	Batch 1	Batch 2	Batch 3	Batch 4
					Bicyclic risperidone	Not detected out	Undetected	Undetected	Not detected out
Cis-nitroxides	0.07％	Not detected out	0.04％	0.03％
					Trans-nitrogen oxides	Undetected	Undetected	Not detected out	Undetected
Impurity A	0.04％	Not detected out	Not detected out	Undetected
					Impurity B	Undetected	Not detected out	Undetected	Not detected out
Impurity C	Not detected out	Not detected out	Not detected out	Undetected
					Impurity D	Undetected	Not detected out	Undetected	Not detected out
Impurity E	Undetected	Not detected out	Not detected out	Not detected out
					Impurity F	Not detected out	Undetected	Undetected	Not detected out
Impurity G	Undetected	Not detected out	Not detected out	Not detected out
					Impurity H	Not detected out	Not detected out	Not detected out	Not detected out
Impurity I	Not detected out	Undetected	Not detected out	Not detected out
					Impurity J	Undetected	Not detected out	Not detected out	Not detected out
Impurity K	Not detected out	Not detected out	Not detected out	Undetected
					Impurity L	Not detected out	Undetected	Not detected out	Not detected out
Impurity M	Undetected	Not detected out	Not detected out	Not detected out
					Other single impurities	Not detected out	0.04％	0.04％	0.04％
Total impurities	0.11	0.04％	0.08％	0.07％

The results show that: the method has high detection capability on the risperidone impurities, controls the risperidone impurities more strictly, and can better ensure the safety of the medication of patients.

From the results of the above examples, it can be seen that the method of the present invention can effectively detect impurities in risperidone capsules; the method has strong specificity, good reproducibility and high accuracy, can ensure the controllability of risperidone impurities, and can be applied to the quality control of risperidone capsules.

The above description is not intended to limit the present invention, and the present invention is not limited to the above examples. Those skilled in the art should also realize that changes, modifications, additions and substitutions can be made without departing from the true spirit and scope of the invention.

Claims

1. A method for detecting 16 impurities in a risperidone capsule is characterized by comprising the following steps:

(1) Preparing a test solution: precisely weighing 16 impurities of a risperidone capsule and a reference substance of the risperidone, wherein the 16 impurities of the risperidone capsule are impurities A, B, C, D, E, F, G, H, I, J, K, L, M, bicyclic risperidone, trans-nitric oxide and cis-nitric oxide, and preparing a mixed solution by using 80% methanol, wherein each 1ml of the mixed solution contains 0.5mg of risperidone, 1 mug of each of the impurities A-M, 2.5 mug of bicyclic risperidone, 2.5 mug of cis-nitric oxide and 1 mug of trans-nitric oxide; the chemical formula of the 16 impurities of the peimine capsule is as follows:

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0-30 min, the volume ratio of acetonitrile is 0%;

30-40 min, wherein the volume ratio of acetonitrile is changed from 0% to 5%;

60-65 min, the volume of acetonitrile is 40%;

65-66 min, the volume proportion of acetonitrile is changed from 40% to 0%;

66-80 min, the volume proportion of acetonitrile is 0%;

2. The method for detecting 16 impurities in a risperidone capsule as claimed in claim 1, wherein the detection wavelength of the ultraviolet detector in step (2) is 275nm.

3. The method for detecting 16 impurities in a risperidone capsule as claimed in claim 1, wherein the column temperature of the chromatographic column in step (2) is 43-47 ℃.

4. The method for detecting 16 impurities in a risperidone capsule of claim 3, wherein the column temperature of the chromatographic column is 45 ℃.

5. The method for detecting 16 impurities in a risperidone capsule as claimed in claim 1, wherein in the step (2), the ratio of water, acetonitrile and trifluoroacetic acid in the mobile phase A is 79: 21: 0.1-81: 19: 0.1.

6. The method for detecting 16 impurities in a risperidone capsule as claimed in claim 5, wherein the ratio of water, acetonitrile and trifluoroacetic acid in the mobile phase A is 80:20:0.1.

7. the method for detecting 16 impurities in a risperidone capsule as claimed in claim 1, wherein the pH of the mobile phase A is 2.6-3.0 in the step (2).

8. The method for detecting 16 impurities in a risperidone capsule of claim 7, wherein the pH of the mobile phase A is 2.8.

9. The method for detecting 16 impurities in a risperidone capsule as claimed in claim 1, wherein the flow rate of the gradient elution in step (1) is 0.9-1.1 ml/min.

10. The method for detecting 16 impurities in a risperidone capsule of claim 9, wherein the gradient elution is at a flow rate of 1.0ml/min.